Plant Physiology Handout

HAWASSA UNIVERSITY PLANT PHYSIOLOGY
Course Description
The course deals with definition, scope, practical aspects of plant physiology and describe how
different disciplines interconnected with plant physiology. the structure and properties of water;
water relations with plant cells and tissues; water relations with soils; osmosis; water movement
into and through plants; the ascent of sap; transpiration; photosynthesis; how plants use the
energy of the sun to assimilate carbon and how they convert that carbon to organic molecules,
how plants grow, develop, and respond to their environment. To explain how growth regulators
affect growth and development of plants. Moreover, how plants develop various mechanisms to
defend stress. In general, the course concerns how plants function to survive.
Course Objectives
At the end of this course, students will be able to:
 Be familiar students with definition, scope and practical aspect of plant physiology
 Acquaint the concept of Soil-Plant-Atmosphere Continuum (SPAC)
 Name different processes in photosynthesis and explain the difference between C3, C4,
and CAM plants
 Differentiate how plants grow and develop and measurements of plant growth
 Recognized how the plant process and functions response to different stress conditions to
understand how plant hormones affect growth and development of plants
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Course Description i
Course Objectives i
CHAPTER 1. INTRODUCTION 1
1.1. Definition and scope of plant physiology 1

1.2. Relation of plant physiology with other disciplines 3
CHAPTER 2. PLANT-WATER RELATIONS 4
2.1. Importance and Properties of Water 4

2.2. Water Movement through Soil- Plant - Atmosphere Continuum (SPAC) Water in the Soil 9
2.2.1. Absorption ofWater by Plants 11
2.2.2. Mechanism of Water Absorption 22
2.2.3. Factors Affecting Water Absorption 23
2.3.1 Types of Transpiration 28
2.3.2. Mechanism of Transpiration 30
2.3.3. Mechanism of Stomata Opening and Closing 31
2.3.4. Significance of Transpiration 32
2.3.5. Factors Affecting Rate of Transpiration 33
CHAPTER 3: PHOTOSYNTHESIS 37
3.1. The Nature of Light 38

3.2. Photosynthetic Pigments 44
3.3. Light Reaction and Dark Reaction 48
3.3.1. Role of Photo-system I and II 52
3.4. Path of Carbon in Photosynthesis 54
3.4.1. C3 Photosynthesis Cycle 55
3.4.2. C4 photosynthesis Cycle 57
3.4.3. CAM (CRASSULACEAN ACID METABOLISM) 59
3.5. Role of C3 and C4 in Crop Productivity 62
3.6. Photorespiration (O2 oxidative photosynthetic carbon cycle) 65
3.7. Factors Affecting Photosynthesis. 66
CHAPTER 4. RESPIRATION 73
4.2. Pentose Phosphate Pathway (PPP) 79

4.3. Fermentation 79
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4.4. TCA (Tricarboxylic acid/Kreb cycle 80

4.5. Mitochondrial Electron transport and oxidative Phosphorylation 82
4.6. Photophosphorylation 86
4.7. Energetic of Glycolysis, TCA, and mitochondrial Electron transport 86
4.8. Factors affecting respiration 87
CHAPTER 5: PLANT GROWTH AND DEVELOPMENT 91
5.1. Definitions of Growth and Development 91

5.2. Processes and Measurements of Growth 104
5.3. Dry Matter Partitioning 107
5.3. Factors Affecting Growth and Development 109
CHAPTER 6: PLANT HORMONES AND GROWTH REGULATORS 112
6.1. Auxins 113

6.2. Gibberellins 125
6.3. Cytokinins (= KININS) 132
6.4. Ethylene 138
6.5. Abscisic Acid (ABA) 142
6.6. Jasmonic Acid 144
CHAPTER 7: STRESS PHYSIOLOGY 147
7.1. Water stress 147

7.2. Temperature stress 149
7.3. Salt stress 151
7.4. Oxygen stress 152
7.5. Mechanism of stress tolerance/resistance by plants 155
REFERENCES 166
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CHAPTER 1. INTRODUCTION
1.1. Definition and scope of plant physiology

The word physiology is derived from two Greek words physis means nature, origin and logos
means speech (to talk about the nature of things). It is the study of mechanical, physical, and
biochemical function of living organisms. Plant physiology is a science to study the law of the
life activity of plant. The study of plant functions, encompassing the dynamic processes of
growth, metabolism and reproduction in living plant. Plant physiology is about how plants use
the energy of sun to assimilate carbon, and how they convert that carbon to stuff of which they
are made. It is about how plants obtain and distribute nutrients and water, how they grow and
develop, how they respond to their environment, how they react to stress, and how they
reproduce. In short, plant physiology is about how plants work.
In essence, plant physiology is a study of the plant way of life, which include various aspects of
the plant lifestyle and survival including: metabolism, water relations, mineral nutrition,
development, movement, irritability (response to the environment), and organization, growth,
and transport processes. Scope The field of plant physiology includes the study of all the internal
activities of plants those chemical and physical processes associated with life as they occur in
plants. This includes study at many levels of scale of size and time. At the smallest scale are
molecular interactions of photosynthesis and internal diffusion of water, minerals, and nutrients.
At the largest scale are the processes of plant development, seasonality, dormancy, and
reproductive control. Major subdisciplines of plant physiology include phytochemistry (the study
of the biochemistry of plants) and phytopathology (the study of disease in plants). The scope of
plant physiology as a discipline may be divided into several major areas of research.
First, the study of phytochemistry (plant chemistry) is included within the domain of plant
physiology. To function and survive, plants produce a wide array of chemical compounds not
found in other organisms. Photosynthesis requires a large array of pigments, enzymes, and other
compounds to function. Because they cannot move, plants must also defend themselves
chemically from herbivores, pathogens and competition from other plants. They do this by
producing toxins and foul-tasting or smelling chemicals. Other compounds defend plants against
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disease, permit survival during drought, and prepare plants for dormancy while other compounds
are used to attract pollinators or herbivores to spread ripe seeds.
Secondly, plant physiology includes the study of biological and chemical processes of individual
plant cells. Plant cells have a number of features that distinguish them from cells of animals, and
which lead to major differences in the way that plant life behaves and responds differently from
animal life. For example, plant cells have a cell wall which restricts the shape of plant cells and
thereby limits the flexibility and mobility of plants. Plant cells also contain chlorophyll, a
chemical compound that interacts with light in a way that enables plants to manufacture their
own nutrients rather than consuming other living things as animals do.
Thirdly, plant physiology deals with interactions between cells, tissues, and organs within a
plant. Different cells and tissues are physically and chemically specialized to perform different
functions. Roots and rhizoids function to anchor the plant and acquire minerals in the soil.
Leaves catch light in order to manufacture nutrients. For both of these organs to remain living,
minerals that the roots acquire must be transported to the leaves, and the nutrients manufactured
in the leaves must be transported to the roots. Plants have developed a number of ways to
achieve this transport, such as vascular tissue, and the functioning of the various modes of
transport is studied by plant physiologists.
Fourthly, plant physiologists study the ways that plants control or regulate internal functions.
Like animals, plants produce chemicals called hormones which are produced in one part of the
plant to signal cells in another part of the plant to respond. Many flowering plants bloom at the
appropriate time because of light-sensitive compounds that respond to the length of the night, a
phenomenon known as photoperiodism. The ripening of fruit and loss of leaves in the winter is
controlled in part by the production of the gas ethylene by the plant.
Finally, plant physiology includes the study of how plants respond to conditions and variation in
the environment, a field known as environmental physiology. Stress from water loss, changes in
air chemistry, or crowding by other plants can lead to changes in the way a plant functions.
These changes may be affected by genetic, chemical, and physical factors.
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1.2. Relation of plant physiology with other disciplines

Plant physiology is a sub-discipline of botany concerned with the function, or physiology, of
plants. Closely related fields include plant morphology (structure of plants), plant ecology
(interactions with the environment), phytochemistry (biochemistry of plants), cell biology, and
molecular biology. Fundamental processes such as photosynthesis, respiration, plant nutrition,
plant hormone functions, tropisms, nastic movements, photoperiodism, photomorphogenesis,
circadian rhythms, environmental stress physiology, seed germination, dormancy and stomata
function and transpiration, both part of plant water relations, are studied by plant physiologists.
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CHAPTER 2. PLANT-WATER RELATIONS
2.1. Importance and Properties of Water

Water plays a crucial role in the life of the plant. For every gram of organic matter made by the
plant, approximately 500 g of water is absorbed by the roots, transported through the plant body
and lost to the atmosphere. Even slight imbalances in this flow of water can cause water deficits
and severe malfunctioning of many cellular processes. Thus, every plant must delicately balance
its uptake and loss of water. This balancing is a serious challenge for land plants. To carry on
photosynthesis, they need to draw CO2 from the atmosphere, but doing so exposes them to water
loss and the threat of dehydration.
A major difference between plant and animal cells that affects virtually all aspects of their
relation with water is the existence in plants of the cell wall. Cell walls allow plant cells to build
up large internal hydrostatic pressures called turgor pressure, which is a result of their normal
water balance. Turgor pressure is essential for many physiological processes, including cell
enlargement, gas exchange in the leaves, transport in the phloem, and various transport processes
across membranes. Turgor pressure also contributes to the rigidity and mechanical stability of
non-lignified plant tissues. In this chapter we will consider how water moves into and out of
plant cells, emphasizing the molecular properties of water and the physical forces that influence
water movement at the cell level. But first we will describe the major functions of water in plant
life.
Water in Plant Life Water makes up most of the mass of plant cells, as we can readily appreciate
if we look at microscopic sections of mature plant cells: Each cell contains a large water-filled
vacuole. In such cells the cytoplasm makes up only 5 to 10% of the cell volume; the remainder is
vacuole. Water typically constitutes 80 to 95% of the mass of growing plant tissues. Common
vegetables such as carrots and lettuce may contain 85 to 95% water. Wood, which is composed
mostly of dead cells, has lower water content; sapwood, which functions in transport in the
xylem, contains 35 to 75% water; and heartwood has slightly lower water content. Seeds, with a
water content of 5 to 15%, are among the driest of plant tissues, yet before germinating they
must absorb a considerable amount of water.
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Plants continuously absorb and lose water. Most of the water lost by the plant evaporates from
the leaf as the CO2 needed for photosynthesis is absorbed from the atmosphere. On a warm, dry,
sunny day a leaf will exchange up to 100% of its water in a single hour. During the plant’s
lifetime, water equivalent to 100 times the fresh weight of the plant may be lost through the leaf
surfaces. Such water loss is called transpiration. Transpiration is an important means of
dissipating the heat input from sunlight. Heat dissipates because the water molecules that escape
into the atmosphere have higher than average energy, which breaks the bonds holding them in
the liquid. When these molecules escape, they leave behind a mass of molecules with lower-
than-average energy and thus a cooler body of water. For a typical leaf, nearly half of the net
heat input from sunlight is dissipated by transpiration. In addition, the stream of water taken up
by the roots is an important means of bringing dissolved soil minerals to the root surface for
absorption.
Properties of Water
Water has a number of unique chemical and physical properties that make it essential for life.
One such property is familiar to everyone: solid water floats on liquid water. Almost all liquids
contract when they get colder and reach a maximum density when they solidify. Water is
different. As water cools, it contracts until it reaches 4 0C, then it expands until it freezes at 0 C.
Water also has a high density and is remarkable in having its maximum density at 4°c instead of
at the freezing point. Even more remarkable is the fact that water expands on freezing, so that ice
has a volume about 9% greater than the liquid water from which it was formed. This explains
why ice floats and pipes and radiators burst when the water in them freezes. Ice is less dense than
water which allows ice cubes to float in a soft drink, icebergs to float in the ocean, and ponds and
lakes to freeze from the top down so that aquatic plants and animals can survive in the unfrozen
liquid below.
Water molecules have a simple structure: two hydrogen atoms bonded to one oxygen atom H2O.
This simple structure is responsible for water’s unique properties. The bond between each
hydrogen atom and the oxygen atom results from a pair of electrons shared between the two
atoms. In water, the electrons in the shared pair are not shared equally between the hydrogen and
oxygen atoms. The oxygen atom has a greater affinity for electrons than does the hydrogen atom,
and the electrons in the O–H bond are more attracted to oxygen. Because electrons have a
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negative charge, the unequal sharing in the O–H bond results in oxygen acquiring a partial
negative charge (-) and hydrogen a partial positive charge (+). The H–O–H bond angle in water
is 104.50, which means that the molecule has a bent shape. This bent geometry and the
accumulation of electrons on the oxygen side of the molecule cause the water molecule to have a
negative charge on one side, the oxygen side, and a positive charge on the other side, the
hydrogen side. Molecules with negative regions and positive regions are called polar molecules.
Water molecules are polar molecules.
Polar molecules are attracted to each other. The attraction results from the negative region of one
molecule, the oxygen atom being drawn to the positive region of another molecule, the hydrogen
atom. Opposites attract, the attractions between water molecules are particularly strong. Oxygen
atoms have a very great affinity for electrons, and so the hydrogen atoms bonded to an oxygen atom
acquire a significant positive charge. These hydrogen atoms are very tiny, so the positive charge is
quite concentrated. This concentrated positive charge enhances the attraction of the hydrogen atoms
in one molecule for the oxygen atom in another molecule. These attractions are represented by the
green lines (highlighted by arrows) in the figure. This attraction is so strong that it has been given a
particular name: hydrogen bonding. The energy associated with hydrogen bonds in water is about
20 kJ·mol-1, which is about 1/10th the strength of a typical shared electron bond within a molecule.
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Polarity of water molecules results in cohesion and adhesion. The strong mutual attraction
between water molecules resulting from hydrogen bonding is also known as cohesion. One
consequence of cohesion is that water has an exceptionally high surface tension, which is most
evident at interfaces between water and air. Surface tension arises because the cohesive force
between water molecules is much stronger than interactions between water and air. The result is
That water molecules at the surface are constantly being pulled into the bulk water. The surface
thus tends to contract and behaves much in the manner of an elastic membrane. Water has
special properties that enable it to act as a solvent and to be readily transported through the body
of the plant. These properties derive primarily from the polar structure of the water molecule.
Water is an excellent solvent, dissolving greater amounts of a wide variety of substances than do
other related substances. This is due to water’s small size and polar nature (i.e., water has a
partial negative charge at the oxygen end and a partial positive charge at the hydrogen end of the
molecule).
Water has the ability to partially neutralize electrical attractions between charged solute molecules or
ions by surrounding the ion or molecule with one or more layers of oriented water molecules, called a
hydration shell. Water provides the medium for the movement of molecules within and between
plant cells and greatly influences the structure of proteins, nucleic acids, polysaccharides, and other
cell constituents. Water has an unusually high specific heat. It takes more energy to raise the
temperature of one gram of water by 10C than any other liquid. Temperature is an expression of the
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amount of kinetic energy in the molecules of a substance; increasing temperature corresponds to

increasing kinetic energy. Again, hydrogen bonds are responsible for the high specific heat of water.
As water is heated, much of the added energy goes to breaking apart hydrogen bonds.
The energy used in breaking hydrogen bonds is not available to increase the kinetic energy of the
water molecules, so the temperature of water does not rise as much as would a liquid with lower
intermolecular forces. Therefore, water must absorb more heat energy to raise its temperature. Water
can absorb large amounts of heat energy before it begins to get hot. Similarly, as water cools, it
releases a great deal of heat. The high specific heat of water is responsible for the ocean’s ability to
act as a thermal reservoir that moderates wings in the Earth’s temperature from day to night and from
winter to summer, and this makes the planet suitable for its diverse inhabitants. Global water
dynamics also greatly influences climates and weather. Each day the sun vaporizes about 1,250 cu
km of water. As this water vapor condenses, heat is slowly released over land areas and bodies of
water. As a result, water is responsible or a massive circulation and distribution of heat over the entire
globe. The high heat capacity of water also prevents large bodies of water from heating or cooling too
rapidly contributing to “lake effects” and maritime climates.
Water has an extremely high surface tension. Cohesion is directly responsible for the unusually
high tensile strength of water. Surface tension can be defined as the force per unit length that
can pull perpendicular to a line in a plane of the surface. It is the surface tension at the
evaporative surface of leaves that generates the physical forces that can pull a stream of water
Through the plant’s vascular system Water also has a definite tensile strength. A substance’s
tensile strength is the maximum tension that it can withstand before breaking. It is the great
cohesive forces between water molecules that allow an appreciable tension to exist in an
uninterrupted water column in a wettable capillary or tube (such as a xylem vessel) for capillary
rise to occur. This is important for the continuous movement of water from the root through the
plant to the surrounding air during transpiration. Water is relatively transparent to visible
radiation. This enables sunlight to reach chloroplasts within the cells of leaves and to reach
plants submerged at appreciable depths in bodies of water.
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2.2. Water Movement through Soil- Plant - Atmosphere Continuum (SPAC) Water
in the Soil
The water content and the rate of water movement in soils depend to a large extent on soil type
and soil structure. Table 1 below shows that the physical characteristics of different soils can
vary greatly. At one extreme is sand, in which the soil particles may be 1 mm or more in
diameter. Sandy soils have a relatively low surface area per gram of soil and have large spaces or
channels between particles. At the other extreme is clay, in which particles are smaller than 2 μm
in diameter. Clay soils have much greater surface areas and smaller channels between particles.
With the aid of organic substances such as humus (decomposing organic matter), clay particles
may aggregate into “crumbs” that help improve soil aeration and infiltration of water.
When a soil is heavily watered by rain or by irrigation, the water percolates downward by gravity
through the spaces between soil particles, partly displacing, and in some cases trapping, air in
these channels. Water in the soil may exist as a film adhering to the surface of soil particles, or it
may fill the entire channel between particles. In sandy soils, the spaces between particles are so
large that water tends to drain from them and remain only on the particle surfaces and at
interstices between particles. In clay soils, the channels are small enough that water does not
freely drain from them; it is held more tightly.
The moisture-holding capacity of soils is called the field capacity. Field capacity is the water
content of a soil after it has been saturated with water and excess water has been allowed to drain
away. Clay soils or soils with high humus content have a large field capacity. A few days after
being saturated, they might retain 40% water by volume. In contrast, sandy soils typically retain
3% water by volume after saturation.
A Negative Hydrostatic Pressure in Soil Water Lowers Soil Water Potential: Like the water
Potential of plant cells, the water potential of soils may be dissected into two components, the
osmotic potential and the hydrostatic pressure. The osmotic potential (Ψs) of soil water is
generally negligible because solute concentrations are low; a typical value might be –0.02 MPa.
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For soils that contain a substantial concentration of salts, however, Ψs is significant, perhaps –
0.2 MPa or lower. The second component of soil water potential is hydrostatic pressure (Ψp).
For wet soils, Ψp is very close to zero. As a soil dries out, Ψp decreases and can become quite
negative. Where does the negative pressure in soil water come from? Water has a high surface
tension that tends to minimize air–water interfaces. As a soil dries out, water is first removed
from the center of the largest spaces between particles. Because of adhesive forces, water tends
to cling to the surfaces of soil particles, so a large surface area between soil water and soil air
develops.
Water Moves through the Soil by Bulk Flow: Water moves through soils predominantly by
bulk flow driven by a pressure gradient. In addition, diffusion of water vapor accounts for some
water movement. As plants absorb water from the soil, they deplete the soil of water near the
surface of the roots. This depletion reduces Ψp in the water near the root surface and establishes
a pressure gradient with respect to neighboring regions of soil that have higher Ψp values.
Because the water-filled pore spaces in the soil are interconnected, water moves to the root
surface by bulk flow through these channels down the pressure gradient.
The rate of water flow in soils depends on two factors: the size of the pressure gradient through
the soil, and the hydraulic conductivity of the soil. Soil hydraulic conductivity is a measure of
the ease with which water moves through the soil, and it varies with the type of soil and water
content. Sandy soils, with their large spaces between particles, have a large hydraulic
conductivity, whereas clay soils, with the minute spaces between their particles, have an
appreciably smaller hydraulic conductivity.
As the water content (and hence the water potential) of a soil decreases, the hydraulic
conductivity decreases drastically (see figure 1). This decrease in soil hydraulic conductivity is
due primarily to the replacement of water in the soil spaces by air. When air moves into a soil
channel previously filled with water, water movement through that channel is restricted to the
periphery of the channel. As more of the soil spaces become filled with air, water can flow
through fewer and narrower channels, and the hydraulic conductivity falls. In very dry soils, the
water potential (Ψw) may fall below what is called the permanent wilting point. At this point
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the water potential of the soil is so low that plants cannot regain turgor pressure even if all water
loss through transpiration ceases. This means that the water potential of the soil (Ψw) is less than
or equal to the osmotic potential (Ψs) of the plant. Because cell Ψs varies with plant species, the
permanent wilting point is clearly not a unique property of the soil; it depends on the plant
species as well.
Figure 1 Root hairs make intimate contact with soil particles and greatly amplify the surface
area that can be used for water absorption by the plant. The soil is a mixture of particles (sand,
clay, silt, and organic material), water, dissolved solutes, and air. Water is adsorbed to the
surface of the soil particles. As water is absorbed by the plant, the soil solution recedes into
smaller pockets, channels, and crevices between the soil particles. At the air–water interfaces,
this recession causes the surface of the soil solution to develop concave menisci (curved
interfaces between air and water marked in the figure by arrows), and brings the solution into
tension (negative pressure) by surface tension. As more water is removed from the soil, more
acute menisci are formed, resulting in greater tensions (more negative pressures).
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2.2.1. Absorption of Water by Plants

The dominant process in water relations of the whole plant is the absorption of large quantities of
water from the soil, its translocation through the plant, and its eventual loss to the surrounding
atmosphere as water vapor. Of all the water absorbed by plants, less than 5 percent is actually
retained for growth and even less is used biochemically. The balance passes through the plant to
be lost as water vapor, a phenomenon known as transpiration. Intimate contact between the
surface of the root and the soil is essential for effective water absorption by the root. This contact
provides the surface area needed for water uptake and is maximized by the growth of the root
and of root hairs into the soil. Root hairs are microscopic extensions of root epidermal cells that
greatly increase the surface area of the root, thus providing greater capacity for absorption of
ions and water from the soil. Water enters the root most readily in the apical part of the root that
includes the root hair zone. More mature regions of the root often have an outer layer of
protective tissue, called an exodermis or hypodermis that contains hydrophobic materials in its
walls and is relatively impermeable to water.
Figure 2 Pathways for water uptake by the root. Through the cortex, water may travel via the
apoplast pathway, the transmembrane pathway, and the symplast pathway. In the symplast
pathway, water flows between cells through the plasmodesmata without crossing the plasma
membrane. In the transmembrane pathway, water moves across the plasma membranes, with a
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short visit to the cell wall space. At the endodermis, the apoplast pathway is blocked by the
Casparian strip. The intimate contact between the soil and the root surface is easily ruptured
when the soil is disturbed. It is for this reason that newly transplanted seedlings and plants need
to be protected from water loss for the first few days after transplantation. Thereafter, new root
growth into the soil reestablishes soil–root contact, and the plant can better withstand water
stress.
Plants absorb water mostly from soil by their roots. As a general rule, a plant's root system is
about as extensive as its shoot system. Water and minerals enter the plant from the soil through
the surface of the fine branches of its roots. Several centimeters of the tip of each root are
covered with numerous root hairs (figure 1). Each root hair is a tubular projection of an
epidermal cell. Root hairs provide the plant with an extensive surface area and maximum access
to the water supply.
Figure 3. Plants have extensive root system to absorb water and minerals
STRUCTURE OF ROOTS
Internally, there are three major parts of a root (Fig 2):
The meristem is at the tip and manufactures new cells; it is an area of cell division and growth.
Behind the meristem is the zone of elongation. In this area, cells increase in size through food
and water absorption. As they grow, they push the root through the soil. The zone of maturation
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is directly beneath the stem. Here, cells become specific tissues such as epidermis, cortex, or
vascular tissue.
A root's epidermis is its outermost layer of cells (Fig 4). These cells are responsible for
absorbing water and minerals dissolved in water. Cortex cells are involved in moving water
from the epidermis to the vascular tissue (xylem and phloem) and in storing food. Vascular
tissue is located in the center of the root and conducts food and water.
Figure 4. cross section of roots
Externally, there are two areas of importance: the root cap and the root hairs (Figure 3). The root
cap is the root's outermost tip. It consists of cells that are sloughed off as the root grows through
the soil. Its function is to protect the root meristem. Root hairs are delicate, elongated epidermal
cells that occur in a small zone just behind the root's growing tip. They generally appear as fine
down to the naked eye. Their function is to increase the root's surface area and absorptive
capacity. Root hairs usually live 1 or 2 days. When a plant is transplanted, they are easily torn off
and also may dry out in the sun.
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Let’s consider how water moves within the root, and the factors that determine the rate of
Water uptake into the root. Water Moves in the Root via the Apoplast, Transmembrane,
and Symplast Pathways
In the soil, water is transported predominantly by bulk flow. However, when water comes in
contact with the root surface, the nature of water transport becomes more complex. From the
epidermis to the endodermis of the root, there are three pathways through which water can flow
Figure 5: the apoplast, transmembrane, and symplast pathways.

1. Apoplastic pathway
The apoplastic movement of water in plants occurs exclusively through the cell wall without
crossing any membranes. The apoplast is the continuous system of cell walls and intercellular air
spaces in plant tissues. The cortex receive majority of water through apoplastic way as loosely
bound cortical cells do not offer any resistance. But the movement of water in root beyond cortex
apoplastic pathway is blocked by casparian strip present in the endodermis.
2. Transmembrane pathway (Fig 5): - The transmembrane pathway is the route followed by
water that sequentially enters a cell on one side, exits the cell on the other side, enters the next in
the series, and so on. In this pathway, water crosses at least two membranes for each cell in its
path (the plasma membrane on entering and on exiting). Transport across the tonoplast may also
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be involved.
3. Symplastic pathway (Fig 5): - The movement of water from one cell to other cell through the
plasmodesmata is called the symplastic pathway of water movement. This pathway comprises
the network of cytoplasm of all cells inter-connected by plasmodermata.
Water potential is the potential energy of water relative to pure free water (e.g. deionized water)
in reference conditions. It quantifies the tendency of water to move from one area to another due
to osmosis, gravity, mechanical pressure, or matrix effects including surface tension. Water
potential is measured in units of pressure and is commonly represented by Ψw. This concept has
proved especially useful in understanding water movement within plants, animals, and soil. Plant
physiologists discuss water relations in terms of water potential (Ψw), an expression of the free
energy of water in a system. In this concept, water moves from a high to a low water potential.
The water potential of pure water is set arbitrarily at 0. In a solution or under tension, the water
potential is lower, a negative value. Plant water transport is readily explained in terms of water
potential.
Water enters a root hair cell from the soil because cell solutes lower the water potential (Ψw). In
the xylem vessels, water is under tension and has a still lower Ψw, and water is pulled from the
endodermis and across the root. Up in the leaf, the bundle sheath cells contain solutes and have a
low Ψw; they pull water from the xylem. Photosynthetic cells in the leaf, with their high sugar
content, have a still lower Ψw and draw water from adjacent cells in a chain extending to the
bundle sheath. As water evaporates from the leaf cells and diffuses out of the leaf, the lowered
Ψw of these cells pulls water from adjacent cells, generating the transpiration pull. Outside the
leaf, if the relative humidity is low, Ψw is low and water diffuses out of the intercellular spaces.
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Components of water potential

Much different potential affect the total water potential and sum of these potentials determines
the overall water potential and the direction of water flow:
ΨW = ΨS + ΨΡ + ΨΟ + ΨM + Ψg
Where:
ΨW is water potential
ΨS is solute potential
ΨP is pressure potential and
ΨO is osmotic potential
ΨM is matrix potential
ΨG gravitational potential
1. Pressure potential
Pressure potential is based on mechanical pressure, and is an important component of the total
water potential within plant cells. Pressure potential is increased as water enters a cell. As water
passes through the cell wall and cell membrane, it increases the total amount of water present
inside the cell, which exerts an outward pressure that is retained by the structural rigidity of the
cell wall. The pressure potential in a living plant cell is usually positive. In plasmolysed cells,
pressure potential is almost zero. Negative pressure potentials occur when water is pulled
through an open system such as a plant xylem vessel. Withstanding negative pressure potentials
(frequently called tension) is an important adaptation of xylem vessels.
2. Solute potential
Pure water is usually defined as having a solute potential (Ψs) of zero, and in this case, solute
potential can never be positive. The relationship of solute concentration (in molarity) to solute
potential is given by the van’t Hoff equation:
Ψs = miRT
Where
m - The concentration in molarity of the solute,
i - The van’t Hoff factor, the ratio of amount of particles in solution to amount of formula units
dissolved,
R - The ideal gas constant, and
T- is the absolute temperature.
17
Water Enters the Cell along a Water Potential Gradient

In this section we will illustrate the osmotic behavior of plant cells with some numerical
examples. First imagine an open beaker full of pure water at 20°C. Because the water is open to
the atmosphere, the hydrostatic pressure of the water is the same as atmospheric pressure (Ψp =0
MPa) There are no solutes in the water, so Ψs= 0 MPa; therefore the water potential is 0 MPa
(Ψw= Ψs + Ψp)
Five examples illustrating the concept of water potential and its components;
(A) Pure water. (B) A solution containing 0.1 M sucrose. (C) A flaccid cell (in air) is dropped in
the 0.1 M sucrose solution. Because the starting water potential of the cell is less than the water
potential of the solution, the cell takes up water. After equilibration, the water potential of the
18
cell rises to equal the water potential of the solution, and the result is a cell with a positive turgor
pressure. (D) Increasing the concentration of sucrose in the solution makes the cell lose water.
The increased sucrose concentration lowers the solution water potential, draws water out from
the cell, and thereby reduces the cell’s turgor pressure. In this case the protoplast is able to pull
away from the cell wall (i.e, the cell plasmolyzes) because sucrose molecules are able to pass
through the relatively large pores of the cell walls. In contrast, when a cell desiccates in air (e.g.,
the flaccid cell in panel C) plasmolysis does not occur because the water held by capillary forces
in the cell walls prevents air from infiltrating into any void between the plasma membrane and
the cell wall. (E) Another way to make the cell lose water is to press it slowly between two
plates. In this case, half of the cell water is removed, so cell osmotic potential increases by a
factor of 2.
Now imagine dissolving sucrose in the water to a concentration of 0.1 M (Figure B). This
addition lowers the osmotic potential (Ψs) to –0.244 MPa (see Table 2) and decreases the water
potential (Ψw) to –0.244 MPa. Next consider a flaccid, or limp, plant cell (i.e., a cell with no
turgor pressure) that has a total internal solute concentration of 0.3 M (Figure C). This solute
concentration gives an osmotic potential (Ψs) of –0.732 MPa. Because the cell is flaccid, the
internal pressure is the same as ambient pressure, so the hydrostatic pressure (Ψp) is 0 MPa and
the water potential of the cell is –0.732 MPa.
What happens if this cell is placed in the beaker containing 0.1M sucrose (see Figure C)?
Because the water potential of the sucrose solution (Ψw= –0.244 MPa; see Figure B) is greater
than the water potential of the cell (Ψw= –0.732 MPa), water will move from the sucrose
solution to the cell (from high to low water potential). Because plant cells are surrounded by
relatively rigid cell walls, even a slight increase in cell volume causes a large increase in the
hydrostatic pressure within the cell. As water enters the cell, the cell wall is stretched by the
contents of the enlarging protoplast. The wall resists such stretching by pushing back on the cell.
This phenomenon is analogous to inflating a basketball with air, except that air is compressible,
whereas water is nearly incompressible.
19
As water moves into the cell, the hydrostatic pressure, or turgor pressure (Ψp), of the cell
increases. Consequently, the cell water potential (Ψw) increases, and the difference between
inside and outside water potentials (ΔΨw) is reduced. Eventually, cell Ψp increases enough to
raise the cell Ψw to the same value as the Ψw of the sucrose solution. At this point, equilibrium
is reached (ΔΨw= 0 MPa), and net water transport ceases. Because the volume of the beaker is
much larger than that of the cell, the tiny amount of water taken up by the cell does not
significantly affect the solute concentration of the sucrose solution. Hence Ψs, Ψp, and Ψw of the
sucrose solution are not altered. Therefore, at equilibrium, Ψw (cell) = Ψw (solution) = –0.244
MPa. The exact calculation of cell Ψp and Ψs requires knowledge of the change in cell volume.
However, if we assume that the cell has a very rigid cell wall, then the increase in cell volume
will be small. Thus we can assume to a first approximation that Ψs (cell) is unchanged during the
equilibration process and that Ψs (solution) remains at –0.732 MPa. We can obtain cell
hydrostatic pressure by rearranging as follows: Ψp = Ψw– Ψs= (–0.244) – (–0.732) = 0.488
MPa.s
Water Can Also Leave the Cell in Response to a Water Potential Gradient
Water can also leave the cell by osmosis. If, in the previous example, we remove our plant cell
from the 0.1 M sucrose solution and place it in a 0.3 M sucrose solution (Figure D), Ψw(solution)
(–0.732 MPa) is more negative than Ψw (cell) (–0.244 MPa), and water will move from the
turgid cell to the solution. As water leaves the cell, the cell volume decreases. As the cell volume
decreases, cell Ψp and Ψw decrease also until Ψw (cell) = Ψw (solution) = –0.732 MPa. From
the water potential equation we can calculate that at equilibrium, Ψp= 0 MPa. As before, we
assume that the change in cell volume is small, so we can ignore the change in Ψs.
.
If we then slowly squeeze the turgid cell by pressing it between two plates (Figure E), we
effectively raise the cell Ψp, consequently raising the cell Ψw and creating a ΔΨw such that
water now flows out of the cell. If we continue squeezing until half the cell water is removed and
then hold the cell in this condition, the cell will reach a new equilibrium. As in the previous
example, at equilibrium, ΔΨw = 0 MPa, and the amount of water added to the external solution
is so small that it can be ignored. The cell will thus return to the Ψw value that it had before the
squeezing procedure. However, the components of the cell Ψw will be quite different.
20
Because half of the water was squeezed out of the cell while the solutes remained inside the cell
(the plasma membrane is selectively permeable), the cell solution is concentrated twofold, and
thus Ψs is lower (–0.732 x 2 = –1.464 MPa). Knowing the final values for Ψw and Ψs, we can
calculate the turgor pressure, using Equation 3.6, as Ψp= Ψw – Ψs = (–0.244) – (–1.464) = 1.22
MPa. In our example we used an external force to change cell volume without a change in water
potential. In nature, it is typically the water potential of the cell’s environment that changes, and
the cell gains or losses water until its Ψw matches that of its surroundings.
One point common to all these examples deserves emphasis: Water flow is a passive process.
That is, water moves in response to physical forces, toward regions of low water potential or low
free energy.There are no metabolic “pumps” (reactions driven by ATP hydrolysis) that push
water from one place to another. This rule is valid as long as water is the only substance being
transported. When solutes are transported, however, as occurs for short distances across
membranes and for long distances in the phloem, then water transport may be coupled to solute
transport and this coupling may move water against a water potential gradient.
For example, the transport of sugars, amino acids, or other small molecules by various
membrane proteins can “drag” up to 260 water molecules across the membrane per molecule of
solute transported (Loo et al. 1996). Such transport of water can occur even when the movement
is against the usual water potential gradient (i.e., toward a larger water potential) because the loss
of free energy by the solute more than compensates for the gain of free energy by the water. The
net change in free energy remains negative. In the phloem, the bulk flow of solutes and water
within sieve tubes occurs along gradients in hydrostatic (turgor) pressure rather than by osmosis.
Thus, within the phloem, water can be transported from regions with lower water potentials (e.g.,
leaves) to regions with higher water potentials (e.g., roots). These situations notwithstanding, in
the vast majority of cases water in plants moves from higher to lower water potentials.
3. Matrix potential: When water is in contact with solid particles (e.g., clay or sand particles
within soil) adhesive intermolecular forces between the water and the solid can be large and
important. The forces between the water molecules and the solid particles in combination with
attraction among water molecules promote surface tension and the formation of menisci within
21
the solid matrix. Force is then required to break these menisci. The magnitude of matrix potential
depends on the distances between solid particles--the width of the menisci and the chemical
composition of the solid matrix. In many cases, matrix potential can be quite large and
comparable to the other components of water potential discussed above. It is worth noting that
matrix potentials are very important for plant water relations. Strong (very negative) matrix
potentials bind water to soil particles within very dry soils. Plants then create even more negative
matrix potentials within tiny pores in the cell walls of their leaves to extract water from the soil
and allow physiological activity to continue through dry periods.
4. Gravity (Ψg): Contributions due to gravity which is usually ignored unless referring to the
tops of tall trees.
2.2.2. Mechanism of Water Absorption

Absorption of water occurs along gradients of decreasing potential from the substrate to the
roots. However, the gradient is produced differently in slowly and rapidly transpiring plants,
resulting in two absorption mechanisms. Active or osmotic absorption occurs in slowly
transpiring plants where the roots behave as osmometers whereas passive absorption occurs in
rapidly transpiring plants where water is pulled in through the roots, which act merely as
absorbing surfaces.
1. Active Absorption: Active transport uses energy to pump molecules against a concentration
gradient. Active transport is carried out by membrane-proteins. Hence different proteins in the
membrane play a major role in both active as well as passive transport. Pumps are proteins that
use energy to carry substances across the cell membrane. These pumps can transport substances
from a low concentration to a high concentration (‘uphill’ transport). Transport rate reaches a
maximum when all the protein transporters are being used or are saturated. Like enzymes the
carrier protein is very specific in what it carries across the membrane. These proteins are
sensitive to inhibitors that react with protein side chains. Water is absorbed due to activities
going on in roots. Absorption of water occurs with the help of energy in the form of ATP, which
is released due to metabolic activities of root cells such as respiration. Absorption takes place
against concentration gradient - even when the concentration of cell sap is lower than that of soil
water.
22
2. Passive Absorption by Transpiring Plants: The force bringing about absorption of water by
transpiring plants originates in the leaves and is transmitted to the roots or the lower end of cut
stems through the sap stream in the xylem. Evaporation of water from leaf cells decreases their
water potential, causing water to move into them from the xylem of the leaf veins. This reduces
the potential in the xylem sap, and the reduction is transmitted through the cohesive water
columns to the roots where the reduced water potential causes inflow from the soil. In this
situation water can be regarded as moving through the plant in a continuous, cohesive column,
pulled by the matric or imbibitional forces developed in the evaporating surfaces of stem and leaf
cell walls. Evaporation of water from the twigs and branches of bare deciduous trees and from
cut flowers also causes upward flow in the xylem.
2.2.3. Factors Affecting Water Absorption

A. Physical factors: The soil and atmosphere are the chief physical factors which determine the
flow rate of water through plant.
Soil factors:
I. Soil water content: The plant roots can easily absorb the soil moisture in between field
capacity and permanent wilting point. When the soil moisture decrease below the wilting point,
plant roots have to exert more pressure and thus rate of absorption decreases. On the other hand,
when the soil is completely saturated with water, then soil temperature and aeration are poor and
this condition also affects the absorption of water.
II) Soil temperature: Soil temperature is known to influence water absorption and ultimately
transpiration to a considerable extent. In many plants, water absorption below a soil temperature
of 10 0C is reduced sharply and 25 0C soil temperature up take of water is slowed down. In most
instances, temperature above 40 0C does not support water absorption and plant can show signs
of wilting. A freezing temperature reduces water absorption because of following causes.
A. Decreased root growth
B. Increased viscosity of water
C. Increased resistance to movement of water in to roots thus is caused by decreased
permeability of cell membrane and the increased viscosity.
23
III. Soil aeration and flooding: Most of crop plants are not able to water while standing under
water logged conditions. The following are the possible reasons of flood injury.
A. Poor availability of oxygen and occurrence of higher CO2 concentration around roots.
B. Accumulation of toxic substances either in the submerged roots or around them.
C. Changes in pattern of ion up take resulting in the accumulation of some toxic ions.
In water logged condition, the availability of oxygen is reduced which affects respiratory actively
of roots. In addition, CO2 concentration is increased and it affects permeability of membranes
and adversely influences water up take. Reduced oxygen also affects root growth adversely.
B. Atmospheric factor: The amount of soil moisture that is available to a plant is determined by
the moisture characteristics of the soil, the depth to which the plant roots extend and the
proliferation or density of the roots. Soil moisture characteristics, such as field capacity and
wilting percentage are peculiar to a soil and are a function of the texture and organic matter.
Little can be done to alter these limits to any great extent. Greater possibilities lie in changing the
characteristic of the plant enabling it extend its rooting system deeper into the soil, thereby
enlarging its reservoir of water. The density of roots proliferation is important.
Water is an unsaturated soil moves very slowly, and only a distance of a few cm. To utilize
effectively the moisture stored in the soil profile, roots must continue to proliferate into
unexploited zones throughout the plants growth cycle. During favorable growing periods, roots
often elongate so rapidly that satisfactory moisture contacts can be maintained even when the
soil moisture content declines. Where transpiration is effected due to the different atmosphere
factors such as wind velocity, humidity, sunlight, etc when temperature and wind velocity are
more sunlight for longer period and humidity are less, under such conditions, transpiration is
more. The increased rate of transpiration results more water uptake.
C. Biological factors: Root system is the plant factor which is directly related to the absorption
of water from soil. Under favorable soil water, potential soil temperature, aeration, and roots
system of the plants strongly influence the uptake of water. When growth of roots (root system)
is more, uptake of water is also more under favorable soil conditions. Root growth is influenced
by soil and more therefore agronomic management practices can help to improve root growth.
24
Other plant factors such as morphology of leaves, stomatal mechanism and growth stage of the
crop influence the rate of transpiration. The increased rate of transpiration results more water
absorption. Good root system has developed during favorable growing periods; a plant can draw
its moisture supply from deeper soil layers. Plants vary genetically in their rooting
characteristics. Vegetable crops such as onions and potatoes have a spare rooting system and are
unable to use all the soil water within the root zone. Forage grasses, sorghum, maize and such
other crops have very fibrous, dense roots. Lucerne has a deep root system. Whether plant is an
annual or perennial is another factor affecting its moisture relations. An annual plant must extend
its roots down into the soil to make availability root depth, and needs only to extend its small
roots and hairs to be able to utilize the entire amount of available soil water.
Plants may be limited in their rooting by factors other than genetic. High water table, shallow
soils and an impermeable formation near the ground surface restrict the depth rooting. Fertility
and salt status of the soil influence the rooting of plants crop management practices, such as
cutting the top growth at different physiological stages and the cultivation and cutting of surface
roots after rooting habits. The rooting pattern of common and crop plants vary widely from soil.
For example, roots of maize crop have been found to extend as deep as 1.5 meters in medium to
textured soils, while in a fine textured soil the crop has a shallower root system.
Effective Root zone: Effective root zone is the depth from which the roots of average mature
plant are capable of reducing soil moisture to the extent that it should be replaced by irrigation. It
is not necessarily to have maximum root depth for ant given plant especially for plants that have
a long taproot. Root development of any crop varies widely with the type of soil and other.
Moisture extraction pattern within root zone: The moisture extraction pattern shows the
relative amounts of moisture extracted from different depths within the crop root zone. It is seen
that about 40 percent of the total moisture used is extracted from first quarter of the root zone, 30
percent from the second, 20 percent from third and only 10 percent from last quarter. This
indicates that the need for making soil moisture measurements at different depths within the root
zone in order to have estimate of soil moisture status.
25
2.3. Transpiration
Transpiration is the evaporation of water from the aerial parts of plants. More than 90% of the
water entering a plant evaporates primarily into the leaf air spaces and then passes through the
stomata into the atmosphere. This evaporative water loss occurs mainly through the stomata
(90%) and to a lesser extent through the cuticle (10%). When stomata are open, gas exchange
occurs freely between the leaf and the atmosphere. Water vapor and oxygen (from
photosynthesis) diffuse out of the leaf while carbon dioxide diffuses into the leaf (fig. 4.2a) . It is
the action of the guard cells that regulates the rate of water lost through transpiration and, at the
same time, regulates the rate of photosynthesis by controlling the CO2 uptake. Each stoma is
surrounded by a pair of guard cells, which have unevenly thickened walls.
The walls of the guard cells that border the stoma are thicker than the outer walls. When guard
cells become turgid they can only expand outward owing to the radial orientation of cellulose
fibrils; this outward expansion of the guard cells opens the stomata. Stomata are generally open
during daylight and closed at night. As long as the stomata are open, both transpiration and
photosynthesis occur, but when water loss exceeds uptake, the guard cells lose turgor and close
the stomata (fig. 4.2b).
Figure 6: Transpiration is the basic driving force behind water movement in the xylem. (a)
When stomata are open, both transpiration and photosynthesis occur as H2O molecules diffuse
out of the leaves and CO2 molecules diffuse in Guard cells. (b) When guard cells are turgid,
stoma are open, and when guard cells are flaccid, stoma are closed.
26
Transpiration pull is a method that water travels through the plant. This path that water travels
may begin in the roots 3 to 6 m or more beneath the surface and travel up the trunk of the tree to
the topmost leaves that may be more than 90 m above the soil. This occurs because there are
continuous tubular pathways of xylem that run throughout the plant, extending from the young
roots up through the stem and branches to the tiny vein lets of the leaves. The water is raised
through the columns of a plant through a combination of factors. Capillary action will allow
water to rise in narrow tubes, and the heights attained are inversely proportional to the diameter
of the tube. Plants also have root pressure, but this is considerably less than needed to raise water
to the top of trees.
The pulling force due to evaporation of water from leaves and stems provides the primary
mechanism for water to reach to the top of trees (this is often referred to as the cohesion-tension
theory). Water molecules are electrically neutral, but are asymmetrical in shape. This results in
the molecules having a very slight positive charge at one end and a very slight negative charge at
the other end. As a result, the molecules are polar. When the negatively charged end of one water
molecule comes close to the positively charged end of another water molecule, weak hydrogen
bonds hold the molecules together. This permits a certain amount of tension to exist.
When water evaporates from the mesophyll cells in a leaf and diffuses out of the stomata
(transpires), the cells involved develop lower water potential than the adjacent cells. Because the
adjacent cells then have correspondingly higher water potential, replacement water moves into
the first cells through osmosis. This continues across the rows of mesophyll cells until a small
27
vein is reached. As transpiration occurs it creates a “pull” or tension on water columns, drawing
water from one molecule to another all the way through an entire span of xylem cells. The
cohesion strength of the water in the columns is usually more than adequate to move the water to
the top of a tree. The passage of water is partly through cell protoplasm and partly through
spaces between cells, between cellulose fibers in the walls, and in the centers of dead cells.
2.3.1 Types of Transpiration

Although a small amount of water vapor may be lost through small openings (called lenticels) in
the bark of young twigs and branches, the largest proportion by far (more than 90%) escapes
from leaves. Indeed, the process of transpiration is strongly tied to leaf anatomy. The outer
surfaces of a typical vascular plant leaf are covered with a multilayered waxy deposit called the
cuticle. The principal component of the cuticle is cutin, a heterogeneous polymer of long-chain
typically 16 or 18 carbons hydroxylated fatty acids. Ester formation between the hydroxyl and
carboxyl groups of neighboring fatty acids forms cross-links, establishing an extensive
polymeric network. The cutin network is embedded in a matrix of cuticular waxes, which are
complex mixtures of long-chain (up to 37 carbon atoms) saturated hydrocarbons, alcohols,
aldehydes, and ketones. Because cuticular waxes are very hydrophobic, they offer extremely
high resistance to diffusion of both liquid water and water vapor from the underlying cells. The
cuticle thus serves to restrict evaporation of water directly from the outer surfaces of leaf
epidermal cells and protects both the epidermal and underlying mesophyll cells from potentially
28
lethal desiccation. The integrity of the epidermis and the overlying cuticle is occasionally
interrupted by small pores called stomata (singular stoma). Each pore is surrounded by a pair of
specialized cells, called guard cells. These guard cells function as hydraulically operated valves
that control the size of the pore. The interior of the leaf is comprised of photosynthetic
mesophyll cells. The somewhat loose arrangement of mesophyll cells in most leaves creates an
interconnected system of intercellular air spaces. This system of air spaces may be quite
extensive, accounting for up to 70 percent of the total leaf volume in some cases. Stomata are
located such that, when open, they provide a route for the exchange of gases (principally carbon
dioxide, oxygen, and water vapor) between the internal air space and the bulk atmosphere
surrounding the leaf.
Because of this relationship, this space is referred to as sub-stomatal space. The cuticle is
generally impermeable to water and open stomata provide the primary route for escape of water
vapor from the plant. Transpiration may be considered a two-stage process: (1) the evaporation
of water from the moist cell walls into the sub-stomatal air space and (2) the diffusion of water
vapor from the substomatal space into the atmosphere. It is commonly assumed that evaporation
occurs primarily at the surfaces of those mesophyll cells that border the sub-stomatal air spaces.
There are two types of transpiration

1. Stomatal transpiration
2. Cuticular transpiration
1. Stomatal transpiration: The diffusion of water vapor from the sub-stomatal space into the
atmosphere is relatively straightforward. Once the water vapor has left the cell surfaces, it
diffuses through the sub-stomatal space and exits the leaf through the stomatal pore. Diffusion of
water vapor through the stomatal pores, known as stomatal transpiration, accounts for 90 to 95
percent of the water loss from leaves.
2. Cuticular transpiration: The remaining 5 to 10 percent is accounted for by cuticular
transpiration. Although the cuticle is composed of waxes and other hydrophobic substances and
is generally impermeable to water, small quantities of water vapor can pass through. The
contribution of cuticular transpiration to leaf water loss varies considerably between species. It is
29
to some extent dependent on the thickness of the cuticle. Thicker cuticles are characteristic of
plants growing in full sun or dry habitats, while cuticles are generally thinner on the leaves of
plants growing in shaded or moist habitats. Cuticular transpiration may become more significant,
particularly for leaves with thin cuticles, under dry conditions when stomatal transpiration is
prevented by closure of the stomata.
A fully grown tree may lose several hundred gallons (a few cubic meters) of water through its
leaves on a hot, dry day. About 90% of the water that enters a plant's roots is used for this
process. The transpiration ratio is the ratio of the mass of water transpired to the mass of dry
matter produced; the transpiration ratio of crops tends to fall between 200 and 1000 (i.e., crop
plants transpire 200 to 1000 kg of water for every kg of dry matter produced).
Transpiration rate of plants can be measured by a number of techniques, including potometers,

lysimeters, porometers, and heat balance sap flow gauges. Desert plants and conifers have
specially adapted structures, such as thick cuticles, reduced leaf areas, sunken stomata and hairs
to reduce transpiration and conserve water. Many cacti conduct photosynthesis in succulent
stems, rather than leaves, so the surface area of the shoot is very low. Many desert plants have a
special type of photosynthesis, termed crassulacean acid metabolism or CAM photosynthesis in
which the stomata are closed during the day and open at night when transpiration will be lower.
2.3.2. Mechanism of Transpiration

The transport of water upward from roots to shoots in the xylem is governed by differences in
water (or osmotic) potential, and these differences account for water movement from cell to cell
or over long distances in the plant. Several factors, including environmental pressure and solute
concentration, contribute to water potential, with water always moving from an area of high
water potential (higher free energy, more water) to lower potential (less free energy, less water).
The process is facilitated by osmosis, root pressure, and the physical and chemical properties of
water. Transpiration creates a lower osmotic potential in the leaf, and the TACT (transpiration,
adhesion, cohesion, and tension) mechanism describes the forces that move water and dissolved
nutrients up the xylem.
30
Water vapor moves from the leaf to the

atmosphere by diffusion through stomata
2.3.3. Mechanism of Stomata Opening and Closing
The mechanism of stomatal opening and closing depends upon the turgidity of guard cells. When
the turgidity increases, the pore opens whereas, it closes when turgidity decreases. Entry of water
takes place by the osmotic pressure, which cause the elongation of guard cell by the radial
orientation of cellulose fibrils. As the water enters the guard cells, a turgidity increase that pulls
the cells causes the opening of stomata.
31
Mechanism for opening of stomata
Because ABA- does not easily pass through
Membranes, under conditions of water

Stress more ABA reaches guard cells
The closing of stomata has the reverse process of opening. During the day time, water enters the
cells due to the less water potential that creates high concentration of solutes. Blue light of day
light, activates proton pumps that is detected by the photoreceptor. By the activation of proton
pumps, protons enter in the guard cells from the cytoplasm. Due to the entry of protons, the
proton motive force is created that opens the voltage operated channels in the membrane that
passes the positive potassium ions into the cell. As the concentration of potassium increases in
the cell, chloride ions enters via chloride-protons symport mechanism, into the cell to balance the
cell’s internal charges. Blue light also induces the production of malate that increases the cell
concentration, supports the increase in turgidity by opening the pores of the stomata.
2.3.4. Significance of Transpiration

Plant growth and development relies on water for transpiration, photosynthesis, and respiration.
The unique ability of water to regulate temperature, dissolves molecules of life, and allows gas
exchange, is essential for all life on earth.
32
Thereby, Transpiration is essential for

Evaporative cooling: - plants are able keep cool when they are in direct sun light through
the evaporation of water that occurs in transpiration. As water changes from the liquid to gas
phase, heat energy is lost and the plant is cooled. Plants rely on transpiration for evaporative
cooling so that despite being exposed to direct sun light, their tissues don’t over heated.
Co2 acquisition: - all the carbon incorporated in to carbohydrate through photosynthesis

comes from atmospheric Co2 entering through the pores in the leaves called stomata. Water loss
through the stomata is a continuous process that occurs as long as stomata are open. Plants are
able to close their stomata to restrict water loss during times of drought or high temperature, but
this directly reduces photosynthetic output because less Co2 enters the leaves.
 Maintaining turgor: - since 90% of plant tissue constitutes water, the structure of plant
tissues depends on cell turgidity and since plant cells are leaky, water needs to be continually
taken up (think of a plant cell like an inflated tire with a puncture). Cell expansion, a driving
force of growth, is also driven by cellular water pressure.
Mineral nutrient uptake: - in addition to carbon assimilation from the air, plants
incorporate minerals dissolved in water taken up from the soil. These are distributed throughout
the plant by way of transpiration process.
2.3.5. Factors Affecting Rate of Transpiration

The rate of transpiration is directly related to the degree of stomatal opening, and to the
evaporative demand of the atmosphere surrounding the leaf. The amount of water lost by a plant
depends on its size, along with the surrounding light intensity, temperature, humidity, and wind
speed (all of which influence evaporative demand). Soil water supply and soil temperature can
influence stomatal opening, and thus transpiration rate.
Environmental factors that affect the rate of transpiration
1. Light: - Stomata of most plants open at sunrise and close in darkness, allowing entry of CO2
needed for photosynthesis during the day time. Light intensity influences the rate of opening and
the final aperture size, bright light causing a wider aperture. Greater light intensity increases the
rate of transpiration because it causes the stomata to open, so increasing evaporation through the
33
stomata. This is largely because light stimulates the opening of the stomata. Light also speeds up
transpiration by warming the leaf.
2. Temperature: Plants transpire more rapidly at higher temperatures because water evaporates
more rapidly as the temperature rises.
Provides the latent heat of vaporisation
Increases the kinetic energy so faster diffusion
Warms the air so lowers the of the air, so w gradient is steeper
High temperatures (30-350C) cause the stomata to close due to water stress or increase in CO2
concentration within the leaf.
3. Humidity: The basis of transpiration is the diffusion of water molecules from an area of high
concentration within the leaf to an area of lower concentration in the atmosphere. Unless the
atmospheric relative humidity is 100%, the air is relatively dry compared with the interior of a
leaf, where the intercellular spaces are saturated with water vapor. As long as stomata are open, a
continuous stream of water vapor transpires from the leaf, creating a pull on the water column
that extends from the leaf through the plant to the soil. When the surrounding air is dry, diffusion
of water out of the leaf goes on more rapidly. Air inside leaf is saturated (RH=100%). The lower
the relative humidity outside the leaf the faster the rate of transpiration as the  gradient is
steeper.
4. CO2 Concentration: - Low concentration of CO2 cause stomata to open and removal of CO2
during photosynthesis by parenchyma and mesophyll cells, results in opening of stomata of most
species in the day. High CO2 concentration causes the stomata to close in the light and dark.
5. Wind (Air Movement):- increase air movement increases the rate of transpiration as it moves
the saturated air from around the leaf so the w gradient is steeper. On hot, dry, windy days the
high rate of transpiration frequently causes the stomata to close early, resulting in a near
shutdown of photosynthesis as well as transpiration. A fine balance must be struck in this
photosynthesis-transpiration dilemma to allow enough CO2 for photosynthesis while at the same
time preventing excessive water loss. Some plants have evolved an alternate pathway for CO 2
uptake at night when rates of transpiration are lower (see CAM Pathway later in this chapter).
Other plants have morphological or anatomical adaptations that reduce rates of transpiration
34
while keeping the stomata open. These physiological and anatomical adaptations are most
common in xerophytes, plants occurring in arid environments.
6. Soil water: - A plant cannot continue to transpire rapidly if its water loss is not made up by
replacement from the soil. When absorption of water by the roots fails to keep up with the rate of
transpiration, loss of turgor occurs, and the stomata close. This immediately reduces the rate of
transpiration. If the loss of turgor extends to the rest of the leaf and stem, the plant wilts. As
water potential decreases (water stress increases), the stomata close.
7. Guard Cell Uptake of K+: When the stomata open, relatively large quantities of K+ move
from the surrounding cells into the guard cells. Light causes a building of K+ in guard cells, as
does CO2 free air. When leaves are transferred to the dark, K+ moves out of guard cells into the
surrounding cells and stomata close. Stomata also close in response to the application of abscisic
acid whose application causes loss of K+ from the guard cells. Blue light under low light intensity
can cause stomatal opening when red light has no effect by acting directly on guard
Mechanism for opening of stomata
8. plant hormones: Cytokine promotes stomata opening while abascicic acid inhibits stomata
opening
Intrinsic Factors Affecting the Rate of Transpiration.
1. Leaf surface area
2. Thickness of epidermis and cuticle
3. Stomatal frequency
4. Stomatal size
5. Stomatal position
35
Ecologists classify plants according to their response to water as follows:

A. Hydrophytes – plants that grow where water is super abundant.
B. Mesophytes – plants that grow where water availability is intermediate
C. Xerophytes – Plants that grow where water is scarce.
Solutes strongly influence water potential, so ecologists further classify plants that are sensitive
to relatively high salt concentrations as GLYCOPHYTES and those that are able to grow in the
presence of high salts as HALOPHYTES. Xerophytes are plants that escape, resist or endure
drought. They can be described as being hardy. Annual plants in the desert escape the drought by
existing as dormant seeds during the dry season.
Succulent species resist drought by storing water in their succulent tissues (cactus). Enough
water is stored and its rate of loss is so extremely low (due to an exceptionally thick cuticle and
stomatal closure during the day time. Non-succulent perennial perennials avoid drought by
having extensive shallow root system capable of absorbing surface moisture after rain storm,
reduction in size of leaf blades (which increases heat transfer by convection, lowers leaf
temperature and thus reduces transpiration, sunken stomata, shedding of leaves during dry
periods and such other factors as heavy pubescence on leaf surfaces.
36
CHAPTER 3: PHOTOSYNTHESIS
Photosynthesis is the single most important chemical process on earth. It is the process by which
plants use solar energy to manufacture food. The term means “putting together with light,” and
the process of photosynthesis uses solar energy to form simple sugars from water and carbon
dioxide gas. Later these sugars are converted into starch, protein, or fat; and we eat them as fruits
and vegetables. Thus photosynthesis changes light energy into food (chemical) energy.
Photosynthesis sustains green plants and as a result all other living things as well. Both directly
and indirectly green plants generate most of the world’s chemical energy. Wood and fossil fuels,
coal, oil and natural gas formed from plants and animals that lived millions of years ago —
provide much of our electricity and heat. Green plants are the source of gasoline that we use to
power buses and cars. Fresh fruits, vegetables and grain, as well as meat from animals that eat
plants, give us the energy to work and play and think.
All of this energy originally came from the sun, and it is available to us only as a result of
photosynthesis. People have dreamed of duplicating this process, and biochemists are still trying
to unravel its complexities. They know that it involves a sequence of chemical changes that takes
place in a millionth of a second. They also know that most chlorophyll molecules and certain
plant pigments act as antennas which receive and absorb solar energy, then transmit it to a pair of
very special chlorophyll molecules that convert it to chemical energy. When the chemical
dynamics of this process are finally understood, people will be closer to the extraordinary goal of
converting sunlight directly to chemical energy. Until that goal is achieved, we remain totally
dependent on green plants for life.
Historical aspects: The study of the process of photosynthesis has attracted the attention of
scientists for over 200 years. It is difficult to mention the contribution of each scientist to the
extensive work done in this field. However, some of the important contributions are as
37
The process of photosynthesis involves the use of light energy to convert carbon dioxide and
water into sugar, oxygen, and other organic compounds. More recently, using radioactive
isotopes, it has been determined that the actual reaction is:
12 H2O + 6 CO2 + Energy C6H12O6 + 6O2 + 6H2O
Water appears on both sides of the equation because water is both used as a reactant and released
as a product. This process is an extremely complex one, occurring in two stages. The first stage,
called the light reactions of photosynthesis, requires light energy. The products of the light
reactions are then used to produce glucose from carbon dioxide and water. Because the
reactions in the second stage do not require the direct use of light energy, they are called the dark
reactions of photosynthesis.
In the light reactions, electrons derived from water are “excited” (raised to higher energy levels)
in several steps, called photo-systems I and II. In both steps, chlorophyll absorbs light energy
that is used to excite the electrons. Normally, these electrons are passed to a cytochrome
containing electron transport chain. In the first photo-system, these electrons are used to generate
ATP. In the second photo-system, excited electrons are used to produce the reduced coenzyme
nicotinamide adenine dinucleotide phosphate (NADPH). Both ATP and NADPH are then used in
the dark reactions to produce glucose.
3.1. The Nature of Light

The energy available on earth is can be traced to the sun (e.g., biomass, wind, and incoming
radiation), cosmic evolution preceding the origin of our solar system (nuclear power), lunar
motion (tidal power), or the earth’s core (geothermal power). Of these, sunlight provides the
most energy for organisms. Sunlight originates inside the sun, where a temperature of
10,000,000 0c fuses hydrogen to form helium and release gamma rays, which then produce
electrons and photons: H+HHe + gamma rays + energy gamma rays + electrons + photons +
energy. Life depends on transforming the energy confined in photons of sun light into chemical
energy before it is transformed into heat. The suns total energy output is about 3.8 trillion
(3,800,000,000,000,000,000,000) megawatts of electricity. Earth intercepts only
about1/2,000,000,000 of this energy; this amounts to the energy equivalent of about 2x10 14 tons
of coal per year. of this,
38
30% is reflected back to space. This what astronauts see while orbiting the earth?
50% is absorbed, converted to heat, and reradiated.
19% powers the hydrological cycle, creates winds, and drives photosynthesis. of this 19%,
only 0.05% to 1.5% is incorporated into plant material. However, this relatively small amount of
energy sustains life. Without photosynthesis, almost all life would quickly disappear from earth.
The amount of sunlight energy at the earth’s surface is about 2 million trillion calories per
second, an amount of energy equivalent to that of 400 million atomic bombs of the size dropped
during world War ΙΙ. Stated another way, the annual amount of energy that strikes the earth as
sunlight is 15,000 times greater than the worlds present supply of energy. This is more than 20
billion times greater than our present rate of energy consumption. Despite this vast resource of
energy, we have learnt relatively little about how to use renewable sources of energy such as
sunlight. These so-called alternative sources of energy differ from oil, gas, and nuclear power
because they are plentiful and free of pollution. Today, renewable forms of energy (e.g.,
hydropower and biomass) supply only 18% of the world’s energy needs, while nuclear power
supplies 4%. The rest of our energy demands are huge. For example, to exert the power
equivalent to the amount of electricity used by USA manufacturers in one week, one worker,
doing average manual labor for fifty 40 hour weeks per year, would have to work for 145, 193,
740 years.
The sun is the earth’s primary source of radiant energy. A significant portion of the radiant
energy from the sun actually gets converted to other forms of energy once it reaches the earth.
For example, the process of photosynthesis by plants converts the radiant energy from the sun to
chemical energy. Both the atmosphere and the surface of the earth absorb large portions of the
sun’s radiant energy and convert it to heat. The resulting heat contributes to the formation of
winds in our atmosphere. Both these winds and the heat of the sun play a critical role in the
earth’s hydrologic cycle. Visible light is the part of electromagnetic radiation that is visible to
human eyes. It has wave length from about 380nm to 760nm. By passing it through a prism
White light gets separated into the different colors (=wavelengths) of light. Longer wavelengths
are towards red colour and shorter wavelengths are towards violet colour. Wavelengths longer
than red are infrared and shorter than violet are ultra violet. Both these are invisible to human
39
eyes. The energy of light is inversely proportional to its wavelength: longer wavelengths have
less energy than shorter ones.
Principles of Light Absorption by Photosynthetic Pigments: Radiation in the visible range

(380 - 760nm wave length) is utilized in the photosynthetic process. Light has both wave and
particle nature. The particle nature of light is usually expressed in terms of quanta or photons i.e.
discrete packets of energy each having a specific associate wave length. Energy level in each
photon is inversely related with wavelength (E α 1/λ). Light is emitted and absorbed in tiny
packets called photons. These exhibit properties of both waves and particles.
Curve A is the energy output of the sun as a function of wavelength. Curve B is the energy that
strikes the surface of Earth. The sharp valleys in the infrared region beyond 760 nm represent the
absorption of solar energy by molecules in the atmosphere, chiefly water vapor. Curve C is the
absorption spectrum of chlorophyll, which absorbs strongly in the blue (about 430 nm) and the
red (about 660 nm) portions of the spectrum. Because the green light in the middle of the visible
region is not efficiently absorbed, most of it is reflected into our eyes and gives plants their
characteristic green color.
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Figure 8. The solar spectrum and its relation to the absorption spectrum of chlorophyll.
Figure 9 Electromagnetic spectrum. Wavelength (λ ) and frequency (ν ) are inversely related.

Our eyes are sensitive to only a narrow range of wavelengths of radiation, the visible region,
which extends from about 400 nm (violet) to about 700 nm (red). Short-wavelength
(highfrequency) light has high energy content; long wave length (low-frequency) light has low
energy content.
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Fundamental principle of light absorption states that any molecule can absorb only one photo of
light at any given time. Absorbed photon excites one electron. Electrons in ground-state orbital
are the one that are usually excited and each excited electron is driven away from its ground-state
to the distance corresponding to the energy exactly equivalent to the energy of photon absorbed.
This excitation energy absorbed (from photons) by photosynthetic pigment molecules is used in
photosynthesis.
Figure 10. Light absorption and emission by chlorophyll. (A) Energy level diagram. Absorption
or emission of light is indicated by vertical lines that connect the ground state with excited
electron states. The blue and red absorption bands of chlorophyll (which absorb blue and red
photons, respectively) correspond to the upward vertical arrows, signifying that energy absorbed
from light causes the molecule to change from the ground state to an excited state. The
downward-pointing arrow indicates fluorescence, in which the molecule goes from the lowest
excited state to the ground state while re-emitting energy as a photon. (B) Spectra of absorption
and fluorescence. The long-wavelength (red) absorption band of chlorophyll corresponds to light
that has the energy required causing the transition from the ground state to the first excited state.
The short-wavelength (blue) absorption band corresponds to a transition to a higher excited state.
Chlorophyll appears green to our eyes because it absorbs light mainly in the red and blue parts of
the spectrum, so only some of the light enriched in green wavelengths (about 550 nm) is
reflected into our eyes. The distribution of electrons in the excited molecule is somewhat
different from the distribution in the ground state molecule (Figure 9) Absorption of blue light
42
excites the chlorophyll to a higher energy state than absorption of red light because the energy of
photons is higher when their wavelength is shorter. In the higher excited state, chlorophyll is
extremely unstable, very rapidly gives up some of its energy to the surroundings as heat, and
enters the lowest excited state, where it can be stable for a maximum of several nanoseconds (10–
9s). Because of this inherent instability of the excited state, any process that captures its energy
must be extremely rapid.
In the lowest excited state, the excited chlorophyll has four alternative pathways for disposing
of its available energy
1. Excited chlorophyll can re-emit a photon and thereby return to its ground state—a process
known as fluorescence. When it does so, the wavelength of fluorescence is slightly longer (and
of lower energy) than the wavelength of absorption because a portion of the excitation energy is
converted into heat before the fluorescent photon is emitted. Chlorophylls fluoresce in the red
region of the spectrum.
2. The excited chlorophyll can return to its ground by directly converting its excitation energy
state
into heat, with no emission of a photon

3. Chlorophyll may participate in energy transfer, during which excited chlorophyll transfers
its energy to another molecule
4. A fourth process is photochemistry, in which the energy of the excited state causes chemical
reactions to occur. The photochemical reactions of photosynthesis are among the fastest known
chemical reactions. This extreme speed is necessary for photochemistry to compete with the
three other possible reactions of the excited state just described.
Chlorophyll and other pigments can remain in an excited state for extremely short period
(usually one-billionth of a second or nanosecond or even less). Excitation by blue or red light
leads to the same level of final energy for photosynthesis because of greater loss of energy
through heat in the former. However, final energy is transferred to adjacent chlorophyll
molecules by Induction. Resonance until energy reaches to reaction center.
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3.2. Photosynthetic Pigments

The energy of sunlight is first absorbed by the pigments of the plant. All pigments active in
photosynthesis are found in the chloroplast. Structures and absorption spectra of several
photosynthetic pigments are shown in Figures 10. The chlorophylls and bacterio-chlorophylls
(pigments found in certain bacteria) are the typical pigments of photosynthetic organisms, but all
organisms contain a mixture of more than one kind of pigment, each serving a specific function.
Chlorophylls a and b are abundant in green plants, and c and d are found in some protists and
cyanobacteria. A number of different types of bacteriochlorophyll have been found; type a is the
most widely distributed. Pigments are chemical compounds that have the ability to absorb certain
wave lengths of light. These are helpful for all autotrophs in carrying out the process of
photosynthesis. Solar light has a range of wavelengths. Each pigment can capture only a specific
wave length of sunlight. To trap maximum solar energy, a variety of pigments are required.
Photosynthetic pigments in plants can be classified into three categories:
1. Chlorophylls (are insoluble in water can be extracted only with organic solvents such as
acetone, petroleum ether and alcohol)
2. Carotenoids; - includes carotenes and xanthophylls. Xanthophylls are also called carotenols
(are insoluble in water can be extracted only with organic solvents such as acetone, petroleum
ether and alcohol)
3. Phycobilins (are soluble in water) It depicts the proportion of light (with different wave
length) absorbed by pigments. Chlorophyll a and b absorb light of violet, blue, orange and red
wavelength (> 90%) and reflect/transmit green wavelength. Carotenoids (β-carotene and lutein)
absorb blue and violet wavelength and transmit/ reflect green, yellow, orange and red wave
lengths and hence give orange or yellow colour appearance. In addition to the light harvesting for
photosynthesis, carotenoids also protect chlorophylls against oxidative destruction by O2 in very
high levels of irradiance in the process mcalled photo-oxidation. High light levels promote
production of free radicals, which react with Chlorophyll in presence of O2 damaging
Chlorophyllleaf bleachingleaf death.
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Figure 11: Absorption spectra of some photosynthetic pigments. Curve 1, bacteriochlorophyll a;

curve 2, chlorophyll a; curve 3, chlorophyll b; curve 4, phycoerythrobilin (pigment in red
algae); curve 5, β-carotene. The absorption spectra shown are for pure pigments dissolved in
nonpolar solvents, except for curve 4. In many cases the spectra of photosynthetic pigments in
vivo are substantially affected by the environment of the pigments in the photosynthetic
membrane.
1. Chlorophyll: Chlorophyll (Greek: ‘Chlor’ means green and ‘phyll’ means leaf) Chlorophyll
consists of two pigments- Chlorophyll a and chlorophyll b. These pigments appear green in
colour. They are present in all photosynthetic autotrophs except bacteria for trapping sunlight.
All chlorophylls have a complex ring structure that is chemically related to the porphyrin-like
groups found in hemoglobin and cytochromes. In addition, a long hydrocarbon tail is almost
always attached to the ring structure. The tail anchors the chlorophyll to the hydrophobic portion
of its environment. The ring structure contains some loosely bound electrons and is the part of
the molecule involved in electron transitions and redox reactions. Because the electrons move
freely, the ring has the potential to gain or lose electrons easily, and thus has the potential to
provide energized electrons to other molecules. This is the fundamental process by which
chlorophyll “captures” the energy of sunlight.
Chlorophyll “a” is the primary pigment for photosynthesis. It is grass-green in colour and
absorbs light energy at 430nm and 662nm wave length. Photosynthesis is possible because of the
presence of porphyrin ring and its ability to pass its energized electrons to molecules which
45
manufacture sugars. A second kind of chlorophyll is chlorophyll “b”, which occurs in all plants,
green algae and some prokaryotes. It is bluish-green in colour and absorbs light energy at wave
lengths 453nm and 642nm. It differs from chlorophyll ‘a’ by having a –CHO group instead of
methyl group.
2. Carotenoids (yellow or orange pigments): Carotenoids are pigments synthesized by plants.

The role of Carotenoids is absorption of light energy and transfer the light energy to chlorophyll
a molecules. They also play a very important role in preventing photodynamic damage within
the photosynthetic apparatus. Photodynamic damage is caused by O2 molecules which is very
reactive and is capable of oxidizing whole range of organic compounds such as chlorophylls and
thereby making them unfit for their normal physiological function. Carotenoids have 40 carbon
atoms. They perform two main functions:
They serve as the harvesting molecules for absorption of light energy.
They protect chlorophyll from photo damage.
They can be classified into two classes:
(i) Xanthophylls: They are yellow pigments. They are similar to carotenes but differ in having
two oxygen atoms in the form of hydroxyl or carboxyl group. The molecular formula is
C40H56O2. In addition, they contain oxygen, which makes them polar. These are found only in
plants and not in animals.
(ii) Carotenes: These are orange coloured pigments. They are unsaturated hydrocarbon shaving
the general formula C40H56. They do not contain oxygen. They are found in fruits and vegetables.
Beta carotene and Lycopene, which give red color to tomato, are common examples of carotene.
3. Phycobilins (red and blue pigment): Phycobilins are water soluble pigments. They are found
in cytoplasm or the stroma of a chloroplast in cyanobacteria (phycocyanin) and red algae
(phycoerythrin). The can absorb red, orange, yellow and green light wave lengths which are not
well absorbed by chlorophyll a. They transmit the light energy to chlorophyll and contribute to
the process of photosynthesis.
Location of photosynthetic pigments in chloroplast: Each chloroplast is bound / enveloped

with double chloroplastic membranes whose main function is to regulate molecular trafficking in
or out of chloroplast. Within the chloroplast, an amorphous, gel-like and enzyme rich material
46
called Stroma fills the entire volume. Embedded throughout stroma are the pigments containing
thylakoids wherein light is used to oxidize the H2O and form ATP and NADPH.
In Figure 11 the chloroplast of higher plants is surrounded by the inner and outer membranes
(envelope). The region of the chloroplast that is inside the inner membrane and surrounds the
thylakoid membranes is known as the stroma. Stroma contains the enzymes that catalyze carbon
fixation and other biosynthetic pathways. The thylakoid membranes are highly folded and appear
in many pictures to be stacked like coins, although in reality they form one or a few large
interconnected membrane systems, with a well defined interior and exterior with respect to the
stroma. The inner space within a thylakoid is known as the lumen.
figure12. Schematic picture of the overall organization of the membranes in the chloroplast
Pigments present in thylakoid membranes are (i) largely two kinds of green molecules:
Chlorophyll a and b and (ii) yellow to orange coloured pigments classified as carotenoids of two
types — pure hydrocarbon carotenes and oxygen containing xanthophylls. Two major green
bands are found in isolated chloroplasts - each containing chlorophyll a and b, xanthopylls, very
little β-carotenoides, light harvesting pigments called Light Harvesting Complex I & II located in
Photosystem (PS) I & II, respectively. Each granum contains approximately 200 units each of PS
I and PSII; however their amount in stroma is variable. PSI is abundantly present in stroma
thylakoids and PSII is abundant in grana. Also present in chloroplasts are DNA, RNA, ribosomes
and many enzymes largely in stroma where transcription and translation occur.
47
3.3. Light Reaction and Dark Reaction
In the Light Dependent Processes (Light Reactions) light strikes chlorophyll in such a way as to
excite electrons to a higher energy state. In a series of reactions the energy is converted (along an
electron transport process) into ATP and NADPH. Water is split in the process, into oxygen,
protons, and electrons in the following manure.
2H2O O2 + 4H+ +4e-
Figure 13: the light and carbon reactions of photosynthesis. Light is required for the generation
of ATP and NADPH. The ATP and NADPH are consumed by the carbon reactions, which reduce
CO2 to carbohydrate (triose phosphates).
In this process, nicotinamide adenine dinucliotide phosphate (NADP) is reduced in to NADPH

and in a coupled process adenosine diphosphate (ADP) is phosphorylated by use of inorganic
phosphate (Pi), forming the energy rich adenosine tri-phosphate (ATP). NADPH and ATP as
well as enzymes bring about the fixation of CO2 in the so-called dark reaction. In this reaction
CO2 is reduced and ATP is split again and NADPH is oxidized. The CO2 gets assimilated and
organic compounds are formed. The enzyme involved in the primary process of CO2 assimilation
is named ribulose diphosphate (biphosphate) carboxylase (Rubisco). This true for the C3 plants,
but for the C4 plants it another enzyme named phosphoenolpyruvate carboxylase (PEPase). The
later enzyme also involved, in CO2 assimilation of certain succulent plants having Crassulacean
metabolism (CAM). The primary process of photosynthesis occurs in the chloroplasts.
Chloroplast occur in every green organs of the plant, i.e. leaves, stems, fruits and flowers. In C3
plants, most of the chloroplasts are located in the mesophyll cells of the leaves. The process
begins this way: -
48
Light from the sun strikes the leaf

A portion of it is reflected back into the atmosphere
A portion of it is absorbed by the leaf
A portion of it is transmitted through it
We are concerned with the portion which is absorbed by the leaf. Three (3) processed are
involved
1. Absorption of photons by chlorophylls within wave length 400-700nm
(PARPhotosynthetically active radiation)
2. Electrons of the chlorophyll molecule get excited following absorption of solar energy of the
light
3. The transfer of the excitation energy to the reaction centers of the photosynthesis These
reactions are called “light reaction” or “Hill reaction” named after the discoverer Robin Hill of
U.K. (won Noble prize for this work). Obviously, these reactions need light. Thus, the main
functions of light reactions are: -
1. Splitting of H2O (water)
2. Production of O2 (oxygen), electrons and protons
3. Production of ATP and NADPH (energy suppliers )
Carbon Reduction or Carboxylation or Carbon Fixation or Calvin Cycle of Photosynthesis

1. The excited energy under light reaction captured by the pigments is transferred to the reaction
centers PSI and PSII, i.e. Photosystem I and Photosystem II. Photosystem II takes place at the
wave length of 680 nm (called P 680) and Photosystem I takes place at the wave length of 700
nm (called P 700).
2. PS I is largely associated with unstacked regions of the thylakoid and PS II is associated with
“Stacked” region of the thylakoid.
Stacked= Grana
Unstacked = stroma membranes
3. In this process protons are transported across the membrane into the thylakoid and lumen. As a
result, the lumen becomes acidified and positively charged.
4. The electrochemical gradient across the thylakoid membrane which represents a “proton
motive force” is subsequently used to produce ATP, the reaction being catalyzed by ATPase.
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5. In non-cyclic electron transport, NADP is the terminal acceptor of electrons from PS I forming
NADPH. In cyclic electron transport, electrons are transferred from PS I to cytochrome in the
lumens, thus contributing to proton extrusion and ATP synthesis. Finally, ATP and NADPH are
used in carbon reduction cycle.
6. CO2 combines with its acceptor RuBP (ribulase 1,5 biphosphate) forming PGA
(phosphoglyceric acid), the reaction being catalyzed by Rubisco (ribulase 1,5 biphosphate
carboxylase).
7. PGA is reduced to triose P in a two step reaction requiring ATP and NADPH.
8. About 1/6th of the triose P is exported and the remaining 5/6th of the triose P is processed to
form 6-ribulose-1-mono-phosphate.
9. 6-ribulose-1-mono-phosphate is converted to RuBP in reaction requiring ATP. RuBP is ready
to accept another CO2.
Figure 13: The Calvin cycle proceeds in three stages: (1) carboxylation, during which CO2 is
covalently linked to a carbon skeleton; (2) reduction, during which carbohydrate is formed at the
expense of the photochemically derived ATP and reducing equivalents in the form of NADPH;
and (3) regeneration, during which the CO2 acceptor ribulose1,5-bisphosphate re-forms.
Finally, 3ATP (molecule) and 2NADPH (molecule) are required per CO2 molecule fixed
Photosynthetically because CO2 is incorporated into ribulase 1,5 biphosphate to yield 2PGA in
C3 plants but 4 or 5 ATP and 2NADPH are required per CO2 molecule fixed in C4 plants.
Remember A mole of photons= 6x1023 photons
2mole photon= 1 joule
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The overall reaction of photosynthesis: -

6CO2 + 12H2O + light C6H12O6 + 6O2 + 6H2O
In the Light Independent Process, carbon dioxide from the atmosphere (or water for
aquatic/marine organisms) is captured and modified by the addition of Hydrogen to form
carbohydrates (general formula of carbohydrates is [CH2O]n). The incorporation of carbon
dioxide into organic compounds is known as carbon fixation. The energy for this comes from
the first phase of the photosynthetic process. Living systems cannot directly utilize light energy,
but can, through a complicated series of reactions, convert it into C-C bond energy that can be
released by glycolysis and other metabolic processes.
Photosystems are arrangements of chlorophyll and other pigments packed into thylakoids. Many
Prokaryotes have only one photosystem, Photosystem II (so numbered because, while it was
most likely the first to evolve, it was the second one discovered). Eukaryotes have Photosystem
II plus Photosystem I. Photosystem I uses chlorophyll a, in the form referred to as P700.
Photosystem II uses a form of chlorophyll a known as P680. Both "active" forms of chlorophyll
a function in photosynthesis due to their association with proteins in the thylakoid membrane.
Photophosphorylation is the process of converting energy from a light-excited electron into the
pyrophosphate bond of an ADP molecule. This occurs when the electrons from water are excited
by the light in the presence of P680. The energy transfer is similar to the chemiosmotic electron
transport occurring in the mitochondria. Light energy causes the removal of an electron from a
molecule of P680 that is part of Photosystem II. The P680 requires an electron, which is taken
from a water molecule, breaking the water into H+ ions and O-2 ions. These O-2 ions combine to
form the diatomic O2 that is released. The electron is "boosted" to a higher energy state and
attached to a primary electron acceptor, which begins a series of redox reactions, passing the
electron through a series of electron carriers, eventually attaching it to a molecule in
Photosystem I.
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3.3.1. Role of Photo-system I and II

Photosynthetic units occur in the form of two distinct groups called photosystems or pigment
systems. Green plants and cyanobacteria possess two photosystems, I and II. But bacteria possess
only one photosystem.
Light reactions or the ‘Photochemical’ phase include light absorption, water splitting, oxygen
release, and the formation of high-energy chemical intermediates, ATP and NADPH. Several
complexes are involved in the process. The pigments are organised into two discrete
photochemical light harvesting complexes (LHC) within the Photosystem I (PS I) and
Photosystem II (PS II). These are named in the sequence of their discovery, and not in the
sequence in which they function during the light reaction. The LHC are made up of hundreds of
pigment molecules bound to proteins. Each photosystem has all the pigments (except one
molecule of chlorophyll a) forming a light harvesting system also called antennae (Figure 14).
These pigments help to make photosynthesis more efficient by absorbing different wavelengths
of light. The single chlorophyll a molecule forms the reaction centre. The reaction centre is
different in both the photosystems. In PS I the reaction centre chlorophyll a has an absorption
peak at 700 nm, hence is called P700, while in PS II it has absorption maxima at 680 nm, and is
called P680.
Figure 14 The light harvesting complex

The Electron Transport
In photosystem II the reaction centre chlorophyll a absorbs 680 nm wavelength of red light
causing electrons to become excited and jump into an orbit farther from the atomic nucleus.
52
These electrons are picked up by an electron acceptor which passes them to an electrons
transport system consisting of cytochromes (Figure 15).
This movement of electrons is downhill, in terms of an oxidation-reduction or redox potential

scale. The electrons are not used up as they pass through the electron transport chain, but are
passed on to the pigments of photosystem PS I. Simultaneously, electrons in the reaction centre
of PS I are also excited when they receive red light of wavelength 700 nm and are transferred to
another accepter molecule that has a greater redox potential. These electrons then are moved
downhill again, this time to a molecule of energy-rich NADP+. The addition of these electrons
reduces NADP+ to NADPH + H+. This whole scheme of transfer of electrons, starting from the
PS II, uphill to the acceptor, down the electron transport chain to PS I, excitation of electrons,
transfer to another acceptor, and finally downhill to NADP+ causing it to be reduced to NADPH
+ H+ is called the Z scheme, due to its characteristic shape (Figure 15). This shape is formed
when all the carriers are placed in a sequence on a redox potential scale.
Figure 15 Z scheme of light reaction

Cyclic and Non-cyclic Photo-phosphorylation: Living organisms have the capability of
extracting energy from oxidisable substances and store this in the form of bond energy. Special
substances like ATP carry this energy in their chemical bonds. The process through which ATP
is synthesised by cells (in mitochondria and chloroplasts) is named phosphorylation.
Photophosphorylation is the synthesis of ATP from ADP and inorganic phosphate in the
presence of light. When the two photosystems work in a series, first PS II and then the PS I, a
process called non-cyclic photo-phosphorylation occurs. The two photosystems are connected
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through an electron transport chain, as seen earlier in the Z scheme. Both ATP and NADPH + H
are synthesised by this kind of electron flow (see Figure 15).
When only PSI is functional, the electron is circulated within the photosystem and the
phosphorylation occurs due to cyclic flow of electrons (Figure 16). A possible location where
this could be happening is in the stroma lamellae. While the membrane or lamellae of the grana
have both PS I and PS II the stroma lamellae membranes lack PS II as well as NADP reductase
enzyme. The excited electron does not pass on to NADP but is cycled back to the PS I complex
through the electron transport chain (Figure 16). The cyclic flow hence, results only in the
synthesis of ATP, but not of NADPH + H+. Cyclic photophosphorylation also occurs when only
light of wavelengths beyond 680 nm are available for excitation.
Figure 16 Cyclic photophosphorylation
3.4. Path of Carbon in Photosynthesis

Let us now see how the ATP and NADPH are used in the biosynthetic phase. We saw earlier that
CO2 is combined with H2O to produce (CH2O)nor sugars. It was of interest to scientists to find
out how this reaction proceeded, or rather what was the first product formed when CO2 is taken
into a reaction or fixed. Just after World War II, among the several efforts to put radioisotopes to
beneficial use, the work of Melvin Calvin is exemplary. The use of radioactive 14C by him in
algal photosynthesis studies led to the discovery that the first CO2 fixation product was a 3-
carbon organic acid. He also contributed to working out the complete biosynthetic pathway;
hence it was called Calvin cycle after him. The first product identified was 3-phosphoglyceric
acid or in short PGA. How many carbon atoms does it have?
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Scientists also tried to know whether all plants have PGA as the first product of CO 2 fixation, or
whether any other product was formed in other plants. Experiments conducted over a wide range
of plants led to the discovery of another group of plants, where the first stable product of CO2
fixation was again an organic acid, but one which had 4 carbon atoms in it. This acid was
identified to be oxaloacetic acid or OAA. Since then CO2 assimilation during photosynthesis
was said to be of two main types: those plants in which the first product of CO 2 fixation is a C3
acid (PGA), i.e., the C3 pathway, and those in which the first product was a C4 acid (OAA), i.e.,
the C4 pathway. These two groups of plants showed other associated characteristics that we will
discuss bellow.
3.4.1. C3 Photosynthesis Cycle
The first identified and principal pathways fixation in chloroplasts utilizes NADPH produced
during non-cyclic electron transport, whereas ATP produced during photophosphorylation serves
as source of energy facilitating the process of reduction. The C in CO2 is the most oxidized form
found in nature. The first stable product formed in a shortest time of 2 seconds is a 3 carbon
compound 3-phosphoglyceric acid (3-PGA) and the substrates are CO2 and 5 C sugar compound.
Calvin and his co-workers then worked out the whole pathway and showed that the pathway
operated in a cyclic manner; the RuBP was regenerated. Let us now see how the Calvin pathway
operates and where the sugar is synthesised. Let us at the outset understand very clearly that the
Calvin pathway occurs in all photosynthetic plants; it does not matter whether they have C3 or
C4 (or any other) pathways (Figure 17). For ease of understanding, the Calvin cycle can be
described under three stages: carboxylation, reduction and regeneration.
1. Carboxylation – Carboxylation is the fixation of CO2 into a stable organic intermediate.

Carboxylation is the most crucial step of the Calvin cycle where CO2 is utilised for the
carboxylation of RuBP. This reaction is catalysed by the enzyme RuBP carboxylase which
results in the formation of two molecules of 3-PGA. Since this enzyme also has an oxygenation
activity it would be more correct to call it RuBP carboxylase-oxygenase or RuBisCO.
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Figure 17: The Calvin cycle proceeds in three stages: (1) carboxylation, during which CO2
combines with ribulose-1,5-bisphosphate; (2) reduction, during which carbohydrate is formed at
the expense of the photochemically made ATP and NADPH; and (3) regeneration during which
the CO2 acceptor ribulose1,5-bisphosphate is formed again so that the cycle continues.
2. Reduction – These are a series of reactions that lead to the formation of glucose. The steps
involve utilization of 2 molecules of ATP for phosphorylation and two of NADPH for reduction
per CO2 molecule fixed. The fixation of 6CO2 molecules and 6 turns of the cycle are required for
the removal of one molecule of glucose from the pathway.
3. Regeneration – Regeneration of the CO2 acceptor molecule RuBP is crucial if the cycle is to
continue uninterrupted. The regeneration steps require one ATP for phosphorylation to form
RuBP. Hence for every CO2 molecule entering the Calvin cycle, 3 molecules of ATP and 2 of
NADPH are required. It is probably to meet this difference in number of ATP and NADPH used
in the dark reaction that the cyclic phosphorylation takes place. To make one molecule of
glucose 6 turns of the cycle are required. Work out how many ATP and NADPH molecules will
be required to make one molecule of glucose through the Calvin pathway. It might help you to
Understand all of this if we look at what goes in and what comes out of the Calvin cycle.
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3.4.2. C4 photosynthesis Cycle

Plants that are adapted to dry tropical regions have the C4 pathway mentioned earlier. Though
these plants have the C4 oxaloacetic acid as the first CO2 fixation product they use the C3
pathway or the Calvin cycle as the main biosynthetic pathway. Then, in what way are they
different from C3 plants? This is a question that you may reasonably ask.C4 plants are special:
They have a special type of leaf anatomy, they tolerate higher temperatures, they show a
response to high light intensities, and they lack a process called photorespiration and have
greater productivity of biomass. Let us understand these one by one. The particularly large cells
around the vascular bundles of the C4 pathway plants are called bundle sheath cells, and the
leaves which have such anatomy are said to have ‘Kranz’ anatomy. ‘Kranz’ means ‘wreath’ and
is a reflection of the arrangement of cells. The bundle sheath cells may form several layers
around the vascular bundles; they are characterized by having a large number of chloroplasts,
thick walls impervious to gaseous exchange and no intercellular spaces. You may like to cut a
section of the leaves of C plants – maize or sorghum – to observe the Kranz anatomy and the
distribution of mesophyll cells.
It would be interesting for you to collect leaves of diverse species of plants around you and cut
vertical sections of the leaves. Observe under the microscope – look for the bundle sheath around
the vascular bundles. The presence of the bundle sheath would help you identify the C4 plants.
Now study the pathway shown in Figure 13.9. This pathway that has been named the Hatch and
Slack Pathway is again a cyclic process. Let us study the pathway by listing the steps. The
primary CO2 acceptor is a 3-carbon molecule phosphoenol pyruvate (PEP) and is present in the
mesophyll cells. The enzyme responsible for this fixation is PEP carboxylase or PEPcase. It is
important to register that the mesophyll cells lack RuBisCO enzyme. The C4 acid OAA is formed
in the mesophyll cells. It then forms other 4-carbon compounds like malic acid or aspartic acid in
the mesophyll cells itself, which are transported to the bundle sheath cells. In the bundle sheath
cells these C4 acids are broken down to release CO2 and a 3-carbon molecule. The 3-carbon
molecule is transported back to the mesophyll where it is converted to PEP again, thus,
completing the cycle. The CO2 released in the bundle sheath cells enters the C3 or the Calvin
pathway, a pathway common to all plants.
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The bundle sheath cells are rich in an enzyme Ribulose bisphosphate carboxylase-oxygenase
(RuBisCO), but lack PEPcase. Thus, the basic pathway that results in the formation of the
sugars, the Calvin pathway, is common to the C3 and C4 plants. Did you note that the Calvin
pathway occurs in all the mesophyll cells of the C3 plants? In the C3 plants it does not take place
in the mesophyll cells but does so only in the bundle sheath cells. All PEPCase is present in MS
cells and Rubisco in BS cells enabling C4 plants to fixing mechanism use both kinds of CO2. The
basic C4 cycle consists of four stages:
1. Fixation of CO2 in mesophyll with a 3-C molecule of PEP to form a 4-C organicacid, such as
OAA (the enzyme that does this is PEP carboxylase). OAA is unstable compound and is
immediately gets converted into malate and or aspartate. Note that this is not the typical fixation
of CO2 by Rubisco, which binds CO2 to the 5-C RuBp.
2. transport of the C4 acids (malate or aspartate) to the bundle sheath cells.

3. Decarboxylation of the C4 acids within the bundle sheath cells to generate CO2and pyruvate.
CO2 is then used in the Calvin cycle--just like regular C3 photosynthesis.
4. Transport of the C3 acid (PEP) back to the mesophyll cells, where the whole process starts all
Over again.
Figure 18: Diagrammatic representation of the Hatch and Slack Pathway The C4 photosynthetic
pathway. The hydrolysis of two ATP drives the cycle in the direction of the arrows, thus pumping
CO2 from the atmosphere to the Calvin cycle of the chloroplasts from bundle sheath cells.
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3.4.3. CAM (CRASSULACEAN ACID METABOLISM)
CAM photosynthesis is a second evolutionary strategy employing a "CO 2 pump" to accumulate

CO2. It is technically also a C4 pathway, because CO2 is fixed first into a 4-C organic acid like
oxaloacetate. This path occurs in a wide variety of plant species, mainly in arid and tropical
regions
• Cacti (barrel, saguaro, prickly pear, teddy bear cholla)
• Crassulaceae (Crassula aquatica, Sedum rosea)
• Bromeliads (pineapples, "air plants"--epiphytes such as Spanish moss)
• Orchids (Vanilla, Phalaenopsis, Dendrobium)
• Agave (Century plant, yucca)
CAM plants are adapted to desert ecosystem with peculiar morphology, anatomy and physiology
to suit this ecosystem. Here initial carboxylation takes place in the dark followed by
decarboxylation and refixation of CO2 in light to economize the water loss through transpiration
Eg. pine aple, vanilla, cacti, agave. In CAM plants, formation of the C4 acids and its
decarboxylation is both temporally and spatially separated. At night, CO2 is captured by PEP
carboxylase in the cytosol, and the malate that forms from the oxaloacetate product is stored in
the vacuole. During the day, the stored malate is transported to the chloroplast and
decarboxylated by NADP-malic enzyme, the released CO2 is fixed by the Calvin cycle.
Theadaptive advantage of CAM is the reduction of water loss by transpiration, achieved by the
stomatal closing during the day. Temporal separation of CO2 uptake from photosynthetic
reactions: CO2 uptake and fixation take place at night, and decarboxylation and refixation of the
internally released CO2 occur during the day.
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The CAM mechanism is similar in many respects to the C4 cycle. In C4 plants, formation of the
C4 acids in the mesophyll is spatially separated from decarboxylation of the C4 acids and from
refixation of the resulting CO2 by the Calvin cycle in the bundle sheath. In CAM plants,
formation of the C4 acids is both temporally and spatially separated.
Why Rubisco and not PEP Carboxylase fixes CO2 in Day Light?
Both Rubisco and PEP Carboxylase are present in CAM plants and both have about equal
affinities for dissolved CO2. Furthermore, cytosolic PEP carboxylase should encounter the
incoming CO2 first before CO2 reaches to chloroplasic Rubisco. However, it is Rubisco that fixes
CO2 in light and not the PEP carboxylase because
• (i) in day light PEP carboxylase in CAM plants is converted into inactive form that has
extremely low affinity to CO2,
• (ii) PEP carboxylase is strongly inhibited by malic acid released from vacuoles in day light,
• (iii) Changes in the activities of still other enzymes that favour CO2 fixation by PEP
carboxylase only occur in darkness.
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CO2 fixation and transpiration loss of water of a typical CAM plants
CAM pathway of CO2 fixation is favoured by factors such as

(i) Hot days with high irradiances,
(ii) Cool nights
(iii) Dry soils
(iv) High salts in soil and
(v) Osmotic drought
Several CAM plant species switch to C-3 mode after a rainstorm or during too hot nights
(opening stomata serves no purpose if night temp is high). C-3 cycle is less costly under high
night T°.
Ecological Significance: The understanding of the potential value of CAM for plant carbon gain
is based largely on studies of desert succulents and upper canopy epiphytes from tropical forests.
When taken together, the phylogeny, biogeography, and physiology of CAM plants provide
compelling evidence for the ecological significance of this photosynthetic pathway in sun
exposed, water limited habitats. Nocturnal CO2, uptake coupled with stomatal closure during the
day maximizes the ratio of carbon gain to water loss. High daytime partia1 pressures of CO 2,
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inside the leaf help to mitigate the photoinhibitory effects of high light common to these
environments. The capacity to maintain a functioning photosynthetic apparatus by refixation of
respired CO2, when stomates are continuously closed during prolonged drought helps CAM
plants to survive in exposed, xeric sites.
3.5. Role of C3 and C4 in Crop Productivity

Plants can adapt to conditions of low water content, high temperature, and bright light intensities
(grow in dry climates of tropics and subtropics). Photosynthetic rate remains higher due to the
absence of photorespiration. C4 plants are about twice as efficient as C3 plants in converting solar
energy into the production of dry matter. At lower temperatures and when light is limiting, rate
of PS will be greater in C3 plants than that of C4 plants. Additional factors such as inactivation of
certain enzymes, reduced enzyme activity and impaired metabolites at low temperatures and the
fact that in some C4 plants, the Rubisco is only 1/3-1/4 of that in C3 plants would mean that at
low temperatures (<15°C), PS rate of C4 may be lesser than that of C3 plants even at saturation
irradiance. However, light saturated rate of PS in C3 plants increase only slightly with increase in
temperature (in the range of 15-30°C) because:
I. CO2 becomes limiting,

II. Photo-respiration is enhanced primarily due to two reasons. Firstly, the rise in temperatures
reduces the solubility of CO2 more than that of O2, reducing CO2/O2 ratio at carboxylation site of
Rubisco. Secondly, temperature also causes direct effect on carboxylation activity of enzyme.
The affinity of Rubisco for CO2 in carboxylation reaction is reduced almost 3-fold as temperature
increases over the range of 15-35 °C depending upon the crop species. In C4 plants,
photorespiration is largely prevented by CO2 concentrating mechanism over the range of
physiological temperatures. These increases in PS rate with increased temperatures are observed
and thus comparatively high optimum temperatures (about 35 °C) in C4 plants as against low
optimum temperature (about 25 °C) in C3 plants. Rate of PS as determined by temperature
regimes could be demonstrated by varying regimes of temperature in growth chambers.
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Water Use Efficiency in C3 and C4 plants: C4 plants have superior water use efficiency
(WUE) (gram of water evapo-transpired per gram of CO2 fixed) than C3 plants. Both CO2 &
water vapour use the same route to diffuse in and out of leaves, respectively. However, for a
given stomatal resistance, more water will be lost than CO2 gained because: (i) Water vapour
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concentration (0.07 mole fraction in air) is almost always greater than that of CO2 (0.00034 mole
fraction in air). (ii) Water (2.4 x 10-5 m-2s-1) has higher diffusion coefficient than CO2 (1.57x10-5
m-2s-1) (iii) CO2 diffusion is impeded by an additional resistance, mesophyll resistance (rm) that
is not encountered by water (as water diffuses out directly from intracellular spaces through
stomata) CO2 intake to carboxylating site encounters boundary, stomatal and mesophyll
resistances as under.
where, [CO2]a, [CO2]i and [CO2]c are CO2 concentrations in air, intra-cellular spaces and at the
site of carboxylation and ra, rs, and rm/rr are boundary, stomatal and mesophyll resistances,
respectively. Hence net photosynthesis rate (Pn) may be expressed as considering gas exchange:
The net carbon assimilation for a leaf can be computed from: • Pn, leaf = gc(Ca -Cc), where gc is
the conductance of the boundary layer and surface (stomata) for CO2 • The reciprocal of a
conductance is resistance
• Pn = [{CO2}a — {CO2}c] / ra + rs + rm
Thus, a decrease in rm for a given stomatal resistance would increase rate of photosynthesis
without affecting the transpiration. In general C4 plants have lower rm (mesophyll resistance)
and have tendency of having higher stomatal resistance (see below Table) favouring a high WUE
Reported minimum values of rm and gaseous resistance in some C3 and C4 crops (top) Water
use efficiency of some C3 and C4 crop species (bottom).
64
It may be inferred from the data in above Tables that;

I. The C4 plants normally have lower rm values than C3 plant and lower values of rm are
attributed to lack of photorespiration in C4 plants,
II. The reason of slightly higher rs values in C4 plants is not clear but fewer stomata per unit
area or smaller stomatal apertures might be responsible for higher rs values,
III. The C4 plants require more light for maximum stomata aperture opening than C3 plants,
IV. High WUE is often important at higher temperatures and
V. Increasing temperatures accentuate the advantage in C4 plants because of favourable effects
on carboxylase over oxygenase activity of rubisco and thus on rm.
3.6. Photorespiration (O2 oxidative photosynthetic carbon cycle)
Let us try and understand one more process that creates an important difference between C3 and
C4 plants – Photorespiration. To understand photorespiration we have to know a little bit more
about the first step of the Calvin pathway – the first CO2 fixation step. This is the reaction where
RuBP combines with CO2 to form 2 molecules of 3PGA that is catalysed by RuBisCO. RuBisCO
that is the most abundant enzyme in the world (Do you wonder why?) is characterized by the fact
that its active site can bind to both CO2 and O2 hence the name. Can you think how this could be
possible? RuBisCO has a much greater affinity for CO2 than for O2. Imagine what would happen
if this were not so? This binding is competitive. It is the relative concentration of O2 and CO2 that
determines which of the two will bind to the enzyme.
In C3 plants some O2 does bind to RuBisCO, and hence CO2 fixation is decreased. Here the RuBP
instead of being converted to 2 molecules of PGA binds with O2 to form one molecule of
phosphoglycerate and phosphoglycolate in a pathway called photorespiration. In the
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photorespiratory pathway, there is neither synthesis of sugars, nor of ATP. Rather it results in the
release of CO2 with the utilization of ATP. In the photorespiratory pathway there is no synthesis
of ATP or NADPH. Therefore, photorespiration is a wasteful process.
In C4 plants photorespiration does not occur. This is because they have a mechanism that
increases the concentration of CO2 at the enzyme site. This takes place when the C4 acid from the
mesophyll is broken down in the bundle sheath cells to release CO2, this results in increasing the
intracellular concentration of CO2. In turn, this ensures that the RuBisCO functions as a
carboxylase minimising the oxygenase activity. Now that you know that the C4 plants lack
photorespiration, you probably can understand why productivity and yields are better in these
plants. In addition these plants show tolerance to higher temperatures. Based on the above
discussion can you compare plants showing the C3 and the C4 pathway? Use the table format
given and fill in the information.
Important of Photorespiration: Under conditions of high light intensity and low intercellular
CO2 concentration (e.g., when stomata are closed because of water stress), it dissipates excess
ATP and reducing power from the light reactions, thus preventing damage to the photosynthetic
apparatus. Arabidopsis mutants that are unable to photorespire grow normally under 2% CO2,
but they die rapidly if transferred to normal air (0.03%). There is evidence from work with
transgenic plants that photorespiration protects C3 plants from photooxidation and
photoinhibition.
3.7. Factors Affecting Photosynthesis.

An understanding of the factors that affect photosynthesis is necessary. The rate of
photosynthesis is very important in determining the yield of plants including crop plants.
Photosynthesis is under the influence of several factors, both internal (plant) and external. The
plant factors include the number, size, age and orientation of leaves, mesophyll cells and
chloroplasts, internal CO2 concentration and the amount of chlorophyll. The plant or internal
factors are dependent on the genetic predisposition and the growth of the plant. The external
factors would include the availability of sunlight, temperature, CO2 concentration and water. As
a plant photosynthesises, all these factors will simultaneously affect its rate. Hence, though
several factors interact and simultaneously affect photosynthesis or CO2 fixation, usually one
factor is the major cause or is the one that limits the rate. Hence, at any point the rate will be
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determined by the factor available at sub-optimal levels. When several factors affect any [bio]
chemical process, Blackman’s (1905) Law of Limiting Factors comes into effect. These states
the following: If a chemical process is affected by more than one factor, then its rate will be
determined by the factor which is nearest to its minimal value: it is the factor which directly
affects the process if its quantity is changed. For example, despite the presence of a green leaf
and optimal light and CO2 conditions, the plant may not photosynthesise if the temperature is
very low. This leaf, if given the optimal temperature, will start photosynthesising.
Light: As light intensity increases, the rate of the light-dependent reaction, and therefore
photosynthesis generally, increases proportionately (straight line relationship). The more photons
of light that fall on a leaf, the greater the number of chlorophyll molecules that are ionized and
the more ATP and NADPH are generated. Light dependent reactions use light energy and so are
not affected by changes in temperature. We need to distinguish between light quality, light
intensity and the duration of exposure to light, while discussing light as a factor that affects
photosynthesis. There is a linear relationship between incident light and CO2 fixation rates at low
light intensities. At higher light intensities, gradually the rate does not show further increase as
other factors become limiting (Figure 19). What is interesting to note is that light saturation
occurs at 10 per cent of the full sunlight. Hence, except for plants in shade or in dense forests,
light is rarely a limiting factor in nature. Increase in incident light beyond a point causes the
breakdown of chlorophyll and a decrease in photosynthesis.
No photosynthesis occurs in a very weak intensity of light. As the intensity of light increases, the
process begins and picks up the pace. A very strong intensity of light causes solarization. During
msolarization, photo-oxidation occurs and if it continues for a few hours, the photosynthetic
apparatus is destroyed. The photosynthesis begins in the morning hour, reaches its peak in the
noon/afternoon and then its rate declines. The value of light at which further increase is not
accompanied by an increase in CO2 uptake is called light saturation point. A plant continues to
respire almost at a uniform rate. During peak hour rate of photosynthesis is about twenty times
faster than that of respiration. However, in the morning as well in the evening (twilight)
photosynthesis equals the rate of respiration. It is called light compensation point.
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Figure 19: Graph of light intensity on the rate of photosynthesis
Carbon dioxide Concentration: CO2 is the major limiting factor for photosynthesis. An
increase in the CO2 concentration increases the rate at which carbon is incorporated into
carbohydrate in the light-independent reaction, and so the rate of photosynthesis generally
increases until limited by another factor. As it is normally present in the atmosphere at very low
concentrations (between 0.03-0.04%), increasing carbon dioxide concentration causes a rapid
rise in the rate of photosynthesis, which eventually plateaus when the maximum rate of fixation
is reached. If the existing atmospheric concentration is increased by 15 to 20 times, the
photosynthetic rate increases, if no other factor are becomes limiting. Still higher concentration
of CO2 is toxic to plant. However, the value of upper limit of CO2 concentration is variable. A
stage in CO2 concentration where there is no net absorption of CO2 by illuminated plant organ is
called CO2 compensation point or threshold value. It is less than 10 ppm for C4 plants and 50 -
100 ppm for
C3 plants.
The C3 and C4 plants respond differently to CO2 concentrations. At low light conditions neither
group responds to high CO2 conditions. At high light intensities, both C3 and C4 plants show
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increase in the rates of photosynthesis. What is important to note is that the C4 plants show
saturation at about 360 μl/L-1 while C3 responds to increased CO2 concentration and saturation is
seen only beyond 450μl/L-1. Thus, current availability of CO2 levels is limiting to the C3 plants.
The fact that C3 plants respond to higher CO2 concentration by showing increased rates of
photosynthesis leading to higher productivity has been used for some greenhouse crops such as
tomatoes and bell pepper. They are allowed to grow in carbon dioxide enriched atmosphere that
leads to higher yields.
Oxygen: The concentration of oxygen in the atmosphere is about 21% volume and it seldom
fluctuates. An increase in oxygen concentration decreases photosynthesis and the phenomenon is
called Warburg effect. The explanation to this problem lies in the phenomenon of
photorespiration. In C3 plants the rate of day (light) respiration is faster than the dark respiration.
This enhanced rate of respiration is called photorespiration. It operates in high light intensity,
high oxygen concentration and high temperature. Here oxygen competes with CO2, for the
oxidation of RuBP to phosphoglycolic acid, thus, reducing the fixation of CO2.
Temperature: The dark reactions being enzymatic are temperature controlled. Although the
light dependent reactions of photosynthesis are not affected by changes in temperature, the light
independent reactions of photosynthesis are dependent on temperature. They are reactions
catalyzed by enzymes. As the enzymes approach their optimum temperatures the overall rate
increases. It approximately doubles for every 10°C increase in temperature. Above the optimum
temperature the rate begins to decrease, as enzymes are denatured, until it stops. The C 4 plants
respond to higher temperatures and show higher rate of photosynthesis while C3 plants have a
much lower temperature optimum. The temperature optimum for photosynthesis of different
plants also depends on the habitat that they are adapted to. Tropical plants have a higher
temperature optimum than the plants adapted to temperate climates. The plants can perform
photosynthesis on a range of temperature. While some cryophytes can do photosynthesis at
35°C, some thermal algae can do this act even at 75°C. Usually the plants can perform
photosynthesis between 10°C - 40°C. The optimum temperature ranges between 25°C - 30°C.
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Water: Even though water is one of the reactants in the light reaction, the effect of water as a
factor is more through its effect on the plant, rather than directly on photosynthesis. Water stress
causes the stomata to close hence reducing the CO2 availability. Besides, water stress also makes
leaves wilt, thus, reducing the surface area of the leaves and their metabolic activity as well.
Water is a reactant in photosynthesis. A plant utilizes less than 1% water from its total absorption
and the rest is transpired. A decrease in water contents cause loss of turgor thereby closing down
the stomata. The effect of water is more indirect than direct.
Chemicals: The chemicals, which act as enzyme inhibitor like all other vital processes, inhibit
photosynthesis also. Such chemicals are cyanides, hydroxyl amine, H2S, CO, iodoacetates etc.
Besides, chloroform and ethers also inhibit photosynthesis. The herbicides dichlorophenyl
dimethyl urea (DCMU) and chlorophenyl dimethyl 'Urea (CMU) are also photosynthetic
inhibitors. They inactivate PS II, thus inhibiting the Hill reaction.
Minerals: Mn2+, CI- Ca2+ ions help in photolysis of water. Mg and Fe in synthesis of
chlorophyll. B and K help in translocation of solutes.
Hormoncs: Auxina, Gibberellin and Cytokinin increase the photosynthesis. ABA decreased it.
Limiting factors: In 1905, when investigating the factors affecting the rate of photosynthesis,
Blackmann formulated the Law of limiting factors. This states that the rate of a physiological
process will be limited by the factor which is in shortest supply. Any change in the level of a
limiting factor will affect the rate of reaction. For example, the amount of light will affect the
rate of photosynthesis. If there is no light, there will be no photosynthesis. As light intensity
increases, the rate of photosynthesis will increase as long as other factors are in adequate supply.
70
As the rate increases, eventually another factor will come into short supply. The graph below
shows the effect of low carbon dioxide concentration. It will eventually be insufficient to support
a higher rate of photosynthesis, and increasing light intensity will have no effect, so the rate
plateaus. If a higher concentration of carbon dioxide is supplied, light is again a limiting factor
and a higher rate can be reached before the rate again plateaus. If carbon dioxide and light levels
are high, but temperature is low, increasing temperature will have the greatest effect on reaching
a higher rate of photosynthesis.
SUMMARY
Green plants make their own food by photosynthesis. During this process carbon dioxide from
the atmosphere is taken in by leaves through stomata and used for making carbohydrates,
principally glucose and starch. Photosynthesis takes place only in the green parts of the plants,
mainly the leaves. Within the leaves, the mesophyll cells have a large number of chloroplasts
that are responsible for CO2 fixation. Within the chloroplasts, the membranes are sites for the
light reaction, while the chemosynthetic pathway occurs in the stroma. Photosynthesis has two
stages: the light reaction and the carbon fixing reactions. In the light reaction the light energy is
absorbed by the pigments present in the antenna, and funnelled to special chlorophyll a
molecules called reaction centre chlorophylls. There are two photosystems, PS I and PS II. PS I
has a 700 nm absorbing chlorophyll a P700 molecule at its reaction centre, while PS II has a
P680 reaction centre that absorbs red light at 680 nm. After absorbing light, electrons are excited
and transferred through PS II and PS I and finally to NAD forming NADH. During this process a
proton gradient is created across the membrane of the thylakoid. The breakdown of the protons
gradient due to movement through the F0 part of the ATPase enzyme releases enough energy for
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synthesis of ATP. Splitting of water molecules is associated with PS II resulting in the release of
O2, protons and transfer of electrons to PS II. In the carbon fixation cycle, CO2 is added by the
enzyme, RuBisCO, to a 5carbon compound RuBP that is converted to 2 molecules of 3-carbon
PGA. This is then converted to sugar by the Calvin cycle, and the RuBP is regenerated. During
this process ATP and NADPH synthesised in the light reaction are utilised. RuBisCO also
catalyses a wasteful oxygenation reaction in C3 plants: photorespiration. Some tropical plants
show a special type of photosynthesis called C4 pathway. In these plants the first product of CO2
fixation that takes place in the mesophyll, is a 4- carbon compound. In the bundle sheath cells the
Calvin pathway is carried out for the synthesis
of carbohydrates.
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CHAPTER 4. RESPIRATION
Two most important prerequisites of life are continuous supply of materials for growth of body
and energy for carrying out various life processes. All systems, from cell to ecosystem, require
energy to work. Only green plants and cyanobacteria can prepare their own food; by the process
of photosynthesis, they trap light energy and convert it into chemical energy that is stored in the
bonds of carbohydrates like glucose, sucrose and starch. It is these complex molecules which are
given the name ‘food’. However, the energy in the food has to be made available to the cells in a
usable form. This is the role of respiration. Respiration is the process by which energy in organic
molecules is released by oxidation. This energy is made available to the living cells in the form
of ATP (Adenosine Tri-Phosphate). The O2 required for respiration is obtained from the
atmosphere. ATP is the energy currency of the cell. We must remember that in green plants too,
not all cells, tissues and organs photosynthesise; only cells containing chloroplasts, that are most
often located in the superficial layers, carry out photosynthesis. Hence, even in green plants all
other organs, tissues and cells that are non-green, need food for oxidation. Hence, food has to be
translocated to all nongreen parts. This chapter deals with cellular respiration or the mechanism
of breakdown of food materials within the cell to release energy, and the trapping of this energy
for synthesis of ATP.
Photosynthesis, of course, takes place within the chloroplasts (in the eukaryotes), whereas the
breakdown of complex molecules to yield energy takes place in the cytoplasm and in the
mitochondria (also only in eukaryotes). The breaking of the C-C bonds of complex compounds
through oxidation within the cells, leading to release of considerable amount of energy is called
respiration. The compounds that are oxidized during this process are known as respiratory
substrates. Usually carbohydrates are oxidised to release energy, but proteins, fats and even
organic acids can be used as respiratory substances in some plants, under certain conditions.
During oxidation within a cell, all the energy contained in respiratory substrates is not released
free into the cell, or in a single step. It is released in a series of slow step-wise reactions
controlled by enzymes, and it is trapped as chemical energy in the form of ATP. Hence, it is
important to understand that the energy released by oxidation in respiration is not (or rather
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cannot be) used directly but is used to synthesise ATP, which is broken down whenever (and
wherever) energy needs to be utilised. Hence, ATP acts as the energy currency of the cell. This
energy trapped in ATP is utilised in various energy-requiring processes of the organisms, and the
carbon skeleton produced during respiration is used as precursors for biosynthesis of other
molecules in the cell.
DO PLANTS BREATHE?
Well, the answer to this question is not quite so direct. Yes, plants require O2 for respiration to
occur and they also give out CO2. Hence, plants have systems in place that ensure the availability
of O2. Plants, unlike animals, have no specialised organs for gaseous exchange but they have
stomata and lenticels for this purpose. There are several reasons why plants can get along
without respiratory organs. First, each plant part takes care of its own gas exchange needs. There
is very little transport of gases from one plant part to another. Second, plants do not present great
demands for gas exchange. Roots, stems and leaves respire at rates far lower than animals do.
Only during photosynthesis are large volumes of gases exchanged and, each leaf is well adapted
to take care of its own needs during these periods. When a cell photosynthesizes, availability of
O2 is not a problem in these cells since O2 is released within the cell.
Third, the distance that gases must diffuse even in large, bulky plants is not great. Each living
cell in a plant is located quite close to the surface of the plant. ‘This is true for leaves’, you may
ask, ‘but what about thick, woody stems and roots?’ In stems, the ‘living’ cells are organised in
thin layers inside and beneath the bark. They also have openings called lenticels. The cells in the
interior are dead and provide only mechanical support. Thus, most cells of a plant have at least a
part of their surface in contact with air. This is also facilitated by the loose packing of
parenchyma cells in leaves, stems and roots, which provide an interconnected network of air
spaces. Respiration is a process by which, energy is derived by the oxidation of various
Photosynthetic products to keep the life process in motion. Respiration is the processes of
cellular oxidation of glucose yielding CO2, H2O and energy as shown below; C6H12O6 + 6H2O +
6O2 → 6CO2 + 6H2O + 686 kcal i.e. Respiration is a reverse process of photosynthesis. A vital
process which occurs in all living cells of the plant but the most actively respiring regions are
growing regions like floral and vegetative buds, germinating seedlings, stem and root apices.
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From the equation the process of respiration appears to be very simple one but it is very
complicated and complex process which takes place with the help of several enzymes into
several steps involving several intermediates and electron carriers before oxygen is reduced by
accepting electrons to form water. The carbons of acetyl-COA are released as CO2.
Respiratory Quotient
The ratio between moles of CO2 released and moles of O2 consumed is called respiratory quotient
(RQ). RQ is useful index of types of substrates used in respiration and the subsequent use of
respiratory energy to support biosynthesis. Some substrates are fully oxidized to CO2 and H2O
others leave some of their intermediate metabolites for synthetic processes for growing cells.
Respiration of [CH2O] n) and proteins releases the same amount of CO2 as O2 consumed. The
RQ for [CH2O] oxidation is 1 & for fat oxidation is < 1(RQ = l8/25 = 0.71) - no complete
breakdown.
Because of the differences in composition of food fuels, different amounts of oxygen are
required to completely oxidize those foods to carbon dioxide and water. Thus, the quantity of
carbon dioxide produced in relation to oxygen consumed varies depending on the substrate
metabolized. This ratio of metabolic gas exchange is termed the respiratory quotient (RQ):
RQ = CO2 produced / O2 uptake
The ratio of hydrogen to oxygen in carbohydrates is equal to that of water. All of the oxygen
consumed by cells is used to oxidize the carbon in the carbohydrate to carbon dioxide. In turn,
the resultant production of carbon dioxide is equal to oxygen consumed and the ratio is 1.00.
C6H12O6 + 6 O2→6 CO2 + 6 H2O
RQ = 6 CO2/ 6 O2= 1.00
Lipids contain considerably fewer oxygen atoms in proportion to atoms of hydrogen and carbon.
Thus, when fat is degraded relatively more oxygen is needed to oxidize fat to carbon dioxide and
water. In the example of palmitic acid, 23O2 molecules are consumed for the oxidation of 16
CO2 molecules:
C16H32O2 + 23 O2 →16 CO2 + 16 H2O
RQ = 16 CO2/ 23 O2 = 0.696 or 0.7
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Significance of Respiration
• It releases energy which is consumed in various metabolic processes essential for plant life and
activates cell division.
• It brings about the formation of other necessary compounds participating as important cell
constituent
• It converts insoluble food into soluble form
• It liberates CO2 and plays a part actively in maintaining the balance of carbon cycle in nature
• It converts stored energy (potential energy) into usable form (kinetic energy).
During the process of respiration, oxygen is utilised, and carbon dioxide, water and energy are
released as products. The combustion reaction requires oxygen. But some cells live where
oxygen may or may not be available. Can you think of such situations (and organisms) where
O2 is not available? There are sufficient reasons to believe that the first cells on this planet lived
in an atmosphere that lacked oxygen. Even among present-day living organisms, we know of
several that are adapted to anaerobic conditions. Some of these organisms are facultative
anaerobes, while in others the requirement for anaerobic condition is obligate. In any case, all
living organisms retain the enzymatic machinery to partially oxidise glucose without the help of
oxygen. This breakdown of glucose to pyruvic acid is called glycolysis.
4.1. Glycolysis
The term glycolysis has originated from the Greek words, glycos for sugar, and lysis for splitting.
The scheme of glycolysis was given by Gustav Embden, Otto Meyerhof, and J. Parnas, and is
often referred to as the EMP pathway. In anaerobic organisms, it is the only process in
respiration. Glycolysis occurs in the cytoplasm of the cell and is present in all living organisms.
In this process, glucose undergoes partial oxidation to form two molecules of pyruvic acid. In
plants, this glucose is derived from sucrose, which is the end product of photosynthesis, or from
storage carbohydrates. Sucrose is converted into glucose and fructose by the enzyme, invertase,
and these two monosaccharides readily enter the glycolytic pathway. Glucose and fructose are
phosphorylated to give rise to glucose 6-phosphate by the activity of the enzyme hexokinase.
This phosphorylated form of glucose then isomerizes to produce fructose-6 phosphate.
Subsequent steps of metabolism of glucose and fructose are same.
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The various steps of glycolysis are depicted in Figure 20. In glycolysis, a chain of ten reactions,
under the control of different enzymes, takes place to produce pyruvate from glucose. While
studying the steps of glycolysis, please note the steps at which utilization or synthesis of ATP or
(in this case) NADH + H+ take place. ATP is utilised at two steps: first in the conversion of
glucose into glucose 6-phosphate and second in the conversion of fructose 6-phosphate to
fructose 1, 6-bisphosphate. The fructose 1, 6-bisphosphate is split into dihydroxyacetone
phosphate and 3-phosphoglyceraldehyde (PGAL).
We find that there is one step where NADH + H+ is formed from NAD+; this is when 3-
phosphoglyceraldehyde (PGAL) is converted to 1, 3-bisphosphoglycerate (BPGA). Two
redoxequivalents are removed (in the form of two hydrogen atoms) from PGAL and transferred
to a molecule of NAD+. PGAL is oxidised and with inorganic phosphate to get converted into
BPGA. The conversion of BPGA to 3-phosphoglyceric acid (PGA), is also an energy yielding
process; this energy is trapped by the formation of ATP. Another ATP is synthesised during the
conversion of PEP to pyruvic acid.
Main points of glycolysis are: It converts one molecule of hexose into 2 molecules of pyruvic
acid. No O2 is used neither CO2 produced. For each hexose, 2 molecules of NADH are produced
by 3-PGAld dehydrogenase. Some of these NADH are used in cytosol to drive various anabolic
reductive processes, whereas some enter into mitochondria. Two molecules of ATP are used
initially in phosphorylating the hexoses catalyzed by hexokinases or phospho-hexokinases. Two
molecules of ATP per molecule of hexose are produced by each phoshpoglycerokinase and
pyruvate kinase thus resulting in total 4 molecules of ATP. Glycolysis involves the conversion
of glucose, via phosphoenolpyruvate (PEP) into malate and pyruvate. These changes occur in
the cytosol. Then, these compounds are transported to mitochondria for krebs cycle to take place,
i.e., further oxidation. Oxidation of one glucose molecule in glycolysis produces two malate
molecules, without a net production of ATP. When pyruvate is the end product, there is a net
production of 2ATP molecules.
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Figure 20: Steps of glycolysis
Hence 2 net molecules of ATP are produced during glycolysis. However, when glycolysis begins
from glucose -1-phosphate, one additional ATP is produced in the process. Chloroplastic
products 3-PGA, 3-PGAld and DHAP (dihydroxy acetone phosphate) are transported to cytosol
and can become substrate for some glycolic reactions. Two molecules of pyruvic acid produced
are transported to mitochondria and oxidized to produce large amount of energy (30 ATP).
Pyruvic acid is then the key product of glycolysis. What is the metabolic fate of pyruvate? This
depends on the cellular need. There are three major ways in which different cells handle pyruvic
acid produced by glycolysis. These are lactic acid fermentation, alcoholic fermentation and
aerobic respiration. Fermentation takes place under anaerobic conditions in many prokaryotes
and unicellular eukaryotes. For the complete oxidation of glucose to CO2 and H2O, however,
organisms adopt Krebs’ cycle which is also called as aerobic respiration. This requires O2
supply.
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4.2. Pentose Phosphate Pathway (PPP)

In the pentose phosphate pathway, also located both in the cytosol and the plastid, the
sixcarbon glucose 6-phosphate is initially oxidized to the five-carbon ribulose-5-phosphate. The
carbon is lost as CO2, and reducing power is conserved in the form of two molecules of another
reduced pyridine nucleotide, NADPH. In the following near-equilibrium reactions, ribulose-5-
phosphate is converted into three- to seven-carbon sugars.
Unlike glycolysis which is predominantly involved in the breakdown of sugars and ultimately in
the production of ATP, the oxidative pentose phosphate pathway plays a more important role in
producing intermediates (i.e., amino acids, nucleotides etc) and NADPH. It is another route
available for the oxidation of sugars in plant cells. It produces NADPH and biosynthetic
intermediates (Pentose – a 5-C cpd). The reactions are carried out by soluble enzymes present in
the cytosol and in plastids.
Summary Rxn of PPP 6-Glucose-6-P + 12NADP+ 7H2O 5-Glucose-6-P + 6CO2 +Pi +

12NADPH +12H+
The net result is the complete oxidation of one glucose-6- phosphate molecule to CO2 with the
concomitant synthesis of 12 NADPH molecules.
4.3. Fermentation
In fermentation, say by yeast, the incomplete oxidation of glucose is achieved under anaerobic
conditions by sets of reactions where pyruvic acid is converted to CO2 and ethanol. The enzymes,
pyruvic acid decarboxylase and alcohol dehydrogenase catalyse these reactions. Other organisms
like some bacteria produce lactic acid from pyruvic acid. The steps involved are shown in Figure
21. In animal cells also, like muscles during exercise, when oxygen is inadequate for cellular
respiration pyruvic acid is reduced to lactic acid by lactate dehydrogenase. The reducing agent is
NADH+H+ which is reoxidised to NAD+ in both the processes. In both lactic acid and alcohol
fermentation not much energy is released; less than seven per cent of the energy in glucose is
released and not all of it is trapped as high energy bonds of ATP. Also, the processes are
hazardous – either acid or alcohol is produced. Yeasts poison themselves to death when the
concentration of alcohol reaches about 13 per cent.
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Figure 21: Major pathways of anaerobic respiration
What then is the process by which organisms can carry out complete oxidation of glucose and
extract the energy stored to synthesise a larger number of ATP molecules needed for cellular
metabolism? In eukaryotes these steps take place within the mitochondria and this requires O2.
Aerobic respiration is the process that leads to a complete oxidation of organic substances in
the presence of oxygen, and releases CO2, water and a large amount of energy present in the
substrate. This type of respiration is most common in higher organisms. We will look at these
processes in the next section.
4.4. TCA (Tricarboxylic acid/Kreb cycle

During the nineteenth century, biologists discovered that in the absence of air, cells produce
ethanol or lactic acid, whereas in the presence of air, cells consume O2 produce CO2 and H2O. In
1937 the German-born British biochemist Hans A. Krebs reported the discovery of the citric
acid cycle also called the tricarboxylic acid cycle or Krebs cycle. The elucidation of the citric
acid cycle not only explained how pyruvate is broken down to CO2 and H2O; it also highlighted
the key concept of cycles in metabolic pathways. For his discovery, Hans Krebs was awarded the
Nobel Prize in physiology and medicine in 1953.
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In the citric acid cycle, pyruvate is oxidized completely to CO2, and a considerable amount of
reducing power (16 NADH + 4 FADH2 equivalents per sucrose) is generated in the process. With
one exception (succinate dehydrogenase), these reactions involve a series of enzymes located in
the internal aqueous compartment, or matrix, of the mitochondrion. Occurs inside matrix of
mitochondria Called tricarboxylic acid cycle, because of the importance of the tricarboxylic
acids: citric acid (citrate) and isocitric acid (isocitrate) as early intermediates. Initial reaction
involves decarboxylation of pyruvate. Pyruvate dehydrogenase exists as a large complex of
several enzymes that catalyze the overall reaction in a three-step process: decarboxylation,
oxidation, and conjugation to CoA. The products are NADH (from NAD), C02 and acetic acid in
the form of acetyl-CoA.
The TCA cycle starts with the condensation of acetyl group with oxaloacetic acid (OAA) and
water to yield citric acid (Figure 22). The reaction is catalysed by the enzyme citrate synthase
and a molecule of CoA is released. Citrate is then isomerised to isocitrate. It is followed by two
successive steps of decarboxylation, leading to the formation of α-ketoglutaric acid and then
succinyl-CoA. In the remaining steps of citric acid cycle, succinyl-CoA is oxidized to OAA
allowing the cycle to continue.
Figure 22: the citric acid cycle
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GDP (Guanosine diphosphate) and GTP (Guanosine tri-phosphate), energy rich
molecules that are structurally and biochemically related to ADP and ATP.
Complete oxidation of one molecules of malate yields 4 molecules of CO2, 5 molecules of
NADH and 1 molecules of FADH2 as well as 1 molecules of ATP. NADH and FADH2
subsequently donate their electrons to the electron transport chain. We have till now seen that
glucose has been broken down to release CO2 and eight molecules of NADH + H+; two of FADH
have been synthesised besides just two molecules of ATP in TCA cycle. You may be wondering
why we have been discussing respiration at all – neither O2 has come into the picture nor the
promised large number of ATP has yet been synthesised.
4.5. Mitochondrial Electron transport and oxidative Phosphorylation

ATP is the energy carrier used by cells to drive living processes, and chemical energy conserved
during the citric acid cycle in the form of NADH and FADH2 (redox equivalents with high
energy electrons) must be converted to ATP to perform useful work in the cell. This O2
dependent process, called oxidative phosphorylation, occurs in the inner mitochondrial
membrane. In this section we will describe the process by which the energy level of the electrons
is lowered in a stepwise fashion and conserved in the form of an electrochemical proton gradient
across the inner mitochondrial membrane. Although fundamentally similar in all aerobic cells,
the electron transport chain of plants (and fungi) contains multiple NAD(P)H dehydrogenases
and an alternative oxidase not found in mammalian mitochondria.
In oxidative phosphorylation, electrons are transferred along an electron transport chain,

consisting of a collection of electron transport proteins bound to the inner of the two
mitochondrial membranes. This system transfers electrons from NADH (and related species)
produced during glycolysis, the pentose phosphate pathway, and the citric acid cycle to oxygen.
This electron transfer releases a large amount of free energy, much of which is conserved
through the synthesis of ATP from ADP and P (inorganic phosphate) catalyzed by the enzyme
ATP synthase. Collectively the redox reactions of the electron transport chain and the synthesis
of ATP are called oxidative phosphorylation. This final stage completes the oxidation of
sucrose. NADH produced in Kreb cycle, glycolysis and in photorespiration are oxidized in
mitochondria to produce energy rich compound, ATP. FADH2, a Kerb cycle product, is also
oxidized to produce ATP in mitochondria. Their oxidation involves O2 uptake and production of
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H2O, but NADH or FADH2 does not directly combines with O2 to form H2O. NADH & FADH2
electrons are transferred through series of intermediate carriers to O2 to form H2O. Hence these
electron carriers constitute Electron Transport System (ETS,) of mitochondria. This O2-
dependent process production of ATP, called oxidative phosphorylation, occurs in the inner
mitochondrial membrane.
Complexes of ETS of Mitocliondria

• Electron Transport System (ETS) of mitochondria is composed of 4 complexes: (i) Complex I:
(NADH- ubiquinone oxido-reductase, (ii) Complex II: Succinat ubiquinone oxido-reductase, (iii)
Complex III: Cytochrome C reductase and (iv) Complex IV: Cytochrome oxidase as illustrated
in Fig. below. These complexes of ETS contain carriers such as flavinmono-nucleotide (FMN),
Fe-S proteins, several flavoproteins similar to ferrodoxins, quinines (especially ubiquinones),
several cytochromes up to 4 Cytochrome b, 2 Cytochrome C and Cytochrome a and a3. Attached
to the complexes is ATP synthase similar to the coupling factor in chloroplast (Fig. below).
Coupling factor: a protein that connects trans-membrane potentials to ATP synthesis in
chloroplasts and mitochondria.
Figure 23: mitochondrial structure

Electron Transport
In ETS, cytochromes and Fe-S proteins transfer and receive one electron at a time, whereas
ubiquinones and flavoproteins transfer and receive two electrons at a time. This property of
ubiquinones and flavoproteins is important in creating a pH gradient from the matrix (about 8.0
pH) across inner membrane to the inter membrane space (about 7.0 pH). This pH gradient drives
the formation of ATP from ADP + Pi accompanied with transport of H from the inter membrane
space to matrix.
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Figure 24: Organization of the electron transport chain and ATP synthesis in the inner
membrane of plant mitochondria.
Step-wise electron transport is described as under:

Major electron transport begins from NADH formed in matrix during TCA. Two electrons (eand
2 H+ are passed to FMN which in turn pass e- to Fe-S protein and accepts no H+ and 2H+ are
transferred to inter membrane space. Reduced Fe-S proteins pass e- (2e- in succession) to UQ
which is reduced to UQH2 By accepting 2H+ from matrix. UQH2 transfers electron to various
Cyt. c and 2H+ are transported into inter-membrane space. Another Fe-S protein then receives
and transfer e- to Cyt.c via Q cycle transporting another pair of H+ into inter-membrane space.
Electrons are further transferred from Cyt.c to Cyt. a/a3 accompanied with transport of another
pair of H+ into inter-membrane space. Release of the e- from terminal carrier cyt. a/a3 is accepted
by O2 to form H2O and reaction is catalyzed by cytochrome oxidase. The chemical energy is
conserved in the form of an electrochemical proton gradient, which is created by the coupling of
electron flow to proton pumping from the matrix to the inter membrane space. This energy is
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then converted into chemical energy in the form of ATP by the F0F1-ATP synthase, also located
in the inner membrane, which couples ATP synthesis from ADP and Pi to the flow of protons
back into the matrix down their electrochemical gradient.
Figure 25: Complexes of ETS of mitochondria, path of electron transport and mechanism of
oxidative phosphorylation.
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4.6. Photophosphorylation
Photosynthetic organisms are able to convert light energy into a chemically useful form
(adenosine triphosphate; ATP) by a process termed photophosphorylation. Photophosphorylation
is performed by membrane-associated protein complexes that serve to capture the light energy
(the reaction centres) and subsequently use the captured energy to make ATP (the ATP synthase
complex). The unifying principle in all photosynthetic organisms (prokaryotic or eukaryotic) is
that they use the captured light energy to initiate electron transfer, and produce a charge
separation across a membrane. Following the charge separation process, secondary electron
transfer reactions occur energetically downhill, with some of the energy being conserved as a
proton electrochemical gradient across the membrane. It is this proton electrochemical gradient
that is the driving force for ATP production.
In oxygenic photosynthetic organisms, two deferent reaction centers, termed photosystem II and
Photosystem I, act in series to transfer electrons from the ultimate electron donor, water, to the
terminal electron acceptor, NADP+. Photosystem II generates a powerful oxidant capable of
extracting electrons from water, generating oxygen and protons. Electrons are transferred from
photosystem II to an organic molecule termed plastoquinone to generate the reduced form,
plastoquinol. Electrons from plastoquinol are then transferred to the cytochrome b6/f complex.
Cytochrome f then reduces plastocyanin, which then transfers its electron to photosystem I. A
second charge separation generates a powerful reductant that eventually transfers electrons to
ferredoxin, and ultimately to NADP+. The proton gradient is generated by water oxidation and
electron transfer between Photosystem II and photosystem I.
4.7. Energetic of Glycolysis, TCA, and mitochondrial Electron transport

 Energetic of the processes can be summarized as:
• Glycolysis: Produces 2 molecules of NADH and net 2 molecules of ATP per molecule of
hexoses
• TCA/Kreb Cycle: Produces 8 molecules of NADH, 2 molecules of FADH2 (ubiquinol) and 2
molecules of ATP per molecule of hexoses
• ETS/Oxidative Phosphorylation: Generates 32 molecules of ATP as under:
• (i) 2 x 2 NADH (from glycolysis) = 4 ATP
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• (ii) 3 x 8 NADH (in TCA) =24 ATP

• (iii) 2 x 2 FADH2 (in TCA) = 4 ATP
 Plants have several mechanisms that lower the ATP yield
 the alternative oxidase (helps to regulate EB in response to a changing environment- a
survival protein that allows plants to cope with the stressful environment)
 The uncoupling protein (has a role in hibernation, b/s the energy is used to generate heat
instead of producing ATP)
 The internal, rotenone-insensitive NADH dehydrogenase, NDin (NADH).
 Mitochondrial respiration is controlled by key metabolites (ADP & Pi).
 Respiration is tightly coupled to other pathways
 The respiratory pathways are central to the production of a wide variety of plant
metabolites such as amino acids, lipids and hormones
 So not all Cs that enter the respiratory pathways are oxidized to CO2
Summary
 Energy is stored in the following ways:
 High energy chemical species (e.g. phosphoanhydride bonds in ATP)
 Redox reactions produce: NADH, FADH2
 Membrane potentials (concentration gradient and voltage difference)
 Utilized in the following ways:
 Chemical synthesis reactions (e.g. protein synthesis, DNA & RNA synthesis)
 Mechanical work (e.g. solute transport & muscle function)
 Electrical work (e.g. ion transport & nerve conduction)
4.8. Factors affecting respiration

• Protoplasmic factors
– Quantity and quality of protoplasm present in the cell
– Quality of respiratory enzymes
Young cells (active protoplasm) respire more rapidly than older cells (protoplasm lesser) Type
and age of the plant
– Differences among members of the plant kingdom

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– Differences among tissues or organs of higher plants

– Differences in age of the plant
Differences in development of ripening fruit

• Substrate availability (concentration)
– Respiration ↑ with the ↑ [substrate] e.g. after photosynthesis
Factors affecting rate of respiration: external
Temperature: Respiration typically increases with temperature. Between 0 and 30°C, the
increase in respiration rate for every 10°C increase in ambient temperature (commonly referred
to as the dimensionless, temperature coefficient, Q10) is about 2. Above 30°C the respiration rate
often increases more slowly, reaches a plateau at 40 to 50°C and decreases at even higher
temperatures. High night temperatures are thought to account for the high respiratory rates of
tropical plants. One of the most commonly applied quantitative measures used to describe the
effect of temperature on a process is the temperature coefficient, or Q10, given by the
expression:
At temperatures between 50C and about 250C, or 300C respiration rises exponentially with
temperature and the Q10 value is approximately 2.0 in many but not all plants. Within this
temperature range, a doubling of rate for every 100C rise in temperature is typical of enzymic
reactions. At temperatures above 300C, the Q10 in most plants begins to fall off as substrate
availability becomes limiting. In particular, the solubility of O2 declines as temperatures increase
and the diffusion rate (with a Q10 close to 1) does not increase sufficiently to compensate. As
temperatures approach 500C to 600C, thermal denaturation of respiratory enzymes and damage
to membranes bring respiration to a halt.
Low temperatures are utilized to retard postharvest respiration rates during the storage of fruits
and vegetables. However, complications may arise from such storage. For instance, when potato
tubers are stored at temperatures above 10°C, respiration and ancillary metabolic activities are
sufficient to allow sprouting. Below 5°C, respiration rates and sprouting are reduced in most
tissues, but the breakdown of stored starch and its conversion to sucrose impart an unwanted
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sweetness to the tubers. As a compromise, potatoes are stored at 7 to 9°C, which prevents the
breakdown of starch while minimizing respiration and germination.
– Other factors not limiting ↑ in temp. (0 – 45 oC) ↑ rate of transpiration
– Further ↑ in temp decreases rate of respiration
• O2 may not get access to the cells fast enough
• Removal of CO2 may not be fast enough (it accumulates)
• Respiratory substrate may become a limiting factor
– At low temp, respiration rate is low (fruits and vegetables stored)
[O2] of atmosphere
As the terminal electron acceptor, oxygen availability is obviously an important factor in
determining respiration rate. The oxygen content of the atmosphere is relatively stable at about
21 percent O2. Under normal circumstances, oxygen is rarely a limiting factor. However,
respiration rates decrease if the atmospheric oxygen concentration is below 5% for whole tissues
or below to 3% for tissue slices. These show that oxygen diffusion through the aqueous phase in
the tissue imposes a limitation on plant respiration. The diffusion limitation imposed by an
aqueous phase emphasizes the importance of the intercellular air spaces found in plant tissues for
oxygen availability in the mitochondria. If there were no gaseous diffusion pathway throughout
the plant, the cellular respiration rates of many plants would be limited by an insufficient oxygen
supply.
Light
Increases in light increases rate of respiration
• PS occurs and hence, more substrate available
• Light increase temperature
• Enhances stomatal opening – exchange of gases facilitated – rate of Respiration
increases
Carbon-dioxide concentration [CO2]
It is common practice in the commercial storage of fruits to take advantage of the effects of
atmospheric oxygen and temperature on respiration, and to store fruits at low temperatures under
2 to 3% oxygen and 3 to 5% CO2. The reduced temperature lowers the respiration rate, as does
the reduced oxygen. Low levels of oxygen are used instead of anoxic conditions to avoid
lowering tissue oxygen tensions to the point that stimulates fermentative metabolism. Carbon
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dioxide has a limited direct inhibitory effect on the respiration rate at a concentration of 3 to 5%,
which is well in excess of the 0.036% (360 ppm) normally found in the atmosphere. The
atmospheric CO2 concentration is increasing rapidly as a result of human activities, and it is
projected to double, to 700 ppm, before the end of the twenty-first century. Increase in [CO2],
rate of respiration falls down. Fruits and vegetables can be preserved in an atmosphere of
increased [CO2].
Water
Water saturation/low O2: Diffusion limitation is even more significant when plant organs are
growing in an aqueous medium. When plants are grown hydroponically, the solutions must be
aerated vigorously to keep oxygen levels high in the vicinity of the roots. The problem of oxygen
supply also arises with plants growing in very wet or flooded soils. Some plants, particularly
trees, have a restricted geographic distribution because of the need to maintain a supply of
oxygen to their roots. For instance, dogwood and tulip tree poplar can survive only in well
drained, aerated soils because their roots cannot tolerate more than a limited exposure to a
flooded condition. On the other hand, many plant species are adapted to grow in flooded soils.
Herbaceous species such as rice and sunflower often rely on a network of intercellular air spaces
(aerenchyma) running from the leaves to the roots to provide a continuous, gaseous pathway for
the movement of oxygen to the flooded roots.
Limitation in oxygen supply can be more severe for trees having very deep roots that grow in
wet soils. Such roots must survive on anaerobic (fermentative) metabolism or develop structures
that facilitate the movement of oxygen to the roots. Examples of such structures are outgrowths
of the roots, called pneumatophores that protrude out of the water and provide a gaseous
pathway for oxygen diffusion into the roots. Pneumatophores are found in Avicennia and
Rhizophora, trees that grow in mangrove swamps under continuously flooded conditions.
Shortage of water reduces rate of transpiration b/c WATER
• Maintains turgidity of the cell
• Provides the medium for many respiratory reactions
• Necessary for many enzymatic reactions
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CHAPTER 5: PLANT GROWTH AND DEVELOPMENT
Figure26. Main processes in growth and development are illustrated above through a flow
diagram indicating the sequence of events.
We know that all plant organs are made up of a variety of tissues. Is there any relationship
between the structures of a cell, a tissue, an organ and the function they perform? Can the
structure and function of these changed? All cells of a plant are descendant of the zygote.
Development is considered as the sum of two process-(1) growth and (2) differentiation. It is
essential to know that the development of a mature plant from a zygote follows the highly
ordered succession events. During this process, a complex body organization is formed that
produce roots, leaves, branches, flowers, fruits, seeds and eventually they die.
5.1. Definitions of Growth and Development

Growth can be defined as an irreversible permanent increase in size of an organ or its parts or
even of an individual cell. Generally, growth is accompanied by metabolic processes (both
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anabolic and catabolic), that occur at the expense of energy. Plant growth is unique because
plants retain the capacity for unlimited growth throughout their life. This ability of the plants is
due to the presence of meristems at certain locations in their body. The cells of such meristems
have the capacity to divide and self-perpetuate. The product, however, soon loses the capacity to
divide and such cells make up the plant body. This form of growth wherein new cells are always
being added to the plant body by the activity of the meristem is called the open form of growth.
Growth as the progressive development of an organism: - new cells are added through the
process of cell division. These cells cause growth in tissues and organs. Physiological growth is
an outcome of metabolism. Anabolic activities are synthesis and catabolic activities are
degrading. Both, anabolic and catabolic activities are interlinked. When, anabolic activities occur
in excess of catabolic activities, growth results.
Characteristics of growth: - in plants, growth is limited to meristematic tissues only. Such a

tissue constitutes shoot apex and root apex. New cells are added there and the cells increase in
size. These newly added cells differentiate in to tissue.
Process of growth: - thus three main activities are involved in the process of growth: (1) cell
divisions of meristematic cells (2) enlargement of newly formed cells (3) cellular differentiation
Primarily and secondary growth: - growth takes place in the stem, root and their sub-branches.
Length increases from time to time in plant organs due to the activities of apical meristems
arranged at their tips. Such a growth is called primarily growth. In the stems and roots of dicot
plants, after the completion of primarily structure of organs, through the activities of lateral
meristems known as cambium, the addition of new and more cells in the girth of concerned
organs occurs, this is called secondarily growth. The intercalary meristem located in the nodal
regions of monocot plants is also responsible for growth.
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Growth Rate: - The increased growth per unit time is termed as growth rate. Initially, the rate of
growth in plants is slow. Then it increases very rapidly. In course time, it again slows down.
Thus, rate of growth can be expressed mathematically. An organism or a part of the organism
can produce more cells in a variety of ways. The growth rate shows an increase that may be
arithmetic or geometrical. In arithmetic growth, following mitotic cell division, only one
daughter cell continues to divide while the other differentiates and matures. The simplest
expression of arithmetic growth is exemplified by a root elongating at a constant rate.
Figure27 Diagrammatic representation of: (a) Arithmetic (b) Geometric growth and (c) Stages
during embryo development showing geometric and arithmetic phases
Look at Figure 28. On plotting the length of the organ against time, a linear curve is obtained.
Mathematically, it is expressed as
Lt= Lo + rt
Where, Lt = length at time‘t’
L0= length at time ‘zero’
r = growth rate / elongation per unit time.
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Figure 28: constant linear growths, a plot of length L against time t
Let us now see what happens in geometrical growth. In most systems, the initial growth is slow
(lag phase), and it increases rapidly thereafter at an exponential rate (log or exponential phase).
Here, both the progeny cells following mitotic cell division retain the ability to divide and
continue to do so. However, with limited nutrient supply, the growth slows down leading to a
stationary phase. If we plot the parameter of growth against time, we get a typical Sigmoid or S-
curve
Phases of growth: - All plants pass through various stages of growth. Growth is being expressed
by means of curve plotted against time. The S-shaped or sigmoid curve is typical of growth
pattern of individual organs of a whole plant and of population of plants. It consists of five
distinct phases:
Lag phase: An initial lag period during which internal changes occur that are preparatory to
growth. The increase in size and weight is very slow or negligible during this period. Lag phase,
characterized with very low rate of dry matter accumulation. It involves crop emergence,
development of root and shoot primordia, crown nodes, crown roots, seminal roots, initial
branching of tap roots in dicots, initial tillering in cereals and initial branching in dicots.
Log phase: Log phase is characterized by rapid accumulation of dry matter in both roots and
shoots. It involves rapid development of crown roots and/or fibrous root system in cereals, rapid
tap root development including root branching, tillering in cereals, branching of stem, initiation
of flowering and flower production, rapid leaf area development, initial pod development. The
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terminal stage of this phase results into maximum phase of crop growth. It is followed by the
phase or log period of growth or the grand period of growth during straight line during this
phase.
Maximum phase of growth: This phase involves redistribution of dry matter in reproductive
organs (grains, pods), tubers, and storage roots. Pod filling, dry matter in grains/ seeds in case of
cereals and legumes, oil in oilseeds, storage proteins, starch accumulation in endosperms,
carbohydrate translocation and accumulation in underground roots and tubers (in case of Irish
potato, sweet potato, cassava, cocoyam, yams etc) leading to the physiological maturity.
Figure28: Growth pattern of annual crops with respect to time. Note that lag, log, maximum and
Senescence phases are quite distinct in annual crops.
Senescence phase: The senescence phase is characterized by a decreasing in growth rate and
change in color (loss of chlorophyll pigment) and Loss of organs like leaf, fruit, and seed. Later,
senescence and death of organism sets in giving rise to another component of the growth curve.
Differentiation refers to qualitative changes that normally accompany growth
Differentiation refers to differences, other than size, that arise among cells, tissues, and organs.
Differentiation occurs when cells assume different anatomical characteristics and functions, or
form patterns. Differentiation begins in the earliest stages of development, such as, when
division of the zygote gives rise to cells that are destined to become either root or shoot. Later,
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unspecialized parenchyma cells may differentiate into more specialized cells such as xylem
vessels or phloem sieve tubes, each with a distinct morphology and unique function. The cells
derived from root apical and shoot-apical meristems and cambium differentiate and mature to
perform specific functions. This act leading to maturation is termed as differentiation. During
differentiation, cells undergo few to major structural changes both in their cell walls and
protoplasm. For example, to form a tracheary element, the cells would lose their protoplasm.
They also develop a very strong, elastic, lignocellulosic secondary cell wall, to carry water to
long distances even under extreme tension. Try to correlate the various anatomical features you
encounter in plants to the functions they perform.
Plants show another interesting phenomenon. The living differentiated cells that by now have
lost the capacity to divide can regain the capacity of division under certain conditions. This
phenomenon is termed as dedifferentiation. For example, formation of meristems interfascicular
cambium and cork cambium from fully differentiated parenchyma cells. While doing so, such
meristems/tissues are able to divide and produce cells that once again lose the capacity to divide
but mature to perform specific functions, i.e., get re-differentiated.
Development is the sum of growth and differentiation

The development of a plant/crop refers to the phasic change during the life cycle from
germination to maturity. It can be considered as a series of discrete periods, each identified by an
accompanying process of change in the structure, size or weight of specific organs. The
fundamental characteristic of developmental processes is that they are discrete. The seed either
germinates or it does not and a leaf primordium is invisible or visible. Hence, they can only be
defined in terms of time, and not in length, volume or weight. The duration of developmental
process is usually measured between events that are detected visually. Development is an
umbrella term, referring to the sum of all of the changes that a cell, tissue, organ, or organism
goes through in its life cycle. Development is most visibly manifested as changes in the form of
an organ or organism, such as the transition from embryo to seedling, from a leaf primordium to
a fully expanded leaf, or from the production of vegetative organs to the production of floral
structures. Embryogenesis, vegetative, and reproductive development are the stages of
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sporophytic development of higher plants. Diagrammatic representation of the sequence of

processes which constitute the development of a cell of a higher plant is given in
Figure 29. It is also applicable to tissues/organs.
Figure 29: Sequence of the developmental process in a plant cell.
The tip of the root is covered by a root cap, which provides mechanical protection for the
meristem as the root grows through the abrasive soil medium. The root cap also secretes
polysaccharides, which form a mucilaginous matrix called mucigel. Mucigel lubricates the root
tip as it moves through the soil. The root cap along with its coating of mucigel is also involved in
perception of gravity by roots. The root apical meristem (RAM) is a cluster of dividing cells
located at the tip of the root just behind the root cap. Each time a cell in the meristem divides,
one of the two daughter cells will be retained to continue cell division while the second daughter
cell proceeds to elongate, thus increasing the length of the root and pushing the root tip through
the soil. In the center of the meristem is a region of slowly dividing cells called the quiescent
zone. Cell divisions responsible for new tissues in the elongation root and regeneration of the
root cap take place around the periphery of the quiescent zone.
Tissues that are derived directly from the root and shoot apical rneristems are called primary
tissues. The stems and roots of woody plants, however, grow in diameter as well. An increase in
diameter results from the activity of a meristem called the vascular cambium. Tissues laid down
by the vascular cambium are called secondary tissues, so the vascular cambium is responsible for
secondary growth. The primary tissue of roots and shoots contains a central core of vascular, or
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conducting, elements. The vascular cambium develops between the xylem and phloem and
produces new xylem toward the inside and new phloem toward the outside. Because of its heavy
cell walls and eventual lignification, xylem is a rigid and long-lasting tissue that eventually
occupies the bulk of most woody stems or trunks. Phloem is a more fragile tissue and with each
year's new growth the previous year's cells tend to be pushed outward and crushed.
The root and shoot apical meristems use similar strategies to enable indeterminate growth
Although it might seem difficult to imagine two parts of a plant more different than a shoot and a
root, certain features of the RAM and SAM and the roles they play in enabling indeterminate
patterns of growth invite comparisons. Each of these structures features a spatially defined
cluster of cells, termed initials that are distinguished by their slow rate of division and
undetermined fate. As the descendants of initials are displaced away by polarized patterns of cell
division, they take on various differentiated fates that contribute to the radial and longitudinal
organization of the root or shoot and to the development of lateral organs.
The development, maturation, and germination of seeds: - The life of an individual plant
begins when an egg nucleus in the maternal organs of a flower is fertilized by a sperm nucleus to
form a zygote. Growth and differentiation of the zygote produces an embryo contained within a
protective structure called a seed. Under appropriate conditions, the embryo within the seed will
renew its growth and will continue to develop into a mature plant.
Seeds bearing embryos are formed in the flowers

Flowers appear to vary enormously in structure, yet all flowers follow the same basic plan. A
generic flower consists of four whorls or circles. The two outermost whorls – the sepals and
petals – are vegetative structures; and the two innermost – the stamens and pistil – are the male
and female reproductive structures, respectively. At the base of the pistil, or female structure, is
the ovary, which contains one or more ovules. Within each ovule, a single large diploid cell,
called the megaspore mother cell, undergoes mitosis to produce four megaspore cells. Only one
megaspore cell survives and that cell undergoes meiotic division to produce an embryo sac with
eight haploid nuclei. Subsequent cell division produces a mature embryo sac in which the eight
nuclei are segregated into seven cells.
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One of those cells is the egg. Another is the large central cell containing two polar nuclei. The
male structures, or stamens, surround the pistil and consist of an anther perched on a stalk, or
filament. In some flowers, the sepals and petals may both be colored. Pollen, containing the
sperm nucleus, is produced in the anthers of the stamens. The female egg cells are produced in
the ovary at the base of the pistil. Pollen is transferred to the stigma or stigmatic surface of the
pistil, where it sends out a pollen tube that grows down the style and delivers the sperm nucleus
to the egg. The anther contains a large number of microspore mother cells, each of which
undergoes meiotic division to form uninucleate, single-celled microspores. The microspores
subsequently become encased in heavy, resistant outer walls and the nucleus divides mitotically,
forming two cells – a tube cell and a generative cell – within the original spore wall. This is the
mature pollen grain.
Mature pollen grains are shed from the anthers and carried to the stigmatic surface of the pistil
by insects, wind, and some other vector. Once the pollen grain lands on the stigmatic surface an
event called pollination the pollen grain takes up water and sends out a pollen tube that grows
down the style of the pistil toward the ovule. The tube nucleus migrates down the pollen tube and
appears to direct its growth. The cell wall of the generative cell breaks down and the generative
nucleus divides once to form two sperm nuclei that follow the tube nucleus down the tube as it
elongates. In the final stage, the elongating pollen tube enters the ovule by growing through the
micropyle (the space between the ends of the surrounding integuments) and releases the two
sperm nuclei into the embryo sac. Ultimately, one of the two sperm nuclei enters the egg cell and
fertilizes the egg cell nucleus to form the zygote. The second sperm nucleus enters the large
central cell and fuses with the two polar nuclei to form a triploid endosperm nucleus. The
endosperm nucleus will go on to form the primary nutritive tissue, or endosperm, for the
developing embryo. The involvement of two sperm nuclei in this way is called double
fertilization, a characteristic unique to the flowering plants or angiosperms.
Seed development is characterized by extensive cell divisions

The development of a seed begins with the fertilized ovule, or zygote. The early stage of seed
development is characterized by extensive cell divisions that form the embryo and, in
endospermic seeds, the tissues that store nutrients that will support the eventual germination of
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the seed and seedling development. The first division of the zygote is usually transverse and
immediately establishes polarity of the embryo. The upper cell is destined to become the embryo
itself while the lower cell produces a stalk-like suspensor that anchors the embryo at the base of
the embryo sac.
Nutrients are stored in endosperm that will support germination and seedling
Throughout the development of the embryo, there is a continuous flow of nutrients from the
parent plant into the endosperm or the cotyledons. In some cases, such as the cereal grains and
most other monocots, the endosperm is retained until maturity and may comprise the bulk of the
seed. These are called endospermic seeds. The endosperm of mature endospermic seeds consists
of cells filled with starch along with protein and some small amounts of lipid. In some monocot
seeds, the endosperm is surrounded by one or more distinctive layers of cells, called the
aleurone. Aleurone cells are distinguished by the presence of numerous protein bodies and are
the source of enzymes needed to mobilize nutrients during germination. Endospermic dicot seeds
have retained a significant amount of endosperm and at maturity the cotyledons are thin, leaf like
structures.
In nonendospermic dicot seeds the cotyledons enlarge at the expense of the endosperm and may
occupy as much as 90% of the seed volume at maturity. Both endosperm and cotyledons contain
large quantities of stored carbon (in the form of carbohydrates, lipids, and protein), mineral
elements, and hormones that support the growth and development of the seedling until it can
establish itself as a photosynthetically competent plant.
Maturation is characterized by cessation of embryo growth and development of desiccation
resistance
Maturation is terminated by a dramatic desiccation in which the water content of the seed is
reduced from 80% or 90% to approximately 5%. Surrounding the mature seed is a hard coat
derived from maternal tissues (the integuments) which surrounded the seed during its
development in the ovary. Comprised of heavy-walled cells and covered with a thick, waxy
cuticule, the seed coat often presents a significant barrier to the uptake of both water and oxygen
by the seed.
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Germination is resumption of embryo growth: Because seeds are severely dehydrated, any
metabolic reactions take place so slowly they are scarcely detectable. Seeds are thus quiescent, or
resting, organs that represent a normal hiatus in the life cycle of a plant. The embryo appears to
be in a state of suspended animation, capable in some cases of surviving adverse conditions for
long periods of time. Resumption of embryo growth, called germination, is dependent upon a
number of factors, but three are especially important: adequate water to re-hydrate the tissues,
the presence of oxygen to support aerobic respiration, and a “physiological” temperature.
Although many seeds will germinate over a wide range of temperatures, the optimum range for
most seeds is 25°C to 45°C. The initial step in germination of seeds is the uptake of water and
rehydration of the seed tissues by the process of imbibition. Like osmosis, imbibition involves
the movement of water down a water potential gradient. Imbibition differs from osmosis,
however, in that it does not require the presence of a differentially permeable membrane and is
driven primarily by surface-acting or matric forces. In other words, imbibition involves the
chemical and electrostatic attraction of water to cell walls, proteins, and other hydrophilic
cellular materials. Matric potential, like osmotic potential, is always negative.
Imbibition of water is followed by a general activation of seed metabolism within minutes of

water entering the cells, initially utilizing a few mitochondria and respiratory enzymes that had
been conserved in the dehydrated state. Renewed protein synthesis is also an early event,
utilizing preexisting RNA transcripts and ribosomes, as existing organelles are repaired and new
organelles are formed. This is followed closely by (1) the release of hydrolytic enzymes that
digest and mobilize the stored reserves, and (2) renewed cell division and cell enlargement in the
embryonic axis. Seeds that store carbon reserves principally in the form of fats and oils will carry
out the synthesis of hexose sugars via gluconeogenesis. In most species, germination is
considered complete when the radicle emerges from the seed coat. Radicle emergence occurs
through a combination of cell enlargement within the radicle itself and imbibition pressures
developed within the seed. Rupture of the seed coat and protrusion of the radicle allows it to
make direct contact with water and nutrient salts required to support further growth of the young
seedling.
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Many seeds will not germinate even though the minimal environmental conditions have been
met. These seeds are said to be dormant and will not germinate until additional conditions have
been met. The most common causes of seed dormancy are the impermeability of the seed coat to
water or oxygen or physiological immaturity of the embryo at the time the seed is shed from the
mother plant. Immature seeds must undergo complex biochemical changes, collectively known
as after-ripening, before they will germinate. After-ripening is usually triggered by low
temperature, a mechanism that appears to ensure that the seed will not germinate precociously in
the fall but will germinate when favorable weather returns in the spring.
The pattern of development from embryo to adult: The first structure to emerge when a seed
germinates is the radicle. The radicle, which is the nascent primary root, anchors the seed in the
soil and begins the process of mining the soil for water and nutrients. As the primary roots
elongates, it gives rise to branch, or lateral, roots. Unlike the situation in the shoot apical
meristem, lateral roots do not originate in the root apical meristem. Lateral root primordial
originate in the pericycle, a ring of meristematic cells that surround the central vascular core, or
stele, of the primary root. The growing lateral root works its way through the surrounding cortex,
either by mechanically forcing its way through or by secreting enzymes that digest the cortical
cell walls. Lateral root primordia arise in close proximity to the newly differentiated xylem
tissue, which allows vascular elements developing behind the growing tip of the lateral root to
maintain connections with the xylem and phloem of the primary root.
Emergence of the radicle is followed by elongation of the shoot axis. It proceeds through a
combination of cell division and enlargement of the cells laid down by the meristem. The rate
and extent of elongation is subject to a variety of controls, including nutrition, hormones, and
environmental factors such as light and temperature. The final height of a shoot is determined by
the rate and extent to which internodes – the sections of stem between leaf nodes – elongate. In
some plants, such as pea (Pisum sativum), elongation occurs primarily near the apical end of the
youngest inter-node. The older internodes effectively complete their elongation before the next
inter-node begins.
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In other plants, elongation may be spread through several internodes, which elongate and mature
more or less simultaneously. Still others exhibit changing rates of elongation with successive
internodes, usually increasing. In some plants, internodes fail to elongate, thus giving rise to the
rosette habit in which all the leaves appear to originate from more or less the same point on the
stem. This rosette habit is common in biennial plants (those that flower in the second year) such
as cabbage and root crops such as carrot (Daucus carota) before they reach the flowering stage.
Failure of inter-node elongation is commonly related to low levels of the plant hormone,
gibberellin, since application of the hormone usually stimulates inter-node elongation in rosette
plants.
Senescence and programmed cell death: The final stage in the development of cells, tissues,
and organs is senescence, an aging process characterized by increased respiration, declining
photosynthesis, and an orderly disassembly of macromolecules. Senescing cells and tissues are
metabolically very active – a number of metabolic pathways are turned off and new pathways,
principally catabolic in character, are activated. Catabolism of proteins, for example, releases
organic nitrogen and sulfur in the form of soluble amines, while nucleic acids release inorganic
phosphate. Chlorophyll is broken down and lipids are converted to soluble sugars via
gluconeogenesis. The products of these pathways are all small, soluble molecules that are readily
exported from the senescing tissue. Senescence thus enables the plant to recover nutrients from
cells or tissue that have reached the end of their useful life and reallocate them to other parts of
the plant that survive or for storage in the roots.
Programmed cell death (PCD) is a specialized type of senescence: Programmed cell death
(PCD) is broadly defined as a process in which the organism exerts a measure of genetic control
over the death of cells. PCD requires energy and is normally regulated by a distinct set of genes.
PCD is essential for normal vegetative and reproductive development. One example is the
development of xylem tracheary elements. In order to function efficiently as a conduit for water
transport, the protoplast of the developing tracheary element must die and be removed at
maturity. PCD also operates in the formation of aerenchyma, a loose parenchymal tissue with
large air spaces. Aerenchyma normally forms in the stems and roots of water lilies and other
aquatic plants. These air spaces, created by a cell death program, provide channels for oxygen
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transport to the submerged portions of the plant. Even corn (Zea mays) and other terrestrial
plants can be induced to form aerenchyma when subject to flooding. PCD is also an important
factor in plant responses to invading pathogens and abiotic stress. When a plant recognizes a
pathogen, for example, host cells in the immediate area of the infection undergo PCD. This
deprives the invading pathogen of living tissue and either slows or prevents it spread.
5.2. Processes and Measurements of Growth

Growth is an irreversible increase in volume or size. Growth is a quantitative term, related only
to changes in size and mass. For cells, growth is simply an irreversible increase in volume. For
tissues and organs, growth normally reflects an increase in both cell number and cell size.
Growth can be assessed by a variety of quantitative measures. Growth of cells such as bacteria
or algae in culture, for example, is commonly measured as the fresh weight, cell number or
packed cell volume in a centrifuge tube. For higher plants, however, fresh weight is not always a
reliable measure. Most plant tissues are approximately 80 percent water, but water content is
highly variable and fresh weight will fluctuate widely with changes in ambient moisture and the
water status of the plant. Dry weight, determined after drying the material to a constant weight, is
a measure of the amount of protoplasm or dry matter (i.e., everything but the water).
Dry weight, determined after drying the material to a constant weight, is a measure of the
amount of protoplasm or dry matter (i.e., everything but the water). Dry weight is used more
often than fresh weight, but even dry weight can be misleading as a measure of growth in certain
situations. Consider the example of a pea seed that is germinated in darkness (Figure 31). In
darkness, the embryo in the seed will begin to grow and produce a shoot axis that may reach 25
to 30 cm in length.
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Figure 31: Changes in fresh and dry biomass of pea seed as it develops into seedling in darkness
(source: Salisbury F.B., Ross C.W., 1992.
Although we intuitively sense that considerable growth has occurred, the total dry weight of the
seedling plus the seed will actually decrease compared with the dry weight of the seed alone
prior to germination. The dry weight decreases in this case because some of the carbon stored in
the respiring seed is lost as carbon dioxide. In the light, this lost carbon would be replaced and
even augmented by photosynthesis but photosynthesis doesn’t operate in darkness. In a situation
such as this, either fresh weight or the length of the seedling axis would be a better measure of
growth. Length, and perhaps width, would also be suitable measures for an expanding leaf.
Length and width would not only provide a measure of the amount of growth, but a length to
width ratio would also provide information about the pattern of leaf growth. There is not any
special universal measure unit to characterize plant growth. It should be obvious that many
parameters could be invoked to measure growth, dependent to some extent on the needs of the
observer. Whatever the measure, however, all attempts to quantify growth reflect a fundamental
understanding that growth is an irreversible increase in volume or size.
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While cell division and cell enlargement normally go hand-in-hand, it is important to keep in
mind that they are separate events; growth can occur without cell division and cell division can
occur without growth. For example, cell division is normally completed very early in the
development of grass coleoptiles and the substantial elongation of the organ that follows is due
almost entirely to cell enlargement. Years ago, it was shown that wheat (Triticum sp.) seeds
could be made to germinate even after having been irradiated with gamma radiation sufficient to
block both DNA synthesis and cell division. The result was a small seedling produced by cell
enlargement alone. Such seedlings generally did not survive more than two or three weeks but,
except for having abnormally large cells, their morphology was more or less normal. On the
other hand, cell division can also proceed without cell enlargement. During the early stages of
embryo development in flowers, for example, a portion of the embryo sac goes through a stage
in which cell division continues to produce a larger number of increasingly smaller cells, with no
overall increase in the size of the embryo sac.
Leaf Area and Leaf Area Index: The leaf area of a crop is a determinant factor in mechanisms
such as radiation interception and water and energy exchange. Thus, accurate measurements of
LAI are essential to understand the interaction between crop growth and environment.
Psychopathological studies require calculating healthy leaf area duration (HAD, days) and
healthy leaf area absorption (MJ m-2). Direct measurement of LAI in crops is commonly carried
out by measuring manually, or with equipment (LAI-2000 Plant Canopy Analyzer), the total leaf
area of the plant in relation to the soil area covered by these leaves. This evaluation is difficult
and time consuming. Direct measurements are generally collected at the end of the growing
season and are prone to very large errors that may lead to inconclusive results in crop growth
analysis. Indirect techniques, which are based on the close coupling between radiation
penetration and canopy structure, are a good alternative to the direct LAT measurement
techniques. The measurement of canopy gap fractions (the fraction of sky visible through the
canopy) at various angles is a particularly powerful approach. Gap fractions have been measured
using fisheye photographs, by traversing a sunward- pointed sensor beneath the canopy, by linear
light sensors and by pushing metal probes through the canopy.
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LAI may also be estimated using an empirical relationship between the maximum width of the
central leaflet and total leaf area using regression equation. The leaf area of common beans can
be calculated the empirical relationship: LA = 2.1371 X (L1.9642) — 2.7013; where LA is leaf
area (cm2) and L is the maximum width of the central leaflet of each leaf (cm). The LAI values
will be obtained by dividing the total leaf area of each plant on each day of assessment by the
area of the soil occupied per plant (0.5mx0.1=0.05 m2). This method is accurate but also very
time consuming, laborious, and difficult to carry out in the field. Use of LAI-2000 Plant Canopy
Analyzer is recommended.
5.3. Dry Matter Partitioning

Plants, in contrast to animals, consist of autotrophic organs that produce sugars and amino acids
from photosynthesis, and heterotrophic organs that consume these products in growth, storage
and reproduction. These metabolically contrasting parts of the plant are often referred to as
source and sink tissues, respectively. Source tissues are net exporters of assimilate. These are
principally mature green leaves, green stems and green seed pods, but also non-photosynthetic
tissues such as germinating seeds, tubers, tap roots, etc. Photosynthesis, either directly or
indirectly, provides the building blocks for all biomass production as well as sink development
and growth. Sink tissues are net importers of assimilate and consist of organs that are rapidly
growing such as meristems and immature leaves and vegetative storage tissues such as tubers,
seeds or roots. Source and sink metabolism are tightly coupled because information on assimilate
availability in each organ is perceived and used to orchestrate gene expression.
Translocation is the movement of dissolved materials throughout the plant. The rate at which,
photosynthetic products such as sucrose move from the leaves to the sink organs controls the rate
of photosynthesis. Plant species that have high Photosynthetic rates also have relatively high
translocation rates. This is consistent with the idea that effective removal of Photosynthetic
products maintains rapid CO2 fixation. Severe infection of leaves by pathogens often so severely
inhibits Ps rates that these leaves become sugar importers instead of sugar exporters. The
adjacent healthy leaves then gradually attain marked increases in Ps rates, suggesting that
enhanced translocation has reduced CO2 fixation.
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Partitioning describes the distribution of new assimilates to growth of various plant parts and to
respiration. Partitioning is subject to a control system whose flexibility is indicated by its
capacity to respond appropriately to different environmental stimuli. Thus shading is likely to
increase partition (relative to the total dry weight) to the laminae whereas water or nutrient stress
improves partition to the roots. Removal of plant parts results in temporarily increased partition
to other similar parts until partition relationship of intact control plants is re-established. All the
vegetative sinks are potentially in competition during the vegetative phase, the dominant sink
being sheath and stem. Yield depends on the translocation of assimilates in most species largely
sucrose from the leaves or other photosynthetic tissues to the parts of economic importance
tubers, seeds, etc. Other parts – roots, stems, young leaves and fruits also rely on the assimilates
produced by the mature leaves. The source and sinks are linked by a transport pathway of
phloem sieve elements. Mature leaves are the primary sources of assimilates, from the current
photosynthesis, but this can be supplemented by the mobilization of stored reserves/organs such
as the stems, which may be described as secondary sources.
In most dicots, cotyledons initially serve as source of reserves, but are subsequently raised above
ground, when they assume a photosynthetic function which supports further growth until the first
true leaves are photosynthetically competent. Storage function is taken over by endosperm from
which mobilized reserves are absorbed by the cotyledons and then translocated to the embryonic
axis. Sucrose from the endosperm is accumulated in the phloem cells of the cotyledons largely
sustain growth of the hypocotyls and roots. Elongation of the hypocotyls raises the cotyledon.
Dry matter partitioning between roots, above ground vegetative growth, and reproductive growth
are usually modified by water deficits. The response depends upon the species, when the stress
occurs, its duration, and its severity. The increase in root-shoot ratio of crops under water deficits
may reflect an increase in the proportion of assimilates allocated to the roots, or a change in the
rate of death or turnover of roots relative to the shoot. An increase in root growth may indicate a
greater density or a greater depth of roots. Soil and root resistances to water uptake are reduced
when root length density increases, thereby permitting higher water flow rates through the plant
and delaying the onset of severe plant water deficits. Whilst partitioning of dry matter to roots
may enhance water uptake, it represents a loss to above ground dry matter production.
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An exception is root crops, such as cassava, where water deficit could be beneficial to economic
yield, provided that the increased proportion of dry matter partitioned to the roots goes mainly to
the tubers. Since photosynthesis is inhibited more than translocation during stress, dry matter
produced before flowering may be transferred from the stem and roots to the grain during grain
filling – “Compensatory translocation”. The amount of pre-anthesis assimilate partitioned to
the grain is dependent on the timing and severity of water deficits as it affects the source-sink
balance.
5.3. Factors Affecting Growth and Development

Plant growth and development are controlled by internal regulators that are modified according
to environmental conditions. The long term climatic conditions for a region determine the type of
vegetation in that region, and regional environmental factors affect growth and development of
the plants. The three most ecologically important environmental factors affecting plant growth
are temperature, water (precipitation), and light.
Light: - Light is the ultimate source of energy and the most important ecological factor affecting
plant growth. It has direct effect on photosynthesis and transpiration. Light in terms of intensity,
quality and periodicity influence the growth very much.
Light intensity: A weak light promotes shortening of internodes and affects expansion of leaf.
Very weak light reduces the rate of overall growth and also photosynthesis due to poor
development of chlorophyll and higher rate of water loss from the plant.
Light quality: The different wavelengths of light have different responses to growth. In blue
violet radiation, the internodal growth is pronounced while green colour light promotes the
expansion of leaves as compared to complete spectrum of visible light. The red light favors the
growth while infra-red and UV is detrimental to growth.
Light duration: There is remarkable effect of the duration of light on the growth. The induction
and suppression of flowering depend on duration
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Temperature: - Temperature, an approximate measurement of the heat energy available from

solar radiation, is an important factor because most plant biological activity and growth occur
within only a narrow range of temperatures, between 0°C and 50°C. High temperatures limit
biological reactions because the complex structures of proteins are disrupted or denatured.
Although respiration and photosynthesis can continue slowly at temperatures well below 0°C if
plants are physiologically “hardened”, low temperatures limit biological reactions because water
becomes unavailable when it is frozen and because available energy is inadequate.
Water requirement: - Water is an essential factor for growth. It is essential for uptake of
nutrients, translocation of nutrients and food materials, regulating transpiration and for various
physiological processes like photosynthesis, respiration and enzymatic activities.
Soil: - is a natural medium that provides anchorage for the plant and supplies water and mineral
nutrients for normal growth. Soil consists of mineral matter, organic matter, air, and water. The
proportion of these four constituents and the types of mineral and organic material determines
soil properties such as soil type, soil pH, and fertility.
Soil type: - Soil is made up in part of mineral particles grouped as sand (0.05 to 2 mm), silt
(0.002 to 0.05 mm), and clay (<0.002 mm). The ratio of these determines soil types, such as clay,
clayey loam, loam, sandy loam, and sand. Loam is composed of sand, silt, and clay in relatively
even amounts, and exerts a greater influence on soil properties than does sand, silt, or clay. Soil
type determines the soil's capacity to store water and nutrients, aeration, drainage, and ease of
field operations. Sandy soils are easily tilled, well-drained and aerated but usually have low
fertility and water-holding capacity. Clayey soils, on the other hand, are more fertile and have
high water retention but are poorly drained and aerated.
Soil pH: - is a measure of the soil's acidity or alkalinity, and it affects the plant indirectly by
influencing the availability of nutrients and the activity of microorganisms. Nutrients are most
available at pH levels between 6.5 and 7.5. Nutrients in the soil may be chemically tied up or
bound to soil particles and unavailable to plants if the pH is outside this range. Individual plants
have pH preferences and grow best if planted in soils that satisfy their pH requirements.
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Soil fertility: - is the inherent capacity of soil to provide plant nutrients in adequate amounts and
in proper balance for the growth of specific plants. A fertile soil is usually rich in nitrogen,
phosphorus, and potassium, and contains sufficient trace elements and soil organic matter that
improves soil structure and soil moisture retention.
Soil salinity: - refers to the presence of excess salts in soil water, which often results from
irrigated agriculture. After the plants take up the water, the dissolved salts from irrigated water
start to accumulate in the soil. Soil salinity is usually measured as electrical conductivity (EC) of
soil solution, and expressed in decisiemens per meter (dS/m). Excess salts generally affect plant
growth by increasing osmotic tension in the soil, making it more difficult for the plants to take up
water. Excessive uptake of salts from the soil by plants also may have a direct toxic effect on the
plants. Soil salinity is most pronounced in arid areas.
Internal factors
1. Growth hormones and their availability
2. Resistance to climatic, edaphic and biological stresses
3. Photosynthetic rate and respiration
4. Assimilate partitioning and nitrogen content
5. Chlorophyll and other pigments
6. Source-sink relationship and enzyme activities
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CHAPTER 6: PLANT HORMONES AND GROWTH REGULATORS
Multi-cellular plants are complex organisms and their orderly development requires an
extraordinary measure of coordination between cells. In order to coordinate their activities, cells
must be able to communicate with each other. The principal means of intercellular
communication within plants are the hormones. Hormones are signal molecules that individually
or cooperatively direct the development of individual cells or carry information between cells
and thus coordinate growth and development. Plant hormones have been the subject of intensive
investigation since auxin was first discovered almost a century ago.
Plant growth regulators or phytohormones are organic substances other than nutrients produced
naturally in higher plants, controlling growth or other physiological functions at a site remote
from its place of production and active in minute amounts. Plant hormones are a group of
naturally occurring, organic substances which influence physiological processes at low
concentrations. The processes influenced consist mainly of growth, differentiation and
development, though other processes, such as stomatal movement, may also be affected. Plant
hormones have also been referred to as ‘phytohormones’ though this term is infrequently used.
A definition of plant hormones with still wider scope has been given by Johannes van Overbeek
(1950). According to him, the plant hormones are defined as “organic compounds which
regulate plant physiological process— regardless of whether these compounds are naturally
occurring and/or synthetic ; stimulating and/or inhibitory ; local activators or substances which
act at a distance from the place where they are formed.”
The plant hormone concept probably derives from observations of morphogenic and
developmental correlations by Sachs between 1880 and 1893. He suggested that
"Morphological differences between plant organs are due to differences in their material
composition" and postulated the existence of root-forming, flower forming and other substances
that move in different directions through the plant. At about the same time Darwin was making
his original observations on the phototropism of grass coleoptiles that led him to postulate the
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existence of a signal that was transported from the tip of the coleoptile to the bending regions
lower down.
The plant growth is controlled by certain chemical substances called the plant hormones. These
are also known as the growth factors, growth substances, plant growth regulators (PGR) or
phytohormones. These can either promote or inhibit the growth of the plant. On the basis of their
function, plant hormones are grouped into six types:
1. Auxin
2. Gibberellin
3. Cytokinin
4. Ethylene
5. Abscisic acid (ABA)
6. Jasmonic Acid
6.1. Auxins
During the latter part of the 19th century, Charles Darwin and his son Francis studied plant
growth phenomena involving tropisms. One of their interests was the bending of plants toward
light. This phenomenon, which is caused by differential growth, is called phototropism. In some
experiments the Darwins used seedlings of canary grass (Phalaris canariensis), in which, as in
many other grasses, the youngest leaves are sheathed in a protective organ called the coleoptile.
Coleoptiles are very sensitive to light, especially to blue light. If illuminated on one side with a
short pulse of dim blue light, they will bend (grow) toward the source of the light pulse within an
hour. The Darwins found that the tip of the coleoptile perceived the light, for if they covered the
tip with foil, the coleoptile would not bend. But the region of the coleoptile that is responsible for
the bending toward the light, called the growth zone, is several millimeters below the tip. Thus
they concluded that some sort of signal is produced in the tip, travels to the growth zone, and
causes the shaded side to grow faster than the illuminated side. The results of their experiments
were published in 1881 in a remarkable book entitled The Power of Movement in Plants. After
Darwin, Boysen-Jenson observed that if the tip of the coleoptile is decapitated, it neither grow
nor bend towards light. The normal bending also occurred if the tip is cut off and replaced with
an intervening block of gelatin. It was thus evident that the influence was due to chemical
diffusing through the block of gelatin.
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There followed a long period of experimentation by many investigators on the nature of the
growth stimulus in coleoptiles. This research culminated in the demonstration in 1926 by Frits
Went of the presence of a growth-promoting chemical in the tip of oat (Avena sativa) coleoptiles.
It was known that if the tip of a coleoptile was removed, coleoptile growth ceased. Previous
workers had attempted to isolate and identify the growth-promoting chemical by grinding up
coleoptile tips and testing the activity of the extracts. This approach failed because grinding up
the tissue released into the extract inhibitory substances that normally were compartmentalized in
the cell. Went’s major breakthrough was to avoid grinding by allowing the material to diffuse out
of excised coleoptile tips directly into gelatin blocks. If placed asymmetrically on top of a
decapitated coleoptile, these blocks could be tested for their ability to cause bending in the
absence of a unilateral light source. Because the substance promoted the elongation of the
coleoptile sections, it was eventually named auxin from the Greek auxein, meaning “to
increase” or “to grow.”
F.W. Went cut off tips of oat coleoptile and placed them on small agar blocks and let them for a
few hours to allow all the chemical influence to diffuse into the agar. Like this, he isolated auxin
in agar block. Then he placed this agar block on the freshly decapitated coleoptiles and observed
that the coleoptiles grew and bent away from the side on which the block was placed. He named
this chemical influence as auxin which is derived from a greek word 'auxein' which means to
increase or to grow. So, Went is credited as the inventor of auxin.
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Auxins are compounds that positively influence cell enlargement, bud formation and root
initiation. They also promote the production of other hormones and in conjunction with
cytokinins, they control the growth of stems, roots, and fruits, and convert stems into flowers.
Auxins were the first class of growth regulators discovered. They affect cell elongation by
altering cell wall plasticity.
Auxins decrease in light and increase where it is dark. They stimulate cambium cells to divide
and in stems cause secondary xylem to differentiate. Auxins act to inhibit the growth of buds
lower down the stems (apical dominance), and also to promote lateral and adventitious root
development and growth. Leaf abscission is initiated by the growing point of a plant ceasing to
produce auxins. Auxins in seeds regulate specific protein synthesis, as they develop within the
flower after pollination, causing the flower to develop a fruit to contain the developing seeds.
Auxins are toxic to plants in large concentrations; they are most toxic to dicots and less so to
monocots.
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Because of this property, synthetic auxin herbicides including 2, 4-D and 2, 4, 5-T have been
developed and used for weed control. Auxins, especially 1- Naphthaleneacetic acid (NAA) and
Indole-3-butyric acid (IBA), are also commonly applied to stimulate root growth when taking
cuttings of plants. The most common auxin found in plants is indole acetic acid or IAA. Indole
Acetic Acid (IAA) is the only naturally occurring auxin in plants.
The synthetic auxins include,
IBA: Indole Butyric Acid
NAA: Naphthalene Acetic acid
MENA: Methyl ester of Naphthalene acetic acid
MCPA: 2 Methyl 4-chloro phenoxy acetic acid
TIBA: 2, 3, 5 Tri iodo benzoic acid
2, 4-D: 2, 4 dichloro phenoxy acetic acid
2, 4, 5-T: 2, 4, 5 – Trichloro phenoxy acetic acid
Natural auxins may occur in the form of either free auxins- which freely move or diffuse out of
the plant tissues readily or bound auxins- which are released from plant tissues only after
hydrolysis, autolysis or enzymolysis.
Distribution of auxin in plants: - In plants, auxin (IAA) is synthesized in growing tips or
meristematic regions from where; it is transported to other plant parts. Hence, the greatest
concentration of auxins is usually found in the growing apices of the plant, i.e., in the coleoptile
tip, in buds and in the growing tips of leaves and roots. However, auxin is found widely
distributed throughout the plant body. In general, it may be stated that where there is active
growth, there is auxin production. The formation of auxin by a mature organ like leaf, however,
suggests that growth may not be the pre-requisite to auxin production.
Thimann (1934) studied the distribution of auxin, in detail, in etiolated Avena coleoptile. He
found that the concentration of auxin drops as one progresses from the coleoptile tip to its base;
the highest concentration being at the tip and the lowest at the base. If one progress further from
the base of the coleoptile along the root, there is a steady increase in auxin content till a
maximum is reached in the root tip. Of the two maximal values, that for the stem tip is much
higher than that for the root tip.
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Transport of auxin in plant: - The main axes of shoots and roots, along with their branches,
exhibit apex-base structural polarity, and this structural polarity is dependent on the polarity of
auxin transport. Soon after Went developed the coleoptile curvature test for auxin, it was
discovered that IAA moves mainly from the apical to the basal end (basipetally) in excised oat
coleoptile sections. This type of unidirectional transport is termed polar transport. Auxin is the
only plant growth hormone that has been clearly shown to be transported polarly, and polar
transport of this hormone is found in almost all plants. Because the shoot apex serves as the
primary source of auxin in the plant, polar transport has long been believed to be the principal
cause of an auxin gradient extending from the shoot tip to the root tip. The auxins are transported
in plants from one organ to the other. The usual direction of auxin transport is downward but
when added to the soil these are absorbed by the roots and carried upward along with
transpiration stream to various plant organs. The prevailing downward movement takes place
through the living phloem cells whereas the upward movement occurs through the dead xylem
elements. The most striking characteristic of auxin movement is its almost strict basipetal (from
apex to base) polarity. Thus, more than one pathway is responsible for the distribution of auxin
in the plant. Both natural auxins (IAA) and synthetic auxins move the same way. The velocity of
auxin transport varies from 26 mm per hour to 6.4 mm per hour (Rajgopal, 1967). These rates are
higher than the rates of diffusion. The velocity of auxin transport is unaffected by temperature,
although the amount of auxin transported is proportional to the temperature. Also, the distance
over which the transport occurs does not influence the velocity.
Destruction / Inactivation of auxin in plants: To be effective developmental signals, hormones
must be short-lived and should not accumulate over time. Auxin catabolism ensures the
degradation of active hormone when the concentration exceeds the optimal level or when the
response to the hormone is complete. Like IAA biosynthesis, the enzymatic breakdown
(oxidation) of IAA involves more than one pathway. On the basis of isotopic labeling and
metabolite identification, two oxidative pathways are probably involved in the controlled
degradation of IAA. In one pathway, the indole moiety of IAA is oxidized to form oxindole-3-
acetic acid (OxIAA) and subsequently, OxIAA-glucose (OxIAA-Gluc). In another pathway,
IAA-aspartate conjugates are oxidized to OxIAA. Auxin is destructed by the enzyme IAA
oxidase in the presence of O2 by oxidation.
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IAA Oxidase
IAA + H2O2 + O2 3-methylene oxindole + H2O + CO2
Rapid inactivation may also occur by irradiation with x-rays and gamma rays. UV light also
reduces auxin levels in plants. Inactivation or decomposition of IAA by light has been called as
photo oxidation.
Mechanism of Action: IAA increases the plasticity of cell walls so that the cells stretch easily in
response to turgor pressure. It has been suggested that IAA acts upon DNA to influence the
production of mRNA. The mRNA codes for specific enzymes responsible for expansion of cell
walls. Recent evidences indicate that IAA increases oxidative phosphorylation in respiration and
enhanced oxygen uptake. The growth stimulation might be due to increased energy supply and it
is also demonstrated that auxin induces production of ethylene in plants.
Functions of Auxin: It was previously thought that the sole function of auxins was to promote
cell enlargement. But the work done in later years has proved them to be deeply associated with
a variety of functions. In some cases they act as a stimulating agent, in others as an inhibitory
agent and in still others as a necessary participant in the growth activity of other phytohormones
such as gibberellins and cytokinins. The various growth processes in which the auxins (both
natural and synthetic) play their role are discussed below:
1. Cell elongation. It is usually considered that cell elongation occurs only in the presence of
auxins and also that the rate of elongation is directly proportional to the amount of auxin applied
provided no other factors are limiting. But relatively high concentrations usually exert inhibitory
effect on this phase of growth. Some commercial weed killers are chemically similar to auxin
and have similar effects. They cause affected plant cells to elongate and the plant grows. The
exact mechanism in not yet understood, but the auxin seems to interfere with DNA transcription
and RNA translation. The amount of auxin applied in a weed killer is far greater than the amount
produced within the plant and so the rate of growth produced is much greater than normal. The
plant cannot sustain this rate of growth: it becomes weakened, unable to reproduce, and then it
dies. The auxins like herbicides have a much greater effect on dicots than on monocots, such as
grasses. This is partly because dicot leaves have a larger surface area than monocot leaves and so
absorb more herbicide.
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Auxins also play a significant role in the elongation of petiole, mid rib and major lateral veins
of the leaves. Thus, adenine favours enlargement in detached leaves of radish and pea. Similarly,
coumarin has been shown to promote expansion of leaves in some plants. An osmotic
equilibrium exists in a cell where the turgor pressure developed is counterbalanced by the wall
pressure acting in opposite direction. Regarding the mechanism of cell elongation, it is thought
that auxins stimulate cell elongation by modifying certain conditions responsible for this
equilibrium (Devlin, 1969).
These modifications include:

A. An increase in osmotic contents of the cell
B. An increase in permeability of the cell to water
C. A decrease in wall pressure
D. An increase in wall synthesis and
E. An inducement of specific RNA and protein synthesis.
2. Cambial activity. In the spring season, the trees exhibit growth by developing buds which
later on open. This is then followed by elongation of the young stems. This resumption of growth
by cambial cells is activated by the auxins which move basipetally in the stems from developing
buds. Snow (1935) has shown that a steady supply of auxin at 1/1,000,000 mg per hour (or of
IAA at 1/500,000 mg per hour) from a gelatin block, upon affixing it to the cut end of a
decapitated shoot of sunflower (Helianthus annuus) seedling, stimulated meristematic activity of
the cambium. The suggestion by Jost (1940) seems to imply that the major function of hormones
that migrate from the developing apex of the epicotyl is to activate the differentiation of
procambial strands.
3. Callus formation and galls. Besides acting as stimulants of cell elongation, the auxins may
also activate cell division. This may be illustrated by applying 1% IAA in lanolin paste to a
debladed petiole of a bean plant. This causes prolific division of parenchyma cells resulting in
the formation of a swelling or callus tissue at a point where the auxin in applied. The amount of
callus tissue formed is directly proportional to the concentration of IAA applied (Ropp, 1950).
Bezerinck, in as early as 1885, investigated the production of cecidomid galls in Poa nemoralis.
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He compared gall production to callus formation and thought of a substance coming from the
larval body as a causative agent. He regarded this action of animal substance as analogous to that
of the inner causes which lead to the formation of roots in normal plants.
4. Rooting of stem cuttings (= Formation of adventitious roots). It is a common observation

that the presence of buds on a cutting favours development of roots when the lower end is dipped
in a suitable rooting medium. Developing buds are effective in accelerating root formation.
Young leaves also favour the initiation of roots on the cuttings. These observations led to the
suggestion that the root formation is favoured by the auxins which are synthesized in the buds
and young leaves and are later translocated to the basal part of the cutting.
5. Apical dominance. It has been generally observed that so long as the apical bud is intact on
the plant, the growth of the lateral buds remains suppressed. Upon removal of the apical bud, the
lateral bud nearest the apical bud establishes its dominance over the remaining buds, causing
them to become inactive again. This inhibitory effect of a terminal bud upon the development of
the lateral buds is called apical dominance and produces a cone-shaped plant. This is why a
gardener keeps on trimming the hedge occasionally in order to obtain a denser growth. Plants
that are tall and unbranched exhibit strong influence of apical dominance than those which are
short and branched. The relation of apical dominance with the auxin supply was first reported by
Skoog and Thimann (1934).They demonstrated that when agar block containing auxin b or IAA
was kept on the decapitated shoot of broad bean (Vicia faba), the lateral buds, as might be
expected, resulted in the usual suppression of growth as if the terminal buds were present. But
when the same decapitated shoot was reheaded with an agar block containing no auxin, these
lateral buds resumed growth. Similar results were also obtained with field-grown tobacco plants
using NAA as the auxin.
Although the exact mechanism behind this growth correlation is not yet known, the best
explanation for this has been furnished by Snow (1939, 40). According to him, under the
influence of auxin, a growth inhibitor is formed that is responsible for the inhibition of growth of
the lateral buds. The growth inhibitor is synthesized in some unknown manner when the auxin
moves in its usual downward direction. The theory lends support from the fact that certain
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growth inhibitors have chemical structure not much different from that of the auxins. It may,
thus, be visualized that such a synthesis of growth inhibitor from the auxin may take place in the
tissues.
The knowledge of apical dominance has been utilized practically in solving the problem of
storage of the potatoes. Potatoes, stored for some time, sprout and become sweet in taste, thus
causing financial loss to the grower as the sweet taste is disliked by its consumers. But spraying
the potatoes with auxins like indole butyric acid (IBA) and NAA would prevent sprouting (or in
other words, would prolong dormancy) by inhibiting the development of buds or ‘eyes’ ; the
effect persisting for as long a period as 3 years. Although such treatment is in the interest of the
breeder but certainly not in that of the consumer.
6. Delay (or inhibition) of abscission of leaves. The abscission of leaves can be delayed or
inhibited by the application of auxins on the surface of the lamina or on the cut surface of a
debladed petiole. The controlling behaviour of the auxins on the abscission was first noted by
Laibach (1933) who showed that the extract of orchid pollinia is capable of preventing the leaf
fall. Since then, enough work has been carried out in this direction. Addicott and Lynch (1955)
have proved conclusively the delaying effect of IAA on the abscission of various plant organs.
As to the mechanism of abscission, it has been suggested that the leaf fall is retarded by the
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basipetal migration of a hormone from the blade to the base of the petiole. Removal of the leaf
blade eliminates the supply of hormone to the abscission zone and thus induces leaf fall. The
correlation between the amount of diffusible auxin and the age of the organ concerned was
demonstrated by Shoji et al in 1951. They found high auxin contents in the young leaf blades of
bean plant as compared to their petioles. But on ageing, the auxin contents in the leaf blade and
the petiole fall almost to the same level.
7. Flowering. A flowering hormone, florigen, is produced in the leaves under correct light and
dark period. It moves first down the petiole and then up the stem to the growing apex where it
causes the development of floral buds in place of vegetative buds. Auxins are useful in
modifying flowering in one of the following ways:
A. Altering earliness- Leopold and Guernsey (1953) conducted experiments on oat, corn,
barley, peas etc., and found that when their seeds were treated with auxins followed by low
temperature treatment at 4°C for 5—15 days, they produced quantitative increase in earliness.
Chakravarti (1955) also produced earliness and greater yield in mustard when auxin was applied
before chilling. No gains were, however, obtained when the auxin was applied after chilling.
B. Inducing flowering- The flowering in pineapple has been successfully promoted in Hawaii
islands by treating the fields with sodium naphthalene acetate at some time during growing
period. In Caribbean areas, however, 2, 4-D is more commonly used. Leopold and Thimann
(1949) have, likewise, found low concentrations of auxins such as a-NAA or IAA as effective in
inducing flowering in barley and Teosinte. Furthermore, 2, 4-D has been successfully used in
control of flowering in sweet potato.
C. Preventing or delaying flowering- Although natural flowering of many species of plants is

inhibited by high endogenous auxin concentrations, attempts to attain this inhibitory effect by
applying auxins exogenously (i.e., from outside) usually fail as the plants are damaged before
effective control is obtained. But experimentally in certain plants like cabbage and celery, the
bolting is prevented by applying p-chlorophenoxypropionic acid during cold periods which
would otherwise normally induce flowering.
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8. Fruiting. Auxins play significant role in fruiting by modifying it in one of the following ways:
A. Fruit setting-Fruit set refers to the changes in the ovary leading to the development of the
fruit. These changes are usually induced after pollination and fertilization the two processes
which are in some way concerned with the release of some stimulus of hormone nature. But the
development of fruit without fertilization, i.e., parthenocarpy (parthenos = virgin; carpos =
fruit), however, is also a common feature in the plant world and henceforth occurs frequently in
nature. Such a parthenocarpic development of fruits nowadays has also been induced artificially.
For example, Yasuda (1934) demonstrated it by application of pollen extracts to cucumber
flowers. An analysis of the extract showed that auxins were present in it. Later, Gustafson (1936,
39) Also observed that ovaries of many plants (orange, lemon, grape, banana, tomato etc.) could
be induced to develop into seedless fruits by application of IAA in lanolin paste to their stigmas.
The various other auxins employed for this purpose are IPA, IBA, α-NAA, phenoxyacetic acid
(POA), α-naphthoxyacetic acid (NOA) etc.
B. Fruit thinning- In many instances, the trees bear extensively large number of fruits. This
causes the trees to fail to produce average number of new flower buds. Such trees, therefore,
have to produce fruits either at alternate years (alternate bearing) or if yearly, the number of
fruits is greatly reduced (infrequent bearing). These trees, obviously, require thinning. Fruit
thinning was, for the first time, done in apple when naphthalene acetic acid applied to flowers
failed to set the fruits and, in fact, caused a decrease in fruit set. It is surprising to note that
naphthalene acetic acid appears to be the only successful auxin which brings about thinning of
fruits. However, other examples of auxins employed for fruit thinning are a-2, 4, 5-
trichlorophenoxyacetic acid for thinning of pears (Griggs, 1951) and p-chlorophenoxyacetic acid
for thinning of grapes (Weaver and Winkle,1952).
C. Control of premature fruit dropping- In many fruit trees, the unripe fruits fall off on
account of the formation of an abscission layer, thus causing serious losses in yield to the
gardeners. This problem has now been successfully overcome in many cases like apples by the
application of auxins which prevent the formation of abscission layer and thus check pre-harvest
drop of the fruits. Besides apples, such as control has also been induced in citrus fruits (like
oranges and lemons) using 2, 4-D and 2, 4, 5-trichlorophenoxyacetic acid as auxins.
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D. Improving fruit quality- The various processes like colouration, softening, sweetening and
ripening are all involved in improving the quality of the fruit. Auxin effects on fruit colouration
are most pronounced in apples where use of 2, 4, 5 trichlorophenoxyacetic acid has greatly
increased red pigments. 2, 4-D when applied to bananas hastened the process of ripening as the
auxin facilitates the conversion of starch to sugars. By injecting 2, 4-D, IBA or maleic hydrazide,
the accumulation of sugars was reported in sugarcane.
9. Increase in respiration. James Bonner (1953), for the first time, recognized that auxins
stimulate the process of respiration. And as such a direct relation between growth due to auxin
treatment and the rate of respiration has been found. Greater the growth, higher is the rate of
respiration. Such knowledge about the behaviour of auxins has been applied in controlling the
development of weeds which grow obnoxiously in the crop fields. This has been successfully
achieved by spraying 2, 4-D which acts as a weed killer. In fact, this hormone, which operates
only on broad-leaved herbaceous plants, increases the rate of respiration so much so that the
plants die of over-oxidation and exhaustion. And fortunately most of the weeds are
broadleaved dicots and the crop plants are usually narrow-leaved monocots which, henceforth,
escape destruction on spraying 2, 4-D. Another hormone 2, 4-dichloropropionic acid (or
dalapon), however, destroys graminaceous weeds.
10. Great weapon of war. Auxins when applied in greater concentrations on enemy crop fields
by air may cause devastation of land and thus form the basis of what is called biological warfare.
The synthetic auxins (such as 2, 4-D, 2, 4, 5-T and NAA) have been widely used since 1940s.
Their use has decreased production costs by reducing the amount of labour and mechanical
weeding, needed to grow and effectively harvest a crop. Unfortunately, the effects of synthetic
auxins have not all been positive. Most of the negative effects can be traced to a defoliant, once
used in Vietnam War, by the code name Agent Orange. Agent Orange was a 1: 1 mixture of 2,
4-D and 2, 4, 5-T that was sprayed throughout the jungles of vietnam and followed by napalm
bombs. This resulted in the destruction of hundreds of square kilometres of Vietnam forests.
11. Auxins as herbicide: - In addition to these applications, today auxins are widely used as
herbicides. The chemicals 2, 4-D and dicamba are probably the most widely used synthetic
auxins. Synthetic auxins are very effective because they are not metabolized by the plant as
quickly as IAA is. Because maize and other monocotyledons can rapidly inactivate synthetic
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auxins by conjugation, these auxins are used by farmers for the control of dicot weeds, also
called broad-leaved weeds, in commercial cereal fields, and by home gardeners for the control of
weeds such as dandelions and daisies in lawns. Auxin in high concentration promotes ethylene
synthesis that causes abscission. Herbicides are enzyme inhibitors (ATP synthesis). Herbicides
denature enzymes. Many different classes of herbicides have been developed, and they act by
blocking amino acid, carotenoid, or lipid biosynthesis or by disrupting cell division. Other
herbicides, such as DCMU (dichlorophenyl dimethyl urea) and paraquat, block photosynthetic
electron flow. Many herbicides, DCMU among them, act by blocking electron flow at the
quinone acceptors of PS II, by competing for the binding site of plastoquinone that is normally
occupied by QB. Other herbicides, such as paraquat, act by accepting electrons from the early
acceptors of PS I and then, a species reacting with oxygen to form superoxide, O-2 that is very
damaging to chloroplast components, especially lipids.
6.2. Gibberellins
The gibberellins are another class of compounds whose minute quantities profoundly stimulate
the growth of many plants.
Discovery: The gibberellins were discovered in an interesting and incidental way. In early part
of the 20th century, Japanese farmers noted that some plants in rice fields were taller, thinner and
paler than the normal plants. They had longer and narrower leaves markedly overgrowing their
unaffected neighbours and were sometimes devoid of fruits too. They named this disease as
“bakanae”, meaning foolish seedlings. Sawada (1912) suggested that the disease is due to a
‘substance’ secreted by a parasitic as Comycetous Fungus, Gibberella fujikuroi (the perfect form,
occurring only occasionally; the imperfect form is Fusarium moniliforme), in infecting the
diseased plants. This suggestion was experimentally supported by Ewiti Kurosawa (1926) who
demonstrated that sterile filtrates of the fungus could initiate symptoms of bakanae disease in
healthy rice seedlings. Later in 1939, Yabuta and Hayashi isolated this growth promoting
substance in crystalline form and named it as gibberellin A, which has now been shown as a
mixture of many growth promoters collectively known as gibberellins.
Definition: A gibberellin (abbreviated as GA, for gibberellic acid) may be defined as a
compound which is active in gibberellin bioassays and possesses a gibbane ring. There are,
however, other compounds (like kaurene) which are active in some of the assays but do not
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possess a gibbane ring. Such compounds have been called gibberellin-like rather than
gibberellins.
Isolation, Distribution and Biosynthesis
About 29 gibberellins were previously isolated and their chemical structures known. These have
been named as gibberellin A1 (GA1), gibberellin A2 (GA2) and so on up to gibberellin A29
(GA29). Of these, Cross et al (1961) have isolated 6 gibberellins from the fungus, Fusarium
moniliforme and designated them as GA1, GA2, GA3, GA4, GA7 and GA9. The subscripts
number shows the order of their discovery. The same year, MacMillan et al isolated 3
gibberellins from bean seeds and named them as GA5, GA6 and GA8. GA10 and GA13 have
been discovered by Mulholland (1963). All these compounds are sometimes referred to as
constituting the gibberellin A series. As of 2002, 125 gibberellins were discovered. Although
the gibberellins were originally isolated from a fungus, but now they have been shown to be
present in almost all the groups of plant kingdom including angiosperms, gymnosperms, ferns,
mosses and algae but are unknown in bacteria. For example, GA1, and GA5 have been isolated
from immature seeds of Phaseolus vulgaris by West and Phinney (1959). Although all the
organs of the flowering plants contain gibberellins, but the highest level has been detected in
seeds. Young leaves and roots are also rich in them. It may, thus, be generalized that rapidly
growing and developing regions of the plant possess higher concentrations of gibberellins.
Distribution of gibberellins in plant

Gibberellins are found in all parts of higher plants including shoots, roots, leaves, flower, petals,
anthers and seeds. In general, reproductive parts contain much higher concentrations of
gibberellins than the vegetative parts. Immature seeds are especially rich in gibberellins (10-100
mg per g fresh weight). In plants, gibberellins occur in two forms free gibberellins and bound
gibberellins. Bound gibberellins usually occur as gibberellin glycosides.
Site of GA biosynthesis & transport
The GAs is mostly synthesized in:
young immature seeds
Young developing leaves
Shoot apex
Roots
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Mature seeds contain almost no GAS. GA biosynthetic enzymes and GAs are specifically
localized in young, actively growing buds, leaves, and upper internodes. Both phloem and xylem
participate in GAs transport (nonpolar).
Physiological Roles: Gibberellins may be regarded as natural phytohormones on account of
their wide range of distribution in plants and specificity of response of individual flowering
plants to the exogenously applied gibberellins. The gibberellins, however, play important roles in
the following processes:
1. Genetic dwarfism. In certain plants, dwarfism is caused by the mutation of a single gene.
Such individuals are called ‘single gene dwarfs’. In these plants, dwarfism is due to shortening of
internodes rather than a decrease in the number of internodes. Application of gibberellins on
such dwarfs causes them to elongate so much as to become indistinguishable from the tall
normal plants. Elongation of the stem, in fact, takes place due to an elongation in the internodes
rather than an increase in the number of internodes. Thus, genetic dwarfism has been
successfully overcome by gibberellin A3 treatment in many single gene dwarf mutants like
Pisum sativum, Vicia faba and Phaseolus multiflorus (Brian and Hemming, 1955). The
gibberellins thus make most plants grow taller by causing the internodes to elongate considerably
(Figure 32). Two views have been put forward regarding the mechanism of control of dwarfism
by gibberellins.
A. It is due to the lack of endogenous gibberellins in dwarf plants or if at all present, they are in
traces as to have no effect.
B. A natural inhibitor is present in those plants which retard growth. And the gibberellin, when
applied, nullifies the effect of this inhibitor.
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Figure 32: Wheat stems, cut lengthwise, to show the modes. The 4 plants on the left, treated
with gibberellic acid, have internode lengths much greater than the intermodes of the two
untreated on the right.
2. Bolting and flowering: In many herbaceous plants, the early period of growth shows rosette
habit with short stem and small leaves. ‘Rosette plants’ are characterized by their profuse leaf
development and retarded internodal growth. But prior to the reproductive phase, there occurs
striking elongation in the internode so that the plant attains 5 to 6 times the original height.
Under short days, the rosette habit is retained while under long days bolting occurs i.e. the stem
elongates rapidly and is converted into polar axis bearing flower primordia. This bolting can also
be induced in such plants by the application of gibberellins even under non-inductive short days.
In Hyoscyamus niger (a long day plant) gibberellin treatment causes bolting and flowering under
non-inductive short days. While in long day plants the gibberellin treatment usually results in
early flowering. In short day plants, its effects are quite variable. It may either have no effect or
inhibit or may activate flowering. By regulating the amount of gibberellin applied, it is also
possible to separate shoot elongation from flowering; with low dosages of gibberellins, the plant
will bolt but not flower.
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It is, therefore, not amazing to find a direct correlation between the amount of gibberellin present
and the habit of the plant, whether rosetted or bolted. Native gibberellin-like substances are
found in higher concentrations in the bolted forms than in the non-bolted ones. This has been
experimentally demonstrated in quite a few plants including the biennial Hyoscyamus niger by
Lang (1957) and the cold-requiring plant Chrysanthemum morifolium and a long-day plant
Rudbeckia speciosa by Harada and Nitsch (1959). As far as the use of gibberellins in agriculture
is concerned, it may be possible to grow cold requiring plants in warm countries and long-day
plants in short-day conditions at lower altitudes Gibberellic acid (GA) hastened flowering and
improved the flower yield in Coriandrum sativum (coriander). This was accomplished by a
decline in starch content and an increase in reducing sugars, as well as enhanced amylase
activity. It was inferred that GA hastened flowering probably through its influence on
carbohydrate metabolism (Amrutavalli, 1979).
3. Light-induced inhibition of stem growth. Light-grown plants reveal suppressed stem growth
than the dark-grown (or etiolated) plants, indicating that light has an inhibitory effect on stem
elongation. But this inhibitory effect of light on stem elongation can be reversed at least in some
plants (like Pisum sativum) by the application of gibberellins on these plants. This clearly
suggests that endogenous gibberellin is the limiting factor in stem elongation. Lockhart (1961)
has given a possible explanation for it. According to him, exposure to light lowers the level of
available gibberellins present in the plant. The lowered available gibberellin contents then, in
turn, decrease the plasticity of cell walls, thus inhibiting stem growth. The theory has, however,
not won the universal support on account of the following drawbacks:
A. Stem elongation is also induced in mustard seedlings, grown in dark, upon application of
gibberellin.
B. In some plants, gibberellin-stimulated stem growth has been found to be partially due to
enhanced cell division and has nothing to do with cell wall plasticity.
C. Germination of the seeds of Lactuca sativa is not only promoted by gibberellins but by red
light too.
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4. Parthenocarpy. Like auxins, the gibberellins are also capable of inducing parthenocarpic
fruit-set. Gibberellins are, in fact, more efficient than the auxins in inducing Parthenocarpy. For
example, Wittwer and Bukovac (1957) have found gibberellin to be about 500 times more
effective than IAA in inducing Parthenocarpy in tomatoes. Moreover, there are cases where
auxins have failed to induce Parthenocarpy while gibberellins are effective, as shown
experimentally for apples (Davison, 1960) and stone fruits (Crane et al, 1960).
Gibberellininduced Parthenocarpy has been reported in many plants such as Cucumis sativus
(cucumber), Solanum melongena (brinjal) and Zephyranthes sp. Whether the production of
parthenocarpic fruits is a direct action of gibberellins or an interaction with the natural auxins of
the plant has not been conclusively proved. Germination of the pollen grains is stimulated by
gibberellins; likewise, the growth of the fruit and the formation of parthenocarpic fruits can be
induced by gibberellin treatment. In many cases example pome and stone fruits where auxins
have failed to induce Parthenocarpy, the gibberellins have proven to be successful. Seedless and
fleshly tomatoes and large sized seedless grapes are produced by gibberellin treatments on
commercial scale.
5. Breaking dormancy of seeds. The light-sensitive seeds (lettuce, tobacco) show poor
germination in dark and on exposure to light their germination starts vigorously. But when these
seeds are treated with GA3, the light requirement is alleviated and they germinate in dark.
6. Breaking dormancy of buds. In temperate areas, the buds produced in winter remain
dormant until the next spring due to very low temperature. The dormancy in such cases is
overcome by gibberellin treatment. Thus, GA3 treatment to birch buds has replaced the light
requirement for breaking dormancy. Gibberellins are also capable of breaking dormancy in
potato tubers.
7. Role in abscission. GA3 treatments have shown accelerated rate of abscission in explants of
bean (Chatterjee and Leopold, 1964) and of Coleus (Gupta and Kaushik, 1969).
8. Stimulation of enzyme activity in cereal endosperm: Yomo (1960) and Paleg (1960)
working independently showed that the gibberellins applied exogenously could stimulate
amylase activity in isolated barley endosperm. It was then shown that it is the aleurone layer of
the endosperm which is sensitive to the gibberellin. Subsequent researches by Paleg (1964) and
Varner (1964) revealed that GA treatment of isolated aleurone can cause release of the enzymes,
amylase and proteinase. Finally, Jacobson and Varner (1967) showed that the two enzymes
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(amylase and proteinase) induced by GA treatment arise through de novo synthesis. de novo is a
Latin phrase, meaning anew. One important function of gibberellins is to cause the synthesis of
the enzyme α -amylase in the aleurone layer of the endosperm of cereal grains during
germination. This enzyme brings about hydrolysis of starch to form simple sugars which are then
translocated to growing embryo to provide energy.
9. Sex expression. Gibberellins are also capable of altering the sex of the flowers. Galun (1959)
could induce maleness by foliar application of GA3 to the female flowers of Cucumis. Also, the
antheridia have been induced to develop in many fern gametophytes by GA3 treatment.
Relationship between Auxins and Gibberellins
Auxins and gibberellins are similar to each other in that both promote cell elongation, flowering
and Parthenocarpy. These, however, differ from each other in many of the physiological
activities. These differences are listed in Table below.
Differences between auxins and gibberellins
Accumulated evidences indicate that auxins and gibberellins act both independently and
together, depending upon the type of plant and the conditions under which the plant grows. The
fact whether the auxins and the gibberellins interact or not is not conclusively proved.
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6.3. Cytokinins (= KININS)

The Cytokinins were discovered in the search for factors that stimulate plant cells to divide (i.e.,
undergo cytokinesis). Auxins and gibberellins, besides inducing cell elongation, also do promote
cell division under certain conditions. But this behaviour of them is an exception rather than a
rule. However, there exist in plants many substances inducing cell division. For example, Van
Overbeek et al (1941) found coconut milk as an active stimulant of cell division. Later, in 1955
Carlos Miller et al isolated a “cell-division-stimulating factor” from yeast DNA. It was named as
kinetin because of its amazing power to stimulate cell division (cytokinesis) in the presence of an
auxin. In subsequent years, many other compounds promoting cell division have been
synthesized. Miller and his associates (1956) have grouped all such compounds including kinetin
under a generic name kinin. D.S. Leetham (1963) of New Zealand proposed the term cytokinins
for such substances. This term is the most acceptable one. Fairley and Kilgour (1966), however,
prefer to use the term ‘phytokinins’ for such substances in order to distinguish them from the
peptide hormones of animal gastrointestinal tract.
Cell division (cytokinesis) promoting hormones; the most active cell division promoting
substances are kinetin, zeatin and zeatin riboside. Cytokinins have effects on many other
physiological and developmental processes, including leaf senescence, nutrient mobilization,
apical dominance, the formation and activity of shoot apical meristems, floral development, the
breaking of bud dormancy, and seed germination. Cytokinins also appear to mediate many
aspects of light-regulated development, including chloroplast differentiation, the development of
autotrophic metabolism, and leaf and cotyledon expansion.
Isolation, Distribution and Biosynthesis: Although kinetin does not occur in nature but other
kinins are found occurring widely, if not universally in plants. The naturally-occurring kinins do
not occur free in nature but are normally bound to a pentose sugar, ribose and sometimes to an
inorganic phosphate, the ribonucleotide. Fruits and endosperm are the richest sources of kinins.
Coconut milk and maize endosperm possess the active substance. Substances with cytokinin
activity have also been reported in tomato juice, in floral extracts of apples and pears and also in
cambial tissues of certain plants. A kinetin like substance is also present in peach embryo and
sunflower root exudates.
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Diphenyl-urea and many of its derivatives have cytokinin activity. Several synthetic cytokinins
are available. These include benzimidazole and 6-benzyladenine. Adenine also has some
cytokinin activity. In angiosperms, the cytokinins are synthesized mostly in roots and probably
originate at the root tips. Whether the shoots also synthesize cytokinins or else receive their
cytokinin requirement from the roots is not certain. Contrary to what was first believed, the
cytokinins are not breakdown products of DNA. Rather, they are made via the mevalonate
pathway, the same pathway used to make gibberellins. Like gibberellins, the cytokinins move
non-polarly in xylem, phloem, and parenchyma cells.
Biosynthesis of Cytokinins: It is assumed that cytokinins are synthesised as in the case of
purines in plants (nucleic acid synthesis). Root tip is an important site of its synthesis. However,
developing seeds and cambial tissues are also the site of cytokinin biosynthesis. Kende (1965)
reported that cytokinins move upwards perhaps in the xylem stream. However, basipetal
movement in petiole and isolated stems are also observed. Seth et al (1966) found that auxin
enhances kinetin movement (translocation) in bean stems. Young organs - seeds, fruits, leaves
and root tips are sites of cytokinin synthesis. Root tips are the major sites of cytokinin synthesis
and are transported to growing meristems, seeds, fruits and leaves through xylem. Sieve tubes
(phloem) transport limited amount of cytokinins.
Physiological Role of cytokinins

1. Cell division: - The most important biological effect of kinetin on plants is to induce cell
division especially in tobacco pith callus, carrot root tissue, soybean cotyledon, pea callus etc.
Kinins are notable for their stimulatory effect on cell division. Using tobacco pith cultures
(Figure 33), Skoog and Miller (1957) found that, in addition to IAA, kinetin is also needed for
growth. The growth response is much more pronounced when both IAA and kinetin are used
together in right ratio of concentrations. When either of them is used alone, a little response is
produced which is due to the presence of small amounts of endogenous kinetin-like substances
and IAA, already present in the tissues.
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Figure 33: Effect of kinetin in stimulation of cell division in tobacco pith cultures
If a mixture of cytokinin and auxin is added to unspecialized cells (Figure 34), they will begin to
differentiate. A high cytokinin to auxin ratio will lead to the formation of shoots, buds and leaves
while a low cytokinin to auxin ratio will lead to root formation. The one treatment followed by
the other provides a means of forming small plantlets. Such in vitro culture methods have
become widely adopted for the rapid propagation of new plant varieties. The technique allows
growers to produce very large numbers of plants, quite rapidly and in a small space. The
alternative conventional propagation methods can take several years and cover a large area of
land.
A. If a small piece of pith from a shoot is placed on agar in aseptic conditions, it will grow into a
mass of unspecialized cells called callus.
B. Plants can be grown from these cells.
The process of cell division completes in 3 steps, viz., DNA synthesis, mitosis and cytokinesis.
Studying the specific influence of IAA and kinetin alone on any of these 3 steps, Patau, Das and
Skoog (1957) found that IAA is involved in the first two steps of cell division (i.e., in DNA
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synthesis and mitosis) and that the last step (i.e., cytokinesis) is controlled by kinetin. It has been
suggested that the adenine moiety of the kinetin molecule is essential for cell division.
Figure 34: Callus formation and production of plants from callus cells
2. Cell elongation. Besides auxins and gibberellins, kinetin also promotes cell elongation. Such
promotion after kinetin treatment has been observed in tobacco pith cultures, tobacco roots and
bean leaf tissues. Since cell elongation induced by kinetin has been well established, the kinetin
should not be regarded as exclusively a cell division factor. Significant cell enlargement has been
observed in the leaves of Phaseolus vulgaris, pumpkin cotyledons, tobacco pith culture, cortical
cells of tobacco roots etc.
3. Counteraction of apical dominance. The auxins emanating from the apical bud inhibit the
growth of lateral buds (apical dominance). Wickson and Thimann (1958) studied the antagonistic
effect of auxin and kinetin in apical dominance using pea stem sections in culture solutions. They
found, as might be normally expected, that the growth of lateral buds is inhibited when the
culture medium contained IAA and is uninhibited when the culture medium does not contain
IAA. They further noted that addition of kinetin, along with IAA, stimulates the growth of lateral
buds. External application of cytokinin promotes the growth of lateral buds and hence
counteracts the effect of apical dominance
The above workers also conducted experiments with entire shoots, i.e., with the apical bud intact.
As long as the apical bud is present, the lateral buds do not develop but removal of the apical bud
leads to the stimulation of growth of the lateral buds. If, however, the intact shoot is soaked in
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kinetin solution, the inhibition of lateral buds is checked to a great extent or, in other words, the
lateral buds tend to develop, although less vigorously, as if the apex of the shoot has been cut off.
The above findings point out towards the possibility of controlling apical dominance by
maintaining a proper balance of concentrations between IAA and the endogenous kinetin-like
substances. Studies conducted in subsequent years by Sachs and Thimann (1964, 67) and
Panigrahi and Audus (1966) also indicated that the cytokinins are strong promoters of lateral bud
growth.
4. Root growth. Kinetin is capable of stimulating as well as inhibiting root development. Skoog
and Miller (1957) found stimulatory effect of kinetin, when applied along with IAA, on root
initiation and development in stem callus cultures. Similarly, kinetins also induced increase in
dry weight and elongation of the roots of lupine seedlings.
5. Shoot growth. The callus tissue of tobacco can be kept in an undifferentiated state so long as
the proper balance of IAA and kinetin is maintained. If, however, the amount of kinetin is
increased, leafy shoots are initiated to develop. Bean seedlings, soaked in kinetin solution, also
showed an increase in dry weight and a marked elongation of stem and petioles.
6. Organogenesis. Cytokinins can cause organogenesis (i.e., the formation of organs) in a
variety of tissue cultures. For instance, Skoog and Miller (1957) observed that tobacco pith callus
can be made to develop either buds or roots by changing the relative concentrations of kinetins
and auxins. High kinetin and low auxin contents result in the production of buds. In reverse
condition (high auxin and low kinetin), however, the roots appear on the pith. The kinins also
stimulate the production of buds in leaf segments of various plants such as Saintpaulia ionantha,
Bryophyllum sp and Begonia sp. In addition to the root and shoot differentiation, the cytokinins
also bring about other morphogenetic responses. These are:
A. Maturation of pro-plastids into plastids
B. Differentiation of tracheids
C. Induction of Parthenocarpy
D. Induction of flowering
7. Breaking dormancy of seeds. Cytokinins are also effective in breaking seed dormancy in
lettuce, tobacco, white clover and carpet grass. Thimann (1963) suggested that the site of
cytokinin action in such cases is the cotyledon. Furthermore, the inhibitory effect of infrared
light on germination of lettuce seeds is also alleviated by kinetin treatment. The seeds of
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parasites such as Striga asiatica require the presence of host plant for germination. But when
treated with kinetin, the seeds germinate even in the absence of their host.
8. Delay of senescence (= Richmond-Lang effect). The term senescence refers to the ageing of
the leaves which is associated with the loss of chlorophyll and the breakdown of proteins.
Richmond and Lang (1957) showed that the senescence in the detached leaves of Xanthium
could be postponed for many days by kinetin treatment. This effect of kinetin in retarding
senescence (or ageing) is known as Richmond-Lang effect. According to Mothes and Engelbracht
(1961), the cytokinins have the ability to attract certain substances including auxins and to
prevent the movement of leaf components out of the treated area. However, the mobilizing effect
of cytokinin may actually induce senescence in others parts of the plant. Osborne (1962)
suggested that the high protein content in kinetin-treated areas is probably due to enhanced
protein synthesis than their breakdown. The protein synthesis, in its turn, is dependent on RNA
synthesis, a process governed by kinetins. It may, however, be emphasized that the cytokinin
induced delay in leaf senescence occurs only in detached leaves; cytokinins have little or no
effect on senescence in attached organs. Leaf senescence is also delayed by the formation of
adventitious roots. As the roots are rich in cytokinins, the transport of these cytokinins from roots
to leaves could account for the delayed senescence.
A correlation between the age of the leaf and the kinetins has been established. Mature leaves of
tobacco respond more vigorously to kinetin treatment in delaying senescence than the young
leaves. Cytokinins are sometimes used commercially to maintain the greenness of excised plant
parts, such as cut flowers. However, their use on edible crops such as broccoli is banned in some
countries. This is possible because any compound like cytokinin that resembles a nucleic acid
component is automatically a suspected carcinogen.
9. Role in abscission. Cytokinins can accelerate as well as retard the process of abscission in leaf
petioles depending on the site of their application (Osborne and Moss, 1963). In explants petioles
of Coleus blumei, accelerated abscission on kinetin application. “
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10. Effects on cotyledons. Cytokinins promote cellular division and expansion in cotyledons.
Cellular expansion results from cytokinin-induced increases in wall plasticity that doesn’t
involve wall acidification. Cytokinins also increase the amount of sugars (especially glucose and
fructose) in cells, which may account for the osmotic influx of water and the resulting expansion
of cytokinin treated cells in cotyledons.
11. Morphogenesis: - It has been shown that high auxin and low kinetin produced only roots
whereas high kinetin and low auxin could promote formation of shoot buds.
12. Accumulation and translocation of solutes: - Plants accumulate solutes very actively with
the help of Cytokinin and also help in solute translocation in phloem.
13. Protein synthesis: - Osborne (1962) demonstrated the increased rate of protein synthesis due
to translocation by kinetin treatment.
14. Other effects: - Cytokinins provide resistance to high temperature, cold and diseases in some
plants. They also help in flowering by substituting the photoperiodic requirements. In some
cases, they stimulate synthesis of several enzymes involved in photosynthesis.
12. Commercial applications: - Cytokinins have been used for increasing shelf life of fruits,
quickening of root induction and producing efficient root system, increasing yield and oil
contents of oil seeds like ground nut. Tobacco plants over expressing the gene for cytokinin
oxidase. The plant on the left is wild type. The two plants on the right are over-expressing two
different constructs of the Arobidopsis gene for cytokinin oxidase: AtCKXJ and AtCKX2. Shoot
growth is strongly inhibited in the transgenic plants
6.4. Ethylene
Discovery: During the 1800s, the city streets of Germany were illuminated by lamps that burned
“illuminating gas”. Soon after these lamps were installed, city residents made a curious
observation: plants growing near the lamps had short thick stems and leaves falling from most of
them. The mystery was solved in 1901 by a Soviet plant physiologist, Dimitry Neljubow who
identified ethylene as the combustion product of “illuminating gas” that was responsible for
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defoliation and stunted growth of plants growing near the lamps. He also showed that only micro
quantities are needed to bring about these effects, i.e., only 0.06 ppm of ethylene. Later in 1910,
an annual report submitted to the Japanese Department of Agriculture recommended that
oranges be not stored with bananas, because oranges released something that caused premature
ripening of the bananas; this “something” was in 1934 identified by R. Gane as ethylene which is
made by plants. Subsequent researches showed that ethylene has all the characteristics which
warrant its inclusion under plant hormones, i.e., it is made in one part of a plant and transported
to another, where it induces a physiological response. Thus was discovered the gaseous plant
hormomes: ethylene. Later, the presence of ethylene was shown in certain fungi (Penicillium
digitatum, Alternaria citri) and in the leaves, flowers and fruits of many higher plants; its
recognition as a natural plant hormone was confirmed by Pratt and Goeschl only in 1969.
Distribution and Biosynthesis: A bioassay is a measurement of the effect of a known or
suspected biologically active substance on living material. All parts of angiospermous plants
produced ethylene but especially large amounts are released into the air by roots, the shoot apical
meristem, nodes, senescing flowers and ripening fruits (for example, the dark flecks on a
ripening banana peel are concentrated pockets of ethylene). Because most ethylene-induced
effects result from ethylene in the air, the effects of ethylene can be contagious: ethylene made
by one “bad” (i.e., overripe) apple can “spoil” (i.e., induce rapid ripening of) an entire bushel of
apples. Ethylene also occurs in minute quantities in city gas and in tail gases of blast furnaces. It
is a volatile gas of peculiar odour and is sparingly soluble in water but a little more in ethanol
and ether. It is inflammable and hence the ignition of a mixture of ethylene with air leads to
explosion.
Physiological Roles
1. Stimulates fruit ripening. The ancient Chinese knew that fruits would ripen faster if in a
room containing burning incense. The factor responsible for this hastened ripening was not heat,
but ethylene released as the incense burned. The stimulation of fruit ripening by ethylene is a
consequence of many ongoing processes such as:
A. the breakdown of chlorophyll and synthesis of other pigments; for example, apples changing
from green to red during ripening,
B. Fruit softening due to breakdown of cell walls by cellulase and pectinase, and
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C. Conversion of starches and acids to sugars.

And ethylene stimulates each of these processes, leading ultimately to fruit ripening. We often
say that ‘one rotten apple spoils the rest of the barrel’ because pieces of fruit near to a rotten one
start to go bad quickly. This is because a damaged on fungus infected fruit starts to produce
ethene. And in a closed Barrel the concentration of ethene is high enough to quickly trigger the
ripening process in neighbouring fruits making them more vulnerable to infection.
Figure 35: The oranges infected with the fungus Penicillium digitatum.
The fungus itself produces large amounts of ethene, which accelerates the postharvest maturation
of oranges, making them more vulnerable to infection. Some fruits (such as tomatoes and apples)
show a conspicuous increase in respiration just before fruit ripening. This increase in respiration
is called a climacteric, and fruits that display it are referred to as climacteric fruits. The
climacteric begins just after a huge increase (up to a 100- fold) in ethylene production. Thus, the
climacteric and fruit ripening are both triggered by ethylene.
Fruit growers often take advantage of ethylene’s capability of stimulating fruit ripening for
making fruits available for sale out-of-season. For instance, many apples are plucked in
September and October when they are green and immature. These are then stored in rooms
containing air that has small amounts (1–3%) of O2 large amounts (5–10%) of CO2 and no
ethylene. As these conditions inhibit protein synthesis, the fruits can be stored without the fear of
their being ripened. When these unripe fruits are needed for sale, producers expose them to
normal air containing 1ppm of ethylene, which is enough to induce climacteric and ripening of
fruits. Thus the “fresh” apples one buys in March are the ones harvested in September/October of
the previous year. This “ripening on demand” is also used in the case of tomatoes, lemons and
oranges. It is for these reasons that, in common parlance, ethylene is known as ‘ripening
hormone’. However, some other fruits (such as grapes and cherries) cannot be ripened by
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ethylene. Such fruits are called non-climacteric and are insensitive to ethylene. Table below lists
the climacteric and non-climacteric type of fruits.
Note that the term ‘climacteric’ can be used either as a noun, as in “most fruits exhibit a
climacteric during ripening” or as an adjective, as in “a climacteric rise in temperature”. The
term ‘non-climacteric’ however, is used only as an adjective.
2. Promotes flowering. Although ethylene inhibits, flowering in most species but induces it in a
few plants including mangoes, pineapples and some ornamentals. The Filipino mango growers
and the Peurto Rican pineapple growers, who knew this effect long ago, set bonfires near their
crops. The fires produced ethylene which initiated flowering of their plants. Nowadays, the
pineapple growers in Hawaii produce pineapple fruits round the year by spraying plants with
ethepon. Ethepon splits under neutral and alkaline conditions to release ethylene.
3. Hastens leaf abscission. Abscission zone in leaves causes the increased production of
ethylene which triggers the breakdown of middle lamella, thus leading to the initiation of
abscission. This effect is also utilized by horticulturists to minimize the harvesting period of such
fruits as cherries, grapes and blueberries. These fruits are sprayed with ethepon to coordinate
abscission, thereby allowing growers to harvest their crops in shorter periods of time.
4. Induces leaf epinasty. When a plant's roots are kept submerged in water for long periods,
water fills the intercellular spaces. Since these spaces are the primary routes of gas exchange
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with the atmosphere, the submerged roots become waterlogged and anaerobic. The symptoms of
water logging (e.g., leaf chlorosis, shorter and thicker shoots and wilting) can also be induced by
placing roots in O2-free air. Because O2 is required to produce ethylene, its synthesis is greatly
checked in roots of waterlogged plants. The small amount of ethylene that is made in these roots
is trapped, where it accumulates and eventually stimulates the activity of enzymes like cellulose
and pectinase. These enzymes break down the cell walls. This leads to the formation of many
intercellular spaces, characteristic of hydrophytes. Concomitantly, the ethylene precursors in the
shoot are also converted to ethylene, which causes parenchyma cells on the upper side of the
petiole to expand and point the leaf down, a physiologic response called epinasty.
5. Controls stem elongation. Mechanical disturbances such as shaking decrease stem
elongation. This effect, which is called thigmomorphogenesis, is mediated by ethylene.
Mechanical disturbances enhance ethylene production several times. Ethylene so produced
causes cells to arrange their cellulose micro-fibrils longitudinally. This lengthwise reinforcement
inhibits cellular elongation, causing cells to elongate radially; this leads to the formation of short
and thick stems. This effect is opposite to that of auxin, which causes cells to orient their
microfibrils transversely, thereby accounting for cellular elongation.
6. Determines sex expression. Both ethylene and gibberellins determine the sex of flowers on
monoecious plants, i.e., plants having male and female flowers on the same individual. As an
instance, cucumber (Cucumis sativus) buds, on treatment with ethylene, become carpellate
flowers, whereas those treated with gibberellins become staminate (%) flowers.
Correspondingly, buds that ultimately become flowers produce more ethylene than do buds that
become male flowers.
Ethylene versus Auxin: IAA stimulates ethylene production, thereby linking the responses of
these two hormones. But ethylene does not account for all the effects induced by applying IAA.
For example, IAA’s stimulation of cellular elongation and the formation of lateral roots occur
independently of ethylene. Similarly, leaf epinasty, decreased elongation of roots and shoots, and
determination of sex are responses to ethylene application rather than IAA.
6.5. Abscisic Acid (ABA)

Discovery: Most of the effects first discovered for plant hormones were stimulatory. For
example, IAA stimulates cellular elongation and cytokinins stimulate cell division. But near the
end of the decade 1940s, Torsten Hemberg of Sweden reported that dormant buds of ash and
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potato contained inhibitors (rather than stimulators) that blocked the effects of IAA. When the
buds germinated, the amount of these inhibitors decreased. Later, Eagles and Wareing (1963)
isolated an inhibitor from the birch (Betula pubescens) leaves held under short day conditions.
When this substance was reapplied to the leaves of birch seedlings, apical growth was
completely arrested. As this substance induced dormancy, they named it as dormin. Later in
1965, Ohkuma et al isolated an inhibitor from cotton fruits and named it abscisin II. The same
year, Cornforth and his associates isolated a growth inhibitor from sycamore and pointed out that
both dormin and abscisin II are identical. Abscisin II is peculiar in that it is effective in much
lower concentration than phenolic inhibitors and is accumulated under short day conditions. This
compound was later named as abscisic acid (abbreviated as ABA) an unfortunate name, because
subsequent research has shown that ethylene rather than abscisic acid controls abscission.
Distribution and Biosynthesis: Abscisic acid occurs in angiosperms and gymnosperms but
apparently not in liverworts. ABA in plants is made from carotenoids. Once synthesized, ABA
moves throughout a plant in xylem, phloem and parenchyma. Like gibberellins and cytokinins,
abscisic acid moves nonpolarly. There are no synthetic abscisic acids.
Physiological Roles
1. Closure of stomata. It is a known fact that during drought, leaves synthesize large amounts of
ABA which causes stomata to close. Thus, ABA acts as a messenger and enables plants to
conserve water during drought. Because ABA-induced closure of stomata occurs within 1 to 2
minutes, this effect probably occurs independently of protein synthesis. As to its mechanism,
ABA probably produces its effect by binding to proteins on the outer surface of the
plasmalemma of guard cells. This renders the plasmalemma more positively-charged thereby
stimulating transport of ions (especially K+) from guard cells to epidermal cells. The loss of these
ions causes water to leave guard cells (via osmosis) which then collapse, thus closing the
stomatal aperture.
2. Delays seed dormancy. In many species, applying ABA delays seed germination. Similarly,
in many other plants, the amount of ABA in their seeds decreases, when seeds germinate. Thus,
it may be inferred that ABA controls seed dormancy in some cases. However, this conclusion
may not be generalized, since germination of many seeds occurs without any changes in the
amount of abscisic acid.
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3. Controls bud dormancy. Bud dormancy was previously thought to be controlled solely by
ABA. But when leaves are treated with radioactive ABA, no radioactivity has been detected in
buds. This suggests that, besides ABA, bud dormancy is probably also influenced by cytokinins
and IAA-induced synthesis of ethylene.
4. Counteracts the effects of other hormones. ABA counteracts the stimulatory /inhibitory
effects of other hormones. For example,
A. ABA inhibits cell growth promoted by IAA.
B. ABA inhibits amylase produced by seed treated with gibberellin.
C. ABA promotes chlorosis that is inhibited by cytokinins.
This may be due to the fact that ABA is a Ca2+ antagonist and its inhibition of the stimulatory
effects of IAA and cytokinin may be due to its interference with Ca2+ metabolism. Although
ABA usually inhibits growth, it is not toxic to plants as are inhibitors of RNA /protein synthesis.
ABA often decreases gene activity, but there are instances of ABA stimulating genes. For
example, ABA stimulates the synthesis of mRNAs for storage proteins in developing wheat
grains.
Other effects
i. Including bud dormancy and seed dormancy
ii. Includes tuberisation
iii. Induces senescence of leaves fruit ripening, abscission of leaves, flowers and fruits
iv. Increasing the resistance of temperate zone plants to frost injury.
Mode of action:
(i) Effect on plasma membrane of roots
(ii) Inhibition of protein synthesis
(iii) Specific activation and deactivation of certain genes (transcription effects)
6.6. Jasmonic Acid

They are named after the jasmine plant in which the methyl ester is an important scent
component. As such they have been known for some time in the perfume industry. There is also
a related hydroxylated compound that has been named tuberonic acid which, with its methyl
ester and glycosides, induces potato tuberization. Jasmonic acid is synthesized from linolenic
acid, while jasmonic acid is most likely the precursor of tuberonic acid.
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Physiological Role: Jasminates play an important role in plant defense, where they induce the
synthesis of proteinase inhibitors which deter insect feeding, and, in this regard, act as
intermediates in the response pathway induced by the peptide systemin. Jamonates inhibit many
plant processes such as growth and seed germination. They promote senescence, abscission,
tuber formation, fruit ripening, pigment formation and tendril coiling. JA is essential for male
reproductive development of Arabidopsis. The role in other species remains to be determined.
Hormonal Interactions: It is very rarely, if ever, that the plant hormones work alone; rather
plant growth and development usually result from interactions of plant hormones (Figure 36).
Figure 36: Hormonal interactions in a plant

Note that numerous hormonal interactions influence plant growth and development. An
intriguing question is what controls the amounts of hormones and thereby their ratios and the
resulting interactions? The amount of hormones is controlled in two ways:
A. Regulation of the rate of synthesis. Many factors influence the rate of hormone production.
For example, day length can stimulate the synthesis of IAA, and the cold temperatures trigger the
synthesis of gibberellins.
B. Regulation of the rate of breakdown or inactivation. Inactivation of a hormone usually
takes place either by its oxidation or by its conjugation with other compound.
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This chart shows the five plant hormones and their functions summary.
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CHAPTER 7: STRESS PHYSIOLOGY
In Both Natural and Agricultural Conditions, plants are frequently exposed to environmental
stresses. Some environmental factors, such as air temperature, can become stressful in just a few
minutes; others, such as soil water content, may take days to weeks, and factors such as soil
mineral deficiencies can take months to become stressful. It has been estimated that because of
stress resulting from climatic and soil conditions (abiotic factors) that are suboptimal, the yield
of field-grown crops in the United States is only 22% of the genetic potential yield (Boyer,1982).
In addition, stress plays a major role in determining how soil and climate limit the distribution of
plant species. Thus, understanding the physiological processes that underlie stress injury and the
adaptation and acclimation mechanisms of plants to environmental stress is of immense
importance to both agriculture and the environment.
The concept of plant stress is often used imprecisely, and stress terminology can be confusing, so
it is useful to start our discussion with some definitions. Stress is usually defined as an external
factor that exerts a disadvantageous influence on the plant. This chapter will concern itself with
environmental or abiotic factors that produce stress in plants, although biotic factors such as
weeds, pathogens, and insect predation can also produce stress. In most cases, stress is measured
in relation to plant survival, crop yield, growth (biomass accumulation), or the primary
assimilation processes (CO2 and mineral uptake), which are related to overall growth.
Throughout their life plants have to adapt to variable environmental conditions. Changes in
photoperiod, light intensity and quality, nutrient abundance and starvation, drought and flooding,
variation in temperature, air and soil pollution and osmotic changes are among the abiotic factors
that can cause stress.
7.1. Water stress

Plants experience water stress either when the water supply to their roots becomes limiting or
when the transpiration rate becomes intense. Water stress is primarily caused by the water
deficit, i.e. drought or high soil salinity. In case of high soil salinity and also in other conditions
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like flooding and low soil temperature, water exists in soil solution but plants cannot uptake it, a
situation commonly known as ‘physiological drought’. Drought occurs in many parts of the
world every year, frequently experienced in the field grown plants under arid and semi-arid
climates. Regions with adequate but non-uniform precipitation also experience water limiting
environments. Since the dawn of agriculture, mild to severe drought has been one of the major
production limiting factors. Consequently, the ability of plants to withstand such stress is of
immense economic importance. The general effects of drought on plant growth are fairly well
known. However, the primary effect of water deficit at the biochemical and molecular levels are
not considerably understood yet and such understanding is crucial. All plants have tolerance to
water stress, but the extent varies from species to species. Knowledge of the biochemical and
molecular responses to drought is essential for a holistic perception of plant resistance
mechanisms to water limited conditions in higher plants.
Effects of water stress on plants: Drought, as an abiotic stress, is multidimensional in nature,

and it affects plants at various levels of their organization. In fact, under prolonged drought,
many plants will dehydrate and die. Water stress in plants reduces the plant-cell’s water potential
and turgor, which elevate the solutes’ concentrations in the cytosol and extracellular matrices. As
a result, cell enlargement decreases leading to growth inhibition and reproductive failure. This is
followed by accumulation of abscisic acid (ABA) and compatible osmolytes like proline, which
cause wilting. At this stage, overproduction of reactive oxygen species (ROS) and formation of
radical scavenging compounds such as ascorbate and glutathione further aggravate the adverse
influence. The partially reduced or activated derivatives of oxygen (O2 1, O2 H2O2, and HO-) are
highly reactive and toxic, and can lead to the oxidative destruction of cells. Drought not only
affects plant water relations through the reduction of water content, turgor and total water, it also
affects stomatal closure, limits gaseous exchange, reduces transpiration and arrests carbon
assimilation (photosynthesis) rates. Negative effects on mineral nutrition (uptake and transport of
nutrients) and metabolism leads to a decrease in the leaf area and alteration in assimilate
partitioning among the organs. Alteration in plant cell wall elasticity and disruption of
homeostasis and ion distribution in the cell has also been reported. Synthesis of new protein and
mRNAs associated with the drought response is another outcome of water stress on plants.
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Under the water stress cell expansion slows down or ceases, and plant growth is retarded.
However, water stress influences cell enlargement more than cell division. Plant growth under
drought is influenced by altered photosynthesis, respiration, translocation, ion uptake,
carbohydrates, nutrient metabolism, and hormones.
7.2. Temperature stress

Mesophytic plants (terrestrial plants adapted to temperate environments that are neither
excessively wet nor dry) have a relatively narrow temperature range of about 10°C for optimal
growth and development. Outside of this range, varying amounts of damage occur, depending on
the magnitude and duration of the temperature fluctuation. In this section we will discuss three
types of temperature stress: high temperatures, low temperatures above freezing, and
temperatures below freezing. Most actively growing tissues of higher plants are tillable to
survive extended exposure to temperatures above 45°C or even short exposure to temperatures of
55°C or above. However, nongrowing cells or dehydrated tissues (e.g., seeds and pollen) remain
viable at much higher temperatures. Pollen grains of some species can survive 70°C and some
dry seeds can tolerate temperatures as high as 120°C.
Most plants with access to abundant water are able to maintain leaf temperatures below 45°C by
evaporative cooling, even at elevated ambient temperatures. However, high leaf temperatures
combined with minimal evaporative cooling causes heat stress. Leaf temperatures can rise to 4 to
5°C above ambient air temperature in bright sunlight near midday, when soil water deficit causes
partial stomatal closure or when high relative humidity reduces the gradient driving evaporative
cooling. Increases in leaf temperature during the day can be more pronounced in plants
experiencing drought and high irradiance from direct sunlight. Temperature stress can result in
damaged membranes and enzymes: Plant membranes consist of a lipid bilayer interspersed with
proteins and sterols, and any abiotic factor that alters membrane properties can disrupt cellular
processes. The physical properties of the lipids greatly influence the activities of the integral
membrane proteins, including H+ pumping ATPases, carriers, and channel-forming proteins that
regulate the transport of ions and other solutes. High temperatures cause an increase in the
fluidity of membrane lipids and a decrease in the strength of hydrogen bonds and electrostatic
interactions between polar groups of proteins within the aqueous phase of the membrane. High
temperatures thus modify membrane composition and structure, and can cause leakage of ions.
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High temperatures can also lead to a loss of the three dimensional structure required for correct
function of enzymes or structural cellular components, thereby leading to loss of proper enzyme
structure and activity. Misfolded proteins often aggregate and precipitate, creating serious
problems within the cell. Temperature stress can inhibit photosynthesis: Photosynthesis and
respiration are both inhibited by temperature stress. Typically, photosynthetic rates are inhibited
by high temperatures to a greater extent than respiratory rates. Although chloroplast enzymes
such as rubisco, rubisco activase, NADP-G3P dehydrogenase, and PEP carboxylase become
unstable at high temperatures, the temperatures at which these enzymes began to denature and
lose activity are distinctly higher than the temperatures at which photosynthetic rates begin to
decline. This would indicate that the early stages of heat injury to photosynthesis are more
directly related to changes in membrane properties and to uncoupling of the energy transfer
mechanisms in chloroplasts.
This imbalance between photosynthesis and respiration is one of the main reasons for the
deleterious effects of high temperatures. On an individual plant, leaves growing in the shade
have a lower temperature compensation point than leaves that are exposed to the sun (and heat).
Reduced photosynthate production may also result from stress-induced stomatal closure,
reduction in leaf canopy area, and regulation of assimilate partitioning. Freezing temperatures
cause ice crystal formation and dehydration: Freezing temperatures result in intra- and
extracellular ice crystal formation. Intracellular ice formation physically shears membranes and
organelles. Extracellular ice crystals, which usually form before the cell contents freeze, may not
cause immediate physical damage to cells, but they do cause cellular dehydration. This is
because ice formation substantially lowers the water potential (Ψw) in the apoplast, resulting in a
gradient from high Ψw in the symplast to low Ψw in the apoplast. Consequently, water moves
from the symplast to the apoplast, resulting in cellular dehydration. Cells that are already
dehydrated, such as those in seeds and pollen, are relatively less affected by ice crystal
formation. Ice usually forms first within the intercellular spaces and in the xylem vessels, along
which the ice can quickly propagate. This ice formation is not lethal to hardy plants, and the
tissue recovers fully if warmed. However, when plants are exposed to freezing temperatures for
an extended period, the growth of extracellular ice crystals leads to physical destruction of
membranes and excessive dehydration.
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7.3. Salt stress

General symptoms of damage by salt stress are growth inhibition, accelerated development and
senescence and death during prolonged exposure. Growth inhibition is the primary injury that
leads to other symptoms although programmed cell death may also occur under severe salinity
shock. Salt stress induces the synthesis of abscisic acid which closes stomata when transported to
guard cells. As a result of stomatal closure, photosynthesis declines and photo inhibition and
oxidative stress occur. An immediate effect of osmotic stress on plant growth is its inhibition of
cell expansion either directly or indirectly through abscisic acid. Excessive sodium ions at the
root surface disrupt plant potassium nutrition. Because of the similar chemical nature of sodium
and potassium ions, sodium has a strong inhibitory effect on potassium uptake by the root. Plants
use both low- and high affinity systems for potassium uptake. Sodium ions have a more
damaging effect on the low-affinity system which has low potassium/sodium selectivity. Under
sodium stress, it is necessary for plants to operate the more selective high-affinity potassium
uptake system in order to maintain adequate potassium nutrition. Potassium deficiency inevitably
leads to growth inhibition because potassium, as the most abundant cellular cation, plays a
critical role in maintaining cell turgor, membrane potential and enzyme activities.
Once sodium gets into the cytoplasm, it inhibits the activities of many enzymes. This inhibition
is also dependent on how much potassium is present: a high sodium/potassium ratio is the most
damaging. Even in the case of halophytes that accumulate large quantities of sodium inside the
cell, their cytosolic enzymes are just as sensitive to sodium as enzymes of glycophytes. This
implies that halophytes have to compartmentalize the sodium into the vacuole, away from
cytosolic enzymes. An important factor in the battle between sodium and potassium ions is
calcium. Increased calcium supply has a protective effect on plants under sodium stress. Calcium
sustains potassium transport and potassium/sodium selectivity in sodium-challenged plants. This
beneficial effect of calcium is mediated through an intracellular signaling pathway that regulates
the expression and activity of potassium and sodium transporters. Calcium may also directly
suppress sodium import mediated by nonselective cation channels.
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7.4. Oxygen stress

Roots usually obtain sufficient oxygen (O2) for aerobic respiration directly from the gaseous
space in the soil. Gas-filled pores in well-drained, well-structured soil readily permit the
diffusion of gaseous O2 to depths of several meters. Consequently, the O2 concentration deep in
the soil is similar to that in humid air. However, soil can become flooded or waterlogged when it
is poorly drained or when rain or irrigation is excessive. Water then fills the pores and blocks the
diffusion of O2 in the gaseous phase. Dissolved oxygen diffuses so slowly in stagnant water that
only a few centimeters of soil near the surface remain oxygenated. When temperatures are low
and plants are dormant, oxygen depletion is very slow and the consequences are relatively
harmless. However, when temperatures are higher (greater than 20°C), oxygen consumption by
plant roots, soil fauna and microorganisms, can totally deplete the oxygen from the bulk of the
soil water in as little as 24 hours.
Flooding-sensitive plants are severely damaged by 24 hours of anoxia. The growth and survival
of many plant species are greatly depressed under such conditions, and crop yields can be
severely reduced. For example, garden pea (Pisum sativum) yields can be halved by 24 hours of
flooding, making garden pea an example of a flooding sensitive plant. Other plants, particularly
species not adapted to grow in continually wet conditions and many crop plants are affected by
flooding in a milder way and are considered flooding tolerant plants. Flooding-tolerant plants
can withstand anoxia (lack of oxygen) temporarily, but not for prolonged periods of more than a
few days. On the other hand, specialized natural vegetation found in wetlands such as marshes
and swamps, and crops such as rice, are well adapted to resist oxygen deficiency in the root
environment. Wetland plants can resist anoxia, and they grow and survive for extended periods
of up to months with their root systems in anoxic conditions. Most of these plants have special
adaptations, which we will discuss here, that permit oxygen available in nearby environments to
reach the tissues held in anoxic conditions. Practically all plants require oxygen when they are
engaging in rapid metabolic activity, and plants can be classified according to the time they can
withstand anoxic conditions in their root environment without demonstrating substantial damage.
Anaerobic Microorganisms Are Active in Water-Saturated Soils: When soil is completely
depleted of molecular O2, the function of soil microbes becomes significant for plant life and
growth. Anaerobic soil microorganisms (anaerobes) derive their energy from the reduction of
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nitrate (NO3-) to nitrite (NO2-) or to nitrous oxide (N2O) and molecular nitrogen (N2). These
gases (N2O and N2) are lost to the atmosphere in a process called denitrification. As conditions
become more reducing, anaerobes reduce Fe3+ to Fe2+, and because of its greater solubility, Fe2+
can rise to toxic concentrations when some soils are anaerobic for many weeks. Other anaerobes
may reduce sulfate (SO4 2-) to hydrogen sulfide (H2S), which is a respiratory poison. When
anaerobes have an abundant supply of organic substrate, bacterial metabolites such as acetic acid
and butyric acid are released into the soil water, and these acids along with reduced sulfur
compounds account for the unpleasant odor of waterlogged soil. All of these substances made by
microorganisms under anaerobic conditions are toxic to plants at high concentrations.
Roots Are Damaged in Anoxic Environments: Root respiration rate and metabolism are
affected even before O2 is completely depleted from the root environment. The critical oxygen
pressure (COP) is the oxygen pressure at which the respiration rate is first slowed by O2
deficiency. The COP for a maize root tip growing in a well stirred nutrient solution at 25°C, is
about 0.20 atmosphere (20 kPa, or 20% O2 by volume), almost the concentration in ambient air.
At this oxygen partial pressure (for a discussion of partial pressures, the rate of diffusion of
dissolved O2 from the solution into the tissue and from cell to cell barely keeps pace with the rate
of O2 utilization. However, a root tip is metabolically very active, with respiration rates and ATP
turnover comparable to those of mammalian tissue. In older zones of the root, where cells are
mature and fully vacuolated and the respiration rate is lower, the COP is often in the range of 0.1
to 0.05 atmospheres. O2 concentrations is below the COP the center of the root becomes anoxic
(completely lacking oxygen) or hypoxic (partly deficient in oxygen). The COP is lower when
respiration slows down at cooler temperatures; it also depends on how bulky the organ is and
how tightly the cells are packed. Large, bulky fruits are able to remain fully aerobic because of
the large intercellular spaces that readily allow gaseous diffusion. For single cells, an O2 partial
pressure as low as 0.01 atmospheres (1% O2 in the gaseous phase) can be adequate because
diffusion over short distances ensures an adequate O2 supply to mitochondria. A very low partial
pressure of O2 at the mitochondrion is sufficient to maintain oxidative phosphorylation.
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In the absence of O2, electron transport and oxidative phosphorylation in mitochondria cease, the
tricarboxylic acid cycle cannot operate, and ATP can be produced only by fermentation. Thus
when the supply of O2 is insufficient for aerobic respiration, roots first begin to ferment pyruvate
(formed in glycolysis) to lactate, through the action of lactate dehydrogenase (LDH). In the root
tips of maize, lactate fermentation is transient because lowered intracellular pH quickly leads to a
switch from lactate fermentation to ethanol fermentation. The shift occurs because of the
different pH optima of the cytosolic enzymes involved.
At acidic pH, LDH is inhibited and pyruvate decarboxylase is activated. The net yield of ATP in
fermentation is only 2 moles of ATP per mole of hexose sugar respired (compared with 36 moles
of ATP per mole of hexose respired in aerobic respiration). Thus, injury to root metabolism by
O2 deficiency originates in part from a lack of ATP to drive essential metabolic processes.
Apparently, active transport of H+ into the vacuole by tonoplast ATPases is slowed by lack of
ATP, and without ATPase activity the normal pH gradient between cytosol and vacuole cannot
be maintained. Cytosolic acidosis irreversibly disrupts metabolism in the cytoplasm of higher
plant cells, as it does in anoxic cells of animals. It is essentially cytosolic acidosis that causes
damage, and the timing and degree to which it is limited are the primary factors distinguishing
flooding-sensitive from flooding-tolerant species.
Damaged O2 Deficient Roots Injure Shoots: Anoxic or hypoxic roots lack sufficient energy to
support physiological processes on which the shoots depend. Experiments have shown that the
failure of the roots of wheat or barley to absorb nutrient ions and transport them to the xylem
(and from there to the shoot) quickly leads to a shortage of ions within developing and expanding
tissues. Older leaves senesce prematurely, because of reallocation of phloem-mobile elements
(N, P, K) to younger leaves. The lower permeability of roots to water often leads to a decrease in
leaf water potential and wilting, although this decrease is temporary if stomata close, preventing
further water loss by transpiration. Hypoxia also accelerates production of the ethylene precursor
ACC (1-aminocyclopropane-1- carboxylic acid) in roots. In tomato, ACC travels via the xylem
sap to the shoot, where, in contact with oxygen, it is converted by ACC oxidase to ethylene. The
upper (adaxial) surfaces of the leaf petioles of tomato and sunflower have ethylene-responsive
cells that expand more rapidly when ethylene concentrations are high. This expansion results in
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epinasty, the downward growth of the leaves such that they appear to droop. Unlike wilting,
epinasty does not involve loss of turgor. In some species (e.g., pea and tomato), flooding induces
stomatal closure apparently without detectable changes in leaf water potential. Oxygen shortage
in roots, like water deficit or high concentrations of salts, can stimulate abscisic acid (ABA)
production and movement of ABA to leaves. However, stomatal closure under these conditions
can be attributed mostly to the additional production of ABA by the older, lower leaves. These
leaves do wilt, and they export their ABA to the younger turgid leaves, leading to stomatal
closure.
Submerged Organs Can Acquire O2 through Specialized Structures: In contrast to flooding
sensitive and flooding-tolerant species, wetland vegetation is well adapted to grow for extended
periods in water-saturated soil. Even when shoots are partly submerged, they grow vigorously
and show no signs of stress. In some wetland species, such as the water lily (Nymphoides
peltata), submergence traps endogenous ethylene, and the hormone stimulates cell elongation of
the petiole, extending it quickly to the water surface so that the leaf is able to reach the air.
Internodes of deep-water (floating) rice respond similarly to trapped ethylene, so the leaves
extend above the water surface despite increases in water depth. In the case of pondweed (
Potamogeton pectinatus), an aquatic monocot, stem elongation is insensitive to ethylene; instead
elongation is promoted even under anaerobic conditions by acidification of the surrounding
water caused by the accumulation of respiratory CO2.
In most wetland plants and in many plants that acclimate well to wet conditions, the stem and
roots develop longitudinally interconnected, gas-filled channels that provide a low-resistance
pathway for movement of oxygen and other gases. The gases (air) enter through stomata or
through lenticels on woody stems and roots, and travel by molecular diffusion, or by convection
driven by small pressure gradients. In many wetland plants, exemplified by rice, cells are
separated by prominent, gas-filled spaces, which form a tissue called aerenchyma, that develop
in the roots independently of environmental stimuli. In a few nonwetland plants, however,
including both monocots and dicots, oxygen deficiency induces the formation of aerenchyma in
the stem base and newly developing roots.
7.5. Mechanism of stress tolerance/resistance by plants

The concept of stress is intimately associated with that of stress tolerance, which is the plant’s
fitness to cope with an unfavorable environment. In the literature the term stress resistance is
155
often used inter-changeably with stress tolerance, although the latter term is preferred. Note that
an environment that is stressful for one plant may not be stressful for another. For example, pea
(Pisum sativum) and soybean (Glycine max) grow best at about 20°C and 30°C, respectively. As
temperature increases, the pea shows signs of heat stress much sooner than the soybean. Thus the
soybean has greater heat stress tolerance. If tolerance increases as a result of exposure to prior
stress, the plant is said to be acclimated (or hardened). Acclimation can be distinguished from
adaptation, which usually refers to a genetically determined level of resistance acquired by a
process of selection over many generations. Unfortunately, the term adaptation is sometimes
used in the literature to indicate acclimation. And to add to the complexity, we will see later that
gene expression plays an important role in acclimation.
Adaptation and acclimation to environmental stresses result from integrated events occurring at
all levels of organization, from the anatomical and morphological level to the cellular,
biochemical, and molecular level. For example, the wilting of leaves in response to water deficit
reduces both water losses from the leaf and exposure to incident light, thereby reducing heat
stress on leaves.
Plants can modify their life cycles to avoid abiotic stress: One way plants can adapt to extreme
environmental conditions is through modification of their life cycles. For example, annual desert
plants have short life cycles: they complete them during the periods when water is available, and
are dormant (as seeds) during dry periods. Deciduous trees of the temperate zone shed their
leaves before the winter so that sensitive leaf tissue is not damaged by cold temperatures. During
less predictable stressful events (e.g., a summer of significant but erratic rainfall) the growth
habits of some species may confer a degree of tolerance to these conditions. For example, plants
that can grow and flower over an extended period (indeterminate growth) are often more tolerant
to erratic environmental extremes than plants that develop preset numbers of leaves and flower
over only very short periods (determinate growth).
Phenotypic changes in leaf structure and behavior are important stress responses: Because of
their roles in photosynthesis, leaves (or their equivalent) are crucial to the survival of a plant. To
function, leaves must be exposed to sunlight and air, but this also makes them particularly
vulnerable to environmental extremes. Plants have thus evolved various mechanisms that enable
them to avoid or mitigate the effects of abiotic extremes to leaves. Such mechanisms include
changes in leaf area, leaf orientation, trichomes, and the cuticle. Turgor reduction is the earliest
156
significant biophysical effect of water deficit. As a result, turgor-dependent processes such as

leaf expansion and root elongation are the most sensitive to water deficits. When water deficit
develops slowly enough to allow changes in developmental processes, it has several effects on
growth, one of which is a limitation of leaf expansion. Because leaf expansion depends mostly
on cell expansion, the principles that underlie the two processes are similar. Inhibition of cell
expansion results in a slowing of leaf expansion early in the development of water deficits. The
resulting smaller leaf area transpires less water, effectively conserving a limited water supply in
the soil over a longer period. Altering leaf shape is another way that plants can reduce leaf area.
Under conditions of water, heat, or salinity extremes, leaves may be narrower or may develop
deeper lobes during development (Figure 37). The result is a reduced leaf surface area and
therefore, reduced water loss and heat load (defined as amount of heat loss [cooling] required
maintaining a leaf temperature close to air temperature). For protection against overheating
during water deficit, the leaves of some plants may orient themselves away from the sun. Leaf
orientation may also change in response to low oxygen availability.
Figure 37: Altered leaf shape can occur in response to environmental changes: leaf from outside
(left) and inside (right) of a tree canopy (source: Taiz L., Zeiger E., 2010).
157
Plants can regulate stomatal aperture in response to dehydration stress: The ability to control
stomatal aperture allows plants to respond quickly to a changing environment, for example to
avoid excessive water loss or limit uptake of liquid or gaseous pollutants through stomata.
Stomatal opening and closing is modulated by uptake and loss of water in guard cells, which
changes their turgor pressure. Although guard cells can lose turgor as a result of a direct loss of
water by evaporation to the atmosphere, stomatal closure in response to dehydration is almost
always an active, energy-dependent process rather than a passive one. Abscisic acid (ABA)
mediates the solute loss from guard cells that is triggered by a decrease in the water content of
the leaf. Plants constantly modulate the concentration and cellular localization of ABA, and this
allows them to respond quickly to environmental changes, such as fluctuations in water
availability.
Plants adjust osmotically to drying soil by accumulating solutes: Osmotic adjustment is the
capacity of plant cells to accumulate solutes and use them to lower Ψw during periods of osmotic
stress. The adjustment involves a net increase in solute content per cell that is independent of the
volume changes that result from loss of water. The decrease in ΨS (= osmotic potential) is
typically limited to about 0.2 to 0.8 MPa, except in plants adapted to extremely dry conditions.
There are two main ways by which osmotic adjustment can take place. A plant may take up
ions from the soil, or transport ions from other plant organs to the root, so that the solute
concentration of the root cells increases. For example, increased uptake and accumulation of K+
will lead to decreases in ΨS due to the effect of the potassium ions on the osmotic pressure
within the cell. This is a common event in saline areas, where ions such as potassium and
calcium are readily available to the plant. The accumulation of ions during osmotic adjustment is
predominantly restricted to the vacuoles, where the ions are kept out of contact with cytosolic
enzymes or organelles.
When ions are compartmentalized in the vacuole, other solutes must accumulate in the cytoplasm
to maintain water potential equilibrium within the cell. These solutes are called compatible
solutes (or compatible osmolytes). Compatible solutes are organic compounds that are
osmotically active in the cell, but do not destabilize the membrane or interfere with enzyme
158
function, as high concentrations of ions can. Plant cells can hold large concentrations of these
compounds without detrimental effects on metabolism. Common compatible solutes include
amino acids such as proline, sugar, alcohols such as mannitol, and quaternary ammonium
compounds such as glycine betaine.
Many plants have the capacity to acclimate to cold temperature: The ability to tolerate freezing
temperatures under natural conditions varies greatly among tissues. Seeds and other partially
dehydrated tissues, as well as fungal spores, can be kept indefinitely at temperatures near
absolute zero (0 K, or -273°C), indicating that these very low temperatures are not intrinsically
harmful. Hydrated, vegetative cells can also retain viability at freezing temperatures, provided
that ice crystal formation can be restricted to the intercellular spaces and cellular dehydration is
not too extreme. Temperate plants have the capacity for cold acclimation – a process whereby
exposure to low but nonlethal temperatures (typically above freezing) increase the capacity for
low temperature survival. Cold acclimation in nature is induced in the early autumn by exposure
to short days and nonfreezing, chilling temperatures, which combine to stop growth. A diffusible
factor that promotes acclimation, most likely ABA, moves from leaves via the phloem to
overwintering stems. ABA accumulates during cold acclimation and is necessary for this
process.
Plants survive freezing temperatures by limiting ice formation: During rapid freezing, the
protoplast, including the vacuole, may super cool; that is, the cellular water remains liquid
because of its solute content, even at temperatures several degrees below its theoretical freezing
point. Super cooling is common to many species of the hardwood forests. Cells can super cool to
only about -40°C, the temperature at which ice forms spontaneously. Spontaneous ice formation
sets the low-temperature limit at which many alpine and subarctic species that undergo deep
super cooling can survive. It may also explain why the altitude of the timberline in mountain
ranges is at or near the -40°C minimum isotherm. Several specialized plant proteins, termed
antifreeze proteins, limit the growth of ice crystals through a mechanism independent of
lowering of the freezing point of water. Synthesis of these antifreeze proteins is induced by cold
temperatures. The proteins bind to the surfaces of ice crystals to prevent or slow further crystal
growth.
159
Cold-resistant plants tend to have membranes with more unsaturated fatty acids: As
temperatures drop, membranes may go through a phase transition from a flexible liquid
crystalline structure to a solid gel structure. The phase transition temperature varies with species
(tropical species: 10-12°C; apples: 3-10°C) and the actual lipid composition of the membranes.
Chilling-resistant plants tend to have membranes with more unsaturated fatty acids. Chilling-
sensitive plants, on the other hand, have a high percentage of saturated fatty acid chains, and
membranes with this composition tend to solidify into a semi-crystalline state at a temperature
well above 0°C. Prolonged exposure to extreme temperatures may result in an altered
composition of membrane lipids, a form of acclimation. Certain transmembrane enzymes can
alter lipid saturation, by introducing one or more double bonds into fatty acids. This modification
lowers the temperature at which the membrane lipids begin a gradual phase change from fluid to
semi-crystalline form and allows membranes to remain fluid at lower temperatures, thus
protecting the plant against damage from chilling.
A large variety of heat shock proteins can be induced by different environmental conditions:
Under environmental extremes, protein structure is sensitive to disruption. Plants have several
mechanisms to limit or avoid such problems, including osmotic adjustment for maintenance of
hydration and chaperone proteins that physically interact with other proteins to facilitate protein
folding, reduce miss-folding and aggregation, and stabilize protein tertiary structure. In response
to sudden 5 to 10°C increases in temperature, plants produce a unique set of chaperone proteins
referred to as heat shock proteins (HSPs). Cells that have been induced to synthesize HSPs
show improved thermal tolerance and can tolerate subsequent exposure to temperatures that
otherwise would be lethal. Heat shock proteins are also induced by widely different
environmental conditions, including water deficit, ABA treatment, wounding, low temperature,
and salinity. Thus, cells that have previously experienced one condition may gain crossprotection
against another.
During mild or short-term water shortage, photosynthesis is strongly inhibited, but phloem
translocation is unaffected until the shortage becomes severe: Changes in the environment may
stimulate shifts in metabolic pathways. When the supply of O2 is insufficient for aerobic
160
respiration, roots first begin to ferment pyruvate to lactate through the action of lactate
dehydrogenase; this recycles NADH to NAD+, allowing the maintenance of ATP production
through glycolysis. Production of lactate (lactic acid) lowers the intracellular pH, inhibiting
lactate dehydrogenase and activating pyruvate decarboxylase. These changes in enzyme activity
quickly lead to a switch from lactate to ethanol production. The net yield of ATP in fermentation
is only 2 moles of ATP per mole of hexose sugar catabolized (compared with 36 moles of ATP
per mole of hexose respired in aerobic respiration). Thus, injury to root metabolism by O2
deficiency originates in part from a lack of ATP to drive essential metabolic processes such as
root absorption of essential nutrients. Water shortage decreases both photosynthesis and the
consumption of assimilates in the expanding leaves. As a consequence, water shortage indirectly
decreases the amount of photosynthate exported from leaves. Because phloem transport depends
on pressure gradients, decreased water potential in the phloem during water deficit may inhibit
the movement of assimilates. The ability to continue translocating assimilates is a key factor in
almost all aspects of plant resistance to drought.
Main Adaptations of Plants to Salt Stress

Halophytes Versus Glycophytes: Based on general tolerance to salt stress, all plants can be
roughly divided into two major groups: a) halophytes, that can withstand even 20% of salts in the
soil and, in most cases, successfully grow in conditions with 2-6% of salts, and b) nonhalophytes
or glycophytes, plants that exhibit various degrees of damage and limited growth in the presence
of sodium salts, usually higher than 0.01%. However, there are great differences in the level of
salt stress tolerance within both the halophytes and nonhalophytes, which include sensitive,
moderately tolerant and very tolerant species. Although halophytes represent only 2% of the
terrestrial plant species, they are present in about half the higher plant families and exhibit a
great diversity of plant forms.
Salt Exclusion: In terms of metabolic energy, use of ions to balance tissue water potential in a
saline environment clearly has a lower energy cost for the plant than use of carbohydrates or
amino acids, the production of which has a significantly higher energy cost. On the other hand,
high ion concentrations are toxic to many cytosolic enzymes, so ions must be accumulated in the
vacuole to minimize toxic concentrations in the cytosol. Because NaCl is the most abundant salt
161
encountered by plants under salinity stress, transport systems that facilitate compartmentation of
Na+ into the vacuole are critical. Both Ca2+ and K+ affect intracellular Na+ concentrations. It is a
very efficient but complex way of preventing massive ion uptake in the root zone, enabling a
lower uptake and accumulation of salts in the upper parts of the plant, especially in the
transpiring organs. Salt exclusion is based upon lower root permeability for ions even in the
presence of high external salinity. It is often found that many glycophytes, when exhibiting
enhanced tolerance to salinity stress, have a greater ability for sodium exclusion from the shoot
and for maintaining high levels of K+. Salt sensitive plants, such as beans and maize are the most
prominent Na+ excluders. The growth reduction in maize was caused by rapid salt accumulation
in older leaves, where a more salt-tolerant cultivar exhibited a higher ability for exclusion of
toxic ions. Additionally, in moderately tolerant crops, such as bread wheat, salt tolerance is
associated with low rates of sodium transport to the shoots and high K+/Na+ discrimination.
Significance of K/Na Selectivity: Uptake and distribution of sodium ions within the root is to a
large extent connected with the effects of potassium, since Na+ efflux in root cortex cells is
stimulated by K influx, which is related to the K/Na root selectivity. The presence of potassium
(and calcium) ions has been shown to decrease Na influx into plant cells. To confer salt stress
tolerance, in many species, the achievement of a high K: Na ratio is more important than simply
maintaining low concentrations of sodium ions.
Ion Retranslocation, Allocation and Leaching: Retranslocation of ions via the phloem is a
potentially important mechanism of preventing salt accumulation in fully expanded leaves, while
at the same time it represents a threat to younger leaves, which are phloem sinks. It seems that
cells of salt sensitive plants lack Na+/H+ transporters at the tonoplast, and therefore, cannot
efficiently sequester any excess of ions into the vacuoles. Instead, they need to rely on sodium
recirculation to exclude sodium salts from the shoot. Export of ions from shoot to root via the
phloem has been demonstrated in beans, maize, barley, cotton, and white lupin. Phloem export
may remove at least 25% of the total Na intake by the leaf, especially when growth decreases
and demands for ions diminish.
162
Besides salt exclusion in the roots, low NaCl levels in leaves can also be achieved by salt
retention in the lower plant parts and also through abscission of old leaves once they accumulate
large quantities of salts. In beans, ions accumulate in the root or in the basal part of the shoot,
from where they are returned to the root system and excreted back into the medium. The
mechanism of intra-plant allocation is also characteristic for many halophytes, which, due to the
limited transpiration, can keep excess of salts within their roots and lower parts of the shoot, thus
preventing ion accumulation in the photosynthetic tissues. The significance of ion leaching from
the leaf apoplast, through the cuticle, by rain, fog or dew is still unclear. Control of the ion
uptake is certainly achieved through restriction of the transpiration. Plants transpire 3070 times
more water than they use for cell growth, which means that solutes will be in the same degree
concentrated by the roots of non-excluder species. It was postulated that partial stomatal closure
observed in Aster tripolium under high salinity is induced by the presence of sodium ions in the
apoplast surrounding the guard cells, causing a reduction in rates of transpiration and increase of
water use efficiency.
Salt Excretion: Salt excretion is also a very efficient way of preventing excessive concentrations
of salts building up in photosynthetic tissues. This mechanism is typical for species that have
developed special features, mostly localized at the leaf epidermis, known as salt glands and salt
hairs (bladders). One of the most obvious signs of salt excretion is the salt crust on leaves and
shoots of those species with salt glands or salt hairs. Glandular structures are usually spread over
the whole surface area of the shoot, though they are most abundant on the leaves. Differentiation
of the salt- excreting structures is generally completed before differentiation of the entire leaf,
indicating the importance of salt glands in survival during the early developmental stages of the
plant. There is still a dilemma about the salt glands, whether they functions to excrete, secrete, or
recrete. There are a lot of similarities in the metabolic principles of ion transport and functioning
of the proton pumps in salt-excreting structures and cells of other tissues. Features that control
free ion transport analogous to the Casparian strips of the endodermis, have also been found in
salt glands (Breckle et al., 1990). The activity of salt glands may be induced by an increase of
external salinity, which has been shown by a 30% increase in the activity of H+ -ATPase isolated
from salt glands. The chemical composition of the secretion present in salt glands (inorganic ions
and organic compounds, such as sugars, amino acids and amines) indicates the possibility of
163
leakage from the protoplasm caused by a disturbed structure of membrane systems of the cell
during the process of excretion. Besides the active pumping of ions into the salt glands from the
palisade and water parenchyma cells, it seems that simultaneous vesicular transport of ions
leading to excretion into the extracellular space and direct salt secretion into the subcuticular area
are operating. Although the apoplastic pathway in solute transport to the gland cell prevails,
symplastic transport, documented through the presence of Cl- in the plasmodesmata of gland
cells of Limonium also operates. Additionally, the significance of vesicle-mediated solute
transport in salt excretion has been suggested.
The efficacy of salt glands has been established by the presence of low concentrations of salts in
leaves and a high K/Na ratio of species possessing such structures, even in conditions of rapidly
increasing external salinity. Altogether, the prevention of excessive salt accumulation within the
plant body is achieved through the following modes, such as: a) control of the salt uptake at the
root level and regulation of Na+ delivery to the shoot by loading of the xylem and retrieval of
ions from the xylem before reaching the shoot), b) K/Na selectivity, c) recirculation of salts via
the phloem, d) allocation of salts within particular parts of the plant, e) ion leakage and
abscission of organs loaded with salts, f) control of transpiration, and g) secretion of ions by salt
excreting structures.
Vacuolar Compartmentation: The high content of salts in aboveground parts of the halophytic
plants is feature associated with efficiency of such plants to deliver ions into the vacuoles. The
ability of plants to transfer ions into the vacuole is dependent on the proportion of highly
vacuolated cells and tissues, as well as the activity of transport systems located at the tonoplast,
which prevent excessive concentration of ions in the cytoplasm. Sequestration of salts into the
leaf and/or shoot vacuoles is typical attribute of dicotyledonous halophytes, coupled with other
physiological adaptations, such as regulation of transpiration (and water regime in general) and
performing of cell metabolism with low potassium concentrations. Therefore, the capacity of
vacuolar compartmentation, which enables effective osmotic adjustment of plants grown in
saline conditions, and liberation of cytosol free from the presence of toxic ions, is one of the key
factors in salinity tolerance. Many halophytic species are characterized by their large vacuoles.
The vacuoles occupy 77% of the mesophyll cells of Suaeda maritima and are capable of
164
accumulating salts to concentrations higher than 500 mM. In the case of salt sequestration into
the vacuole, potassium ions and organic solutes should be accumulated in the cytoplasm in order
to achieve and maintain the osmotic and ionic balance between these two compartments.
Succulence: The feature of succulence is characteristic for many dicotyledonous halophytes, and
represents a very efficient way of mitigation of adverse osmotic and toxic effects of ions through
dilution. In natural conditions, the content of salts in vegetative organs of halophytes increases
with ageing, and might reach toxic levels. Therefore, halophytic species tend to lower such
unfavorable salt concentrations by increasing of water content in their tissues and/or by enhanced
growth. Morpho-anatomical alterations of succulent halophytes include increase of cell volume,
especially of spongy and water parenchyma, increase of leaf thickness and decrease in number of
stomata. The significance of phenotypic plasticity in adaptations to salt stress was documented in
experiments with Plantago coronopus, where salt treatment lead to increased leaf thickness and
leaf dry matter percentage.
165
REFERENCES
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London.
Berrie, M.M. (1977). An introduction to the botany of major crop plants. Botanical Science
series, London.
Bilgarami, K.S., Srivastava, L.M., Shreemali, J.L. (1979). Fundamentals of Botany. Vikas
Publishing house, PVT LTD, New Delhi.
Coulter,M.C. and Dittmar, H.T. (1959). The story of the plant kingdom. University of Chikago,
USA.
Devlin, R.M. (1975). Plant Physiology, 3rd ed. Affiliated East-West Press PVT LTD, New
Delhi, India. 93
Devlin, R.M. and Witham, F.H. (1986). Plant physiology, 4th ed. CBS Publishers and
distributors,
Shahadara Delhi, India.
Eames, A.J. and Mackdamiels, L.H. (1972). An introduction to plant anatomy. McGraw Hill,
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Esau, K.(1965). Plant Anatomy. 2nd ed.
Lawrence, H.H. (1951). Taxonomy of vascular plants. Mackmilan Company, New Delhi.
Nabors, M.W. (2004). Introduction to botany. Benjamin Commings, USA.***
Nobel, P.S. (1983). Biophysical plant physiology and ecology. W.H. Freeman and Company,
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. Noggle, G.R. and Fritz, G.J. (1983). Introdictory Plant Physiology 2nd ed. Prentice-Hall of
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Pandey, S.N. and Chadha, A. (1996). Plant anatomy and embryology. Vikas Publishing house,
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Rost, T.L., Barbour, G.L., Weier, T.E. and Stocking, T.E. (1984). Botany: A brief introduction
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Teize, L. and Zeiger, E. (1997). Plant physiology. The Benjamin/ Cumming Publishing
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166

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HAWASSA UNIVERSITY PLANT PHYSIOLOGY

 Acquaint the concept of Soil-Plant-Atmosphere Continuum (SPAC)

1.1. Definition and scope of plant physiology 1

2.1. Importance and Properties of Water 4

3.1. The Nature of Light 38

4.2. Pentose Phosphate Pathway (PPP) 79

4.4. TCA (Tricarboxylic acid/Kreb cycle 80

5.1. Definitions of Growth and Development 91

6.1. Auxins 113

7.1. Water stress 147

1.1. Definition and scope of plant physiology

1.2. Relation of plant physiology with other disciplines

CHAPTER 2. PLANT-WATER RELATIONS

2.1. Importance and Properties of Water

amount of kinetic energy in the molecules of a substance; increasing temperature corresponds to

2.2.1. Absorption of Water by Plants

Figure 4. cross section of roots

Figure 5: the apoplast, transmembrane, and symplast pathways.

Components of water potential

Water Enters the Cell along a Water Potential Gradient

2.2.2. Mechanism of Water Absorption

2.2.3. Factors Affecting Water Absorption

2.3.1 Types of Transpiration

There are two types of transpiration

Transpiration rate of plants can be measured by a number of techniques, including potometers,

2.3.2. Mechanism of Transpiration

Water vapor moves from the leaf to the

2.3.3. Mechanism of Stomata Opening and Closing

Mechanism for opening of stomata

Because ABA- does not easily pass through

Membranes, under conditions of water

2.3.4. Significance of Transpiration

Thereby, Transpiration is essential for

Co2 acquisition: - all the carbon incorporated in to carbohydrate through photosynthesis

2.3.5. Factors Affecting Rate of Transpiration

Mechanism for opening of stomata

Ecologists classify plants according to their response to water as follows:

12 H2O + 6 CO2 + Energy C6H12O6 + 6O2 + 6H2O

3.1. The Nature of Light

Principles of Light Absorption by Photosynthetic Pigments: Radiation in the visible range

Figure 9 Electromagnetic spectrum. Wavelength (λ ) and frequency (ν ) are inversely related.

into heat, with no emission of a photon

3.2. Photosynthetic Pigments

Figure 11: Absorption spectra of some photosynthetic pigments. Curve 1, bacteriochlorophyll a;

2. Carotenoids (yellow or orange pigments): Carotenoids are pigments synthesized by plants.

Location of photosynthetic pigments in chloroplast: Each chloroplast is bound / enveloped

3.3. Light Reaction and Dark Reaction

In this process, nicotinamide adenine dinucliotide phosphate (NADP) is reduced in to NADPH

Light from the sun strikes the leaf

Carbon Reduction or Carboxylation or Carbon Fixation or Calvin Cycle of Photosynthesis

The overall reaction of photosynthesis: -

3.3.1. Role of Photo-system I and II

Figure 14 The light harvesting complex

This movement of electrons is downhill, in terms of an oxidation-reduction or redox potential

Figure 15 Z scheme of light reaction

Figure 16 Cyclic photophosphorylation

3.4. Path of Carbon in Photosynthesis

3.4.1. C3 Photosynthesis Cycle

1. Carboxylation – Carboxylation is the fixation of CO2 into a stable organic intermediate.

3.4.2. C4 photosynthesis Cycle

2. transport of the C4 acids (malate or aspartate) to the bundle sheath cells.

3.4.3. CAM (CRASSULACEAN ACID METABOLISM)

CAM photosynthesis is a second evolutionary strategy employing a "CO 2 pump" to accumulate