COURSE: SEED SCIENCE AND TECHNOLOGY

COURSE CODE: BSA 4201

CREDIT UNITS: 4
COURSE INSTRUCTOR: DR. OPIO PETER

1
INTRODUCTION

Importance of seeds in Agriculture

Seed or planting materials of a crop or a plant is a very important component of crop production
as good seed interacts with the good environment around it and determines the health of the crop
or plant. If the environment is conducive and the seed is poor, production is likely to be less.
Hence, farmers must use good and healthy seeds. The importance of seed in agriculture are as
follows;

❖ Carrier of new technology that is, for breeding purposes.

❖ Basic tool for a secure food supply; direct or indirect source of food, industry and essential
products.
❖ The principle means to obtain crop yields in less favorable production area
❖ Medium for rapid rehabilitation of agriculture after natural disasters.
❖ Seed can act as a source of income to farmers

Seed physiology

Concepts of Plant biology vs Seed technology

Plant Biology
At the highest level, Plant Biology can be divided into two major categories: first, Plant Biology
that is basic foundational research, and second, applied plant sciences that builds on the
discoveries of basic science and develops leading-edge technologies through innovation and
translation into agriculture. Within the basic areas of Plant Biology, four major categories
emerge as: (1) plant anatomy, (2) plant morphology and evolution, (3) plant physiology, and (4)
plant molecular and cellular biology. Within the applied plant sciences, there are many more
categories that have been grouped into major subfields, including vegetable, fruit and nut
crops; floriculture; horticulture; postharvest physiology; agronomy and weed science; plant
pathology and plant breeding; and genetic modification of crops, along with broader areas of
investigation, such as plants and the environment; tropical crops and ecosystems; forestry;
sustainability and production systems; and conservation biology of crop plants.

Seed technology
In a narrow sense, seed technology can be can be defined an aspect in crop science that
comprises of techniques of seed production, seed processing, seed storage, seed testing and
certification, seed marketing and distribution and the related research on these aspects.

2
On the other hand, Cowan (1973) defined seed technology as “that discipline of study having to
do with seed production, maintenance, quality and preservation”.

Furthermore, Feistritzer (1975) defined seed technology as the methods through which the
genetic and physical characteristics of seeds could be improved. It involves such activities as
variety development, evaluation and release, seed production, processing, storage and
certification.
Thus, seed technology is essentially an inter disciplinary science which encompasses broad range
of subjects. In its broadest sense, seed technology includes the development of superior crop
plant varieties, their evaluation and release, seed production, seed processing, seed storage, seed
testing, seed certification, seed quality control, seed marketing and distribution and research on
seed physiology, seed production and seed handling based upon modern botanical and
agricultural sciences.
History
Seed physiology; Seed physiology, especially seed germination, has intrigued the human mind
since antiquity, partly out of curiosity, partly because of practical reasons. Theophrastus could be
called the father of seed physiology since he already described many of the facts and problems
which up to this day are being investigated by seed physiologists. Most of the other ancient
authors writing about natural history or agriculture (Columella, Varro, Pliny the elder, Virgil)
added little to Theophrastus' all-encompassing knowledge of seed physiology. The 1700 years
from the first century A.D. to the end of the 18th century was a "black out" (Johansen, 1951) of
seed physiology. At the beginning of the 19th century people like Schleiden, Amici, Brown and
Hofmeister first understood structure and function of ovule, embryo sac and pollen tube whereas
fertilization and double fertilization were only detected at the end of the 19th century by
Strasburger, Nawaschin and Guignard. The physiology and biochemistry of seed maturation
were first inves- tigated by Sachs and Pfeffer, two great masters of modem plant physiology. In
modem times our detailed knowledge of the effect of external factors on seed germination is due
to the many authors working at the end of the 19th and the beginning of the 20th centuries
(Evenari, 1984)

Seed technology; The history of agricultural progress from the early days of man has been the
history of seeds of new crops and crop varieties brought under cultivation. In the early days it
was achieved through the cultivation of indigenous but useful plants and those taken through
introductions. Later through the well-known techniques of selection, hybridization, mutation,
polyploidization and plant biotechnology the scientists made available many new and better
varieties. However, to the farmer all this scientific research would be of little value unless he gets
seeds, which are genetically pure, high germination percentage and vigour, high purity, sound
health etc., When the farmers do not get seeds possessing these qualities the yields they obtain
may not be as expected. The pace of progress in production therefore, will largely depend upon

3
the speed with which we are able to multiply and market good quality seeds of high yielding
varieties.

Quality seed and its importance in Agriculture

What is seed? In broad sense, seed is a material which is used for planting or regeneration
purpose. However scientifically, Seed is a fertilized matured ovule together covered with seed
coat and this can be referred to as a propagating material i.e., part of agriculture, sericulture,
silviculture and horticultural plants used for sowing or planting purpose.
Structurally, true seed is a fertilized matured ovule, consisting of an embryonic plant, a store of
food and a protective seed coat, a store of food consists of cotyledons and endosperm.

From the seed technology point of view, seed can be regarded as sexually produced matured
ovule consisting of an intact embryo, endosperm and or cotyledon with protective covering (seed
coat). It also refers to propagating materials of healthy seedlings, tuber, bulbs, rhizome, roots,
cuttings, setts, slips, all types of grafts and vegetative propagating materials used for production
purpose.
Thus, seed is the most vital and crucial input for crop production, one of the ways to increase the
productivity without adding appreciably to the extent of land now under cultivation is by
planting quality seed.

Seed Quality attributes

Thompson (1979) defined seed quality as a multiple concept comprising several components and
their relative importance in different circumstances and laid much emphasis on; Analytical purity
/ physical purity, Species purity / Genetic purity, Freedom from weeds, Germination percentage,
Seed vigour and health, Seed Moisture content, Seed size, weight and specific gravity. Seed
quality characters: A good seed should have the following quality characters.
1. Improved variety: It should be superior to the existing variety i.e. the yield should be higher by
20-25% than the existing variety or it should have some desirable attributes like disease
resistance, drought resistance, salt tolerance etc., with good yield potential.
2. Genetic Purity: The seed should be true to type. The seed should possess all the genetic
qualities / characters, which the breeder has placed in the variety, genetic purity has direct effect
on the yields. If there is any deterioration, there would be proportionate decrease in the yield or
performance.
3. Physical Purity: Physical purity of a seed lot refers to the physical composition of the seed
lots. A seed lot is composed of pure seed, inert mater, broken seeds, undersized seeds, soil and
dust particles weed seeds, OCS etc. Higher the content of pure seed better would be the seed
quality. Pure seed together with germination gives the planting value of the seed lot.

4
4. Seed germination and vigour: Seed germination refers to the ability of a seed when planted
under normal sowing conditions to give rise to a normal seedling. Seed vigour refers to the sum
total of all seed attributes that give effective plant stand in the field. Higher germination
percentage and vigour gives adequate plant population and uniform growth, which have
profound effect on, yield and determine the planting value of the seed.
5. Freedom from weeds and other crop seeds: This is an extension of physical purity described
earlier. There are certain weed species, which are very harmful to the crop and once established
they are difficult to eradicate. An absolute freedom from seed of such species is highly desirable
and is one of the important criteria for determining the planning quality of seeds.
6. Seed health: Seed health refers to the presence or absence of disease organisms or insect pests
on the seed. The quality of a seed lot depends on its health; hence the seed should be free from
seed borne disease and insect pests.
7. Seed moisture: The seed moisture is the most important factor in determining the seed
germination and viability during storage. At high seed moisture content there is high incidence of
pest attack and at moisture content above 16% seed get heated and the viability is lost. Hence the
seed should be stored at safe moisture levels of 11-13% 8. Seed size, weight and specific gravity:
Seed size, weight and specific gravity has been found to have positive correlation with seed
germination and vigour in many crops. Therefore, the seed should be bold with high specific
gravity.
8. Seed Colour: The colour of the seed often reflects the condition during seed maturation. The
farmers from ancient times have regarded good normal shine as invariable quality guides. The
colour and shine deteriorates only when the weather conditions are adverse during maturation or
when insects infest the crop or when it is handled badly. The seed lots having high genetic
purity, high germination and with a minimum amount of inert matter, weed seeds and other crop
seeds and are free from diseases is said to be of high quality and if it is lacking of these it is said
to be of low quality.

5
Seed quality aspect

Fig 1. The interaction of various seed quality attributes and other factors which lead to good
crop yields per unit area.

Seed vs Grain
The distinction between seed and grain is vital, being of seminal importance to agriculture. A
seed, strictly speaking, is an “embryo” a living organism embedded in the supporting or the food
storage tissue. The seed pertains to material (seed, fruit or vegetatively propagating material)
meant for saving for planting purposes, the essential function being the reproduction.
A grain on the other hand, includes cereals and pulses meant for human consumption. However,
in most parts of Uganda, due to lack of access to improved good quality seed, farmers have
resorted to use grain as planting material.

Differences between scientifically produced seed and the grain (used as seed)
S/N Seed (Scientifically produced) Grain (used as seed)
1 It is the result of well-planned seed program It is the part of commercial produce
saved for sowing or planting purposes
2 It is the result of sound scientific knowledge, No such knowledge or effort is
organized effort, investment on processing, required
storage and marketing facilities.
3 The pedigree of the seed is ensured. It Its varietal purity is unknown
can be related to the initial breeder’s seed
4 During production, effort is made to rogue out No such effort is made. Hence, the
off-types, diseased plants, objectionable weeds purity and health status may be
and other crop plants at appropriate stages of inferior
crop growth which ensures satisfactory seed
purity and health.

6
 5 The seed is scientifically processed, treated and The grain used as seed may be
packed and labeled with proper lot identity. manually cleaned. In some cases, prior
to sowing it may also be treated. This
is not labeled
6 The seed is tested for planting quality namely, Routine seed testing is not done
germination, purity, admixture of weed seeds and
other crop seeds, seed health and seed moisture
content.
7 The seed quality is usually supervised by an There is no quality control.
agency not related with production (seed
certification agency)
8 The seed has to essentially meet the “quality No such standards apply here. The
standards”. The quality is therefore, well known. quality is non-descript and not known.
The labels, certification tags on the seed
containers serve as quality marks.

Benefits of using quality seeds

1. They are genetically pure (true to type).

2. The good quality seed has high return per unit area as the genetic potentiality of the crop can
be fully exploited.
3. Less infestation of land with weed seed/other crop seeds.
4. Less disease and insect problem.
5. Minimization of seed/seedling rate i.e., fast and uniform emergence of seedling.
6. They are vigorous, free from pests and disease.
7. They can be adopted themselves for extreme climatic condition and cropping system of the
location.
8. The quality seed respond well to the applied fertilizers and nutrients.
9. Uniform in plant population and maturity.
10. Crop raised with quality seed are aesthetically pleasing.
11. Good seed prolongs life of a variety.
12. Yield prediction is very easy.
13. Handling in post-harvest operation will be easy.
14. Preparations of finished products are also better.
15. High produce value and their marketability.

7
SEED FORMATION
The seed formation process
Seed formation begins with the combination of a male and female gamete: a process known as
fertilization. Fertilization, or syngamy, can occur when both male and female gametophytes are
fully mature. This usually occurs in a dual fusion process known as double fertilization (Figure
2.1). When the pollen grain lands on the stigma, it germinates by sending out a pollen tube,
which grows down the style, through the micropyle and into the embryo sac, with the tube
nucleus closely following the tube apex downward. The tube nucleus soon degenerates, but the
two pollen sperm cells enter the embryo sac, one fusing with the diploid (2N) polar nucleus to
form a triploid (3N) endosperm nucleus and the other fusing with the egg cell to form a diploid
(2N) zygote, or fertilized egg. The process of fertilization is very important because it not only
results in the formation of a seed but also dictates the level of genetic diversity present in the
zygote. Fertilization in angiosperms typically occurs either by self- or cross-fertilization.

Self-Fertilization; self-fertilization occurs when pollen from the anthers of a flower is transferred
to the stigma of the same flower, resulting in fertilization. In most cases, this happens when
flowers do not open until the pollination and fertilization of the flower is complete.
Cross-Fertilization; cross-fertilization occurs when pollen from one flower is transferred to the
stigma of another flower to cause fertilization. The flowers can be on the same or different
plants. In most agricultural crops, cross-fertilization occurs by two principal methods: wind
(anemophily) and insects (entomophily). Unlike self-fertilization, where progeny are genetically
similar, cross-fertilization results in progeny that are more dissimilar.

Fig 2.1. Double fertilization in an angiosperm ovule : (A) pollen tube its two sperm cells and
tube nucleus approaching the microphyle, (B) sperm cells approaching egg and polar nuclei, (c)
they migrate to the egg cell and central (polar) cell resulting in: sperm cell (n) + egg cell (n)
→zygote (2n) →embryo (2n); and sperm cell (n) + central cell (2n) → endosperm (3n) (double
fertilization has occurred) (From Purves et al.,1998).

8
This evolutionary approach produces a population of individuals that are more adaptable to a
wide array of environmental conditions.

Embryogenesis and embryo development

Embryonic development represents an important reproductive phase of sexually reproducing
plant species. The fusion of egg and sperm produces the plant zygote, a totipotent cell that,
through cell division and cell identity specification in early embryogenesis, establishes the major
cell lineages and tissues of the adult plant. The subsequent morphogenesis phase produces the
full-sized embryo, while the late embryogenesis maturation process prepares the seed for
dormancy and subsequent germination, ensuring continuation of the plant life cycle.

Embryogenesis in dicot model species; Arabidopsis thaliana

❖ After fertilization in A. thaliana, the nucleus of the zygote moves to the apical pole and the
zygote divides asymmetrically to produce a small apical cell that will generate the entire
embryo, with the exception of the suspensor, hypophysis, and root cap, which are derived
from the larger basal cell. Cell division in these two lineages produces the 2- to 4-cell-stage
embryo at~48 h after pollination (HAP), the 8-cell embryo at~60 HAP, the 16-cell
dermatogen embryo at~66 HPA, the 32-cell globular embryo at~72 HAP, and the~500-cell
heart-stage embryo~120 HAP (Jurgens and Mayer,1994).
❖ The suspensor (S) facilitates the transport of nutrients from the endosperm to the embryo and
undergoes programmed cell death (PCD) as the embryo matures. Its uppermost cell produces
the root meristem. During development the embryo progresses through the globular, heart
and torpedo stages to reach maturity
❖ The epidermal lineage is specified at the dermatogen stage, while the ground and vascular
tissue lineages, as well as shoot apical meristem (SM) and root apical meristem (RM), are
produced at the globular stage (Fig 3).
❖ Cotyledons (embryonic leaves, the first lateral organs) are produced at the heart stage, when
all major pattern elements of the Arabidopsis embryo have been established.
❖ After pattern formation, the embryo undergoes a period of morphogenesis where growth of
the organs that were established by the heart stage lead to an embryo with fully organized
SM and RM, hypocotyl (embryonic stem), and elongated cotyledons which fold over as the
embryo grows inside the seed coat (Fig 3).
❖ After pattern formation and morphogenesis, embryos of Arabidopsis and closely related
oilseed crop species undergo maturation, culminating in the desiccated seed (Goldberg et al.,
1994).
❖ Maturation stages include the mid-heart, torpedo, bent cotyledon, mature green, and dry seed.
During early maturation, also known as ‘‘seed filling,’’ embryos experience rapid cellular
division and expansion, turning green starting at the mid-heart stage, and also produce and
accumulate seed storage proteins (cruciferins/12S globulins and arabins/2S albumins).

9
❖ Starting at the early torpedo stage, the oils and waxes that form the cuticle of the epidermis
are produced. Storage of lipids, carbohydrates, and FAs during late embryogenesis is critical
for supporting the subsequent energy demands of germination (Baud and Lepiniec 2010).
Late maturation also involves the loss of water (desiccation) and the establishment of seed
dormancy (Leprince et al., 2017)

Fig. 3. Stages of development of a dicot embryo from the fertilized egg cell (zygote), as typified
by Arabidopsis thaliana. Initially this cell divides to form an apical and a basal cell (not shown);
the former divides to become the embryo (EP) and the latter the suspensor (S), as is evident at
the preglobular stage. The protoderm (Pd) develops into the epidermal layer (Ed) of the mature
embryo, the ground meristem (Gm) into the storage parenchyma (P), the procambium (Pc) into
the vascular tissue (V) and the hypophysis (Hs) into the root and shoot meristems (RM, SM). A,
axis; C, cotyledons. (From Bewley et al., 2000.)

Embryogenesis in monocot model species (Maize, rice and wheat)

❖ Zygotes of grasses, such as maize, rice, and wheat, are also polarized, with the nucleus at the
apical pole. Unlike Arabidopsis, the zygote does not elongate after fertilization (Chen et al.,
2017; Xiang et al., 2019).
❖ In rice, maize, and wheat the first transverse division of the zygote forms a two-cell embryo,
where the basal cell transforms into a large vesicular cell and continued division of the apical
cell gives rise to the quadrant, octant, and dermatogen stages (known as proembryo stages),
followed by the transition-stage embryos (Xiang et al., 2019). At the transition stage,
corresponding to about 8 days after pollination (DAP) in maize and 4 DAP in rice, the
adaxial–abaxial axis of the embryo becomes obvious, as the coleoptile primordium begins to
protrude from the adaxial region of the embryo, while the scutellum arises from the abaxial
side.
❖ The coleoptile protects the developing SM, while the scutellum is equivalent to the dicot
cotyledon (Fig. 4). The SM develops on the adaxial side and will produce several embryonic
leaves during seed development, while differentiation of the RM defines the basal pole of the
10
 embryo. In rice, the embryo reaches its mature shape when organ differentiation is complete
at 7–8 DAP, although morphogenesis continues as the embryo enlarges and shoot and root
meristems are completely surrounded by the protective coleoptile and coleorhiza,
respectively. In grasses, the early vegetative stages of the embryonic seedling are
incorporated into the embryo before dormancy (Fig. 4) (Itoh et al., 2005, 2016; Vernoud et
al., 2005).
In summary, generally in Poaceae (monocots), after the first division of the zygote, the basal cell
does not divide but forms the terminal cell of the suspensor (Black et al., 2006). The other few
suspensor cells and the embryo are produced from the axial cell. After further divisions a
globular-shaped embryo is formed (Fig. 4). It elongates to a club-like shape with radial
symmetry and then becomes bilaterally symmetrical through differentiation of the absorptive
scutellum and coleoptile, which covers the first foliage leaf. During the coleoptile stage of
development, the embryo axis and suspensor are established. Later, during the leaf stage, the
shoot and root meristems are defined and leaf primordia differentiate. The embryonic cells cease
to divide during maturation, the embryo desiccates and becomes quiescent. Mature graminaceous
embryos also contain a specialized thin tissue, the coleorhiza, which covers the radicle.

Fig. 4. Stages of development of a monocot embryo, such as in rice. (a) The fertilized egg cell
(zygote) undergoes (b) mitosis to form an axial (A) and a basal (B) cell. (c) A multicellular
proembryo is produced from the former, and then (d) an early coleoptile (Cp) stage. Further cell
divisions result in (e) a late coleoptile stage when the shoot meristem (Sm) is visible. (f) By the
first leaf (1st) stage other tissues are differentiated including the scutellum and the radicle, and
by (g) the second leaf (2nd) stage differentiation is more advanced. In the mature embryo (h) all
of the tissues are evident, with the addition of a third leaf (3rd), root meristem (Rm) and the
structure covering this, the coleorhiza (Cr). En, endosperm; Su, suspensor (Black et al., 2006).

11
Endosperm and perisperm development
The endosperm is a short-lived but essential constituent of seeds in flowering plants. It is one of
the twin products of the double fertilization event, which evolved to support and nourish its
sibling, the embryo. In cereals, the endosperm stores the seed reserves and represents a major
source of food and industrial raw materials for mankind. As a result, most early research about
endosperm development was focused on cereals.

Arabidopsis thaliana has emerged as an ideal plant model species for understanding the genetic
and epigenetic mechanisms of endosperm development and differentiation. As many dicot crops
such as soybean and peas, the endosperm in Arabidopsis is transient and is consumed by the
embryo during seed maturation; however, the basic organization and development of endosperm
are conserved in most flowering plants. The endosperm development in Arabidopsis shares
similar features with cereals, including an initial phase of intense nuclear division leading to
a syncytium (coenocytic phase) followed by cellularization, a unique cytokinesis process.

Endosperm development in A. thaliana is of the nuclear type, that is, the first and several
following divisions of the primary endosperm do not form separate cells but the free nuclei are
distributed throughout the embryo sac (as opposed to the cellular type where each nuclear
division of endosperm is associated with the formation of daughter cells, and the mixed helobial
type of endosperm development). So, the initial stage in the development of endosperm is
characterized by cell proliferation, leading to formation of a syncytium composed of numerous
nuclei within the cytoplasm, enclosed in a single common membrane. In A. thaliana this process
lasts until the heart stage (Fig. 5) when three regions of endosperm are first identified: the central
peripheral, micropylar peripheral nuclei embedded in a common cytoplasm surrounding the
suspensor, and the chalazal pole of a multinuclear nodule. The cellularization of the syncytial
mass begins at about the 5th day after pollination (Adams et al., 2000).

Fig. 5. Diagram of a seed containing a heart stage embryo and cellularizing endosperm. (From
Adams S., Vinkenoog, R., Spielman, M., Dickinson, H.G., Scott, R.J., 2000. Parent-of-origin
effects on seed development in Arabidopsis thaliana require DNA methylation. Development
127, 2493–2502.)

12
Fig. 6. Endosperm and embryo development, Capsella bursa-pastoris. A. Very young seed,
showing immature embryo (composed of suspensor and embryo proper), early endosperm, and
seed coat. B. Close-up of young embryo, showing basal suspensor cells and terminal, actively
dividing cells of globular embryo proper. C. More mature seed, showing embryo with two
cotyledons (Adapted from Simpson, 2010).

The cellularization of the endosperm begins from the area surrounding the embryo.
Besides serving as a storage of nutrients and hormones for the embryo, it is suggested that
endosperm is also involved in the determination of the size of the seed and fruit (Bouariky,
2010).

Perisperm; In some species the nucellar region of the ovule gives rise to a storage tissue called
the perisperm. It is usually present in mature seeds together with the endosperm, in various
proportions, locations and shapes (Black et al., 2006). A perisperm is present in some monocots
(e.g. Zingiberaceae) as well as certain dicots (e.g. Piperaceae, Nymphaeaceae, Cactaceae,
Amaranthaceae, Chenopodiaceae). In a few species it is the major source of the storage reserves,
the endosperm being absent (e.g. Quinoa and Yucca spp.) (Bewley and Black, 1994).

DIFFERENCE BETWEEN MONOCOT AND DICOT EMBRYO

S/N MONOCOTS DICOTS
1 Only one cotyledon attached to the There are two cotyledons attached to an
embryonal axis embryonal axis
2 Plumule is lateral. Plumule occur distally
3 A single cotyledon occupies terminal Cotyledons occur laterally
position
4 The envelope of plumule is called Coleoptile absent
coleoptile
5 Coleorrhiza is a protective sheath of Coleorrhiza absent

13
 radicle
6 A single cotyledon called scutellum is Scutellum absent
present
7 Relatively small suspensor Suspensor is larger

14
The Seed coat
The seed coat (testa) is a maternal tissue that surrounds the embryo, endosperm and perisperm (if
present). It protects the internal structures from biotic stresses and against desiccation and
mechanical injury, assists in gas exchange and water uptake, provides a conduit for nutrients for
the embryo and endosperm during their development, and may be a means for seed dispersal. Its
structure influences seeds’ viability, longevity and germinability/dormancy. Sometimes the seed
coat can serve as a useful characteristic for taxonomy (e.g. in the Fabaceae).

During seed development the integuments, which are an initial seed coat tissue, undergo
differentiation into layers containing specialized cells. Some cell layers in the seed coats may
accumulate large quantities of compounds, such as mucilage, phenolics or pigments (Moise et
al., 2005). In the Brassicaceae and Fabaceae, some cell layers do not undergo any significant
differentiation and remain parenchymatous (they are often crushed at maturity), while others
undergo a slight thickening of the cell wall and become collenchymatous. Some cell layers
undergo extensive secondary thickening of parts of the cell walls and become sclerified (palisade
layers).

The seed coat of the Brassicaceae consists of four distinct layers: (i) the outermost, or epidermal
layer of the outer integument, most frequently one cell thick, which may or may not contain
mucilage; (ii) a subepidermal layer (in the centre of the outer integument; in some species
absent), one or more cells thick, typically parenchymatous, although it may be collenchymatous
or sclerified; (iii) the palisade layer (the innermost layer of the outer integument), often
characterized by thickened inner tangential and radial walls; and (iv) the pigment layer (derived
from the inner integument), formed of parenchyma cells compressed at maturity (Moise et al.,
2005).

In the Fabaceae the outermost layer of the seed coat is a waxy cuticle of variable thickness
(Moise et al., 2005). The next layer is the epidermis consisting of a single layer of palisade cells
(macrosclereids) that are elongated perpendicular to the surface of the seed. Inside the palisade
layer is an hourglass cell layer (Fig. 7), which is composed of thick-walled osteosclereids. The
innermost multicellular layer is composed of partially flattened parenchyma.

Besides its functional aspects, the seed coat can also develop characteristics or specialized
structures that assist dispersal (Black et al., 2006). In some angiosperm species the coat becomes
fleshy and brightly coloured, which attracts animals, mainly birds. It is called the sarcotesta and
is typical of 17 dicot and monocot families, e.g. Annonaceae, Cucurbitaceae, Euphorbiaceae,
Liliaceae, Magnoliaceae and Palmae. Coats of species such as flax (Linum usitatissumum)
develop a mucilaginous epidermis, which facilitates the passage through a dispersing animal’s
intestines. The aril, a specialized outgrowth of the ovule or funiculus, can create a fruit-like
structure attractive to fruit-eating birds (e.g. longan (Dimocarpus longan), akee (Blighia sapida),
15
lleuque (Prumopitys andina)). Wind-dispersed seeds often possess wings (e.g. Asphodelaceae,
Bignoniaceae, Cucurbitaceae, Hippocrateaceae). Also, trichomes (hairs) produced by the seed
coat, funiculus or placenta assist in wind dispersal (e.g. Populus and Gossypium spp.).

The seed coat may be undifferentiated or degraded, especially when mechanical protection of the
seed is achieved by the fruit wall (pericarp, e.g. Poaceae) or inner region (endocarp, e.g.
Apiaceae, Fagaceae, Urticaceae) or in small, wind-dispersed seeds of dehiscent fruits (e.g. in
Orchidaceae) (Black et al., 2006).

FIG. 7. Schematic diagrams illustrating the general organization of the seed coat of Arabidopsis
thaliana and soybean (G. max L. Merrill). a, A. thaliana. The seed coat at the torpedo stage. mu,
Mucilaginous epidermal cells; p, palisade layer with thickened inner cell walls; pa,
parenchymatous cells; en, endothelium layer. The mucilaginous cells and the palisade layer
comprise the outer integument while the inner integument consists of parenchymatous cells and
the endothelium layer. Adapted from Beeckman et al. 2000; b, Soybean. A mature seed coat. p,
Palisade layer; h, hourglass cells; pa, partially crushed parenchyma; a, aleurone; em, crushed
endosperm The palisade layer and the hourglass cells comprise the outer integument while the
inner integument consists of parenchymatous cells. Adapted from Miller et al. (1999).

16
Developmental depiction of fruit growth
Initiation of fruit development is defined as the time of ovule fertilization and, in A. thaliana,
marks the morphological transformation of the ovary to form a silique with two separate locules.
Fertilization occurs at anthesis, when the ovules are mature and the stigma is receptive for pollen
germination. Pollen grains germinate on the stigma and the pollen tubes grow into the ovary
through the transmitting tract (Kandasamy, Nasrallah, & Nasrallah, 1994).

As fertilization of the ovules occurs, signals from the developing embryos and seeds are
exchanged with the plant to initiate fruit elongation and differentiation of specific tissues along
the carpel margins (Fig. 8).
The cells of the valves and the replum divide and expand primarily along the longitudinal axis,
contributing to the lengthening of the fruit (Eldridge et al., 2016). In the first phase after
fertilization, the fruit grows mainly due to cell division, and subsequently by cell expansion.

Fig. 8. Fruit development: developmental stages and tissue definitions. Left: A. thaliana fruit at
different developmental stages, from fertilization to dehiscence. Right: schematic representation
of tissue organization in the A. thaliana fruit and the genes controlling the identity of the
different tissue domains.

Concomitant with valve elongation, the valve margin starts to differentiate and will form narrow
files of lignified cells as well as a layer of separation cells (Fig. 8). Both cell layers will be
fundamental for seed dispersal when the fruit reaches maturity and dries out. At that point in
development, the valves detach from the replum, releasing the brown and mature seeds to fall
from the replum in a process called fruit dehiscence (Fig. 8).

Simple fruits can be classified as dry or fleshy, dehiscent or indehiscent, although this
classification can vary considerably, taking different shapes and sizes into account. Development
and final fruit size depend on (i) genotype, (ii) number of cells within the ovary prior to
fertilization, (iii) number of successful fertilizations that occur in the ovary, and (iv) the number

17
of cells composing the fruit (Srivastava & Handa, 2005). Moreover, seeds themselves are known
to impact the development of fruits. Indeed, size and shape of many fruits are being determined
by seed number and distribution within the ovary.

Both dry and fleshy fruits undergo the developmental phases of fruit set, fruit growth,
maturation, and ripening. Fleshy fruits are believed to have evolved from dry fruits, and a high
level of conservation exists between the genetic and molecular circuits that guide the
development of fruits in both classes (Knapp, 2002; Seymour et al., 2013). Fleshy fruits are
classified as climacteric if ripening happens concomitantly with an increase in respiration an
ethylene biosynthesis, and non-climacteric if these events do not occur.

18
Seed structure
The seed functions as the reproductive unit of the Spermatophyta (seed plants), and links the
successive generations. Other important seed functions concern dispersal and survival under
cold, dry or other inclement conditions. There is an immense diversity in the internal and
external structure of seeds. These differences are, to an appreciable extent, related to dispersal
and germination strategies, and may involve the size and position of endosperm and embryo,
structure, texture, and color of seed coat, and the shape and dimensions of the seed as a whole.
The minute seeds of orchids may weigh as little as 0,000002 g each. The seeds of the
legume Mora oleifera are possibly the heaviest, and the weight of each exceeds 1,000 g,
whereas a one-seeded fruit of the palm Lodoicea maldivica weighs more than 20,000 g. The
seed consists of three components: embryo, endosperm (sometimes perisperm), and seed-coat.
Both endosperm and embryo are the products of double fertilization, whereas the seed-coat
develops from the maternal, ovular tissues.

The embryo contains the precursor tissues for leaves, stem, and roots. The endosperm and
cotyledons—seed leaves—act as the food reserves for the growing embryo. The embryo contains
a double set of chromosomes, one set from each parent. Fertilization of the haploid egg by the
haploid sperm gives rise to the zygote, which develops into the embryo.

The endosperm is a feature common to most flowering plants, and it is created during the process
of double fertilization. Here, two sperm enter into each ovule. One sperm fertilizes the egg; the
other fertilizes the central cell, producing the endosperm. Conifers and other gymnosperms do
not undergo double fertilization, and therefore do not have a true endosperm.

Seed structure differs between monocots and dicots, two types of flowering plants (fig. 9).
Monocots, such as corn, have a single large cotyledon called the scutellum, which directly
connects to the embryo vascular tissues. The endosperm acts as the food reserve. During
germination, the scutellum absorbs enzymatically-released food materials and transports them to
the developing embryo.
The monocot embryo is surrounded by two protective sheaths. The first, the coleoptile, covers
the young shoot. The second, the coleorhiza, encases the young root. Both structures facilitate
soil penetration after germination.
Dicot seeds may be endospermic or non-endospermic. In endospermic dicots, such as tomatoes,
the food reserves are present in the endosperm. During germination, the cotyledons absorb the
enzymatically-released food material from the endosperm and transport it to the growing
embryo.

19
Fig. 9. Schematic drawing of dicot and monocot seed

20
Apomixis and polyembryony
In angiosperms, double fertilization is the common reproductive pattern in which two sperms
combine with an egg cell and a central cell. The former develops into an embryo, the embryonic
form of a new plant, and the latter develops into an endosperm whose function is to support the
early growth of this new plant (Amphimixis- the megaspore mother cells undergo mitosis and
meiosis to form an embryo sac with 8 nuclei, and then form embryos under the double
fertilization). By contrast, seed formation also occurs in apomictic plants but without
fertilization, hence, Apomixis (Greek. Apo-without, mixis-mixing) is a mode of reproduction
which does not involve formation of zygote through gametic fusion. Generally, in plants
apomixis commonly mimics sexual reproduction without fertilization. In apomictic plants, the
genetic information of the offspring is consistent with the female parent, but the process of
formation is different.

There are two main forms of apomixis (Fig. 10): (1) sporophytic and (2) gametophytic.
Sporophytic apomixis involves the direct formation of an embryo from a somatic cell in the
ovule. In gametophytic apomixis the embryo sac is obtained via apospory and diplospory. In
both cases, the unreduced egg cell develops parthenogenetically into an embryo. At the same
time, endosperm development in apomictic species may be independent of fertilization
(autonomous endosperm formation) or may need the fusion of the polar nuclei with a pollen
nucleus (pseudogamy) (Koltunow and Grossniklaus, 2003).

Diplospory involves the megaspore mother cells undergoing mitosis to form unreduced embryo
sac, and the apomictic initiation cells originate from the location of the megaspore mother cells
and eventually develop into embryos. The diploid egg as well as the diploid cells of the embryo
sac can grow into normal embryos and formation of embryo directly from diploid egg without
fertilization is called diploid parthogenesis, e.g Rubus, Apple, Poa. Meanwhile for Apospory, the
aposporous initial cells (AI) near the megaspore mother cells form an unreduced embryo sac
undergoes three rounds of mitosis and eventually develop into embryos. In facultative apomixis
in sexual plants, the appearance and differentiation of archesporial cells occur at much the same
time. During the development of the aposporous embryo sac, the nucellus cells close to the
megaspores gradually become larger and form an aposporous initial. After two mitoses, these
develop into an aposporous embryo sac. In a study of the ovule of Poncirus trifoliate,
aposporous initials were not evident prior to megaspore formation but, in one ovary, an
archesporial cell was observed with nearby enlarged nuclear cells—possibly representing an
aposporous initial.

21
Fig. 10. Amphimictic and apomictic embryo formation. (a) Amphimixis, the megaspore mother
cells undergo mitosis and meiosis to form an embryo sac with 8 nucleus, and then form embryos
under the double fertilization; (b) Diplospory, the megaspore mother cells undergo mitosis to
form unreduced embryo sac, and the apomictic initiation cells originate from the location of the
megaspore mother cells and eventually develop into embryos; (c) Apospory, the aposporous
initial cells (AI) near the megaspore mother cells form an unreduced embryo sac undergoes
three rounds of mitosis and eventually develop into embryos; (d) Sporophyte apomixis, the
apomictic initiation cells are derived from ovules, which rapidly divide and invade the embryo
sac, producing one or more embryos, and sporophyte apomixis can coexist with amphimixis
(adapted from Fei et al., 2019).

22
Polyembryony is more common than might be expected in sexually reproducing species. In
general, it is associated with apomixis. The emergence of polyembryony is thought to be a result
of apomixis. Polyembryony is the phenomenon of forming multiple embryos in one ovule. There
are many species exhibiting polyembryonal behavior including Mango (Mangifera indica L.)
(Litz et al. 1982), Citrus unshiu (Nakano et al. 2012) (Fig. 12) and Clidemia hirta (L.) (Mendes-
Rodrigues and Oliveira 2012). The occurrence of polyembryony is believed to be caused by
apomixis and generally occurs through adventitious embryo development of nucellar cells. The
existence of polyembryony in a species is usually associated with one in which there is a high
probability of apomixis. Indeed, the presence of polyembryony is considered indicative of
apomixis. Sporophytic apomixis encompasses polyembryony (Adventive embryony or
sporophytic budding, fig. 10). Here, an embryo directly develops directly from a diploid cell i.e.
nucellus and integument other than egg. This commonly occurs in citrus and opuntia specie to
give rise to polyembryony. There may be more than one egg cell in an embryo sac or more than
one embryo sac in an ovule. All the egg cells may get fertilized. Synergids and antipodal cells
may also form embryos. In gymnosperms polyembryony can also occur due to cleavage of
growing embryo and this is called cleavage polyembryony.

Occurrence of polyembryony due to fertilization of more than one egg is called simple
polyembryony. Formation of extra embryos through sporophytic budding is called adventive
polyembryony (Fig. 11).

23
Fig. 11. Overview of sexual and asexual seed development in citrus. Major processes of seed
development are depicted during the formation of monoembryonic and polyembryonic seeds.
Sexual reproduction pathways including meiosis, mitosis, and double fertilization occur in both
monoembryonic and polyembryonic seeds. In apomictic ovules, nucellar cells may possess the
ability to form nucellar embryos, which is histologically invisible, when the megaspore is divided
into the binuclear embryo sac. Several nucellar embryo initial cells (purple) emerge and develop
into adventitious embryos in the nucellus tissue (pink) surrounding the sexual embryo sac. In the
depicted adventitious embryogenesis pathway, parts of the nucellar embryo initial cells begin to
divide and launch embryogenesis when the zygote remains dormant after fertilization. Multiple
adventitious embryos develop within the ovule forming a polyembryonic seed. The
monoembryonic seed contains only the zygotic embryo (adapted from Zhang et al., 2018)

24
Fig. 12. Polyembryony and seedling emergence in apomictic citrus (adapted from Kishore et al.,
2012)

Importance of apomixis
In agriculture, apomixis has the potential to transform plant breeding, allowing new varieties to
retain valuable traits (such as high yields, tolerance to drought) through asexual reproduction.
The production of seeds without sexual union is considered the holy grail of plant biology.
❖ Hybrid varieties of seeds can be produced which provide higher and better yield.
❖ Adventive embryos are better clones than cuttings
❖ Embryos so formed are generally free from infections.
❖ Apomixis prevents the loss of specific characters in the hybrid. Also, it is a cost-effective
method for producing seeds.
❖ Apomixis is important for crop yield improvement.
❖ Polyembryony is significant in the formation of identical twins by cells of the embryo sac
other than the egg. Embryos develop from additional embryo sacs in an ovule

25