005 Allied III - Plant Pathology, Biofertilizers and Biopesticides - III Sem

STUDY MATERIAL FOR B.
SC MB
PLANT PATHOLOGY, BIOFERTILIZERS & BIOPESTICIDES
SEMESTER - III, ACADEMIC YEAR 2020-21
UNIT CONTENT PAGE Nr
I PLANT DISEASES AND ITS CONTROL 02
II HISTOPATHOLOGY OF PLANT 11
III PLANT BACTERIAL DISEASES 17
IV BIO-FERTILIZERS 35
V BIO-PESTICIDES 48
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STUDY MATERIAL FOR B.SC MB
UNIT - I
PLANT PATHOLOGY
Plant pathology (also phytopathology) is the scientific study of diseases in plants caused
by pathogens (infectious organisms) and environmental conditions (physiological factors).
Objectives of Plant Pathology:-

It deals with the cause, etiology, resulting losses and control or management of the
plant diseases.
The objectives of the Plant Pathology are the study on:

a. The living entities that cause diseases in plants;
b. The non-living entities and the environmental conditions that cause disorders in plants;
c. The mechanisms by which the disease causing agents produce diseases;
d. The interactions between the disease causing agents and host plant in relation to overall
environment
e. The method of preventing or management the diseases and reducing the
losses/damages caused by diseases.
Importance of the plant diseases

Globally, enormous losses of the crops are caused by the plant diseases. The loss can
occur from the time of seed sowing in the field to harvesting and storage.
Important historical evidences of plant disease epidemics are Irish Famine due to late
blight of potato (Ireland, 1845), Bengal famine due to brown spot of rice (India, 1942) and
Coffee rust (Sri Lanka, 1967). Such epidemics had left their effect on the economy of the
affected countries.
Definition and Terms:-

1. Parasite: An organism living upon or in another living organism (the host) and obtaining
the food from the invading host.
2. Pathogen: An entity, usually a micro-organism that can cause the disease.
3. Biotroph: A plant pathogenic fungus that requires living host cells i.e. an obligate
parasite.
4. Hemibiotroph: A plant pathogenic fungus that initially requires living host cells but after
killing the host cell grows on the dead and dying cells.
5. Necrotroph: A pathogenic fungus that kills the host and survives on the dying and dead
cells.
6. Pathogenicity: The relative capability of a pathogen to cause disease.
7. Pathogenesis: It is a process caused by an infectious agent (pathogen) when it comes in

contact with a susceptible host.
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8. Virulence: The degree of infectivity of a given pathogen.
9. Infection: The initiation and establishment of a parasite within a host plant.
10. Primary infection: The first infection of a plant by the over wintering or over summering
of the pathogen.
11. Inoculum: That portion of pathogen which is transferred to plant and cause disease.
12. Invasion: The penetration and spread of a pathogen in the host.
13. Colonization: The growth of a pathogen, particularly a fungus, in the host after infection
is called colonization.
14. Inoculum potential: The growth or threshold of fungus available for colonization at
substratum (host).
15. Symptoms: The external and internal reaction or alterations of a plant as a result of
disease.
16. Incubation period: The period of time between penetration of a pathogen to the host
and the first appearance of symptoms on the plant.
17. Disease cycle: The chain of events involved in disease development.
18. Disease syndrome: The set of varying symptoms characterizing a disease are collectively
called a syndrome.
19. Single cycle disease (Monocyclic): This type of disease is referred to those caused by the
pathogen (fungi) that can complete only one life cycle in one crop season of the host
plant. e.g. downy mildew of rape seed, club root of crucifers, sclerotinia blight of brinjal
etc.
20. Multiple cycle disease (Polycyclic): Some pathogens specially a fungus, can complete a
number of life cycles within one crop season of the host plant and the disease caused by
such pathogens is called multiple cycle disease e.g. wheat rust, rice blast, late blight of
potato etc.
21. Alternate host: (a) Plants not related to the main host of parasitic fungus, where it
produces its different stages to complete one cycle (heteroecious). (b) Collateral host:
The wild host of same families of a pathogen is called as collateral host.
22. Predisposition: The effect of one or more environmental factors which makes a plant
vulnerable to attack by a pathogen.
23. Physiologic race: One or a group of micro organisms similar in morphology but
dissimilar in certain cultural, physiological or pathological characters.
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24. Biotype: The smallest morphological unit within a species, the members of which are
usually genetically identical.
25. Symbiosis: A mutually beneficial association of two or more different kinds of
organisms.
26. Mutualism: Symbiosis of two organisms that are mutually helpful or that mutually
support one another.
27. Antagonism: The counteraction between organisms or groups of organisms.
28. Mutation: An abrupt appearance of a new characteristic in an individual as a result of an

accidental change in genes present in chromosomes.
29. Disease: Any deviation in the general health, or physiology or function of plant or plant
parts, is recognized as a disease.
30. Cop Damage: It is defined as any reduction in the quality or quantity of yield or loss of
revenue resulting from crop injury.
31. Deficiency: Abnormality or disease caused by the lack or subnormal level of availability
of one or more essential nutrient elements.
Causes of plant diseases:

Plant diseases are caused by pathogens. Hence a pathogen is always associated with a
disease. In other way, disease is a symptom caused by the invasion of a pathogen that is able to
survive, perpetuate and spread. Further, the word “pathogen” can be broadly defined as any
agent or factor that incites ‘pathos or disease in an organism or host.
In strict sense, the causes of plant diseases are grouped under following categories:
1.Animate or Biotic Causes:

Pathogens of living nature are categorized into the following groups.
a. Fungi
b. Algae
c. Bacteria
d. Phanerogams
e. Phytoplasma
f. Protozoa
g. Rickettsia-like organisms
h. Nematodes
2.Mesobiotic causes:
These disease incitants are neither living or non-living, e.g.
 Viruses
 Viroides
Inanimate or Abiotic causes:

In true sense these factors cause damages (any reduction in the quality or quantity of
yield or loss of revenue) to the plants rather than causing disease.
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The causes are:

Deficiencies or excess of nutrients (e.g. ‘Khaira’ disease of rice due to Zn deficiency)
a. Light
b. Moisture
c. Temperature
d. Air pollutants (e.g. black tip of mango)
e. Lack of oxygen (e.g. hollow and black heart of potato)
f. Toxicity of pesticides
g. Improper cultural practices
h. Abnormality in soil conditions (acidity, alkalinity)
Effect of Pathogen on the Plants:-

During the course of pathogenesis, normal activities of the infected host plant undergo
malfunction. Consequently, morphological and physiological changes occur.
Morphological or structural changes:

Physiological malfunctioning of the host cells causes disturbances in chemical reaction
which ultimately lead to some structural changes viz., overgrowth, phyllody, sterile flowers,
hairy roots, witches broom, bunchy top, crown gall, root knot, leaf curling, rolling, puckering
etc.
Physiological changes:
Disintegration of the tissues by the enzymes of the pathogen.
Effect on the growth of the host plant due to growth regulators produced by the
pathogen or by the host under the influence of the pathogen.
Effect on uptake and translocation of water and nutrients:

a. Abnormality in respiration of the host tissues due to disturbed permeability of cell
membrane and enzyme system associated with respiration.
b. Impairing the phenomenon of photosynthesis due to loss of chlorophyll and destruction
of leaf tissue.
c. Effect on the process of translation and transcription.
d. Overall reproduction system of the host.
DEFINITION OF DISEASE CYCLE:

The chain of events that leads to the development of a disease is called the disease
cycle – which may be different to the pathogen’s life cycle.
The incidence and severity of the majority of plant diseases vary on a distinct cyclic
basis. Each cycle includes two alternating phases; the parasitic phase and the survival or over
summering phase.
The disease cycle consists of a number of distinct steps:

1. The pathogen moves to and makes contact with the host surface;
2. It establishes itself on the host and penetrates plant tissues;
3. It initiates infection (early stages of proliferation);
4. It colonizes the host;
5. It reproduces and spreads inside the host;
6. It escapes from the host, and travels to, and infects, a new host.
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DEFINITION OF PATHOGENICITY:
The ability of infectious casual agent (fungus, bacterium, phytoplasma, virus) to produce
disease in a host organism is called pathogenicity.
This means that when a micro-organism that is pathogenic to a plant enters that plant, it
causes a greater or less degree of deviation from the plant’s normal functioning, or its normal
physiological processes, of sufficient duration to cause disturbance to the plant, leading to the
onset of disease symptoms, or the cessation of vitality.
In practical terms, a micro-organism is pathogenic to a plant if it damages that plant and

the pathogenicity is the capacity of a micro-organism to damage a host plant. The pathogenicity
of a micro-organism is a state or quality (a “character”, according to Shaner et al., 1992) of the
micro-organism, and as such is not measurable. Pathogenicity usually refers not to a single
micro-organism but to a group (e.g. pathovar, species or genus).
Pathogenic micro-organism must enter, colonise and damage its host, all the factors
that enable a micro-organism function in this manner should be considered factors of
pathogenicity.
Pathogenicity would then be a broad term that would also include virulence, pathogen
entry, and pathogen spread throughout the host.
SYMPTOMS OF PLANT DISEASE:

A disease manifests itself in the form of some typical external and internal changes in
the host plant.
Such visible changes, abnormalities or signs which serve to recognize the disease in the
lost plant are called symptoms of the disease.
The symptoms of plant diseases are of following 4 types:

a. Necrosis,
b. Hyperplasia,
c. Hypoplasia,
d. Gummosis and exudations.
Necrotic symptoms:
Death of the host cells, tissues and organs induced by a pathogen is called necrosis. The
necrotic areas are called lesions.
The various necrotic symptoms are:

(a) Streaks or strips:
These are elongated narrow lesions.
(b) Spots:
These are minute circular or sub-circular lesions of various colours such as brown, white,
dark, orange or red e.g. leaf spot in the blight of tea due to Glomerella cingulata and Fungal
members of Phacidiales or Doihideales cause raised black spots called tar-spots, Cystopus
candidus cause white spots in the cabbage. In some cases, the dead tissue of leaf snots shed,
leaving circular perforations called shot-holes.
(c) Blight:
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It involves rapid discoloration followed by death of a plant organ which gives a burnt
appearance, e.g. Blight ‘of Wheat, maize.
(d) Canker:
These are spreading sunken lesions in the stems, surrounded by raised margin. When
canker encircles the entire branch, cause the death of distal part. This happens in Pink disease
of rubber caused by Pellicularia salmoni color or in apple trees attacked by Physalospora mutila.
(e) Rot:
The affected tissues die and decompose to a great extent. For example, on the basis of
organization involved, they are root rot, stem rot, leaf rot, bud rot, fruit rot etc. On the basis of
type of decomposition they are soft rot, dry rot, wet rot and black rot.
(f) Damping off:

The stem of seedling attacked near the ground level so that the seedlings collapse.
Pythium and Rhizoctonia are important fungi causing damping-off of seedlings in chili, tobacco,
mustard, tomato, castor, cucurbits etc.
(g) Burn, Scald or Scorch:

Brownish lesions in succulent organs due to high temperature.
(h) Die-Back:
Progressive death of the affected part from the tip downwards.
(i) Anthracnoses:
The sunken lesions with raised margins on fruits, pods, chilies etc.
(II) Hypoplasia (Atrophy):

This is a symptom of many diseases where the growth of plants arrest causing stunting
or dwarfing e.g. Suppressed floral buds in mustard is due to Peronospora brassicae. In some
cases internodes fail to elongate causing the growing of foliage which gives a rosette forms.
This is called Resetting, e.g., groundnut rosettes (a viral disease).
(III) Hyperplasia and Hypertrophy:

Excessive growth of host tissues caused due to hyperplasia and hypertrophy. In
hyperplasia the overgrowth is due to increased cell division and cell number e.g. corn, smut.
But in hypertrophy the overgrowth of organ is due to increase in size of individual cells, e.g.
corn galls, witches brooms of Cherry etc. In mustard plants Cystopus candidus causes
hypertrophy of floral parts.
Some symptoms of abnormal growth are:

(a) Galls:
These are abnormal fleshy or woody outgrowths, when it is small its called as warts or
tubercles, when large, called as knots, e.g. Potato wart caused by Synchytrium endobioticum,
mustard root- galls by Urocystis brassicae.
(b) Intumescences:
Those are wart, like swellings of epidermal cells of leaves due to environmental factors.
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(c) Curl:
Leaves become puckered or crinkled, e.g. peach leaf-curl due to Taphrina defeormans.
(d) Hairy root:

It is are cluster of fibrous roots that appear like root hairs.
(e) Blotch:
Superficial growth on fruit
(IV) Discolouration symptoms:

The yellowing of green parts due to lack of light is called etiolation. But, when it
happens due to mineral deficiency, low temperature or infection of pathogens is called
chlorosis. When leaves appear transparent due to lack of any pigment is called albinism. Colour
conversion to red, purple or orange is called chromosis. Sometimes due to viral or fungal
infection yellow and green patches found irregularly called mosaic. The discolouration
surrounding leaf veins is called vein banding and the discolouration along leaf veins is called
vein clearing.
(V) Gummosis and exudations:

The production of a clear, amber-colour exudates on the surface of the affected parts to
a plant, which later sets into a solid mass insoluble in water, is known as gummosis. Gummosis
is common in cherry, peach and citrus trees. Reddish exudate is a common feature of stems
bleeding disease of coconuts caused by Ceratostomella paradoxa.
(VI) Other Symptoms:

i. Phyliody or Virecence:
Transformation of floral organs to vegetative organs, e.g. in peal millet flowers convert
into leave, due to downy mildew.
ii. Rusts:
Coloured pustules on host surface due to fungal spores.
iii. Smuts:
These are the affected areas releasing charcual-like dusty mass of spores.
iv. Mildews:
These are the coloured superficial patches on the host surface due to fungal infection.
When the superficial patches appear cottony or downy called downy mildews and when dusty
or powdery called powdery mildew appears.
v. Wilting:
It is withering or drooping of whole plant due to loss of turgidity. It generally caused due
to excessive transpiration, injuring to root system, toxins of pathogens etc.
vi. Witches brooms:

The woody branches of infected tree become swollen from which upward turned shoots
and small leaves arise which give broom – like appearance e.g. witches brooms of deodar plant
caused by Peridermium cedri.
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INVASION
Invasiveness is the ability of a pathogen to invade tissues.
Invasiveness encompasses,
(1) mechanisms for colonization (adherence and initial multiplication),
(2) production of extracellular substances ("invasins"), that promote the immediate
invasion of tissues
(3) ability to bypass or overcome host defense mechanisms which facilitate the
actual invasive process.
COLONIZATION
The first stage of microbial infection is colonization: the establishment of the pathogen
at the appropriate portal of entry. Pathogens usually colonize host tissues that are in contact
with the external environment. Sites of entry in human hosts include the urogenital tract, the
digestive tract, the respiratory tract and the conjunctiva. Organisms that infect these regions
have usually developed tissue adherence mechanisms and some ability to overcome or
withstand the constant pressure of the host defenses at the surface.
Bacterial adherence to mucosal surfaces.

In its simplest form, bacterial adherence or attachment to a eucaryotic cell or tissue
surface requires the participation of two factors: a receptor and a ligand. The receptors so far
defined are usually specific carbohydrate or peptide residues on the eucaryotic cell surface. The
bacterial ligand, called an adhesin, is typically a macromolecular component of the bacterial cell
surface which interacts with the host cell receptor. Adhesins and receptors usually interact in a
complementary and specific fashion with specificity comparable to enzyme-substrate
relationships and antigen-antibody reactions.
Dissemination of plant diseases:

Dissemination is the spread of plant pathogens within the general area in which it is
established is termed as their dissemination or dispersal or transmission.
Methods of dissemination
Different methods of dissemination of the plant pathogens with in a crop season as well
as to the next season after its survival are:
i. Direct methods
ii. Indirect methods.
 In direct transmission, the dispersal takes place along with the seeds and vegetative
plant parts used for propagation.
 Indirect transmission may be active/autonomous or brought about passively by
different agencies like wind, water, animals or human beings.
Direct methods:
It is further divided into:
Adherent transmission
Germinative transmission
Vegetative transmission
a. In adherent type, the pathogen propagules are carried over the surfaces of
seed or other propagative materials
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b. In germinative type, the plant pathogens are carried through the seed or other
propagules internally as in case of loose smut of wheat and barley.
c. In vegetative transmission, a large number of fungal, bacterial, viral and
phytoplasmal plant pathogens are carried in the vegetative plant parts used as
seeds such as tubers, cuttings, runners, grafts etc.
Indirect methods
a. Dissemination by Insects
b. Dissemination by Animals Other Than Insects
c. Dissemination by Air Currents
d. Dissemination by Water
e. Dissemination by Exporting and Importing of Commodities
f. Dissemination by Natural Root Grafting
g. Dissemination by Shooting Out of Spores
h. Dissemination through Pollen Grains
i. Dissemination through Other Media
PERENNATION
Perennation is the ability of organisms, particularly plants, to survive from
one germinating season to another, especially under unfavourable conditions such as drought
or winter. It typically involves development of a perennating organ, which stores
enough nutrients to sustain the organism during the unfavourable season, and develops into
one or more new plants the following year. Common forms of perennating organs are storage
organs (e.g. tubers, rhizomes and corm), and buds. Perennation is closely related
with vegetative reproduction, as the organisms commonly use the same organs for both
survival and reproduction.
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UNIT - II
HISTOPATHOLOGY OF PLANT
Adjustment is probably, one of the most important virtue of a system that ensures it
survival, be it host or parasite. On planet earth, the green plants (autotrophs) constitute the
only biological system capable of converting solar energy (electro-magnetic radiations) into
chemical energy.
Plants as a biological system resist this exploitation, at all levels and by all means. The co
evolution, forced by co-existence with pathogen, has led to development of defence
mechanism in plants. Thus, resistance against any 'deleterious act' has become a natural and
universal response of plant system. The resistance against parasites/pathogen is the heritable
trait of plants by virtue of which they resist attack by parasites/pathogens or their activities.
The defence mechanism(s) has ensured the survival of plants in spite of living amongst
some of the potentiality devastating pathogens in addition to abiotic stresses. Plants have also
developed ability to resist/tolerate various abiotic stresses. Pathogenesis and Host Response
Analysis most of the host parasite relationships reveals the pattern of pathogenesis, the plants
on their part, do exhibit defence mechanisms (structural and chemical) as soon as challenged
by the pathogen. The moment pathogen propagules come in contact with host surface. the
plants due to hereditary characters have several naturally occurring physical and chemical
barriers (pre-existing) resisting penetration, and if at all the penetration occurs, the host reacts
by different means resulting in formation of physical and chemical barriers. These two
conditions are discussed in picture below:
DEFENCE MECHANISM:
Induced or active plants have to face the wide variety of pathogens (enemies) standing
at a place. Thus a strategically designed pre-existing (structural and biochemical) defence
mechanism in plants exists. The real value of this system has not been critically examined. It
appears that these pre-existing defence mechanisms help plants in warding-off most of
microbes as non-pathogens. But it does not seem to be sufficient.
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The induced/active defense mechanism in plants may operate at different levels

a. Biochemical defence
b. Defence at cellular level
c. Defenses at tissue level
The activation or induction of defence mechanism may be both specific and non-specific
type. Several structural changes are known to be induced by a range of biotic or abiotic
elicitors. These dynamic defence mechanisms prevent further colonization or spread of
pathogen. Active defence in plants involves cellular defenses that rely upon preformed
surveillance systems are encoded by resistance genes. The receptor-proteins are strategically
located in cell membrane to detect the pathogen or factor translocated by pathogens. The
ability of plant to mount an active defence response is again under genomic control.
Inducible structural defenses:

Even after the establishment of infection in plant cells, the host defence system tries to
create barriers for further colonization of tissues. This may be at various levels.
Lignifications:
Lignified cell wall provide effective barrier to hyphal penetration. They also act as
impermeable barrier for free movement of nutrient causing starvation of pathogen.
Following are examples.
a. Radish: Peronospora parasitica, Alternaria japonica
b. Potato: Phytophtora infestans
c. Wheat: Septoria nodorum
d. Cucumber: Cladosporium cucumerium, Colletorichum lagenarium
e. Carrot: Botrytis cineria
Cork layer
In several plants the infected cells are surrounded by suberized cells. Thus, isolating
them from healthy tissue. Corky layer formation is a part of natural healing system of plants. eg.
common scab of potato and rot of sweet potato are good examples.
Abscission layers:
It is a gap between host cell layers and devices for dropping –off older leaves and
mature fruits. Plant may use this for defence mechanism also. I.e. To drop-off infected or
invaded plant tissue or parts, along with pathogen. Shot holes in leaves of fruit trees is a
common feature.
Tyloses:
The tyloses are formed by protrusion of xylem parachymatous cell walls, through pits,
into xylem vessels. The size and number of tyloses physically block the vessel. The tyloses are
inductively formed much ahead of infection, thus blocking the spread of pathogen. It suggests
biochemical elicitors and movement of tyloses inducing facto (TIF) up the stem. eg. Sweet
potato: Fusarium oxysporum f. sp. Batatas.
Gum deposition:
The gums and vascular gels quickly accumulate and fill the intercellular spaces or within
the cell surroundings the infection thread and haustoria, which may starve or die.
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Induced biochemical changes:

The induced biochemical changes in host plants are the last line of host defence. This
may condition a plant or plant tissue from susceptible to resistant to immune status as per their
genetic potential. The role of bio chemical factor in host defence is based on the following four
attributes:
1. The substance is associated with protection against disease at the site where protection
occurs.
2. The substance can be isolated from the host showing protection against the disease.
3. Introduction of isolated substance to the appropriate susceptible host confers
protection.
4. The nature of protection so induced resembles that of the natural agents of a resistant
plant.
HYPERSENSITIVE RESPONSE (HR)

Hypersensitive response (HR) is a mechanism used by plants to prevent the spread
of infection by microbial pathogens. HR is characterized by the rapid death of cells in the local
region surrounding an infection and it serves to restrict the growth and spread of pathogens to
other parts of the plant. It is analogous to the innate immune system found in animals, and
commonly precedes a slower systemic (whole plant) response.
HR is triggered by the plant when it recognizes a pathogen. The identification of a
pathogen typically occurs when a virulence gene product, secreted by a pathogen, binds to, or
indirectly interacts with the product of a plant resistance (R) gene (gene for gene model). R
genes are highly polymorphic, and many plants produce several different types of R gene
products, enabling them to recognize virulence products produced by many different
pathogens.
The cells surrounding the lesion synthesize antimicrobial compounds,
including phenolics, phytoalexins, and pathogenesis related (PR) proteins, including β-
glucanases and chitinases. These compounds may act by puncturing bacterial cell walls; or by
delaying maturation, disrupting metabolism, or preventing reproduction of the pathogen in
question.
Host processes usually targeted by bacteria include; alterations to programmed cell
death pathways, inhibiting cell wall-based defenses, and altering plant hormone signaling and
expression of defense genes.
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SYSTEMIC ACQUIRED RESISTANCE (SAR)

Induced resistance (cross protection) in plants is a phenomenon of significance, which
has not been properly exploited for plant disease management, probably because of our poor
understanding. Induced resistance, localized or systemic, may be specific. The signal molecule,
that propagates the resistance to distant places are vital in systemic induced resistance. The
resistance is induced in manner comparable to immunization in mammals but the mechanism
differs.
The resistance may be induced due to any of the following:
a. Accumulation of PR proteins
b. Activation of lignin synthesis
c. Enhanced peroxidase activity
d. Suitable changes in plant metabolism
Principle of induced resistance

Induced resistance is a phenomena where a lead treated with certain chemicals or
inoculated with pathogen’s avirulent strain produce a signal compounds that is transported
systemically throughout the plant and activities its defence mechanism (making the entire plant
resistant to subsequent infection) without its own physical presence at the site. The picture
below explains a hypothetical mode to explain induction of SAR.
Toxic substances produced:

Rapid production/suitable modifications and/or/ accumulation of chemicals toxic to
pathogen up to effective concentrations is an important component of overall active defence
strategy of plants. Slow production or accumulation or low levels of similar chemicals have
reported in susceptible host plants also.
Plantibodies:
A plantibody is an antibody that is produced by plants that have been genetically
engineered with animal DNA encoding a specific human antibody known to neutralize a
particular pathogen or toxin. A plantibody is produced by insertion of genes encoding
antibodies into a transgenic plant. The plantibodies are then modified by intrinsic plant
mechanisms (N-glycosylation).
Phenolic compounds:
The phenolic compounds, viz., chlorogenic acid caffeic acid and oxidation products of
floretin, hydroquinone hydroxyquionones and phytoalexins are main toxic chemical produced
to inhibit pathogen or its activities. Some of these are performed toxic chemicals while others
may be de novo synthesized or modified to more toxic forms. The enzymes involved in chemical
pathways are present in host cell (pre-existing).
Phytoalexins:
Most common response of plants to stress, biotic (phytoalexins/insects) or abiotic
(wounding), is the production and accumulation of substrates that can inhibit the growth and
activities of the biotic factors or may help in healing process. Muller and Borger proposed the
concept of phytoalexins in their study on hypersensitive reaction of potato to avirulent
P.infestans strains. Phytoalexins are antibiotics produced in plant pathogens interactions or as
result response to injury or other psychological simulation.
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Quinones:
Plants sometimes produce compounds during normal growth that inhibit the
development of pathogens. Phytoanticipins may be excreted into the external environment
(e.g. rhizosphere or phylloplane), accumulate in dead cells or they may be sequestered in
vacuoles in an inactive form. The dead cells of brown onion skins contain the quinones catechol
and protocatechuic acid, which inhibit germination of spores of the smudge pathogen.
Active defense to pathogens

Induction of host resistance, structural or biochemical seems to be universal I plants.
Active defense responses have been reported against all classes of pathogens (fungi, bacteria,
viruses and nematodes). Active defense response may lead to incompatible host-pathogen
interaction.
The oxidative burst

a. Membranes are also the sites where the oxidative burst occurs.
b. The term 'oxidative burst' was first used to describe a rapid increase in respiration
observed in neutrophils involved in the immune response.
c. This increased level of respiration is now known to be due to the generation of reactive
oxygen species, especially hydrogen peroxide and the superoxide anion (Oz-), through
the addition of electrons to 02 catalysed by the membrane-bound enzyme, NADPH
(oxido reductase).
d. The first burst rapidly follows wounding and inoculation, while a much larger burst in
incompatible interactions immediately precedes hypersensitive cell death.
e. An oxidative burst has been described in a range of plant-fungal and plant bacterial
interactions.
f. The rapid oxidative burst generates levels of reactive oxygen species that initiate
membrane lipid peroxidation and cell death.
g. The oxidative burst in plants is associated with the release of local and systemic signals
that trigger gene expression and the oxidative cross-linking of host cell wall
components.
h. Levels of reactive oxygen species accumulate at the infection court that is sufficient to
kill micro-organisms in vitro.
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UNIT - III
WHITE RUST OF CRUCIFERS
White rust or white blisters disease is one of the common diseases of crucifer crops. It is
worldwide in distribution occurring in all the areas wherever crop is cultivated. Both wild and
cultivated varieties are attacked.
The disease affects a large number of crucifer crops of economic importance like
Mustard, Cress, Rape, Radish, Cabbage, Cauliflower, turnip etc. In India the disease is reported
on Mustard, Rape, Eruca sativa, turnip. Cauliflower and Cleome viscosa.
Effect of White Rust Disease:

Although considered unimportant in proportion to its widespread occurrence, the
disease may cause serious damage in certain areas and the losses may be up to 25 percent
when the floral parts get malformed and seeds are not formed at all.
The disease in association with downy mildew disease of crucifers caused by

Peronospora parasitica cause severe damage to the crop
Symptoms of White Rust Disease:

The disease affects all the aerial parts of the plant, the roots are not attacked.
Symptoms may appear as a result of two types of infection: Local and Systemic.
In case of local infection, isolated spots or pustules appear on leaves or stems or

inflorescence. The pustules are of variable size, measuring 1 -2 mm in diameter and are raised
shiny white areas.
These may arise in close proximity and coalesce to form large irregular patches. Usually,
the pustules appear in circular or concentric arrangement with one or two central areas. The
host epidermis ruptures exposing white powdery mass consisting of spores of the fungus.
Pustules occurring on leaves are usually confined to the lower surface only.
In systemic infections, young stems and inflorescence are infected. The fungus becomes
systemic in these parts and the affected tissues are stimulated to various types of deformities.
The most prominent is Hypertrophy of the affected parts. Due to Hypertrophy and Hyperplasia
of floral parts, these show swellings and distortion.
The peduncle and pedicel may become enormously thickened upto 12-15 times, the
normal diameter. Floral parts become fleshy, swollen, green or violet in colour, the stamens
falling off early.
The petal may turn green sepal like and stamens and carpels are also converted to
swollen leaf like structures. The ovules are usually atrophied as also the pollen grains resulting
in total sterility. Pustules may also appear on these parts. However, the affected parts are full
of oospores and starch.
When the systemic infection has taken early, the growth of the entire plant is checked,
stunted and only small leaves may be formed.
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The stem and the axis of the inflorescence may get twisted appearing in a zigzag
sequence. Normal dormant buds are stimulated and grow into lateral shoots.
Causal Organism:
The causal organism Albugo Candida (Lev.) Kunze or Cystopus candidus Lev. is an
obligate parasite.
Disease Cycle:
The primary infection occurs due to oospores perennating in the soil or due to mycelium
perennating on perennial hosts. These serve as primary inoculum when the environmental
conditions are favourable.
Oospores germinate in presence of water to form a vesicle in which a large number of

zoospores are formed. These zoospores swim in a film of water and land on the suitable host,
germinate by germ tubes, enter the host and establish infection. The mycelium in the host is
intercellular with globose haustoria.
Soon the mycelium after absorbing nutrients and food materials from the host,
accumulates below the lower epidermis. Conidiophores, which are clavate, and formed at the
tip of hyphae, begin to produce conidiosporangia in basipetal succession. The pressure of these
breaks open the lower epidermis and white rust symptoms become apparent on the leaves.
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The conidiosporangia produced during early phase of the growing season cause
secondary infection in the host. These are blown away by wind or any other agency, land on the
host surface and germinate to form zoospores.
The zoospores germinate by formation of germ tubes which enter the host and cause
secondary infection. If the conditions are favorable, this is repeated.
When the conditions become unfavourable or during the later phase of the growing
season, the fungus begins sexual reproduction producing oospores. These oospores, being
thick-walled, can withstand the unfavourable conditions.
During harvesting of the crop, the diseased hypertrophied portions of the plant are
generally left in the field where they perennate waiting for the favorable conditions to return
back.
Control Measures of White Rust Disease:

The disease may be controlled by the following methods:
a. Clean cultivation and destruction of weed should be practiced.
b. Crop rotation will avoid the soil borne primary inoculum.
c. Spraying with 0.8 percent Bordeaux mixture or Dithane M-45 (0.2%) may be undertaken
to check the spread of the disease.
d. Disease resistant varieties be preferred.
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LATE BLIGHT OF POTATO

Introduction
Late blight is a serious fungal disease of potatoes. It is worldwide in its distribution. It
occurs in potato growing areas of the world. Winter is the main potato growing season in India.
It is followed by hot summer months in the plains. The drought and high temperature kill the
fungus in the soil.
The late blight epidemics are thus rare in the plains in India. It is destructive to the crop
grown in the rainy season. The disease occurs annually in the cooler Himalayan regions
extending from Assam to Kashmir at an altitude of 6,000 ft. or more as the crop is grown in the
rainy season.
Moreover, the temperature during the day is never above 22°-23°C which is favourable
for the appearance of disease. The crops grown in the plains have been usually free from the
epidemics of late blight because the chief predisposing factors (temperature and moisture) that
render potato plants susceptible to disease are absent during the period of their growth.
The temperature is high for the development of the disease. Now it has established
itself in the Indo- Gangetic plain and occurs annually in the states of Punjab, Uttar Pradesh,
Bihar, and W. Bengal. The disease is also destructive to tomatoes.
Effects of Late Blight:

The damage caused by the disease is frequently very high. Severe damage to the foliage
shortens the growing season (Fig. 22.5). Consequently the tubers remain small and reduced in
weight.
They are produced in smaller numbers. This results in the reduced yield. In severe cases
of infection there is complete loss of the crop, Infection also results in the decay of tubers in
the field and storage.
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Symptoms of Late Blight:

The disease first appears on the tops of the plants generally after the blossoming period
but mostly in the month of January. It may appear as well at any time during the growth period
of the plant. The conditioning factor is the favorable environment.
The disease makes its appearance as small, dead, brownish to purplish black areas or
lesions. These appear on the tips and margins of the leaflets, rachis, petiole and stem. Under
favourable conditions (low temperature and high humidity) the lesions rapidly increase in size
involving the whole surface of the leaf.
The disease generally first attacks the leaves, and petioles near the ground and the
lesions appear on the lower surface of the leaflets on individual plants and then spreads
upwards.
Finally, a rapid and general blighting of foliage occurs. The blighted leaves curl and
shrivel in dry weather. Under moist conditions they decay and emit a characteristic offensive
odour.
Examination of the lesions on the lower surface of the leaf on a dew morning reveals a
delicate growth of the fungus parasite in the form of whitish powdery bloom. It consists of
sporangiophores and sporangia (Fig. 22.7 E) of the pathogen pushing out through the stomata.
The sporangia serve to spread the disease in the growing season.
Potato tubers are often infected in the field after the tops have been blighted. They get
separate infections while in the hill. There is brownish discoloration of the skin of those parts of
the tubers which lie nearest the surface of the soil.
These dry rot spots remain firm and extend to about half an inch below the surface.
During storage, the bacteria assist to set in the wet rot phase. In cool and dry conditions the
progress of the disease is slower and the wet rot phase is generally checked.
Under moist conditions hyaline mycelial hyphae and sporangiophores push out through
the lenticels and appear on the surface of infected tubers.
Causal organism and over-wintering:

The causal organism is Phytophthora infestans (Mont.) De Bary. The mycelium is
aseptate conenocytic, hyaline and branched. The hyphae are both intercellular and
intracellular.
They form rudimentary haustoria in the host leaf cells but in the tubers the haustoria
are more common and elaborate (club-shaped, hooked or spirally twisted). According to De
Bary (1876), the mycelium overwinters in the infected tubers.
Disease Cycle
The infected tubers (A) are generally considered as the main source of primary infection
in India. The survival of the fungus in the soil in the Indian climatic conditions in any form
appears remote. According to the widely held view, the fungal parasite overwinters as a
dormant mycelium in the infected tubers.
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It becomes activated at the time of germination of the diseased seed tubers among the
planting stock or waste tubers in dump heaps or infected tubers remaining in the ground after a
previous crop. The activated mycelium invades the healthy sprouts (B).
The second view is that the thick-walled resting oospores which are found in abundance
in the infected tubers are the important overwintering structures. They play a significant role as
the source of primary infection.
At the planting time, the resting oospore germinates. The germ tube after emergence
usually ends in a terminal sporangium. The contents of the latter divide to form zoospores. The
released zoospores invade the healthy sprouts and bring about infection.
According to some, the sexual phase seems to play no role in the life history of the
pathogen. The infected sprouts emerge above ground and produce shoots which contain the
mycelium (C).
It grows and ramifies in the intercellular spaces absorbing nutrition by putting haustoria
into the host cells (D). Under suitable conditions of temperature and humidity, the mycelium
pushes out hyahne, branched, indeterminate sporangiophores through the stomata of the host
leaves (E).
The thin-walled, ovoid or lemon-shaped sporangia, each with an apiculate tip, are borne
singly at the tips of sporangiophores or their branches. As the sporangium reaches maturity,
the supporting hyphal branch immediately below it swells slightly and continues to grow
turning the attached sporangium to the side.
The elongation of the branch proceeds and a new sporangium is formed. The process is
repeated. A fertile branch or sporangiophore is thus characterized by 9 or 10 such swellings
occurring at intervals.
Each nodular swelling marks the point where the sporangium was borne. The mature
sporangia are readily detached and spread by splashing rain or air currents to new potato
plants (F1 and a).
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On reaching a suitable host (potato), the sporangia germinate on the leaves (F).
Germination is influenced by moisture and temperature conditions.
(a) Indirect Germination:

In cool moist weather the sporangia function as zoosporangia (F 1-3). The optimum
temperature for the formation of zoospore is 12ºC (54°F). In the indirect germination the
protoplasmic contents of sporangium divided to form a number of (usually 8) biflagellate
zoospores (F3).
They are liberated in a group through terminal pore formed by rupture of the apical
papilla. The released zoospores, after a brief period of activity in rain water or dew come to
rest. Each retracts its flagella and secretes a wall around it.
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The clothed zoospores (cyst) then germinates by pushing out a germ tube or infection
thread (F4). The zoospores germinate rapidly at 12° to 15°C. Cool and moist nights are thus
favourable for the formation and germination of zoospores. The germ tubes show rapid growth
at 21°C . After infection they grow best at a slightly higher temperature.
(a) Direct Germination:

Under dry and warmer conditions no zoospores are formed. The sporangium functions
as a conidium (Fa). It directly puts out a germ tube or infection thread (F b). The optimum
temperature for this direct germination of sporangia is about 24° or ’25°C.
The indirect method of germination of sporangia by the formation of zoospores in a

terrestrial late blight fungus is an instance of retention of an ancestral primitive character which
was normally used by its aquatic ancestor.
(b) Spread of the Disease (Secondary Infection):

The infection thread produced on the surface of the host leaf in either of the two above-
mentioned methods enters the host tissue (leaves or stem). It makes its entry occasionally
through the stoma but more often it penetrates directly through the cuticle by a penetration
hypha arising from an appresorium (F4).
The lower surface of the leaf is more susceptible than the upper. The infected leaves
produce another crop of sporangia. These are carried by wind to the healthy plants which are
thus infected. This constitutes secondary infection. The process is repeated.
As a result the disease spreads during the growing season over large tracts under potato
cultivation. The disease spreads quickly when cool and wet nights alternate with warm moist
days. Low temperature and high humidity favour the spread of the disease.
Field infection of Potato Tubers:

The tubers get separate infections (G). It is caused by zoospores produced in foliage
lesions (blighted tops) or present in the contaminated soil. Sporangia and zoospores come in
contact with the tubers in two ways.
Firstly, by contact freshly lifted healthy and wounded tubers with diseased haulms and
contaminated soil.
Secondly, during crop growth, the zoospores and sporangia washed down the stems
into the soil by rain come in contact with the tubers.
Tuber infection is dependent on the germination of sporangia, release and motility of

zoospores. The released zoospores have to move through soil to the infection sites. The longer
the zoospores continue to swim and greater their number, the greater are the chances of
infection. The germ tubes gain entrance through the eyes, wounds and lenticels.
According to Sato (1979), wet cool soil promotes infection but wet warm soil lowers it
because cool water at 16°C or below 12-14°C favours indirect germination of sporangia and
prolongs motility of zoospores. The sexual phase seems to play no significant role in the life
history of the pathogen.
The severity of late blight infection is governed by environmental conditions.
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The chief among them are:

a. Night temperature below dew point for 4 or more hours,
b. Minimum temperature 10°C or slightly above,
c. Mean cloudiness not below O.8°C on the next day,
d. Rainfall during next 24 hours, at least 0.1 mm.
Control Measures of Late Blight:

Various methods of control of the disease are known.
1. Selection of Seed (Planting) Tubers:

The seed tubers should be free from the disease. This requires strict seed tuber
inspection at the cutting time. This measure will eliminate direct infection.
2. Storage of Tubers at 40ºF or below:

Storage of potato tubers in cold storage rooms reduces or even checks the progress of
the rot.
3. Growing Disease Resistant Varieties:

Considerable success has been achieved in the perfection of resistant varieties of potato
at the potato breeding station, Simla. Growing these will provide an increasing opportunity to
combat the disease.
4. Use of Fungicides:
Resistance alone has not effectively checked the disease. Therefore the complete
control of blight is accomplished by the application of protectant fungicides. These are applied
before infection for effective control in two ways namely by spraying or dusting as follows:-
4a. Spraying:
The best method of control is the timely and repeated foliage spray schedule with
copper fungicides such as Perenox, Blitox-50 and Fytolan. Dithane Z-78, and Dithane M-22 have
proved more effective than the copper fungicides. The spraying should start when the plants
are 8 inches tail.
It should continue until the harvest time at 10 days’ interval. Both the surfaces of foliage
should be properly protected by adequate spraying delivered with a considerable force in the
form of fine mist.
Mancozeb and Chlorothalonil are the major fungicides which are presently used. Both
fungicides inhibit sporangial and spore germination but has little effect on the mycelium in the
leaf tissue.
4b. Dusting:
Some people claim that dusting the foliage with copper-lime dust is a more effective
control measure. Dusting is done in the morning when the plants are wet with dew.
5. Sanitation:
Destruction or proper disposal of potato tuber refuge from pits and store houses IS
another practical measure to reduce the incidence of disease.
6. Tuber treatment before storage:
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The tubers should be dipped in 1: 1,000 mercuric chloride solution for 90 minutes
before storage. Before use they should be washed.
7. Avoidance of injuries to tubers at harvest is also important.

8. In cool humid areas killing of foliage a few days before harvest proves beneficial. This is
accomplished by spraying with herbicides or flame throwers or by the use of mechanical vine
beaters.
ERGOT OF RYE
Ergot is a fungal disease caused by fungi of the genus Claviceps. Species in this genus
are unique in that they only infect ovaries of the host plants; no other part of the plant is
infected. There are approximately 40 species of Claviceps with C. purpurea (Fries ex Fries)
Tulasne being the species of greatest concern. Although C. purpurea has a very broad host
range, including approximately 400 grass species, the most economically important of these is
rye.
Symptoms and Signs

The first obvious sign of ergot infection is appearance of ‘honeydew’, a sticky yellow
sugary solution consisting of host sap and conidia between the affected glumes of the rye. This
is secreted by the infected plant ovary, which eventually is replaced by a purplish-black
sclerotium, commonly referred to as an ergot. The size of the sclerotium depends on the host
plant; it is generally 1 to 5 times larger than the host seed. Thus, the largest ergots (1-5 cm, 0.4-
2 inches) are found in large-seeded plants such as cereal rye. The sclerotium consists of a
whitish mycelial tissue containing storage cells and a dark pigmented outer cortex that protects
the fungal mycelia from desiccation, UV light and other adverse environmental conditions.
Pathogen Biology
Sexual reproduction
Sexual reproduction involves the fusion of female ascogonia and male antheridia,
karyogamy to form diploid nuclei, which is followed by meiosis to return to the haploid state.
This results in the production of sexual fruiting bodies called perithecia that contain multiple
asci, each of which contain eight filiform ascospores that are up to 2 µm in diameter and 60-70
µm long. Ascospores are ejected from the asci and perithecia into the air in an explosive
manner (to a height of 7-15 cm) and are disseminated by air currents or by insects. Each
sclerotium produces up to one million ascospores. Only those ascospores that land on a host
stigma or ovary can cause infection. The stigma of a grass flower is large and featherlike to trap
windborne pollen. This same feature traps the airborne ascospores. Ascospores, which are the
primary (initial) inoculum, germinate and infect the ovary within 24 h.
Asexual reproduction
A germinated ascospore produces a long filamentous hypha that colonizes the ovary of
the host plant flower. As the hyphae grow longer and more numerous, they are called a
mycelium. Fungal mycelium produces numerous asexual conidia (secondary inoculum) from
palisade conidiophores within a sweet, yellowish, mucilaginous substance called honeydew.
This stage is also called the sphacelia or honeydew stage. The honeydew attracts insects to the
wind-pollinated flowers. Insects contaminated with conidia may visit healthy flowers where
new infections are initiated. Splashing raindrops also aid in the dispersal of the conidia. Conidia
from ergot-infected wild grasses, particularly in fence rows, can be the primary inoculum in
cereal and grass seed production fields. Over time, hyphae consume the entire ovary and
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hyphal threads become thicker and interwoven. Meanwhile, conidia and honeydew production
ceases. Mutual pressure of the ever-growing hyphae causes production of dense mass of
compact tissue called pseudoparenchyma, which eventually develops into a hard, dark colored
sclerotium, or ergot.
Disease Cycle and Epidemiology
Disease Cycle
The sclerotium (or ergot) is the survival or overwintering structure of C. purpurea. At
harvest, a fraction of sclerotia is harvested along with the grain and another fraction falls on the
ground. Sclerotia lie dormant until they are exposed to favorable weather conditions. Four to
eight weeks of 0 – 10°C temperatures is required for vernalization of the sclerotia before
germination. Sclerotia germinate in the spring, just prior to flowering in the cereals and grasses,
and give rise to a stroma (stromata plural), formed in mushroom-like fashion on stipes with
spherical capitula. It is within these stromata that sexual reproduction occurs.
Epidemiology
Claviceps purpurea is common in temperate climates in which the cold period required
for sclerotial germination is met. In warmer climates, such as the southeastern U.S., sclerotia
are colonized by other fungi and do not survive well.
Rainfall or high soil moisture is required for stroma formation and ascospore
production. In cereal grains and many of the grasses, resistance to infection develops after
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fertilization. Thus, conditions that delay or interfere with pollination, such as cool, wet weather,
can increase the period of susceptibility.
Conidia are an important means of secondary spread. Any transfer of honeydew from
infected to healthy flowers can lead to infection. Secondary spread can occur through any
means that moves conidia to healthy flowers, including rain splash, insects, head-to-head
contact, and moving equipment.
Disease Management
Prevention is the best management strategy, but once ergot is observed in the field,
there is not much a farmer can do to control the disease.
Cultural Practices
Cleaning seed:
Even though this is labor intensive and expensive, most ergots can be removed from
grain by gravity-based cleaning equipment or flotation in brine (e.g., a 20% salt solution).
Planting clean seed:

There are no effective seed treatments against ergot, so only ergot-free seed should be
planted. Use of certified seed can greatly reduce the risk of planting ergot infected seed.
Ensure uniform stands:

Only seed with a good germination percentage should be used. Seed should be sown at
a consistent depth, following a balanced fertilizer program and other recommended agronomic
practices. Non-uniform stands within a region can result in a prolonged flowering period, which
in turn increases the spread of the disease within the field or from one field to another.
Late tillers and side shoots flowering outside the pollination period of main crop are
more affected by ergot than the main shoots, especially when the crop stand is thin due to poor
growing conditions. This is because they do not receive enough pollen and they still produce
honeydew even after the main crop is mature, thus contributing considerably to the secondary
spread.
The best crop husbandry practices against ergot are those that ensure a well-developed,
well-nourished crop stand. Thus, adequate seed density and fertilization are very important.
Adequate copper fertilization is known to play a role in the control of ergot in wheat. Evidence
also exists that copper deficiency produces small anthers and enhances pollen sterility. If the
flower stays open for long without pollination, it is susceptible to ergot infection by spores of
the fungus for a longer time.
Sanitation:
Wild, weedy grasses and cereal volunteers that are within or outside the field should be
eradicated before heading. These volunteers could be the first source of ergot inoculum from
overwintered sclerotia or as a source of honeydew if produced prior to crop flowering.
Crop rotation:
As ergots (sclerotia) do not usually survive for more than one year, rotation with non-
susceptible host plants is a viable management tactic for annual crops.
Deep plowing:
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Ascospores from stromata cannot be discharged into the air if sclerotia are buried in
the soil through deep plowing. Although deep plowing buries the ergots, many cereal crops are
now grown with "no-till" practices in which the new crop is seeded directly into the stubble
from the previous crop to reduce soil erosion. Crop rotation is even more important when deep
plowing is not practiced.
Harvesting techniques:
Ergot is often more severe around the edges of a field, because spores are transported
from roadside grasses by wind and active insects. Prior to harvest, fields should be scouted to
determine heavily infected areas and the plants in those areas should be harvested separately.
Field burning:
Ergot is one of the most important diseases in grass seed production. As many of the
grass species are perennial, tillage and crop rotation are not management options.
Post-harvest field burning has been practiced in the northwestern U.S. to manage ergot and
other diseases and pests since the 1940s, but environmental concerns have resulted in
legislative restrictions to burning.
Chemical Control
Chemicals have been applied to seed or soil to inhibit production of ascospores from
sclerotia but are not economical. Recently, sterol-inhibiting fungicides have been applied at the
onset of flowering to prevent infection in Kentucky bluegrass seed production, but such
treatments are not usually economical in cereal production.
STEM RUST OF WHEAT

Host: Tritlcum Vulgare
Pathogen: Puccinia Graminis tritici.
The disease occurs in all wheat growing countries of the world. In India it appears at
different times of the year in different parts of the country. It appears in the month of March in
Northern India. In Southern and Peninsular India it appears very early in the 4th week of
November.
Effects and Importance of Stem Rust Disease:

The stem rust is a serious threat to wheat in India. It causes serious damage in moist
areas and moist season. The nature and extent of damage varies from slight to almost complete
failure of the crop.
Symptoms on the Grain Host:

Elongated pustules or streaks appear chiefly on the stem and leaf bases, leaf sheaths
and even on the glumes. These are the uredia or uredosori. Each uredosorus contains a mass of
reddish brown or rusty red, one-celled, binucleate uredospores.
They are exposed by the rupture of the leaf epidermis. The torn epidermis forms a
collar-like structure around the oblong sorus. The uredospores are produced by the dikaryotic
or secondary mycelium parasitizing the tissue of the host.
This spore stage of the parasite is called the red rust or summer stage. Later in the
season the sori turn black. This is due to the appearance of two- celled, black teleutospores or
teliospores in the place of uredospores in the old uredia.
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When the grains are almost ripe, new and independent oblong to linear teleutosori
make their appearance on the stem of the host. They are smooth and dark brown or black in
colour. The overlying epidermis is ruptured and the spores are exposed.
The stems at this stage are dry and cracked. The teleutosori contain masses of black
two-celled spores called the teleutospores. There is no other marked abnormality in the
appearance of the plants. However, the plants look sickly and fail to develop normal ears in the
case of a severe attack.
Causal organism:
The causal agent of this disease is Puccinia graminis tritici Erikss. and Henn. (P. graminis
Pers.,). It is a heteroecious parasite which completes its disease cycle in two hosts namely
wheat and barberry (Berberis) or Mahonia.
Disease Cycle
Eriksson on the basis of his observations in cereal rusts, especially Puccinia glumarum
proposed the “mycoplasm theory”. He held that on the onset of winter, the fungal hyphae
degenerated in the host plant. The fungus cytoplasm mingled with that of the host protoplasm
in the cell.
As soon as the winter was over, the mycoplasm (fungal cytoplasm) migrated into the
intercellular spaces of the host. There the mycoplasm reforms the hyphae and haustoria. The
mycoplasm theory was vehemently opposed.
It was not widely accepted because of opposition and thus fell by the wayside. The
commonly held view is that normally teleutospores are the overwintering structures (A). But in
the plains in India uredospores constitute the primary inoculum.
They are carried from the distant high altitude hills by wind. After the usual resting
period the teleutospores germinate in situ (on wheat stem and stubbles in the field). Each cell
produces a short promycelium (epibasidium) into which the synkaryon migrates (B).
Each synkaryon undergoes meiosis in the promycelium or the epibasidium. Segregation

of the sexual strains takes place. Walls are laid between the haploid nuclei so that each
promycelium or epibasidium becomes septate and four-celled (B).
Each cell of the basidium produces a single, uninucleate haploid basidiospores at the
end of a sterigma. Of the four basidiospores thus produced two are of plus strain and two
minus strain.
The basidiospores are disseminated by air currents. While floating in the air they may
chance to fall on young barberry leaves. If temperature and moisture conditions are favourable
the basidiospores germinate.
Each basidiospore develops a germ tube or a primary hypha (C). It penetrates the cuticle
directly and brings about infection of the new host. Within the host tissue the primary hypha
branches freely to form a monokaryotic or haplomycelium.
The hyphae constituting it ramify in the intercellular spaces between the mesophyll
cells. The cells are uninucleate. The nuclei in the mycelium are either of plus strain or minus
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strain depending upon the nature of the germinating spore. The mycelium feeds and grows
vigorously.
Eventually it enters the reproductive phase and forms thick mats of hyphae here and
there beneath the upper and lower epidermis. The hyphal mats beneath the upper epidermis
function as primordia of spermogonia.
Those beneath the lower epidermis function as primordia of aecidia or aecia. In about a
week’s time the primordia beneath the upper epidermis produce small flask-shaped fruiting
bodies called the spermagonia.
They are embedded in the tissue in orange yellow spots on the upper surface of the
leaves of barberry bush (E). Each spermagonium opens to the outside through a small aperture
called an ostiole which projects above the surface of the leaf.
The spermagonium contains three types of hyphal threads:

1. Periphyses:
These are slender sterile, hyphae guarding the ostiole and projecting through it.
2. Spermatiophores:
These are numerous, fine, elongated hyphae arising from the interior of the swollen
portion of the spermagonium. They abstrict small, hyaline spermatia at their tips in succession.
The abstricted spermatia lie free in the cavity of the spermagonium.
3. Receptive or Flexuous Hyphae:

These are the fine, hair-like hyphal threads seen interspersed between the periphyses.
They extend out through the ostiole and project much beyond the periphyses.
The contents of the spermagonium are entirely plus or minus according as the
spermagonium has developed from a plus or a minus mycelium. The spermatia emerge in a
viscous sugary liquid through an ostiole to the leaf surface along with the flexuous hyphae.
Sexual union or spermatisation, as it is called, takes place between spermatia of one

strain and flexuous hyphae of the other strain. The intervening walls between the spermatium
and the flexuous hypha dissolve at the point of contact.
The spermatium nucleus passes into the receptive or flexuous hypha through the pore.
The spermatium nucleus now passes down the receptive hypha through the septal pores and
reaches the basal cell which becomes binucleate or dikaryotic.
The dikaryotic cell develops into a secondary or a dikaryotic mycelium. The transference
of spermatia from leaf to leaf is the work of insects. They are attracted by the nectar and visit
one spermagonium after another.
Meanwhile the hyphal mats beneath the lower epidermis develop into spherical masses
of cells. These are known as the protoaecidia. By this time the secondary mycelium formed
from the dikaryotic cells at the base of the receptive hypha reaches the young aecidium.
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Its cells mingle with the haploid tissue of young aecidium. As a result a palisade-like
layer of binucleate cells is formed at the base of aecidium. These binucleate basal cells produce
binucleate aecidiospores in terminal chains. The wall of the aecidium splits open.
The aecidium now assumes a cup-shaped form (E). The lower epidermis also ruptures
and the aecidiospores are now exposed. They are unable to reinfect barberry.
The aecidiospores (F) are binucleate and are carried by air currents to the wheat host.
Here the aecidiospores germinate each by putting out a germ tube or an infection hypha which
enters the host tissue through a stoma (G).
The tip of the infection hypha swells to form an appressorium which covers the mouth
of a stoma. The contents of the aecidiospore migrate into the appressorium. A narrow, peg like
infection hypha emerges from the appressorium, passes through the stomatal opening and
enters the substomatal cavity to form a vesicle.
From the substomatal vesicle arise hyphae which proceed intercellularly into the
parenchymatous tissue of the host leaf to form the intercellular mycelium (H). The hyphae send
small round or branched haustoria into the host cells.
About ten days after the infection of the grain host (wheat) rusty red and powdery
masses of uredospores appear on the stem and the leaves in oblong to circular sori (I). They
appear in the months of February to March.
The uredospores are shortly stalked, oblong, echinulate structures with four
equatorially arranged germ pores on the outer wall (J). These spores are binucleate. Being
exposed they are easily carried by the wind to other wheat plants.
Here, within few hours each uredospore germinates in the surface moisture provided by
rain or dew. It develops a germ tube (K) which enters the host tissue through a stoma. Within a
week’s time provided the weather conditions are favourable, the new dikaryotic mycelium
produces a new crop of uredospores.
They are disseminated in the same way. In this way the disease is spread rapidly and
widely during the growing period. The uredospores are the only spores in the life cycle which
can reinfect the host plant on which they are produced.
When the grain is almost ripe, black teleutospores begin to appear in the uredosori.
Soon after the teleutospores develop in new and independent dark brown or black teleutosori
or teliosori (A). The teleutospores are two-celled, thick-walled structures. They are the resting
spores.
The part of the life cycle which is passed on the grain host or the wheat plant represents
the dikaryophase (H-L). During this phase each cell of the mycelium, each uredospore and each
cell of teleutospore has a pair of nuclei called the dikaryon.
One of these nuclei is of plus strain and the other of minus strain. Towards maturity plus
and minus strain nuclei in each cell of the teleutospore fuse to form a synkaryon or the fusion
nucleus. The mature teleutospores thus represent the reduced diplophase (A).
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Puccinia graminis tritici is a polymorphic species as it produces a succession of different

types of spores. However, investigations carried out by late Dr. K.C. Mehta (1923-40) in India
revealed that there is no local source of infection in the plains.
The uredospores perish in the high summer temperature of the plains. There are no
barberry bushes in the plains and thus the teleutospores which are unable to survive the high
summer temperatures following the harvest remain ineffective.
Hence, the sexual stage comprising the spermagonium and aecidia is cut out from the
life cycle. The actual disease cycle is completed with uredospores alone. The teleutospores are
produced but they are non-functional.
This eliminates the chances of perennation of the rust on the alternate barberry host in
the hills. This has been confirmed by the fact that the races of P. graminis found on the
barberry bushes on the hills are different from those occurring on the cereals in the plains.
In spite of the above-mentioned facts, there is annual recurrence of the rusts in the
plains in India. It is due to the fact that the uredospores remain viable on the hills. The summer
temperature at an altitude of 5,000 ft. and above is quite congenial for their survival.
There at different altitudes, the uredospores over summer on the out of season wheat
plants, stubbles, and grass hosts in the uredial stage. These serve as primary inoculum for the
wheat crops when sown. The infection thus starts from the hills.
The uredospores are wafted down from the higher altitudes to the foot hills. From there
the infection is carried to the crops in the plains.
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Control Measures of Stem Rust Disease:

1. Cultivation of Rust Resistant Varieties:
The cultivation of varieties immune to the rust disease is an important means of
combating the disease. Some rust resistant varieties of wheat are available in India. Np 710, Np
718 and Np 770 find favour with the farmers.
The newly bred hybrids Np 822, Np 823 and Np 825 have given good results. They
possess high degree of rust resistance. In addition they are high yielders. The recently
introduced dwarf Mexican wheat varieties such as Sonora 64 and Lerma Rojo are almost
completely resistant to black rust.
2. Eradication of Barberry:
Formerly it was believed that the eradication of the less important alternate host
barberry is a possible means of eliminating the disease. We now definitely know that the
control of stem rust by this method is not possible in India.
The uredospores which are able to survive on stray and self-sown wheat plants on the hills
serve as an inoculum.
3. Use of fungicides including antibiotics:

Despite ceaseless efforts to control wheat rusts through resistant varieties, much
success has not been achieved. Practically no variety is resistant for a long period due to
emergence of new physiological races.
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UNIT - IV
BIOFERTILIZERS
Biofertilizers are defined as preparations containing living cells or latent cells of efficient
strains of microorganisms that help crop plants’ uptake of nutrients by their interactions in the
rhizosphere when applied through seed or soil.
AZOTOBACTER
Azotobacter is a genus of free-living diazotrophic bacteria whose resting stage is a cyst.
It is primarily found in neutral to alkaline soils, in aquatic environments, and on some plants. It
has several metabolic capabilties, including atmospheric nitrogen fixation by conversion to
ammonia.
Growth of Azotobacter
Usually Azotobacter is grown on a solid medium free of nitrogen. After some times (6
months) old growth of Azotobacter is transferred to a fresh solid medium to renew the growth.
This procedure is repeated periodically so that the culture can be maintained in good condition.
Production:
Mother culture:
A pure growth of any organism on a small scale is called as a mother culture. Mother
culture is always prepared in a conical flask of 500 or 1000 ml. Capacity and then this mother
culture is used for further production.
For this purpose, one litre conical flasks are taken to which 500 ml of broth of nitrogen free
medium is added and these flasks are then plugged with non-absorbent cotton, sterilized in an
auto slave for 15-20 minutes at 75 lbs pressure for 15 minutes. Flasks are then inoculated with
mother culture with the help of inoculating needle aseptically. The flasks are transferred to
shaker and shaking is done for 72-90 hours so as to get optimum growth of bacteria in broth.
Bacteria are multiplied by binary method i.e. cell division. After about 90 days, the number of
per milliliters comes to about 100 crores. Total growth of bacteria in this broth means starter
culture or mother culture, which should carefully be done, since further purity of biofertilizer or
quality of biofertilizer depends upon how mother culture is prepared.
i. Production on a large scale:

Azotobacter is multiplied on a large scale by two ways viz. Fermenter and Shaker. The
fermenter is most automatic and accurate method of multiplication of any micro-organism. In
this method, the medium is taken in a fermenter and then sterilized. After this pH of the
medium is adjusted and 1% mother culture is added. In order to get an optimum growth of the
Azotobacter required temperature and oxygen supply is adjusted so that concentrated broth is
made. This concentrated broth of the culture is then mixed with a carrier previously sterilized
and bio-fertilizers is prepared. Depending upon the demand and supply suitable fermenter is
selected.
In the 2nd method i.e. shake method, a suitable medium is prepared transferred to
conical flask of suitable capacity. These flasks are then sterilized in an autoclave at 15 lbs
pressure for 15 minutes. Each flasks is inoclulated with 10 ml mother culture and they are
transferred to shaker for multiplication where they are kept for 72-90 hours. This broth is mixed
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with a suitable carrier previously sterilized. Thus biofertilizer is prepared, filled in plastic bags
and stored in cool place.
Selection of carrier:
A carrier is nothing but a substance which has high organic matter, higher water holding
capacity and supports the growth of organism. In order to transport the biofertilizer and
becomes easy to use the suitable carrier is selected. Generally Lignite cool, compost and peat
soil are suitable carriers for Azotobacter. Out of these carriers lignite is most suitable for this
organism, since it is cheaper, keeps organism living for longer period and does not lower the
quality of bio-fertilizers.
The lignite comes in clouds and hence it is ground in fine powder by grinding machine.
Its finesses should be 250-300 mesh. The pH of the carrier is adjusted to neutral by adding
CaCO3. The lignite naturally has a variety of micro-organism and hence it is sterilized in
autoclave at 30 lbs. Pressure for 30 minutes. After this the broth is mixed with lignite 1:2
proportion by following method.
Galvanized trays are sterilized and used. To these trays, previously sterilized lignite is
transferred and broth is then added (lignite2: broth 1) and mixed properly. Trays are then kept
one above the other for 10-12 hours for allowing the organism to multiply in the carrier. This
mixture is then filled in plastic bags of 250 g or 500 g capacity. Plastic bags are properly. Trays
are then kept one above the other for 10-12 hours for allowing the organism to multiply in the
carrier. This mixture is then filled in plastic bags of 250 g or 500 g capacity. Plastic bags are
properly sealed. All the required information such as name of biofertilizer, method of use expiry
date, etc. is printed on plastic bags. In this way biofertilizer is ready to sell or use. If biofertilizer
is used immediately then bags are stored in cool place otherwise they should be stored in cold
storage in order to keep biofertilizer in good quality.
As per ISI standards, one gram of biofertilizer immediately after it is prepared should
have one crore cells of bacteria and 15 days before expiry date one gram of biofertilizer should
have 10 lakh bacteria. If biofertilizer is stored at 15-20 0C then it will remain effective for 6
months. However, at 0 to 4 0C (cold storage) the bacteria will remain active for 2 years. The
storage periods are decided after testing the biofertilizer for that particular storage conditions,
such temperature and humidity.
Use of Biofertilizer:
A plant needs nitrogen for its growth and Azotobacter fixes atmospheric nitrogen non-
symbiotically. Therefore, all plants, trees, vegetables, get benefited. However, especially
cereals, vegetables, fruits, trees, sugarcane, cotton, grapes, banana, etc. are known to get
addition nitrogen requirements from Azotobacter. Azotobacter also increases germination of
seeds. Seeds having less germinating percent if inoculated can increase germination by 20-30%.
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Field Application:
Seed inoculation
On the basis of efficiency of Azotobacter, other micro-organisms present in the soil,
benefits obtained from biofertilizer and expenditure it has been fixed to use Azotobacter - bio-
fertilizer at the rate of 250 g biofertilizer for 10-15 kg. If one knows this proportion then take a
definite quantity of seed to be inoculated. The required quantity of fresh biofertilizer is secured
and a slurry is made by adding adequate, quantity of water. This slurry is uniformly applied to
seed, seed is then dried in shed and sown. Some stickers are used in order to adher biofertilizer
to seeds. Viz. Jaggery or gum arabia.
Seedling inoculation:
This method of inoculation is used where seedlings are used to grow the crop. In this
method, seedlings required for one acre are inoculated using 4-5 packets (2-2.5 kg). For this, in
a bucket adequate quantity of water is taken and biofertilizer from these packets is added to
bucket and mixed properly. Roots or seedlings are then dipped in this mixture so as to enable
roots to get inoculum. These seedlings are then transplanted e.g. Tomato, Rice, Onion, Cole,
Crops, flowers.
Self inoculation or tube inoculation:

In this method 50 litres of water is taken in a drum and 4-5 kg of Azotobacter
biofertilizer is added and mixed properly. Sets are required for one acre of land are dipped in
this mixture. Potato tubers are dipped in the mixture of biofertilizer and planting is done.
Soil application:
This method is mostly used for fruit crops, sugarcane, and trees. At the time of planting
fruit tree 20 g of biofertilizer mixed with compost is to be added per sappling, when trees
became matured the same quantity of biofertilizer is applied.
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In sugarcane after two to three months of planting i.e. before earthing up 5-6 kg of
biofertilizer per acre is applied by mixing with compost or soil. Although, Azotobacter fixes
nitrogen non-symbiotically, it also fixes atmospheric nitrogen in the rhizospere region i.e. soil
around the seedlings or trees. Biofertilizer applied to seed or seedlings bacteria remain around
seeds or seedlings and use organic carbon for their metabolism. When seeds are germinated or
seedlings set in soil they leave or exude root exudates which become food of these bacteria.
They grow on these substances which include sugars, organic acids, amino acids and fix
atmospheric nitrogen most efficiently. Nitrogen so fixed by these bacteria becomes available to
plants after dead and degradation of bacterial cells.
AZOSPIRLLIUM
Azospirillum is a Gram-negative, microaerophilic, non-fermentative and nitrogen-fixing
bacterial genus from the family of Rhodospirillaceae. Azospirillum bacteria can promote plant
growth.
It belongs to bacteria and is known to fix the considerable quantity of nitrogen in the
range of 20- 40 kg N/ha in the rhizosphere in non- non-leguminous plants such as cereals,
millets, Oilseeds, cotton etc.
The efficiency of Azospirillium as a Bio-Fertilizer has increased because of its ability of

inducing abundant roots in several pants like rice, millets and oilseeds even in upland
conditions.
Considerable quantity of nitrogen fertilizer up to 25-30 % can be saved by the use of

Azospirillum inoculant.
The genus Azospirillum has three species viz., A. lipoferum, A. brasilense and
A. amazonense. These species have been commercially exploited for the use as nitrogen
supplying Bio-Fertilizers.
One of the characteristics of Azospirillum is its ability to reduce nitrate and denitrify.
Both A. lipoferum and A. brasilense may comprise of strains which can actively or weakly
denitrify or reduce nitrate to nitrite and therefore, for inoculation preparation, it is necessary to
select strains which do not possess these characteristics.
Azospirllium lipoferum present in the roots of some of tropical forage grasses such as
Digitaria, Panicum, Brachiaria, Maize, Sorghum, Wheat and Rye.
Mass Production of Cyanobacterial Biofertilizers (Anabaena, Nostoc)

For outdoor cultivation of cyanobacterial biofertilizers, the regional specific strain
should be used. In such practices, a mixture of 5 or 6 regionally acclimatized strains of
cyanobacteria e.g. species of Anabaena, Aulosira, Cylindrospermum, Gloeotrichia, Nostoc,
Plectonema, Tolypothrix etc. are generally used as starter inoculum.
The following methods are used for mass cultivation:

(a) Cemented tank method,
(b) Shallow metal troughs method,
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(c) Polythene lined pit method and

(d) Field method.
The polythene lined method is most suitable for small and marginal farmers for the
preparation of bio-fertilizer. In this method, small pits are prepared in field and lined with thick
polythene sheets.
a. Prepare the cemented tank, shallow trays of iron sheets or polythene lined pits in an
open area. Width of tanks or pits should not be more than 1.5 m. This will facilitate the
proper handling of culture.
b. Transfer 2-3 kg soil and add 100 g superphosphate. Water the pit to about 10 cm height.
Mix lime to adjust the pH. Add 2 ml of insecticide to protect the culture from
mosquitoes. Mix well and allow to settle down soil particles.
c. When water becomes clear, sprinkle 100 g starter culture on the surface of water.
d. When temperature remains around 35-40°C during summer, optimum growth of

cyanobacteria is achieved. The water level is always maintained about 10 cm during the
period.
e. After drying, the algal mass (mat) is separated from the soil that forms flakes. During
summer about 1 kg pure algal mat per m 2 area is produced. It is collected, powdered,
and packed in polythene bag and supplied to the farmers after sealing the packets.
f. The algal flakes can be used as starter inoculum again.
MASS CULTIVATION OF AZOLLA

The aquatic heterosporus fern contains endophytic cyanobacterium, Anabaena azollae
in its leaf cavity. There are number of species of Azolla, namely A. caroliniana, A. filiculoides, A.
maxicana, A. nilotica, A. pinnata and A. rubra which are used as biofertilizer especially for
paddy. For mass cultivation of Azolla, microplots (20 m 2) are prepared in nurseries in which
sufficient water (5-10 cm) is added.
For profuse growth of Azolla, 4-20 kg P2O5/ha is also amended. Optimum pH (8.0) and
temperature (14-30°C) should be maintained. Finally, microplots are inoculated with fresh
Azolla (0.5 to 0.4 kg/m2). An insecticide (Furadon) is used to check the insect’s attack. After 3
weeks, the mat of Azolla is ready for harvest and the same microplot is inoculated with fresh
Azolla to repeat the cultivation.
Azolla mat is harvested and dried to use as green manure.
There are two methods for its application in field:

(a) Incorporation of Azolla in soil prior to rice cultivation, and
(b) Transplantation of rice followed by water draining and incorporation of Azolla.
However, reports from the IRRI, Manila (Philippines) revealed that growing of Azolla in
rice field before rice transplantation increased the yield equivalent to that obtained from
30 kg/ha nitrogen as urea or ammonium phosphate.
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Production of Mycorrhizal Bio-Fertilizer:

A mycorrhiza is a symbiotic association between a fungus and a plant. The term
mycorrhiza refers to the role of the fungus in the plant's rhizosphere, its root system.
Mycorrhizae play important roles in plant nutrition, soil biology and soil chemistry.
In a mycorrhizal association, the fungus colonizes the host plant's root tissues,
either intracellularly as in arbuscular mycorrhizal fungi (AMF or AM), or extracellularly as
in ectomycorrhizal fungi. The association is sometimes mutualistic. In particular species or in
particular circumstances mycorrhizae may have a parasitic association with host plants.
Methods of inoculum production of mycorrhizal fungi differ with respects to their

nature, depending upon types i.e., ectomycorrhizal or endomycorrhizal.
(i) Ectomycorrhizal Fungi:

In this case, the basidiospores, chopped sporocarps, sclerotia, pure mycelial culture,
fragmented mycorrhizal roots or soil from mycorhizosphere region can be used as inoculum.
The inoculum is mixed with nursery soil and seeds are sown thereafter.
Institute for mycorrhizal Research and Development, USA and Abbot Laboratories, USA
have developed a mycelial inoculum of Pisolithus tinctorius in a mycelial vermiculite-peat moss
substrate with trade name ‘MycoRhiz’ which is commercially available on large quantities.
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(ii) VA Mycorrhizal Fungi:

An arbuscular mycorrhiza (plural mycorrhizas, a.k.a. endomycorrhiza) is a type
of mycorrhiza in which the symbiont fungus (AM fungi, or AMF) penetrates the cortical cells of
the roots of a vascular plant forming arbuscules.
VA mycorrhiza can be produced on a large scale by pot culture technique. This requires
the host plant mycorrhizal fungi and natural soil. The host plants which support large scale
production of inoculum are sudan grass, strawberry, sorghum, maize, onion, citrus, etc.
The starter inoculum of VAM can be isolated from soil by wet sieving and decantation
technique. VAM spores are surface sterilised and brought to the pot culture. Commonly used
pot substrates are sand: soil (1:1, w/w) with a little amount of moisture.
There are two methods of using the inoculum:

(a) Using a dried spore-root-soil to plants by placing the inoculum several centimetres
below the seeds or seedlings,
(b) Using a mixture of soil- roots, and spores in soil pellets and spores are adhered to
seed surface with adhesive.
Commercially available pot culture of VA mycorrhizal hosts grown under aseptic
conditions can provide effective inoculum. Various types of VAM inocula are currently
produced by Native Plants, Inc (NPI), Salt Lake City.
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RHIZOBIUM
Rhizobium is a genus of Gram-negative soil bacteria that fix nitrogen. Rhizobium species
(Rhizobium leguminosarum) form an endosymbiotic nitrogen-fixing association with roots
of legumes. The bacteria colonize plant cells within root nodules, where they convert
atmospheric nitrogen into ammonia using the enzyme nitrogenase.
Rhizobium are kept in different specialization groups. Inoculum of different strains are
prepared separately and cultivated on large scale, as required. Strains of Rhizobium sp. are
grown in Yeast Extract Mannitol (YEM) broth in a small or large container as needed.
Isolation of Rhizobium:
Sample Preparation
a. Carefully uproot a leguminous plant (sweetpeas, beans etc)
b. Gently wash the roots using sterilized water to remove any soil, mud etc.
c. Using a clean knife/blade, carefully remove/detach the nodules (which are visible to the
naked eye) from the roots and place them in a container with clear water
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d. After washing the nodules, place them in a container with a solution of merculic
chloride for a few minutes - this helps disinfect and remove any microorganism on the
surface of the nodule
e. Wash the nodules using alcohol and then wash with clean water several times
f. Place the nodules in a test-tube and add a few drops of water (distilled water)
g. Using a glass rod, crush the nodules to release the bacteria into the water
Culture Procedure
a. Gently pour mannitol-agar medium into 3 sterile test-tubes
b. Add a few drops of the sample (rhizobium water) into the test-tubes using sterilized
droppers
c. Mix the contents thoroughly
d. Pour the contents into different Petri dishes and incubate at 45 degrees Celsius
e. Turn the Petri dish when the medium solidifies
f. Allow the culture to incubate for about 3 days
Methods of Cultivation
Following are the steps of mass cultivation of Rhizobium.
a. sterilize the growth medium and inoculate with broth of mother culture
prepared in advance,
b. incubate for 3-4 days at 30 - 32°C,
c. test the cultures for its purity and transfer to a large fermenter, wait for 4-9 days
for bacterial growth (for good bacterial growth make the device for its aeration),
d. allow to grow the bacteria either in a large fermenter containing broth or in
small flasks as per demand,
e. check the quality of broth,
f. blend the broth with sterile carrier e.g. peat, lignite, farmyard manure and
charcoal powder,
g. pack the culture in polyethylene bags and keep at 25°C,
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h. check the quality of carrier culture,

i. store at 4°C in a controlled-temperature room
j. supply to farmers.
During the blending of broth a variety of carriers are used, for example, peat, lignite,
farmyard manure, charcoal powder, etc. In India powdered farmyard manure and charcoal
powder are good carrier and an alternative to peat and lignite. Good quality of carrier culture is
that which contains sufficient amount of rhizobial cells i.e. 1000 x 106 to 4000 x 106 rhizobia/g
carrier. Seed inoculation with aqueous suspension of carrier culture during sowing has revealed
the luxuriant nodulation and good yield of crops.
Methods of seed inoculation with rhizobial culture

The steps of seed inoculation with rhizobial culture.. Dissolve 10 per cent sugar
or gur (jaggery) in water by boiling it for some time. Leave the content to cool down. Gum
arabic solution (10%) may also be added to the solution. This serves as sticker
for Rhizobium cells to seeds. Mix this carrier based culture of Rhizobium to form the inoculum
slurry. For one hectare, 400 g charcoal based culture would be sufficient for mixing the seeds.
Transfer the inoculum slurry on seeds and mix properly. The number of rhizobial cells/ seed
should be between 105 to 106. Spread the seeds in shade for drying on cement floor or plastic
sheets.
While using rhizobial cultures, certain precautions are taken into account. For example,
use of culture before expiry date, use of small amount of pesticides when required, immediate
sowing of seeds after mixing, etc. Seeds must be stored at 4°C when not used immediately to
protect the rhizobial cells.
Pelleting
When soil has the adverse conditions such as dryness, acidity, excess fertilizers and
pesticides, etc., the rhizobial cells are protected by adopting special method of inoculation. One
of these methods is pelleting i.e. preparation of pelleted seeds. This method involves the
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procedure as described earlier. High amount of gum arabic (40%) or carboxymethylcellulose

(20%) is added to the inoculum slurry before mixing with seeds. Finally, pelleting agent is mixed
when inoclated seeds are moist. The commonly used pelleting agents are calcium carbonate,
rock phoshate, charcoal powder, gypsum and betonite.
PHOSPHOBACTERIA
Phosphorous is such an important macronutrient which is very often present in the soil
in unavailable form. Many soil bacteria particularly those belonging to the genera Bacillus and
Pseudomonas posses the ability to bring insoluble phosphates in the soluble forms by secreting
organic acids. These acids lower the pH and bring about the dissolution of bound forms of
phosphorous. These bacteria are commonly known as phosphobacteria.
They can be applied either through seed or soil application. Phosphorus, both native in
soil and applied to inorganic fertilizers becomes mostly unavailable to crops because of its low
level of mobility and solubility and its tendency to become fixed in soil. The phosphate
solubulizing bacteria are life forms that can soil. The phosphate solubulizing bacteria are life
forms that can help in improving phosphate uptake of plantain on different ways
Phosphate solubilizing bacteria are beneficial bacteria capable of solubilizing inorganic

phosphorus from insoluble compounds. P-solubilization ability of rhizosphere microorganisms is
considered to be one of the most important traits associated with plant phosphate nutrition.
Many different strains of these bacteria have been identified including Pantoea
agglomerans , Microbacterium laevaniformans and Pseudomonas putida strains are highly
efficient insoluble phosphate solubilizers.
Isolation of Phosphate solubilizing bacteria

Isolation of Strains for isolation of Phosphate Solubilizing Microbes, 1g rhizosphere soil
was suspended in 100ml of distilled water. An aliquot (100 l) from decimal dilutions was
inoculated on Picovskaya’s medium by pour plate technique and incubated at 30 oC. Colonies
showing phosphate solubilizing zone around them were considered as PSM. Single colonies
appearing on Picovskaya’s agar plates were transferred in liquid broth of Picovskaya’s and on
agar slants for further study.
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Mass cultivation:
Inoculum preparation for phosphorus biofertilizer

Prepare appropriate medium specific to the bacterial inoculant in 250 ml, 500 ml, 3 liter
and 5 liter conical flasks and sterilize. The media in 250 ml flasks are inoculated with an efficient
bacterial strain under aseptic conditions. Keep the flasks at room temperature in a rotary
shaker incubator (200 rpm) for 5–7 days. Observe the flasks for growth of the culture and
estimate the population, which serves as the starter culture. Using the starter culture (at log
phase) inoculate the larger flasks (500 ml, 3 liter and 5 liter) containing medium, after obtaining
growth in each flask. The above medium is prepared in large quantities in a fermenter,
sterilized well, cooled and kept ready.
The medium in the fermenter is inoculated with the log-phase culture grown in the 5-
liter flask. Usually 1–2% inoculum is sufficient; however, inoculation is done up to 5%,
depending on the growth of the culture in the larger flasks. The cells are grown in the
fermenter by providing aeration (passing sterile air through a compressor and sterilizing agents
like glass wool, cotton wool, acid etc.) and giving continuous stirring. The broth is checked for
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the population of the inoculated microorganism and contamination, if any, during the growth
period. The cells are harvested with a population load of 10 9 cells ml-after the incubation
period. There should not be any fungal or any other bacterial contamination at a 10-6 dilution
level. It is not advisable to store the broth after fermentation for periods longer than 24 hours.
Even at 4°C, the number of viable cells begins to decrease.
Field application:
It can be used for all crops, including paddy, millets, oilseeds, pulses and vegetables.
The methods recommended for application are:
1. Seed treatment;
2. Seedling dipping;
3. Soil application.
SIDEROPHORE ACTIVITY
Siderophores are organic compounds with low molecular masses that are produced by
microorganisms and plants growing under low iron conditions. The primary function of these
compounds is to chelate the ferric iron [Fe(III)] from different terrestrial and aquatic habitats
and thereby make it available for microbial and plant cells.
Microorganisms produce a wide range of siderophores. Most of the bacterial

siderophores are catecholates (i.e. enterobactin), and some are carboxylates (i.e. rhizobactin)
and hydroxamates (i.e. ferrioxamine B). There are also certain types of bacterial siderophores
that contain a mix of the main functional groups (i.e. pyoverdine)
Fe is an essential micronutrient for plant growth. Under conditions of Fe deficiency,

graminaceous plants (e.g. barley and wheat) have developed an efficient strategy for acquiring
Fe from insoluble sources .These plants secrete Fe(III)-chelating compounds called
phytosiderophores that form specific strong complexes with Fe(III). The phytosiderophores are
hexadentate ligands that coordinate Fe(III) with their amino and carboxyl groups. When the
phytosiderophore is released to the rhizosphere, it chelates Fe from the soil by forming Fe(III)–
phytosiderophore complexes that can be subsequently transported across the root plasma
membrane. This called siderophore activity.
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UNIT - V
BIOPESTICIDES
Biopesticides are certain types of pesticides that are derived from natural materials like
plants (Botanical origin), bacteria, fungi and virus (Microbial origin) and certain minerals.
Generally, biopesticides are made of living things, come from living things, or they are
found in nature. They tend to pose fewer risks than conventional chemicals. Very small
quantities can be effective and they tend to break down more quickly, which means less
pollution.
Some biopesticides are targeted in their activity, often working on a small number of
species. However, users need more knowledge to use biopesticides effectively. This is because
they are often most effectively used as part of an Integrated Pest Management approach.
BACTERIAL BIOPESTICIDES
Bacillus thuringiensis (Bt) is a rod shaped, gram-positive bacteria that forms a spore and
is found in the soil.
Classification of this bacterium includes Bacteria (domain); Eubacteria (kingdom);

Firmicutes (phylum); Bacilli (class); Bacillales (order); Bacillaceae (family). Bt was isolated in
1901 and named in 1911.
It was used as a commercial biopesticide for the first time in the United States in 1958.
It is placed in IRAC group 11, microbial disruptors of insect midgut membranes.
Bt is toxic to caterpillars, some fly larvae, and some beetle larvae but not toxic to other
organisms. A few strains of Bt are available in products used in the United States. Bt var.
kurstaki is toxic to lepidopteran (butterfly, skipper, and moth) larvae; Bt var. aizawai is toxic to
wax moth larvae; Bt var. israelensis is toxic to mosquito, midge, fungus gnats, and blackfly
larvae; Bt var. galleriae is toxic to larvae of May or June beetles (white grubs); Bt var.
tenebrionis (or var. San Diego) is toxic to Colorado potato beetle, elm leaf beetle, and willow
leaf beetle larvae. However, Bt var. tenebrionis does not kill all leaf beetles.
Bt strains are very specific to the insects they kill. Therefore, identification of the
injurious insect is very important.
The correct strain must be applied to susceptible insects. Applications of Bt to insects

that are not susceptible will be ineffective. Bt is most effective against young larvae and usually
does not kill adults or other stages of an insect.
Insects must eat Bt for it to be effective, and good coverage is important. Some insects
do not eat the outside of the plant part they attack and applications of Bt on the surface of the
plant are ineffective against them.
Bt is rapidly deactivated by ultraviolet radiation. Applications made in the evening, on

cloudy, or on rainy days last longer.
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However, heavy rains wash Bt off the plant. Applications become inactivated in one to a
few days and may need to be reapplied in 3 to 7 days. Applications for leaf beetles may be
effective for only one day.
Applications of Bt do not result in continuous management of insects by reproduction of

bacterial cells, and Bt is applied similar to chemical insecticides. Once a solution of Bt is
prepared it should be used immediately; especially, if the water used to make the solution has a
pH greater than 7 (basic).
The effectiveness of Bt may be reduced after two or three years of storage. Dry
formulations last longer than liquid formulations. Bt products should be stored out of sunlight
and in cool, dry conditions.
A crystalline toxin and spore is usually produced by Bt cells. The toxin is called a delta
endotoxin. Bt products usually contain the toxin and spores (environmental resistant stage of
the bacterium) but some products do not contain spores. Spores may become bacterial cells
inside the insect. Once the insect eats the Bt the delta endotoxin is activated in the insect’s gut
by enzymes and alkaline (basic) conditions of the gut.
A specific pH is required to activate the endotoxin. The endotoxin disrupts the cell walls
of the gut. Bacterial cells enter the body of the insect. Infected insects stop feeding in a few
hours and die in a few hours to weeks (frequently 2-3 days).
Different strains of Bt have different endotoxins and kill different insects. The endotoxin
is not activated in the gut of humans.
In summary, Bt is a microbial biopesticide that is very specific to certain insects. It

causes insects to stop feeding in a few hours and usually kills insects in a few days. It must be
eaten and kills larvae. It does not last long on the plant, may require frequent applications, is
considered organic, and is not toxic to beneficials.
Page 49 of 56
Agrobacterium tumefaciens
Agrobacterium tumefaciens causes crown gall disease of a wide range of dicotyledonous
(broad-leaved) plants, especially members of the rose family such as apple, pear, peach, cherry,
almond, raspberry and roses.
The disease gains its name from the large tumour-like swellings (galls) that typically
occur at the crown of the plant, just above soil level. Although it reduces the marketability of
nursery stock, it usually does not cause serious damage to older plants.
The unique mode of action of A. tumefaciens has enabled this bacterium to be used as a
tool in plant breeding. Any desired genes, such as insecticidal toxin genes or herbicide-
resistance genes, can be engineered into the bacterial DNA and thereby inserted into the plant
genome. The use of Agrobacterium not only shortens the conventional plant breeding process,
but also allows entirely new (non-plant) genes to be engineered into crops.
The story of Agrobacterium goes even further than this, making it one of the most
interesting and significant bacteria for detailed study. For example, there is a highly
effective biological control system for this disease - one of the first and most successful
examples of biological control of plant disease.
A. tumefaciens is a Gram-negative, non-sporing, motile, rod-shaped bacterium, closely

related to Rhizobium which forms nitrogen-fixing nodules on clover and other leguminous
plants. Strains of Agrobacterium are classified in three biovars based on their utilisation of
different carbohydrates and other biochemical tests. The differences between biovars are
determined by genes on the single circle of chromosomal DNA. Biovar differences are not
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particularly relevant to the pathogenicity of A. tumefaciens, except in one respect: biovar 3 is

found worldwide as the pathogen of grave vines. But this is almost certainly because biovar 3
has been spread around the world in vegetative cuttings of vines, not by natural mechanisms.
Most of the genes involved in crown gall disease are not borne on the chromosome
of A. tumefaciens but on a large plasmid, termed the Ti (tumour-inducing) plasmid. In the same
way, most of the genes that enable Rhizobium strains to produce nitrogen-fixing nodules are
contained on a large plasmid termed the Sym (symbiotic) plasmid. Thus, the characteristic
biology of these two bacteria is a function mainly of their plasmids, not of the bacterial
chromosome.
Agrobacterium tumefaciens is found commonly on and around root surfaces - the

region termed the rhizosphere - where it seems to survive by using nutrients that leak from the
root tissues. But it infects only through wound sites, either naturally occurring or caused by
transplanting of seedlings and nursery stock.
Biological control of crown gall

In general, bacterial diseases of plants are very difficult to control owing to the lack of
effective chemicals. Antibiotics could be used, but they are expensive and, in any case, the
compounds that are valuable for human therapy are not allowed to be used in agriculture. The
most effective alternative is the use of copper, which is potentially phytotoxic.
However, for the nopaline-producing strains of A. tumefaciens there is a highly effective

biocontrol system, discovered and developed by Allan Kerr in Australia. It has been used in
Australia since 1973 - the first commercial biological control agent for any plant disease. It is
now used world-wide, and marketed by several companies under a range of trade names (e.g.
"Galltrol").
Kerr discovered this biocontrol system by isolating non-pathogenic strains

of Agrobacterium radiobacter from disease sites and testing their ability to compete with
pathogenic strains in mixed inoculations. Several non-pathogenic strains helped to reduce
infection, but one strain in particular, A. radiobacter strain K84, completely prevented disease
when added to wound sites at a 1:1 ratio with cells of A. tumefaciens.
This strain is the one that is marketed globally. It is supplied commercially on agar plates
or in a peat substrate, and it is used by suspending the bacterial cells in water, then dipping
seeds, seedlings or cuttings in this suspension before planting. It acts only as a preventative
treatment, not to cure infections, so it is applied at a high population level to protect any
wound sites against pathogenic invasion.
Trichoderma viridae
Trichoderma species are free-living fungi that are common in soil and root ecosystems.
They are highly interactive in root, soil and foliar environments, and produce a variety of
compounds that induce localized and systemic resistance responses in plants. Trichoderma
have long been recognized as biocontrol agents for the control of plant diseases and for their
ability to enhance root growth and development, crop productivity, resistance to abiotic
stresses, and uptake and use of nutrients.
Trichoderma is a very effective biological mean for plant disease management especially
the soil born. It is a free-living fungus which is common in soil and root ecosystems. It is highly
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interactive in root, soil and foliar environments. It reduces growth, survival or infections caused
by pathogens by different mechanisms like competition, antibiosis, mycoparasitism, hyphal
interactions, and enzyme secretion.
Benefits of Trichoderma
Disease Control:
Trichoderma is a potent biocontrol agent and used extensively for soil born diseases. It
has been used successfully against pathogenic fungi belonging to various genera, viz. Fusarium,
Phytopthara, Scelerotia etc.
Plant Growth Promoter:

Trichoderma strains solubilize phosphates and micronutrients. The application of
Trichoderma strains with plants increases the number of deep roots, thereby increasing the
plant's ability to resist drought.
Biochemical Elicitors of Disease:

Trichoderma strains are known to induce resistance in plants. Three classes of
compounds that are produced by Trichoderma and induce resistance in plants are now known.
These compounds induce ethylene production, hypersensitive responses and other defense
related reactions in plant cultivars.
Transgenic Plants:
Introduction of endochitinase gene from Trichoderma into plants such as tobacco and
potato plants has increased their resistance to fungal growth. Selected transgenic lines are
highly tolerant to foliar pathogens such as Alternaria alternata, A. solani, and Botrytis cirerea as
well as to the soil-borne pathogen, Rhizectonia spp.
Bioremediation:
Trichoderma strains play an important role in the bioremediation of soil that are
contaminated with pesticides and herbicides. They have the ability to degrade a wide range of
insecticides: organochlorines, organophosphates and carbonates.
Biocontrol mechanisms of Trichoderma:

The Trichoderma may suppress the growth of the pathogen population in the
rhizosphere through competition and thus reduce disease development. It produces antibiotics
and toxins such as trichothecin and a sesquiterpine, Trichodermin, which have a direct effect on
other organisms. The antagonist (Trichoderma) hyphae either grow along the host hyphae or
coil around it and secrete different lytic enzymes such as chitinase, glucanase and pectinase
that are involved in the process of mycoparasitism. Examples of such interactions are T.
harzianum acting against Fusarium oxyporum, F. roseum, F. solani, Phytophthara
colocaciae and Sclerotium rolfsii. In addition, Trichoderma Enhances yield along with quality of
produce. Boost germination rate. Increase in shoot & Root length Solubilizing various insoluble
forms of Phosphates Augment Nitrogen fixing. Promote healthy growth in early stages of crop.
Increase Dry matter Production substantially. Provide natural long term immunity to crops and
soil.
Method of application:
Seed treatment:
Mix 6 - 10 g of Trichoderma powder per Kg of seed before sowing.
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Nursery treatment:
Apply 10 - 25 g of Trichoderma powder per 100 m2 of nursery bed. Application of neem
cake and FYM before treatment increases the efficacy.
Cutting and seedling root dip:

Mix 10g of Trichoderma powder along with 100g of well rotten FYM per liter of water
and dip the cuttings and seedlings for 10 minutes before planting.
Soil treatment:
Apply 5 Kg of Trichoderma powder per hector after turning of sun hemp into the soil for
green manuring. Or Mix 1kg of Trichoderma formulation in 100 kg of farmyard manure and
cover it for 7 days with polythene. Sprinkle the heap with water intermittently. Turn the
mixture in every 3-4 days interval and then broadcast in the field.
Plant Treatment:
Drench the soil near stem region with 10g Trichoderma powder mixed in a liter of water.
Beauveria
Beauveria bassiana is a fungus that grows naturally in soils throughout the world and
acts as a parasite on various arthropod species, causing white muscardine disease; it thus
belongs to the entomopathogenic fungi.
It is being used as a biological insecticide to control a number of pests such

as termites, thrips, whiteflies, aphids and different beetles. Its use in the control of bedbugs
and malaria-transmitting mosquitos is under investigation.
In culture, B. bassiana grows as a white mould. On most common cultural media, it

produces many dry, powdery conidia in distinctive white spore balls. Each spore ball is
composed of a cluster of conidiogenous cells.
The conidiogenous cells of B. bassiana are short and ovoid, and terminate in a narrow
apical extension called a rachis. The rachis elongates after each conidium is produced, resulting
in a long zig-zag extension. The conidia are single-celled, haploid, and hydrophobic.
White muscardine disease

The insect disease caused by the fungus is a muscardine which has been called white
muscardine disease.
When the microscopic spores of the fungus come into contact with the body of an
insect host, they germinate, penetrate the cuticle, and grow inside, killing the insect within a
matter of days.
White mold emerges from the cadaver and produces new spores. A typical isolate of
B. bassiana can attack a broad range of insects; various isolates differ in their host range. The
factors responsible for host susceptibility are not known.
Mode of action
This entomopathogenic fungi, Beauveria species attack their host insects
percutaneously. The infection pathway consists of the following steps: (1) attachment of the
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spore to the insect cuticle, (2) spore germination on cuticle, (3) penetration through the cuticle,
(4) overcoming the host immune response , (5) proliferation within the host, (6) saprophytic
outgrowth from the dead host and production of new conidia.
Biocontrol
Beauveria bassiana can be used as a biological insecticide to control a number of pests
such as termites, whiteflies, and many other insects. Its use in the control of malaria-
transmitting mosquitos is under investigation.
As an insecticide, the spores are sprayed on affected crops as an emulsified suspension

or wettable powder or applied to mosquito nets as a mosquito control agent.
As a species, Beauveria bassiana parasitizes a very wide range of arthropod hosts. Some
strains do have a wide host range and should, therefore, be considered nonselective biological
insecticides. These should not be applied to flowers visited by pollinating insects
Phytophthora palmivora
Phytophthora palmivora is an oomycete that causes bud-rot of palms, fruit-rot or kole-
roga of coconut and areca nut. These are among the most serious diseases caused
by fungi and moulds in South India.
Phytophthora root rot of papaya seedlings is most serious during rainy periods. Under
waterlogged conditions, P. palmivora may attack roots of papaya older than three-months of
age, the time at which they become resistant to the pathogen under normal conditions.
Therefore, Phytophthora root rot may occur on papaya at any age in poorly drained areas.
Waterlogged conditions appear to weaken the defense mechanism of papaya roots against
invasion by the pathogen. Mobility of zoospores of P. palmivora under such conditions also may
contribute to the severity of the disease due to their attraction by papaya roots.
Favorable temperature is also a contributing factor to the severity

of Phytophthora diseases because of its effect on growth and sporulation of the
pathogen. Phytophthora palmivora has an optimum temperature for growth of 30 °C, a
maximum temperature of 36 °C and a minimum temperature of 12 °C. The pathogen produces
the most sporangia at 25 °C but no sporangia are produced at temperatures higher than 35 °C
or lower than 15 °C.
One common symptom of P. palmivora is fruit rots which are found in papaya, citrus,
coconuts, durian, and cacao. Root rots are another symptom of P. palmivora and can be seen
in red maples, citrus, papaya, mango, durian, and black pepper.
Another symptom is the presence of cankers which are found in red maple,
papaya, rubber, mangos, and cacao. Bud rots can also be seen in papaya and coconuts infected
with P. palmivora. Bud rots are also found in Palmyra palms and coconut palms. Collar rots are
found on citrus, mango, and black pepper infected with P. palmivora.
Rain and wind are the two major factors in the epidemiology of Phytophthora fruit rot
of papaya. Rain splash is needed for liberation of sporangia of P. palmivora from the surface of
infected fruit into the atmosphere and for projection of the soil inoculum into air.
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Control:
P. Palmivora is an oomycete the simplest management technique is to control the
amount of water present in the soil. Techniques for controlling moisture include: monitored
watering, pruning to increase airflow and decrease humidity in the soil, as well as making sure
that areas where potential hosts are planted are not prone to flooding, oftentimes this includes
planting on an incline. Other means of cultural control for P. Palmivora include mulching to
reduce the number of spores released via rain splash, complete removal of infected host plants
and materials, and in some cases the use of companion crops. Companion crops are planted in
the same fields as the host plant and are used to divert some of the pathogen away from the
hosts, an example being planting bananas and avocados in the same field.
Chemical control methods for P. palmivora include: protectant fungicides such as

the Bordeaux mixture, phosphonates which control the mycelial growth of the
pathogen, dithiocarbamates such as Mancozeb, and phenylamides which control the spread of
the pathogen from the roots of the host. Host resistance is also a method of controlling the
pathogen, resistant plants generally have thicker cuticles which inhibits the ability of the
pathogen to enter the host.
NUCLEAR POLYHEDROSIS VIRUS (NPV)

Nuclear Polyhedrosis viruses are the Baculoviruses. These viruses are excellent for
species-specific, narrow spectrum insecticidal applications. They show no negative impacts on
plants, mammals, birds and fish or even on non-target insects. This is especially desirable when
beneficial insects are being conserved when an ecologically sensitive area is being treated.
The virions are rod-shaped, 40-70 nm X 250-400 nm, comprising a lipoprotein envelope
around a protein capsid containing DNA-protein core. Baculoviruses are large, circular and
double-stranded DNA genomes ranging from 81.7 to 178.7 kb. They are pathogenic to
Lepidoptera, Hymenoptera, and Diptera
Mass production of NPV:

a. Nuclear Polyhedrosis Virus (NPV) is host specific for Spodoptera litura and Helicoverpa
armigera. NPV is a stomach poison.
b. NPV is effective when it is ingested by the larvae.
c. The mass production of NPV of H. armigera and S. litura is same except H. armigera is
reared in individual vial to avoid cannibalism.
d. The NPV of these pests can be produced by using both natural as well as artificial diet
contaminated with the respective NPV.
e. The third instar larvae of these pests either collected from field or reared in the
laboratory are allowed to feed the contaminated diet.
f. Keep larvae hungry for 24 hours before inoculation of virus. Larvae get infected with
virus within 3-4 days and hang themself upside down in tubes.
g. These infected larvae are collected in a beaker containing water for 3-4 days for
purification. The putrefied larvae are crushed with the help of mixer cum grinder by
adding some water.
h. This solution is filtered with the help of muslin cloth and add more water, if required.
The filtered extract solution is centrifuged (30,000 RPM) to separate the polyhedral of
NPV from the solution. The polyhedral is separated from the water and is ready to use.
Page 55 of 56
Mode of action:
Virus after ingested by larva, the occlusion body dissolves in alkaline gut juice (pH 9.0-
10.5) and virus particles released in gut. The virus particles are attached to the peritrophic
membrane lining the midgut. The lipoprotein membrane surrounding the virus fuses with
plasma membrane of the gut wall cells and liberates nucleocapsids into the cytoplasm. The
nucleotide transports virus DNA into the nucleus of the cell and virus gene expression begins.
The virus multiplies rapidly and eventually fills the body of the host with virus particles. These
virus particles become occluded late in life cycle. After the death of larvae, they release massive
amount of occlusion bodies in environment which further infect other larvae.
Page 56 of 56

005 Allied III - Plant Pathology, Biofertilizers and Biopesticides - III Sem

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005 Allied III - Plant Pathology, Biofertilizers and Biopesticides - III Sem

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STUDY MATERIAL FOR B.

UNIT CONTENT PAGE Nr

I PLANT DISEASES AND ITS CONTROL 02

III PLANT BACTERIAL DISEASES 17

Objectives of Plant Pathology:-

The objectives of the Plant Pathology are the study on:

Importance of the plant diseases

Definition and Terms:-

2. Pathogen: An entity, usually a micro-organism that can cause the disease.

6. Pathogenicity: The relative capability of a pathogen to cause disease.

7. Pathogenesis: It is a process caused by an infectious agent (pathogen) when it comes in

8. Virulence: The degree of infectivity of a given pathogen.

9. Infection: The initiation and establishment of a parasite within a host plant.

12. Invasion: The penetration and spread of a pathogen in the host.

17. Disease cycle: The chain of events involved in disease development.

27. Antagonism: The counteraction between organisms or groups of organisms.

28. Mutation: An abrupt appearance of a new characteristic in an individual as a result of an

Causes of plant diseases:

1.Animate or Biotic Causes:

Inanimate or Abiotic causes:

The causes are:

Effect of Pathogen on the Plants:-

Morphological or structural changes:

Effect on uptake and translocation of water and nutrients:

DEFINITION OF DISEASE CYCLE:

The disease cycle consists of a number of distinct steps:

In practical terms, a micro-organism is pathogenic to a plant if it damages that plant and

SYMPTOMS OF PLANT DISEASE:

The symptoms of plant diseases are of following 4 types:

The various necrotic symptoms are:

(f) Damping off:

(g) Burn, Scald or Scorch:

(II) Hypoplasia (Atrophy):

(III) Hyperplasia and Hypertrophy:

Some symptoms of abnormal growth are:

(d) Hairy root:

(IV) Discolouration symptoms:

(V) Gummosis and exudations:

(VI) Other Symptoms:

vi. Witches brooms:

Bacterial adherence to mucosal surfaces.

Dissemination of plant diseases:

The induced/active defense mechanism in plants may operate at different levels

Inducible structural defenses:

Induced biochemical changes:

HYPERSENSITIVE RESPONSE (HR)

SYSTEMIC ACQUIRED RESISTANCE (SAR)

Principle of induced resistance

Toxic substances produced:

Active defense to pathogens

The oxidative burst

Effect of White Rust Disease:

The disease in association with downy mildew disease of crucifers caused by

Symptoms of White Rust Disease:

In case of local infection, isolated spots or pustules appear on leaves or stems or

Oospores germinate in presence of water to form a vesicle in which a large number of

Control Measures of White Rust Disease:

LATE BLIGHT OF POTATO

Effects of Late Blight:

Symptoms of Late Blight:

Causal organism and over-wintering:

(a) Indirect Germination:

(a) Direct Germination:

The indirect method of germination of sporangia by the formation of zoospores in a