A Practical manual on

PRINCIPLES OF INTEGRATED DISEASE MANAGEMENT

Credits: 3(2+1)

Course Code: PPA-201

B.Sc. (Ag.) IInd Year

Semester: IIIrd

Compiled By:
Dr. Sandeep Kumar
Assistant Professor
Department of Agriculture,
Mobile No. 8318420900

Buddha P.G. College Ratsia Kothi- Deoria (U.P)

Objectives: Methods of diagnosis of plant diseases

Plant disease diagnosis is designed to recognize the primary disease causing agents. The
majority of plant diseases can be diagnosed by a relatively, straight forward procedure involving
an evolution of background information and a microscopic and often microscopic examination of
the diseased plant.

Steps in disease diagnosis:

The steps in disease diagnosis are as follows:

A. Obtain background information on host and diseases.

B. Obtain a good sample of disease plants.
C. Examine plants and describe symptoms and signs of disease.

A. Obtain background information from the grower:

Clues to the identification of Max diseases can be found in the background information
following information is necessary for the diagnosis of plant disease. For example was the
problem.

a) Under development of tissues or organs: Such symptoms as stunting of plant shortened

internodes inadequate development of roots malformation of leaves inadequate production
of chlorophyll and other pigment and failure of fruits and flower to develop.
b) Over development of tissues or organs: Examples include galls on root, stem, leaves
witches brooms and profuse flowering.
c) Necrosis or death of plant parts: such as wilts or diebacks. Other example include shoot
or leaf blights leaf spots and fruits rots.
d) Alteration of normal appearance: like mosaic patterns of light and dark green on leave
and altered coloration in leaves and flowers.

B. Obtain good sample of disease plants:

Always obtained fresh sample in various stage of disease development and entire plants
when feasible. For some diseases such as leaf, spot or fruits those parts of the plant may be
sufficient. Sometimes it is necessary to go directly to where the plant is growing to make the
diagnosis example garden tree or bushes.

C. Examination of diseased plants and isolation of biotic agent:

Make a through external and internal macroscopic examination of the diseased plants. Be
observant describe the symptoms. Are there leaf spot malformed tissue lesions or cankers
discolored stems or vascular tissue stunted plant rotted roots? Exactly what parts of the plant
are affected? Look for evidence of the pathogens such as sclerotia, fungal growth galls white
orange or black powdery substance or nematodes. Remember that disease tissue is usually
rapidly colonized by saprophytes or secondary microorganism which can confuse the diagnosis
especially when you are looking for signs. That is why samples fresh sample are necessary.
Objectives: Methods of Plant Disease Measurement

It is often desirable rather necessary to know the exact amount of disease occurring in a crop
as it would help in ascertaining the economic importance (losses) of the disease as well as the
impact of the treatment employed for the management of the disease concerned disease offers the
following for measurement.

1. Disease incidence (field basis): It is defined as number or proportion of plant that are
diseased in a population (0 to 1) or as percentage (0 to 100) of diseased entities within a
sampling unit.
Number of infected plants
Disease Incidence (%) = x 100
Total number of plants
2. Disease prevalence (area basis): occurrence of a disease in a particular area is known as
disease prevalence.
3. Disease severity (single plant basis): The proportion of area or amount of plant tissue that
is diseases in a plant is known as severity, it is also called intensity.
4. Yield loss: Defined as the measurable reduction in quantity and quality of yield.

Methodology for computation of Disease severity/Disease intensity

or Disease index

1. Collect a simple of 10 plants or leaves randomly and without any preconceived idea from
the crop in question.
2. Use the disease rating scale already assigned for the concerned and scores each leaf for
disease severity by assigning a particular grade or class. Record the number of leaves in
each grade in frequency table
3. Based on the distribution of leaves in frequency table calculate disease index. For each
grade multiply the number of leaves with grade value and get the sum of all grades.
4. Put the value in the given formula and compute degrees index (%) for different treatments.

Disease severity following Kumar et al. (1998) double digit scale based on per cent
blighted area on the flag and flag-1 leaf at flowering, soft dough and hard dough stages. The disease
score of each selected plant were recorded twice.

Table- The double digit scale, based on per cent blighted area on the flag leaf and one leaf
just below given by Kumar et al. (1998).
 A double digit* scale for appraising blight severity
S. Severity** Rating
No. Range of
Flag leaf Flag-1 leaf Disease response value
1. 0 0-1 Immune (I) 00-01

2. 1-2 2-4 Resistant (R) 12-24

3. 3-4 4-6 Moderately Resistant (MR) 34-46

4. 5-6 6-8 Moderately susceptible (MS) 56-68

5. 7-8 8-9 Susceptible (S) 78-89

6. 9 9 Highly susceptible (HS) 99

* First and second value respectively, represents per cent blighted area on the flag leaf
and flag-1 leaves.
** Values 1,2,3,4,5,6,7,8, and 9 respectively correspond to 10,20,30,40,50,60,70,80 and
90 per cent blighted area.
Fig.: Assessment key of Kumar et al. (1998) double digit scale based on per cent blighted area
on the flag and flag -1 leaf 0-9 double digit (dd) scale.

The recording of spot blotch was done in 0-9 double digit scale at dough stage. The scoring
of the disease incidence was done on the top two leaves, i.e. flag leaf and one below it of two
digits, while the first indicates the score of flag leaf, the second digit gives the score of next (flag-
1) leaf.

Disease Index or Per cent Disease Index (PDI) was calculated by the following formula:

Sum of all numerical rating x 100

PDI=
Total number of leaf examine or graded x Maximum grade

In each plot 10 randomly selected plants were scored.

Question: Calculate the PDI of 10 plants randomly selected having two leaf selected i.e. flag and
flag-1 leaf in spot blotch of wheat disease. The disease score of each selected plant
were recorded is-
23, 23, 24, 24, 23, 23, 23, 23, 23, 23

Solve:

Given,

Total number of leaf examine or graded = 2

Total number of plant = 10

Maximum grade = 9

Then,

Sum of Ist digit = Total no. of Ist digit/ Total plants

= 20/10 = 2.0

Sum of IInd digit = Total no. of IInd digit/ Total plants

= 32/10 = 3.2

Total sum of numerical rating = 2.0 + 3.2 = 5.2

Using formula,

Sum of all numerical rating

PDI (%) = x 100
Total number of leaf examine or graded x Maximum grade

5.2
PDI (%) = x 100
2x9

520
PDI (%) = = 28.88
18
Question: Calculate the Disease incidence (DI) in a field, a total of 10 plants were selected, out
of which 5 plants turned out to be infected or sick.

Solve:

Given,

Total number of plants = 10

Number of infected plants = 5

Disease incidence = ?

Using formula,
Number of infected plants
Disease Incidence (%) = x 100
Total number of plants

5
Disease Incidence (%) = x 100
10

= 50%
Objective- To study about the mass multiplication of Trichoderma and Pseudomonas

Materials required

• Rice/Wheat/ Sorghum/ Maize

• Mother Culture
• Flask or 8" x 12 " plastic bag
• Cotton
• Rubber band
• Heating system (gas/electric heater)
• Fresh Water
• Autoclave
• Inoculation needle

(A) Mass multiplication of Trichoderma on seed

Procedure:

➢ Take 200g of Rice/Wheat/Sorghum/Maize seed in the flask or poly pack and add 200 ml
of fresh water in the flask (if grains contain dust then wash it twice before adding fresh
water).
➢ Sorghum seeds and wheat seed were socked separately in water for 12 hours and then
excess water was removed.
➢ The flask mouth is closed by cotton plug and tightly with the help of rubber band.
➢ Cover the cotton plug with a paper using rubber band.
➢ The material was sterilizing them in an autoclave at 121.6ºC for an hour.
➢ Take out the sterilized flask after releasing the steam and keep them in isolation chamber.
➢ The flasks were inoculated with bit of 10 days old culture (Trichoderma) grown on potato
dextrose agar.
➢ The flasks were placed in an incubator at 210C or in room temperature for 10-12 days.
➢ The entire grain based medium will turn green due to sporulation of Trichoderma.
(B)Mass multiplication of Trichoderma on Molasses yeast medium

Preparation of mother culture:

Molasses yeast medium is prepared as detailed below:
Molasses : 30 g
Yeast :5g
Distiller water : 1000 ml
The medium is prepared and dispensed into conical flasks and sterilized at 15 lb
pressure for 15 minutes in an autoclave. After the medium is cooled it is in inoculated with
10 days old fungal disc of T. viride and then incubated for 10 days for fungal growth. This
serves as mother culture.

Mass multiplication:

Molasses yeast medium is prepared in fermentor and sterilized as described earlier.

Then after the medium is cooled, the mother culture is added to the fermentor @ 1.5 lit /
50 lit of the medium and incubated at room temperature for 10 days. Then the incubated
broth containing the fungal culture is used for commercial formulation preparation using
talc powder.

(C) Mass production of Pseudomonas

Preparation of mother culture

Mother culture is prepared by using the king’s B medium
Peptone : 20.0 g
K2HPO4 : 1.5 g
Mg SO4 : 1.5 g
Glycerol : 10 ml
 Distilled water : 1000 ml

The above broth is dispersed into conical flasks and autoclaved at 15 lb pressure
for 15 minutes and cooled and inoculated with a loop of P. fluorescens and incubated for
2 days.

Mass multiplication:

The kings B medium is prepared and poured into the fermentor and sterilized at 15
lb pressure for 15 minutes. After the broth has cooled below the mother culture of P.
fluorescens is added to the king’s B medium in the fermentor at the rate of 3 lit for 40 lit
of the broth. Then it is incubated in the fermentor for 2 days with frequent mixing of the
broth by operating the stirrer. Then the broth containing the bacterial growth is collected
in plastic buckets and used for mixing with talc powder for commercial formulation.

Precautions

• Do not open the cotton plug until use.

• Keep it in a cold place (refrigerator preferably after sporulation)
• Avoid direct sunlight until use
Objective- To study about the identification, nature of damage and management of plant
disease

(I)Brown Leaf Spot of Rice

Brown leaf spot disease caused by the fungus Helminthosporium oryzae

Symptoms

✓ Occur in nursery as well as main field and causes blight of seedlings.

✓ Also called as sesame leaf spot or Helminthosporiose or fungal blight.
✓ Leaf spotting is very common, Isolated brown, round to oval (resemble sesame seed).
✓ Seed also infected (black or brown spots on glumes spots are covered by olivaceous velvety
growth).
✓ Infection also occurs on panicle neck with brown colour appearance.

Management:

✓ As disease is seed borne,Use disease free seeds.

✓ Removal of alternate & collateral hosts.
✓ Growing Resistant varieties like ADT-44, PY-4, CORH-1, CO-44, CAUVERY,
BHAVANI, TPS-4 and Dhanu.
✓ Seed treatment with Pseudomonas fluorescens @ 10g/kg of seed followed by seedling dip
@ of 2.5 kg or products/ha dissolved in 100 litres and dipping for 30 minutes.
 ✓ Since the fungus is seed transmitted, a hot water seed treatment (53-54°C) for 10-12
minutes may be effective before sowing.
✓ Spray Mancozeb (2.0g/lit) or Edifenphos (1ml/lit) - 2 to 3 times at 10 - 15 day intervals.

(II)Panama wilt of Banana:

Panama wilt disease caused by the fungus Fusarium oxysporum f. sp. cubense

The disease also known as banana Covid or banana cancer or banana epidemic. The disease
first discovered by Bancroft in 1874 in Australia (Panama Island). The disease introduced in India
1920 in Bombay.
Symptoms:
✓ Yellowing of older leaves starting from margin to midrib of the leaves.
✓ The leaf hangs between the pseudostem while, the middle of lamina is still green.
✓ Fungus blocks the vascular system and causes wilting.
✓ Discoloration of vascular vessels as red or brown streaks.
✓ The leaves break near the base and hang down around pseudostem after 4-6 weeks.
Longitudinal splitting of pseudostem.
✓ The cut pseudostem smells like rotten fish.

Etiology or Pathogen:
✓ The pathogen is facultative saprophyte.
✓ The fungus mycelium is septate and branched.
✓ Fungus produces three types’ conidia: Micro, Macro and Chlamydospores.
✓ Micro conidia - Single celled or rarely 1-2 septate, elliptical or oval.
✓ Macro conidia - Sickle or curved shaped, 3-5 septate and tapering at both ends.
✓ Chlamydospores - Thick walled, spherical to oval, yellowish in color and found in chain.
Management
✓ Use disease free and healthy planting materials.
✓ Apply crop rotation 3-5 years.
✓ Grow resistant verities Poovan, Robusta and Grand naine.
✓ Avoid growing of susceptible cultivars viz., Rasthali, Monthan, Red banana and
Virupakshi.
✓ Corm/ rhizomes treatment with Carbendezim @ 3 ml/ lit. water.
✓ Bio-control with Pseudomonas fluorescens or Trichoderma viride application to the soil
along with FYM.