MASS PRODUCTION OF ENTOMOPATHOGENIC FUNGUS TO KILL THE FIELD CROP PESTS

MASS PRODUCTION OF ENTOMOPATHOGENIC FUNGUS TO KILL THE FIELD CROP PESTS

Dr. Indira A. Ghonmode and Dr. S. B. Kharbade

Department of Agricultural Entomology, Mahatma Phule Krishi Vidyapeeth,

College of Agriculture, Karad-415114 (Maharashtra) India

Corresponding Author: Indira. Ghonmode, Assistant Professor, Section of Agril. Entomology, College of Agriculture, Karad-415114 (Maharashtra State) India. Mobile: 7276066627,E-mail: indirabhojne@yahoo.co.in

­­­­­­­­­­­­­­­­­­­­____________________________________________________________________________

ABSTRACT: Entomopathogenic fungus are of four types which include Beauveria, Metarhizium,  Lecannicilium lecanii, Nomurea rileyi The term entomopathogenic refers to any member of the kingdom that can infect insects pest and other terrestrial arthropods, such as mites, ticks, and spiders. Amongst all Metarrhizium anisopleae proved considerably effective showing highest mortality of sucking as well as lepidopteron and coleopteran pests. These findings indicated that Metarrhizium were prominently cheaper in rate followed by other fungus when compared. This shows that significant biopesticide under Biological Control against various field crop pests.

Key words:  Beauveria, Metarhizium,  Lecannicilium lecanii, Nomurea rileyi

­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­­______________________________________________________________________________

INTRODUCTION

M.anisopliae is a green muscardine fungi, it detected from 300 species of insects. It is effectively active against chysomelid, curculionid, and scarabid from coleoptera, pyrilla, desert locust, grass hoppers, termies and some caterpillar. Earlier, it was successfully used for control of sugarcane pyrilla, wheat cockchafer beetle, locust and rhinoerous beetle on coconut.

 

CLASSIFICATION

Common name: Green muscardine fungi.

Kngdom                                                : fungi

Phylum                                                  : Ascomycota

Class                                                      : Sordariomucetes

Order                                                     :hypocreal

Family                                                    :clavicipitaccac

Genus                                                    : Metarrhizium

Species                                                   : anisopliae

First discovered by ilyailichmetschnikaft

 

MODE OF ACTION

 

Entomopathogenic fungi Metarrhizium anisopliae produces spores which infect their host insect by germinating on its surface and then growing into its body, death of insect take place between 4-10 days. The most common portal entry of fungi is through the alimentary tract is also reported. The development of fungal infections in terrestrial insect is highly influenced by humidity which is being vital for the germination of fungal spores and transmission with the most entomopathogenic fungi, diseases development involves.

1. Attachment of the inactive unit i.e. spore or 200 spores to insect cuticle.
2. Germination of fungal spores on cuticle.
3. Penetration of the cuticle, either directly by germ tube or by aspersorium.
4. Multiplication of the hyphal bodies in the haemocoel.
5. Death of insect.
6. Gowth in mycelial phase with invasion at host range.
7. Penetration of hyphal from the interior through cuticle to the exterior of the insects.
8. Production of infective units on the exterior of insect.

INPUT DETAILS

For mother culture

Material required

1. dist.water:1000ml
2. dextrose :20gm
3. agaragar :15gm
4. yeast :10gm
5. Infested insects for conidia for inoculation.

Equipments

• Petri plate
• Test tube
• Autoclave
• Needle
• Laminar air flow
• Measuring cylinder
• Weighing balance
• Spirit/ethyl alcohol

 Mass production

1. Rice grain -25kg
2. Distilled water -28 Lit
3. Yeast -150 gm
4. Mother culture

 Equipments

• Glass bottles
• Cotton plugs
• Aluminium foil
• Grinder
• Autoclave
• Laminar air flow
• Iron racks

 SUMMARY OF ACTUAL WORK PERFORMED

1. Prepared culture media for culture of fungus.
2. Inoculated the media with mother culture and produce 940 kg of
3. After full growth of fungus on rice culture and liquid broth,mixed with talk after grinding.
4. 940 kg of Metarrhizium anisolpiae was prepared and packed.

PROCEDURE OF PRODUCTION IN DETAIL

Preparation of mother culture

1. Isolated the infested insects and brought into the laboratory.
2. Media preparation
3. Took 20 gm dextrose+ agar agar 15 gm +10 gm in 1000 ml distilled water.
4. Put it to the conical flasks and then plug the flasks with cotton.
5. Also sterilized the petridishes and conical flasks at the same time(containing media)
6. Later on shifted it to the laminar air flow(culture was sterilized previously) and allowed to cool it and sterilized under uv light.
7. Then under aceptic condition poured the media from conical flasks into petri plates and test tubes.
8. Sterilized the needle on the flame and sterilized the hands with ethyl alcohol to avoid contamination.
9. Then inoculated the spores from from body of infested insects to the petriplates under aseptic condition and also carried out the same in test tubes.
10. Later on shifted it t the refrigerator.

• Procedure for mass production of Metarrhizium anisopliae.

1. Washed the bottles of glass by distilled water.
2. Then weigh 100 gm of rice grains on electronic weighing balance.
3. Then poured them in the glass bottles.
4. Added 50 ml distilled water.
5. Then added little amount of yeast solution to it.
6. Yeast solution was prepared by taking 5 gm of yeast in 100 ml of distilled water in conical flask.
7. Then plugged the necks of glass bottles tightly with cotton plug.
8. Shacked the content to mix well and kept over a night for fermentation.
9. On the next day, autoclaved the bottles at 121 ⁰c for 20 minutes.
10. Then allowed the bottles to cool and shifted it to the laminar flow.
11. Sterilized it in the uv light.
12. Inoculated the medium with mother culture of fungus with sterilised needle.
13. Kept the bottles for 3 weeks on racks for growth of culture.
14. Covered the cotton pug with aluminium foil.
15. Precaution should be taken that contamination should be avoided in every step.
16. Then after full growth on fungus on grains, I is poured in the tray and allowed to areate.
17. Then they were grinded in grinding machine.
18. Then placed the produce in plastic bags and was ready for application.

Flow chart for Mycoinsecticides

 

 

Washing and cleaning of culture bottle

 

 

Take 100 g Grain ( Jowar/ Rice)

 

 

 

Add 50 ml distiled water

 

 

Add  1 %  yeast extract solution  in 1 ml

 

 

Mixed well with yeast with grain and finally plugged the bottles tightly

 

 

 

Autoclave the bottles a 121^O c for 15 minutes

 

Keep Autoclave bottle in laminar flow and allowed to cool under UV light for sterilization

 

 

Inoculated the media with pure culture

 

 

 

Kept this bottle for incubation

 

 

 

After 15 days harvesting of spores

 

Spores Count under Microscope

 

 

Packaging and store in cool and dry place, CFU count

 

Ready for field application