BIOPESTICIDE- FUNGAL Beaveriasp,Metarrihizium sp,Verticillum sp

soil and agricultural microbiology

1. BIOPESTICIDE FUNGAL M.AYISHA RAFIYA 24SPMB01 ST .MARYS COLLEGE (AUTON0MOUS )

2. BIOPESTICIDE Biopesticides are pesticides derived from natural sources such as microorganisms, plants, or minerals that help in controlling pests in an eco-friendly manner. They are an alternative to chemical pesticides and are usually specific to target pests, reducing harm to humans, animals, and the environment. Types of Biopesticides Microbial Biopesticides Contain microorganisms like bacteria, fungi, viruses, or protozoa . Example: Bacillus thuringiensis (Bt) for controlling caterpillars,Beauveria bassiana for insect pests . Plant- Incorporated Protectants (PIPs) Genes from plants or microbes incorporated into crops to produce pest-resistant plants.

3. Example: Bt cotton producing proteins toxic to specific insects . Biochemical Biopesticides Naturally occurring substances that control pests by non-toxic mechanisms. Examples: insect pheromones, neem extracts, or plant essential oils . Advantages Target-specific, reducing non-target effects. Environmentally safe and biodegradable. Can reduce pest resistance compared to chemical pesticides. Safe for humans, animals, and beneficial insects like pollinators. Limitations Often slower acting than chemical pesticides. Sensitive to environmental conditions (sunlight, temperature, pH). May have a short shelf life. . Sometimes require repeated application for effectiveness.

4. Applications Agriculture: controlling insect pests, weeds, and plant pathogens. Forestry and horticulture: eco-friendly pest management. Integrated Pest Management (IPM): used along with cultural and mechanical methods for sustainable pest control . Fungal Biopesticides Fungal Biopesticides are a type of microbial biopesticide that use entomopathogenic or pathogenic fungi to control insect pests, plant pathogens, or weeds. They are natural, eco-friendly alternatives to chemical pesticides .

5. FUNGAL BIOPESTICIDES Fungal biopesticides are fungi that infect or inhibit pests (insects, nematodes, or plant pathogens), leading to their death or reduced activity. Mechanism of Action: The fungal spores attach to the pest’s body. Germinate and penetrate the pest’s cuticle. Grow inside the pest, producing toxins or enzymes. Kill or weaken the pest over time .Examples of Fungal Biopesticides: Beauveria bassiana – kills many insect pests .Metarhizium anisopliae – effective against locusts, beetles, and termites. Trichoderma species – act as biocontrol agents against fungal plant pathogens. Paecilomyces species – used against whiteflies and aphids.

6. Advantages :Environmentally safe and biodegradable. Specific to target pests; minimal effect on humans, animals, and beneficial insects. Can be used in organic farming .Limitations: Slow acting compared to chemical pesticides. Sensitive to environmental conditions like UV light, temperature, and humidity. Requires careful storage and handling

7. Beaveria sp

8. Beaveria sp Scientific Classification Kingdom : Fungi Division : Ascomycota Class : Sordariomycetes Order : Hypocreales Family : Cordycipitaceae Genus : Beauveria Species : B. bassiana

9. The species is named after the Italian entomologist  Agostino Bassi , who discovered it in 1835 in  silkworms  ( Bombyx mori ). Bassi performed the first infection experiments, and determined the fungus to be the cause of the muscardine disease, which then led to carriers transmitting it by airborne means. Later the same year, the fungus was named  Botrytis bassiana  by  Giuseppe Gabriel Balsamo-Crivelli .  The species epithet honours Bassi, the discoverer: In 1911  Jean Beauverie  did further study and the next year  Jean Paul Vuillemin  made it the  type species  of his new genus,  Beauveria,  a name which honors Beauverie. The name  B. bassiana  has long been used to describe a  species complex  of morphologically similar and closely related  isolates . Rehner and Buckley have shown that  B. bassiana  consists of many distinct lineages that should be recognized as distinct phylogenetic species and the genus  Beauveria  was redescribed with a proposed type for  B. bassiana  in 2011. The species was formerly also known as  Tritirachium shiotae,  among other synonyms.

10. Beaveria sp Scientific Name: Beauveria bassiana (most common species) Type: Entomopathogenic fungus (infects insects) Habitat: Soil and decaying plant material; naturally found worldwide. Form: Usually applied as spores (conidia) in biopesticide formulations .Morphology Mycelium: White, cottony growth on culture media Spores: Small, round, asexual spores (conidia) produced on conidiophores. Growth: Grows well at moderate temperatures (25–30°C) on artificial media .Mode of Action Spores attach to the insect’s cuticle. Germinate and produce enzymes (chitinases, proteases) that degrade the exoskeleton. Fungal hyphae penetrate into the insect body.Fungus grows inside, producing toxins that kill the insect. Infected insects often become mummified and covered with white fungal growth.

11. Target Pests Aphids, whiteflies, thrips Beetles, caterpillars, locusts Termites and other soil-dwelling pests Advantages as a Biopesticide Specific to insects; safe for humans, animals, and plants. Can be integrated into Integrated Pest Management (IPM). Environmentally friendly and biodegradable .Limitations Slow-acting compared to chemical pesticides. Sensitive to UV light, temperature extremes, and desiccation. Requires proper formulation and storage for effectiveness

12. Ecology 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. Afterwards, a 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. Beauveria bassiana  parasitizing the  Colorado potato beetle  has been reported to be, in turn, the host of a mycoparasitic fungus  Syspastospora parasitica . This organism also attacks related insect-pathogenic species of the  Clavicipitaceae .

13. Toxicity The fungus rarely infects humans or other animals, so it is generally considered safe as an insecticide. However, at least one case of human infection by  B. bassiana  has been reported in a person with a suppressed immune system. Additionally, the spores may exacerbate  breathing difficulties . Uses 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.

14. 1. Mass Production A. Solid-State Fermentation (SSF): Substrate examples: Rice, wheat, sorghum, maize, or agro-industrial residues (e.g., bagasse , bran). Process: Substrate is cleaned, moistened (40–60% moisture), and sterilized. Inoculated with Beauveria sp. spores or mycelial culture. Incubated at 25–28°C for 10–14 days. Spores are harvested by sieving or blending with carriers. Advantages: High sporulation , easy to handle, low-cost. low-cost. B. Submerged Fermentation ( SmF ):Medium: Nutrient-rich liquid with carbon (glucose, sucrose) and nitrogen sources (yeast extract, peptone). Process:Inoculate sterile liquid medium with fungal culture. culture. Aerobic fermentation with agitation (to supply oxygen).Harvest spores or biomass after 5–7 days. Advantages: Faster production, controlled conditions, less contamination risk.

15. C. Formulation Types: Wettable powders (WP): Mixed with inert powders, diluted in water for spraying. Granules (GR): Applied to soil, slowly release spores. Oil-based formulations: Protect spores from UV and desiccation. Emulsifiable concentrates: Can mix with water or oils for foliar application 2. Quality Control Viability Tests: Germination percentage of spores on artificial media. Pathogenicity Tests: Test on target pests to confirm infectivity. Purity Tests: Microscopic examination to rule out contamination. Moisture Content: Ideally below 8–10% for long-term storage. Shelf-Life Studies: Check spore survival over months under various storage conditions.Particle Size & Flowability : Ensures easy application in spraying equipment.

16. 3. Field Application Timing & Conditions: Best applied during morning or evening to avoid sunlight. High humidity favors spore germination and infection. B. Application Methods: Foliar Spray: Suspension sprayed on leaves to control aphids, whiteflies, and caterpillars. Soil Treatment: Granules mixed into soil to control termites, grubs, and root-feeding pests. Seed Treatment: Seeds coated with spores to protect seedlings. Combination with Other IPM Methods: Use with pheromone traps, neem extracts, or predator insects for integrated pest control. C. Dosage:Typical application: 1–5 × 10^12 spores per hectare (varies with crop and pest). D. Safety & Environmental Considerations: Non-toxic to humans, animals, and beneficial insects. Biodegradable; does not accumulate in soil or water

17. Metarrhizium sp

18. Kingdom ; Fungi ( Mycota ) Division ; Ascomycota Subdivision ; Pezizomycotina Class ; Sordariomycetes Subclass ; Hypocreomycetidae Order ; Hypocreales Family ; Clavicipitaceae

19. Metarrhizium sp General Information Scientific Name: Metarhizium anisopliae , also includes M. brunneum , M. acridum (used for locust control) Type: Entomopathogenic fungus Habitat: Soil, plant debris, insect cadavers Role: Natural regulator of insect populations, widely used in biological pest control 2. Morphology & Growth Mycelium: White initially, turns greenish as spores develop Spores (Conidia): Green, cylindrical or slightly oval, produced on conidiophores Optimal Growth: 25–30°C, pH 5–6, aerobic conditions Reproduction: Asexual via conidia; sexual stage ( teleomorph ) exists in some species 3. Mode of Action Spores attach to insect cuticle using adhesive substances..

20. Germinate and penetrate through cuticle using enzymes: Proteases: degrade proteins Chitinases : degrade chitin Lipases: degrade cuticular lipids Fungus multiplies inside hemolymph , producing toxins. Infected insect dies within 5–14 days depending on species and environmental conditions. Cadaver often turns greenish due to spore proliferation, releasing more spores into soil or crops . 4.Target Pests Locusts and grasshoppers: M. acridum is highly effective Soil pests: White grubs, root-feeding beetles, termites Crop pests: Aphids, thrips , caterpillars, beetlesEffective against pests resistant to chemical insecticides.

21. 1. Mass Production A. Solid-State Fermentation (SSF) Substrates: Rice, wheat, sorghum, barley, or agro-wastes Process: Moist substrate inoculated with spores, incubated 10–14 days, harvested by sieving or blending Advantages: High spore yield, low cost, easy handling B. Submerged/Liquid Fermentation ( SmF ) Nutrient-rich liquid medium (carbon + nitrogen sources) Aerobic fermentation with agitation for 5–7 days Produces spores or mycelial biomass for formulation C. Formulations Wettable powders (WP) for foliar spray Granules (GR) for soil application Oil-based suspensions to protect spores from UV Emulsifiable concentrates for easy mixing and spraying

22. 2.Quality Control Spore viability: % of germinating spores Pathogenicity test: Ability to kill target pests Purity check: Free from contaminating microbesMoisture content: <10% for long shelf lifeShelf -life evaluation: Months under controlled storage Particle size & flowability : Ensures uniform application. 3. Field Application A. Methods Foliar spray: For insects on crops (aphids, whiteflies, caterpillars) Soil application: Granules for root-feeding insects Seed coating: Protects seedlings from early pest attack Aerial spraying: Used for locusts and large-area infestations.

23. B. Conditions for Application Apply in cool, humid conditions for better spore germination Avoid direct sunlight during spraying (UV-sensitive spores) Combine with Integrated Pest Management (IPM) practices C. Dosage Typical application: 1–5 × 10^12 spores/ha, depending on crop and pest D. Advantages in Field Use Specific to target insects Biodegradable, eco-friendly Reduces pesticide resistance Compatible with organic farming E. Limitations Slower action than chemical pesticides Sensitive to environmental extremes (UV, temperature, desiccation) Requires proper storage and

24. Verticillum sp Kingdom ;Fungi Phylum ; Ascomycota Class ; Sordariomycetes Subclass ; Hypocreomycetida Family; Plectosphaerellaceae genus Verticillium .

25. Verticillum sp

26. Verticillum sp 1. General Information Scientific Name: Verticillium lecanii (syn. Lecanicillium lecanii ) Type: Entomopathogenic fungus Habitat: Soil, leaf surfaces, insect cadavers; widely distributed in temperate and tropical regions Role: Biological control agent for soft-bodied insect pests, especially in vegetables, cotton, and ornamental plants 2. Morphology & Growth Mycelium: Septate , white, cottony initially; may turn cream or yellowish Conidia (asexual spores): Small, single-celled, elliptical or ovoid, produced in clusters Conidiophores: Branched or simple, giving rise to conidia Growth Conditions: Optimal at 20–25°C, high relative humidity (70–90%), pH 5–63.

27. 3.Mode of Action Spores attach to the insect cuticle via adhesion Germination and penetration occur using enzymes ( chitinases , proteases) Fungus proliferates inside the insect, producing toxins that paralyze and kill Infected insect may develop fungal growth externally, releasing new spores Infection is generally host-specific, mostly targeting aphids, whiteflies, and thrips . 4. . Target Pests Aphids ( Myzus persicae , Aphis gossypii ) Whiteflies ( Bemisia tabaci ) Thrips ( Frankliniella spp.) Soft-bodied insects feeding on leaves, stems, or flowers 5. Mass Production Techniques A. Solid-State Fermentation (SSF)Substrates: Rice, wheat, sorghum, or agro-residues like brand

28. Steps: Sterilize and moisten substrate (40–60% moisture) Inoculate with fungal culture Incubate 10–14 days at 20–25°C Harvest spores, mix with carrier for formulation Advantage: High spore yield, low-cost production B. Submerged Fermentation ( SmF ) Growth in liquid nutrient medium with carbon (sucrose, glucose) and nitrogen sources Aerobic agitation to supply oxygen Spores or mycelium harvested after 5–7 days C. Formulations Wettable powder (WP) for foliar spray Oil-based suspensions to protect against UV and desiccation Granules (GR) for soil application.

29. Quality Control Spore viability: % germination under laboratory conditions Pathogenicity test: Infection efficiency against target insects Purity check: Absence of contaminating microorganisms Moisture content: Ideally <10% for long shelf life Shelf-life stability: Months under controlled storage Particle size & dispersibility : Ensures uniform application . Field Application Methods Foliar spray: Most common method, applied to crops infested with aphids, whiteflies, or thrips Seed treatment: Coating seeds with spores protects seedlings Soil application: Granules applied to soil target root-feeding insects Environmental conditions: Apply in cool, humid conditions; avoid direct sunlight Dosage:Typically 1–5 × 10^12 spores/ha, depending on crop and pest density

30. Advantages Specific to target pests, safe for humans, animals, and beneficial insects. Biodegradable, environmentally safe Can reduce dependence on chemical insecticides Compatible with organic farming and IPM programs Limitations Slower acting compared to chemical pesticides. Sensitive to UV, desiccation, and high temperature Requires proper storage and formulation for effectiveness. Advanced Mechanism of Action Adhesion: Conidia attach to the insect cuticle via hydrophobic interactions and sticky substances. Penetration: Germ tubes secrete cuticle-degrading enzymes such as chitinases , proteases, and lipases. Colonization: Hyphae grow inside the insect body ( hemocoel ), consuming nutrients and producing toxins.

31. . Death & Sporulation : Host insect dies in 3–10 days (depending on species and environment). Fungal mycelium grows externally, producing new conidia that spread to other hosts . Host specificity: Primarily targets soft-bodied insects (aphids, whiteflies, thrips ), minimizing non-target effects. 2 . Formulation of Verticillium sp . Biopesticides Wettable Powder (WP): Dried spores mixed with inert carriers; diluted in water before spraying . Oil Suspensions: Protect spores from UV light and desiccation; increase field persistence . Granules (GR): Applied to soil for root-feeding pests; release spores gradually . Encapsulation /Advanced Formulations: Use of polymer coatings or beads to enhance shelf life, stability, and slow release in the field. 3. Mass Production (Industrial Scale) Solid-State Fermentation (SSF):Substrates: Rice, wheat, sorghum, bran, agro-residues Moisture: 40–60%, incubated 10–14 days at 20–25°C Harvested spores are blended with carriers for application.

32. Submerged/Liquid Fermentation ( SmF ) Nutrient broth culture in aerated fermenters Faster production and controlled environmental parameters Spores harvested after 5–7 days for formulation 4. Quality Control Measures Viability: % germination on nutrient media Pathogenicity test: Confirm infectivity against target pest species Purity: Free from bacterial or fungal contaminants Moisture content: ≤10% for long-term storage Shelf-life stability: Evaluated under varying storage conditions Dispersibility & particle size: Ensures effective spraying and application 5. Field Application Strategies Foliar Spray: Most common; targets aphids, whiteflies, and thrips Seed Treatment: Coating seeds to protect seedlings from early pest infestation

33. Soil Application: Granules target root-dwelling pests Timing: Early morning or evening to avoid UV damage; high humidity and moderate temperatures enhance effectiveness Integration with IPM: Can be combined with: Beneficial insects (ladybugs, parasitoids) Neem -based products or botanical insecticides Cultural practices (crop rotation, trap crops)6 crops) 6. Advantages of Verticillium sp. Biopesticides Specificity to soft-bodied insects, minimizing non-target effects Biodegradable and environmentally friendly Reduces reliance on chemical insecticides Compatible with organic farming and IPM systems Can help manage pesticide-resistant pest populations.

34. 7. Limitations & Challenges Slower action compared to synthetic insecticides Sensitive to environmental stress: UV radiation, extreme heat, and low humidity Requires careful storage and handling May need repeated applications under high pest pressure Field performance depends on environmental conditions 8. Commercial Use Examples Vertalec (UK/Europe ) – used against whiteflies and aphids in greenhouse crops Lecanicillium -based products – widely used in Europe, North America, and Asia for horticultural crops

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