Biopesticides in Integrated Pest Management

Division of Agricultural Chemicals 1

Agrochemicals for food & nutritional security: BIOPESTICIDES IN IPM

Prithusayak MondalDivision of Agricultural Chemicals

Roll No. 4944

Chairperson : Dr. Anupama SinghSeminar Leader : Dr. Seeni Rengasamy

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Per capita land availability

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Problem of food security

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Division of Agricultural Chemicals 41/10/2011


GREEN REVOLUTION

RICE WHEAT PULSES ALL FOOD GRAINS

Demand production

104.

2

98.8

83.6

77.4

243.

3

239.

3

Press Information Bureau, 27-10-2008

Production and Demand of Food grains in 2011-2012 (million tonnes)

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Bacteria Fungi

VirusesNematodes

Attack to Crops

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Food plants of the world are damaged by more than 10,000 species of insects, 30,000 species of weeds, 100,000 diseases (caused by fungi, viruses, bacteria and other microbes) and 1000 species of nematodes (Hall, 1995; Dhaliwal et al., 2007)

Insects

Weeds


Estimation of crop losses caused by insect pests to major agricultural crops in India

Dhaliwal et al., 20101/10/2011


Role of PesticidesCrop production without pesticide is unimaginable

To ensure better production at harvest against unpredictable losses caused by plant diseases & pests To improve both quality & quantity of food

To decrease the extent of vector born & other diseases in humans & animals

“Complete ban on agrochemicals use in agriculture might result in 50% reduction in global food production and 4 to 5 times increase in food prices” Nobel Laureate Norman Borlaug

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Risk Associated With Chemical PesticidesToxicity to plants

Toxicity to mammalsToxicity to aquatic creatures

Toxicity to beneficial organisms

High persistence of residues

• Indiscriminate use leads to the Three sad R’s :Resistance, Resurgence and Residues

• Elimination of Natural enemies of pests

• Upsetting the ecological balance

• Environmental degradation/Pollution

• Enters food chain and lead to Bio-Accumulationand Bio-Magnification

As a result of The misuse and overuse of pesticides crop losses have consistently shown an increasing trend (Dhaliwal and Koul, 2010)

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New form of pesticide

Low residual toxicity

Environmentally safe

Host specific in action

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Active ingredient- Living organisms

1st Biopesticide discovered in the year 1835

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Biopesticides are used to control pests, pathogens, and weeds by a variety of means

Microbial biopesticides may include a pathogen or parasite that infects the target

Alternatively, they might act as competitors or inducers of plant host resistance

BI PESTICIDE


Bio means involving life or living organisms

Pesticide includes substance or mixture of substances intended for preventing, destroying or controlling any pest

Biopesticide refers introduction of any living organism such as microorganism including bacteria , fungi , nematodes viruses, protozoa and parasitoids and predators that controls pests by biological non-toxic means e.g. Trichoderma sp., Bacillus thuringiensis, Beauveria etc.

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All the living organisms, which are cultivated in the laboratory on large scale & used and exploited experimentally for the control of harmful organisms are called biopesticides

Division of Agricultural Chemicals 14Global biopesticides & synthetic pesticides market, 2003-

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Locked Horns:Synthetic pesticides Vs. Bio-pesticides

(Source : agriculture Today. Nov. 2005)

Factors Synthetic Pesticides Bio-pesticides

Cost effectiveness Cheap but increasedspraying cost

Costlier but reduced number of applications

Persistence and residualeffect

High Low

Knockdown effect Immediate Delayed

Handling and Bulkiness Easy but danger andHazardous

Bulky : Carrier basedEasy : Liquid formulation

Pest resurgence More Less

Effect on Beneficial flora More harmful Less harmful

Target specificity Mostly broad spectrum Mostly host specific

Nature of control Curative Preventive

Shelf life More Less

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The market share of bio-pesticide is only 2% as compared to synthetic pesticide

Division of Agricultural Chemicals Woo et al., 2010

MICROBIAL PESTICIDEActive ingredient : Microorganism (Fungi, bacteria, virus, nematode etc.)

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List of registered microbial products by CIB

Name of microbes TypeBacillus sp. Bacteria

Trichoderma sp. Fungi

Pseudomonas fluorescens Bacteria

Gliocladium sp. Fungi

Beauveria bassiana Fungi

Verticillium lecanii Fungi

Metarhizium anisopliae Fungi

Nomuraea rileyi Fungi

Nuclear Polyhedrosis Viruses Virus

Granulosis Viruses Virus

Courtesy: http://www.cibrc.nic.in1/10/2011

MICROBIAL PESTICIDE


CharacteristicsStorable

EconomicalEasy to produce

Safe & acceptableConvenient to apply

Virulent against target pest

Advantages

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High degree of specificityCompatible with chemical pesticides

Easy to apply & aid growth through outNo adverse effect on non-target organisms

Absence of residue build-up in the environmentRelatively cheaper by 50% as compared to chemical pesticides

(Narayanasamy, 1995)


Bio-pesticides

Entomopathogenic FungiFungal Antagonists

Bacterial AntagonistsEntomopathogenic Bacteria

Parasites & Predators

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Moore & Prior, 1993


Entomopathogenic FungiEntomopathogenic fungi are fungi that can act as parasites of insects and

kill or seriously disable them

Mode of Action

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Entomopathogenic fungi in insect control


BeauveriaBeauveria bassiana most commonHabitat: FoliageInsect Host: White flies, beetles & caterpillars (including Helicoverpa sp.)Dose: 2 treatments made at 15-day intervals with 1.5 kg/ha concentrated product of B. bassiana (3.0 × 109 conidia)Treatment:i) Foliar spray: 400-500 g in ½ bigha (5g/L of water)ii) Soil drench: 250-500 g/3 bighaHealth impact: It causes granulosis disease in human ear

Grasshoppers killed by B. bassianaBeauveria bassianaCultures of B. bassiana

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MetarhiziumMetarhizium anisopliae var. anisopliae & var. major

Habitat: Foliage

Insect host: Frog hoppers, beetles

Dose: Aerial treatment at 50 l/ha with 6 × 1011 to 1.2 × 1012 conidia/l of water

Conidia

Different cultures of M. anisopliaeCockroach killed by M. anisopliae

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Verticillium

Verticillium (Cephalosporium) lecanii

Habitat: Glasshouse foliage

Insect host: Aphids, whiteflies & scales

Dose: 41 × 107 active spores/g either undiluted or as a 10% concentration (diluted with talc or water)

Whitefly scale infected with V. lecanii

Cultures of Verticillium lecaniiConidia1/10/2011


Fungal Antagonists Principal fungi: Gliocladium virens & Trichoderma sp. Trichoderma sp. mainly T. harzianum & T. viride Habitat: Soil Effective against: damping-off & wiltParasitize Rhizoctonia & SclerotiumInhibit growth of Pythium, Phytophthora & Fusarium

T. harzianum T. virideDisease: T. harzianum causes green mold in cultivated button mushrooms & T. viride causes green mold rot of onion

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Mode of action Direct parasitism or lysis (lytic enzymes like chitinase, cellulase & glucanase) & death of

the pathogen

Direct toxic effects on the pathogen by antibiotic substances released by the antagonist

Mycoparasitism by a Trichoderma strain on the plant pathogen Pythium

Competition with pathogen for food

Indirect toxic effects on the pathogen by volatile substances released by the metabolic activities of the antagonist

Cultures of Trichoderma harzianum

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The aim of investigations was to confirm the effect of Trichoderma harzianum on Rhizoctonia solani and make a possibility for its usage in tobacco production

T. harzianum was applied before and after sowing including a fungicide Top M (0.1%)At additional treatment with Trichoderma after use of fungicide, had a better result than fungicide alone

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The influence of T. harzianum on intensity of disease attack Artificial inoculationNatural inoculation

The best results have shown by a variant with T. harzianum applied on a soil beforesowing and further application at certain intervals any time in a growing season oftobacco seedlings

Additional treatment with T. harzianum after a fungicide Top M is advantageous to the situation with a disease, so, it may be applied with this fungicide treatment


Bacterial Antagonists• Pseudomonas sp. are gram negative, aerobic, rods that are inhabitants of wide

range of soil, water & plant surfaces

• P. fluorescens recognized by fluorescent pigment called ‘pyoverdines’

• Bio-control abilities of strains depend on aggressive root colonization, induction of systemic resistance in the plant & production of diffusible or volatile antifungal antibiotics

• Antibiotics with bio-control properties include – phenazines, hydrogen cyanide, 2,4-diacetylphloroglucinol, pyoluteorin, pyrrolnitrin, lipopeptides etc.

Phenazin

2,4-diacetylphloroglucinol

pyoluteorin

pyrrolnitrin

Lipopeptide

Hydrogen cyanide1/10/2011


Mode of Action

Control of diseases• Different strains of P. fluorescens extensively used in bioremediation of various organic compounds & bio-controls of pathogens in agriculture

• P. fluorescens found effective in controlling fungal pathogens such as wilt/root rot, Fusarium oxysporum f. sp. Cubense, Pythium sp., R. solani, R. oryzae, S. rolfsii & bacterial pathogens like Xanthomonas citri & P. solanacearum in field tests

• Bacterial preparations widely used in organic spice cultivation of southern India

Theories include -• Induction of systemic resistance – resist attack by true pathogen

• Competition with other (pathogenic) soil microbes, e.g. siderophores

• Production of compounds (antibiotics) antagonistic to other soil microbes

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Entomopathogenic Bacteria

• Bacillus thuringiensis (Bt), a Gram-positive, motile, rod shaped bacterium produces a parasporal crystal composed of one or more proteins

• The strains of Bt characterized so far affect members of 3 insect orders: Lepidoptera (butterflies and moths), Diptera (mosquitoes & biting flies), and Coleoptera (beetles)

• EPA registered Bt products include B.t. israelensis (Diptera)—frequently used for mosquitoes B.t. kurstaki (Lepidoptera)—frequently used for gypsy moth, spruce budworm, and many vegetable pests B.t. sandiego and tenebrionis (Coleoptera)—frequently used for leaf beetle, Colorado potato beetle

B.t. kurstaki is the most commonly used Bt formulation

Bacillus thuringiensis

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Mode of Action

Bacillus thuringiensis strains produce crystalline proteins (called δ-endotoxins)

Caterpillar consumes the Bt spore (diagram 1) & crystalline toxin-treated leaf

The Bt crystalline toxin (diamond shapes in diagram 2) binds to gut wall receptors, and the caterpillar stops feeding

Within hours, the gut wall breaks down, allowing spores (oval tube shapes) and normal gut bacteria (circular shapes) to enter body cavity, where the toxin dissolves

The caterpillar dies in 24 to 48 hours from septicemia, as spores and gut bacteria proliferate in its blood (diagram 3)

Treatments:Dose: i) 100 – 150 g/ bigha for field crops.ii) 150-200 g /bigha for orchards.

Method: The powder is first mixed with small quantity of water to prepare a uniform suspension. Then the required quantity of water is added and thoroughly mixed before spray.

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Laboratory assays were done to evaluate the effect of Bacillus thuringiensis, neem seed kernel extract (Azadirachta indica), Vitex negundo leaf extract, & applied separately or together, on nutritional indices of the rice leaf-folder Cnaphalocrocis medinalis

Bt biopesticide & other 2 botanical pesticide suppressed feeding and larval growth and low concentrations affected the larval performance

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Division of Agricultural Chemicals(Nathan et al. ,2005)

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The combined effect of these resulted in a considerable decrease in nutritional indices indicating strong deterrence


• Bt is considered to be “practically nontoxic” to humans and other vertebrates

• It can cause a “very slight irritation” if inhaled & can cause eye irritation

• Bt is not carcinogenic, mutagenic, or teratogenic

• Bt does not persist in the brains, lungs, or digestive systems of animals, including humans

• Bt has been found in fecal samples of exposed greenhouse workers, no gastrointestinal symptoms were associated with its presence

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Human Health & Safety


• Bt appears to be a normal component in the feces of vegetable-consuming animals, where it apparently causes no problem

• Like the active bacterial ingredient, the inert ingredients in Bt formulations have also been studied and modified for safety

• Granular and microcapsule formulations reduce the inhalation hazard

• Volatile agents associated with some Bt formulations do not appear to constitute a significant health hazard.

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Human Health & Safety…


Environmental Impacts• No danger has been found to aquatic communities accidentally exposed to Bt or to non-target organisms including beneficial insects, amphibians, fish, and mammals

• Few reports of Bt lethality upon non-target organisms, such as leaf-feeding caterpillars

• Clay soils may bind the bacterial toxin, increasing its environmental persistence and possible toxicity to non-target species

• Newer formulations employ preservatives, like sorbitol, that are safer than the xylene used decades ago

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Phytonematode management through bacteria

Bacteria Genus/species Target nematode Mode of action References

Parasitic bacteria

Pasteuria penetrans, P. thornei

Phytonematodes Parasitism Bekal et al.(2001), Bird et al. (2003)

Opportunistic bacteria

Brevibacillus laterosporus, Bacillus nematocida

Free living & Phytonematodes

Parasitism Niu et al. (2006),Tian et al. (2007)

Rhizobacteria Bacillus sp., Pseudomonas sp.

Meloidogyne sp., Heterodera sp.

Interfering with recognition, production of toxin, nutrient competition, plant growth promotion

Marleny et al. (2008), Meyer (2003)

Crystal forming bacteria

Bacillus thuringiensis (Cry 5,6,12,13,14,21)

Trichostrongylus colubriformis, Caenorhabditis elegans

Cry proteins cause damage to the intestines of nematodes

Kotze et al.(2005), Wei et al. (2003)

Endophytic bacteria

Root knot nematode,Cyst nematode

Rhizo-bacterial & endophytic bacterial mode of action

Sturz et al. (2004), Compant et al. (2005)

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Nuclear polyhedrosis virus (NPV)A) NPV (Helicoverpa): It is highly effective on H. armigera, pest of

cotton,gram, pea, pigeon pea, tomato, cabbage, ground nut, millets, oilseeds & roses

B) NPV (Spodoptera): It is highly effective against S. litura caterpillar, pest of cotton, gram, pigeon pea, cabbage, tomato, chillies & oilseeds

Treatments: Dose: 250 – 500 LE/ha

Method: i) Shake the bottle properly and prepare a solution @ 1 ml/litre of waterii) Spray the solution 2-3 times at 10-15 days intervaliii) Spray preferably in the evening and on young larval stages or on sighting of egg laying

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Enhancing food security by the local production of microbial bio-pesticides against insect crop pests:

African armyworms as a case study

2 types of application studiedA) Aerial spray of SpexNPVB) Ground spray of SpexNPV & OP pesticide Diazinon

separately

(Wilson et al., 2008)

SpexNPV = Spodoptera exempta Nucleo polyhedrovirus

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Division of Agricultural Chemicals 41(Wilson et al., 2008)1/10/2011




Commercial bio-pesticides for the control of plant pathogens

Microorganisms Trade Name Pathogens/ Diseases

Bacteriophages of Xanthomonas sp. and Pseudomonas syringae pv. Tomato

Agriphage™ Bacterial spot in pepper & tomatoes & bacterial speck in tomatoes

Pseudomonas syringae strain ESC 10

Bio-Save® 10LP3 Ice inducing bacteria & biological decay

Pantoea agglomerans strain E325

Bloomtime, Biological™ 3

Fire blight( Erwinia amylovora)

Bactericides

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Microorganisms Trade Name Pathogens/ DiseasesStreptomyces lydicus WYEC 108

Actinovate®AG, Actinovate®SP

Soiborne pathogens: Pythium sp., Rhizoctonia sp., Phytophthora sp., Fusarium sp.Foliar pathogens: Alternaria sp., Peronospora sp.

Bacillus subtilis GB03

Kodiak® Concentrate Rhizoctonia, Fusarium, Alternaria, Aspergillus /Phoma. root rot, damping off, crown rot

Trichoderma harzianum Rifai strain KRL-AG2

T-22™HC, Plant Shield®, T-22™. Planter Box, Serenade® MAX™

Fusarium, Pythium & Rhizoctonia/ Root rot, powdery mildew

Bacillus pumilus QST 2808

Ballad® Plus Cercospora sp./ Rust, powdery mildew,, and brown spot

Fungicides

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Types of bio-control agents

Names of bio-control agents

Target species

PARASITOIDS Trichogramma chilonis Brinjal shoot and fruit borer, shoot borers of cotton, sugarcane, rice

T. brasiliensis and T. pretiosum (egg parasitoids)

tomato fruit borer

PREDATORS Cryptolaemus montrouzieri (Austrtralian ladybird beetle)

several species of mealy bugs and soft scales

Chrysoparla sp. (green lacewing bug)

aphids, white flies

Parasitoids & Predators


Few examples of bio-controlMuscodor albus strain QST 20799 acts as bio-fumigant & controls bacteria and soil borne pest by releasing volatile toxin

Aspergillus flavus strain AF36 can act as bio-fungicide for cotton. Unlike other strains it will not produce carcinogenic ‘Aflatoxin’

Pasteuria sp. acts as bio-nematicide & controls microscopic worms & other nematodes that feed on plant roots

Cydia pomonella granulosis virus acts as bio-insecticide & controls codling moth in fruits like apples & pears

Phytophthora palmivora acts as bio-herbicide & controls milkweed (Asclepias sp.) in citrus orchards

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Division of Agricultural Chemicals

Path AheadMore studies needed to determine the environmental effects on the fate of bio-agents

New technologies such as micro encapsulation of bio-control agents may be of high priority in enhancing their potential

Integration of bio-pesticides with botanical pesticides has a lot of potential in pest management

Integration of bio-pesticides with chemical pesticides as part of Bio-intensive Integrated Pest Management (BIPM)

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ConclusionMicrobials such as bacteria, fungi, viruses are the major bio-pesticides being studied mostly to develop alternatives to chemicals

The no. & growth rate of bio-pesticide showing an increasing marketing trend in past few decades

Bio-pesticides are host specific & bio-degradable resulting in least persistency of residual toxicity

Bio-pesticides саn mаkе vital contributions tο IPM & can greatly reduce conventional pesticides, while crop yield remains high

Bio-pesticides having lesser health hazard provides an important alternative in the search for an environmentally sound and equitable solution to the problem of food security

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“Life is not living, but being in health.”

- Latin poet Martial


Education

Biopesticides in Integrated Pest Management