An entomopathogenic fungus seminar was presented covering the role of these fungi in insect control. The key points are: Entomopathogenic fungi such as Beauveria, Metarhizium, Lecanicillium, and Nomurae act as parasites to insects, killing or disabling them. They are effective against many agricultural and forest pests like beetles, caterpillars, whiteflies, and grasshoppers. These fungi penetrate the insect cuticle, grow internally, and produce toxins that ultimately kill the host. They have potential for use in reducing chemical pesticide use in agriculture.

1. MASTER’S SEMINAR
Presented by
Ankita
M.Sc (Plant Pathology)
1st year
2016-17

2. ROLE OF
ENTOMOPATHOGENIC
FUNGI IN INSECT
CONTROL

3. INTRODUCTION
•An entomopathogenic fungus is a fungus that can
act as a parasite of insects and kills or seriously
disables them (Singkaravanit et al., 2010).
•It is a form of microbial control
•Here virulence is caused by contact and action is
through penetration(Nadeau et al., 1996)
•Main aim of insect control is to keep the population
of insect below economic threshold level(ETL)
•They are effective against eggs, larvae,
intermediate stages and adults of a variety of
insects including locusts, grasshoppers,
mosquitoes, and others

4. Impact of
Entomopathogenic
fungi

5. ENTOMOPATHOGENIC FUNGI
(Moore et al.,1996)
homoptera
Lepidoptera
Diptera
Hemiptera
Coleoptera
Orthoptera
Dermaptera
Hymenoptera

6. Deuteromycetes
/fungi
imperfecti
Zygomycetes Ascomycetes Basidiomycetes
Beauveria Coelomomyces Cordyceps Septobasidium
Metarhizium Entomophthora - -
Lecanicillium Massospora - -
Paecilomyces - - -
Hirsutella - - -
Nomurae - - -
EXAMPLES OF SOME IMPORTANT
ENTOMOPATHOGENIC FUNGI

7. MODE OF ACTION OF FUNGUS
• Entry of fungi is through the integuments. (Wang et al.,
2005; Cho et al., 2006)
•Infective unit- spore (conidium).
•Germination of conidia and formation of appresoria.
•Penetration of cuticle by enzymatic as well as
mechanical action(Hajek and St.Leger,1996).
•Complete invasion.
•Production of conidiophores by erumpent of cuticle.
•Death of the host by obliteration of tissues as well
through the toxins produced.

8. Healthy insect’s cuticle
Mycelium formation
Insect covered
with mycelium
Spore formation
and death of the
insect
Spore discharge
conidium
Germination of conidiaAppresorium formation and
formation of infection peg
mycelium LIFE CYCLE OF FUNGUS

9. SYMPTOMS SHOWN BY
INSECTS ON INFESTATION BY
FUNGUS
•Loss of appetite
•Attempt to climb higher up
•General/partial paralysis
•Discolored patches on integuments
•Body hardens and the insect is in upright on its leg at
the time of death
•Specifically we uses the term “Mycoses” for such
changes in insects and can be seen in – Lepidoptera,
Hemiptera, Hymenoptera , Coleoptera and Diptera

10. Enzymes produced
1.Chitinase
2.Chitosanase
3.Chitobiase
4.Lipases
The successfulness of infection was directly
proportional to secretion of exoenzymes
(Khachatourians, 1996).
5.Phospholipase
6.Proteases
7.Peptidases

11. PATHOGENECITY OF
ENTOMOPATHOGENIC FUNGI
The extensive transcriptomic and genetic study
(Freimoser et al., 2003; Cho et al., 2007) of
entomopathogenic fungal infection process has
revealed that the genes governing following
characters has important role in pathogenecity -
•Chitinases(Fang et al., 2005),
•Guanine nucleotide-binding proteins and its
regulator(Fang et al.,2007),
•Adhesin which helps in attachment of spore(Wang &
St. Leger, 2007),
•A perilipin-like protein(Wang & St. Leger, 2006).
•Subtilisin protease(St. Leger et al., 1996)

12. TOXIN PRODUCED
•Toxin produced are the by products of metabolism and
are not primarily used by the producer for killing the
insect .
•Entomogenous fungi are known to produce “Destruxins”
and “Aflatoxins”
•They are the only mycotoxins detected in the insect
body at advance stages of infection in sufficient
quantities to cause death.

13. Tritrophic interaction
•Tritrophic effects-reason behind the success of
entomopathogens(Cory J,S.& Ericsson J,D.2009)
•Plant-mediated effects on fungal
entomopathogens.
•Plants could affect fungal entomopathogens
either directly or indirectly-
1. Direct effects means anything emanating from or
produced by the plant that influences fungal
infection of the insect
2. Indirect effects could occur before or after
exposure of the insect to fungus, and would
include factors that alter insect condition.

14. •Potential direct plant-mediated effects
might include:
(1) Plant exudates affecting the
conidia directly,
(2) Herbivore-induced plant volatiles
affecting sporulation or germination,
(3) Leaf topology and surface chemistry, influencing
the rate of spore acquisition by the host insect,
(4) Plant architecture altering spore persistence,
and
(5) Leaf modifications of microclimate and thus
spore germination.

15. •Indirect effects-
(1)Plant quality, either allelochemicals or nutrients,
altering insect condition (e.g. immunity) and thus
disease resistance,
(2) nutritional quality altering insect morphology (e.g.
cuticle depth) which would influence the infection
process,
(3) changes in insect growth rate, which might alter the
exposure of the insect to fungal entomopathogens, and
(4) Plant structure altering insect behaviour, and thus
fungal encounter rate.
E.g..- Leaf surface wax- Metarhizium anisopliae-
Mustard beetles (Phaedon cochleariae) on various
crucifers- Dewaxing and Surface leachates increased
germination and virulence. (Inyang et al.,1999)

16. 3.IMPORTANT
ENTOMOPATHOGENIC
FUNGI

17. 1.Beauveria bassiana

18. Beauveria spp-
• White muscardine fungus
•Discovered in 1835 as cause of the Muscardine Disease
of domesticated Silkworms(Furlong & Pell, 2005;
Zimmermann, 2007) .
• Got high host specificity
• Pest of agricultural and forest including the Colorado
potato beetle, the codling moth, and several genera of
termites, American bollworm (Thakur et al., 2010).
•Can be isolated from insect cadavers or from soil in
forested areas by using media as well as by baiting soil
with insects(Beilharz et al., 1982).

19. AGAINST
•Termites
•Thrips
• Whiteflies
•Aphids
•Grasshoppers
•Beetles
•Caterpillars
•Silkworms

20. SILK WORM LARVA

21. GRASSHOPPER

22. Spodoptera litura

23. BEETLE CICADA

24. 2.Metarhizium anisopliae

25. Metarhizium spp.-
•Earlier it was first isolated from infected larvae of
the wheat cockchafer Anisopliae austriaca in 1879
and named as Entomphthora anisopliae .
•Renamed as M. anisopliae by Sorokin in 1883
(Tulloch, 1976)
•Causes green muscardine disease.
•Pathogenic to a large number of agricultural and
forest insect species.(Ferron, 1978)

26. •Grubs of Coconut rhinoceros beetle
•Grasshopper
•Rice BPH
•Sugarcane Pyrilla
•Bollworm
AGAINST

27. RHINOCEROS BEETLE GRUB

28. GRASS HOPPER

29. RICE BPH

30. 3.Lecanicillium muscarium

31. Lecanicillium lecanii (Zimm.) Zare & W.Gams(2001)
•Previously known asVerticillium
•The fungus appears to have been first observed in
Ceylon (Sri Lanka) about 1861, on diseased Lecanium
coffeae
•Widely distributed fungus
•Controls whitefly and several aphids species,
including the green peach aphids (Myzus persicae) for
use in the greenhouse chrysanthemums( Hamlen et
al., 1979).
•Fungus attacks nymphs and adults of white fly and
stuck to the leaf underside by means of a filamentous
mycelium (Nunez et al., 2008).

32. GREEN SCALE
Effective against-
•Coffee green scale
•Other Hemipterans

33. Nomurae rileyi

34. Nomuraea sp.-
• Nomuraea rileyi can cause epizootic death in insects.
• Lepidopteran including Spodoptera litura and some
belonging to Coleoptera are susceptible to it(Ignoff,
1981) .
• The host specificity of N. rileyi and its ecofriendly
nature encourage its use in insect pest management.
•Although, its mode of infection and development have
been reported for several insect hosts such as
Trichoplusia ni, Heliothis zea, Bombyx mori,
Pseudoplusia includans

35. Larva of Spodoptera infested with Nomurae

36. Paecilomyces fumosoreus

37. Paecilomyces sp.-
•Paecilomyces fumosoroseus also called “Yellow
Muscardine” (Nunez et al., 2008) .
•Effective over Bemisia and Trialeurodes spp. in both
greenhouse and open field environments.
•Grows extensively over the leaf surface under humid
conditions that helps it to spread rapidly through
whitefly populations( Wraight et al., 2000) .
•Best for controlling the nymphs of whitefly (Kim et
al.2002).
•These fungi cover the body of whitefly with mycelial
threads and stick them to the underside of the leaves.
•The nymphs show a “feathery appearance” (Nunez et
al., 2008) .

38. • Trichoplusia ni
• Heliothis zea
• Bagrada cruciferarum
• Bombyx mori
• Anticarsia gemmatalis.
AGAINST

39. Bagrada cruciferarum infested by Paecillomyces fumosoroseus

40. Hirsutella thomsoni

41. Hirsutella thompsonii –
•Originally isolated from an eriophyid mite in Tamil
nadu
•Effective on Eriophyid mites, particularly the coconut
mite (Aceria guerreronis Keifer)
•Major crop use is in coconut plantations, but can be
used in palmyrah palm and in arecanut.
•Widespread in nature
•Not pathogenic to non-target species.
•Not shown any adverse effects on the environment

42. It is specific to the eriophyid mites
1.Coconut mite
2.Citrus rust mite
AGAINST

43. CITRUS RUST MITE

44. Coconut mite infected with Hirsutella thomsonii

45. Formulation of fungal propagules
and Usage
• They are being used in different forms like -
a. Dust
b. Mixed with water
c. Mixed with Oil
• Best method for is application of active ingredient
with oil
The formulation of propagules of fungal
entomopathogenic fungi are guided by
•Improved product shelf-life
•Bio-control efficacy,
•Physical characteristics of the product for application
For e.g.- Control of insect pests of the phylloplane -
spore suspensions are applied as spray application

46. Entomopathogenic fungi
product available in market
Product name Firm
Bio-guard rich Plantrich chemicals & biofertilizers
ltd.
Bio-power T.Stanes &company Ltd.
Racer Agri life
beavera Jai biotech industries
Brigade Kan biosys pvt Ltd.
Mycojal Pest control pvt. Ltd.
Bio-be-ba Microplex- hosted by nagarjuna
agro chemicals
Baba Multiplex bio tech pvt. Ltd.
Metabeave R b herbal agro
Beauveria bassiana

47. Product Firm
Bio-magic T.Stanes &company Ltd
Biomet rich Plantrich chemicals & biofertilizers
ltd.
Pacer Agrilife , India
Kalichakra International panacea Ltd
Cropmet Microplex- hosted by nagarjuna
agro chemicals
Metrocid Sri biotech laboratories India Ltd.
Metaz Jai biotech
Metarhizium Multiplex biotech pvt. Ltd.
Jasmeta Shri ram solvent extraction pvt Ltd.
Biostorm Varsha bioscience & technology
Metarhizium anisopliae

48. Product Firm
Bio-catch T.Stanes &company Ltd
Biovert rich Plantrich chemicals &
biofertilizers ltd.
Mealikil Agrilife , India
Vertimust Jai biotech industries
Biograde - v Kan biosys pvt. Ltd.
Vertifire - L International panacea Ltd.
Cropfit Microplex- hosted by nagarjuna
agro chemicals
Lecanicillium muscarium

49. Product Firm
Paci hit rich Plantrich chemicals & biofertilizers
Ltd.
Mysis Varsha bioscience & technology
Nematox Sri biotech laboratories India Ltd.
Paecillomyces spp.

50. •In India more than 60% of pesticide is used in agriculture
•Among chemical pesticides(Wahab,2003,2005,2009,2015)
1. Insecticides=60%
2. Fungicides + bactericides=20%
3. Herbicides =17%
4. Others =3%
• Majority of chemical pesticide not only kill the pest but also kill
many beneficial insects and non-target animals of ecosystem
(Abhilash and Singh,2009)
• Posing serious threat to human health (Maroni et al., 2006,
Wahab,2004)
• High risk to field workers and consumers
• Resistance, resurgence and persistence of insect
PRESENT SCENARIO

51. USEFULNESS IN PRESENT
SCENARIO
•By going for this method we can reduce our dependency on
chemicals.
•We can have favourable diversity of flora and fauna near
crop region by keeping non target organisms.
•No problem of biomagnification.
•No pollution of water table .
•They have shown induced systemic resistance in the host
plant (Vega, 2008 )

52. Fungus Pest & Crop Field efficacy Reference
Beauveria
bassiana
Coffee berry
borer,
Hypothenemus
hampei
Spray of B. bassiana spore
suspension(1×107
spores/ml) containing 0.1%
sunflower oil and 0.1per
cent wetting agent during
monsoon
reduced 50-60per cent
berry borer incidence in
Coorg, Karnataka
Anon.
(2001)
Sunflower:
Helicoverpa
armigera
Spray of oil suspension of
B. bassiana (200mg/l) in
Andhra Pradesh
Devi and
Hari (2009)
Green gram:
White grubs
Soil application @5×1013
conidia/ha effective
control achieved in Assam
Bhattachary
ya et
al.(2008)
Biological Control of pests using entomopathogenic fungi
in India

53. Fungus Crop and pest Field efficacy Reference
Metarhizium
anisopliae
Coconut:
Rhinoceros
beetle, Oryctes
rhinoceros
Spraying of Spores in its
breeding sites 5X1011
spores/m3 to the compost
pits and
manure heaps
Anon.
(2000)
Potato White
grub
Brahmina sp.
Soil application @ 5x1013
conidia/ha along with
chlorpyrifos 20 EC@ 200 g
a.i./ha resulted in the
highest tuber yield (155
q/ha) in HP.
Bhagat et
al. (2003)
Soyabean: white
grub
Holotrichia
longipennis
Soil application formulation
applied @ 5x1013
conidia/ha, 61.50per cent
reduction in grub
Population in MP
Pandey
(2010)

54. Sr. No Treatment Grub mortality (%)
1 Steinernema carpocapsae (80
IJ/cm²)
34.9
2 Heterorhabditis indica (80
IJ/cm²)
45.9
3 Beauveria bassiana (10 ⁶
conidia/cm2 )
34.0
4 Metarahizium anisopliae (10⁶
conidia/cm²)
74.4
5 Chlorpyriphos (0.06%) 86.4
6 Control( Untreated ) 8.5
Evaluation of entomopathogenic fungi and EPNs
against Apple root borer Dorysthenes hugelii at
YSPUHF, Solan
(Anonymous:Annual progress report (2012-13) of AICRP
on biological control of crop pest and weed)

55. Fungus Crop and pest Field efficacy Refere
nce
Lecanicilli-
um lecanii
Coffee green
scale,
Coccus viridis
Spraying spores@ 16 X 106
spores/ml along withTween-80
twice at 2 weeks interval caused
97.6 per cent mortality of the pest
Jayaraj
(1989
Citrus green
scale
Coccus virids
Spraying of spore (2x106spores/ml)
along with 0.005 per cent
Quinalphos and 0.05per cent
Teepol was found highly effective
killing 95.58 percent and
97.55percent scales in coffee and
citrus
respectively
Singh
(1995)
Indian mustard
and
Rapeseed:
Mustard aphid,
Lipaphis
erysimi
Spray @ (106spores/ml). There was
a significant reduction in aphid
infestation at 10 DAS
Rana
et al.
(2002

56. Fungus Crop and pest Field efficacy Reference
Nomurae rileyi Castor:
Spodoptera litura
in AP
Spraying of spore
(10x1010
spores/ml) along
with 0.02per cent
Tween-80
Vimala Devi and.
Prasad (1997)
Soybean:
Spodoptera
litura,
Helicoverpa
armigera,
Thysonoplusia
orichalcea
N. rileyi spores
spraying @ 2X106
/ml twice at10
days interval
during Kharif in
North Karnataka
was cheaper than
insecticidal
treatment and cost
effective
Lingappa et al.
(2002)

57. ADVANTAGES
Harmless to other forms of life
High degree of specificity
Compatible with many chemical insecticide
Resistance to microbes is less likely to develop
Self sustaining so economical
Easy application
Aesthetically acceptable

58. DISADVANTAGES
Specificity is disadvantageous as in some cases only
single insect is not involved
Totally dependent on environment, so uncertainty is
always there
Not quick results ,as establishment takes time

59. CONSTRAINTS
•A hi-tech venture terms of safety and sustenance
•Viability and virulence of fungal inoculum are the pre-requisite
threshold for their efficacy (Doust & Roberts, 1983).
•An array of abiotic stresses like temperature (Rangel et al., 2005a),
UV radiations (Rangel et al., 2006a), humidity (Lazzarini, 2006),
edaphic factors and nutrient source (Shah,2005) affect negatively the
field use of entomopathogens as bio-control agents
•Soil temperature is a major factor, that affects the success or failure
in the establishment and production of fungal inoculums (Thomas &
Jenkins, 1997)

60. Eg. 1) M. anisopliae has temperature tolerance upper limit as 37-
40ºC (Thomas & Jenkins, 1997).
2) B. bassiana on the other hand can survive up to a maximum
temperature of 37 ºC (Fargues et al., 1997)..
Environmental factors affect pathogenicity as well as mode of
virulence of entomopathogenic fungi (Hasan, 2014).
•They should be tolerant to the soil temperature but also have to
survive through thermoregulatory defense response of the host insect
•Dry heat exposure causes DNA damage leading to depurination
resulting into mutation (Nicholson et al., 2000)
•Wet heat i.e. heat in conjunction with high humidity causes protein
denaturation and membrane disorganization.

61. The application of entomopathogenic fungi for insect control is
increasing largely because of
•Greater environmental awareness
•Food safety concerns
•The failure of conventional chemicals due to an increasing
number of insecticide resistant species.
•They provides us significant and selective insect control.
CONCLUSION