Entomopathogenic fungi: Biological control of arthropod pests in ...
New Strategies for Using Entomopathogenic Fungi to · 2016. 5. 3. · New Strategies for Using...
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New Strategies for Using Entomopathogenic Fungi to
Benefit Agriculture
Stefan Jaronski USDA Agricultural Research Service,
Northern Plains Agricultural Research Lab Sidney MT USA
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• Earliest reports of
insect disease:
• 700 BC: China
• 322 BC- Aristotle
– “Historia
Animalium”
• 29-32 BC- Virgil
– “Georgica”
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data, Courtesy BPIA
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Courtesy Bill Stoneman, Biopesticide Industry Alliance
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Courtesy Bill Stoneman, BPIA
One reason: Fewer chemicals, more expensive
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Chromobacterium subtsugae
Paenibacillus lentimorbis
Pasteuria penetrans
Paranosema locustae
Lagenidium giganteum
Beauveria bassiana
Metarhizium anisopliae
Isaria fumosorosea
Current US Microbial Insecticides
Granulosis Virus
NucleoPolyhedrosis Virus
Bacillus popilliae
Bacillus thuringiensis kurstaki
B thur. aizawai
B thur. israelensis
Bacillus sphaericus
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Agraquest
Agrivir
Anatis
Bioprotection
Arvesta
Becker
Microbials
Biotepp
Certis
JABB
LAM Intl
Marrone
BioInnovations
M&R Durango
Novozymes
Biologicals
Planet Natural
Reuter
Troy Biosciences
Valent Biosciences
U.S. Companies
with microbial products
AgBiochem
Arysta Lifescience NA
AZ Cotton Research &
Protection Council
Bayer Crop Science
Bioworks
Circle One Global Inc.
Growth Products Ltd
Jet Harvest Systems
Montana Microbial Products
Myco-Forests Corp
Mycologic Inc.
Natural Industries Inc.
Nufarm
NW Agricultural Products
Premier Horticulture Inc
Prophyta
Sylvan Bioproducts
Verdera Oy
OmniLytics
Bayer BASF Syngenta Monsanto
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B. brongniartii
5%
B. bassiana
40%
M. anisopliae
39%
H. thompsonii
1%
M. acridum
3%
I. fumosorosea
6%
L. longisporium
2%I. farinosus
1%
L. muscarium
3%
Mycoinsecticides: 110 active, commercial products in 2006 ~200 in 2017
Faria and Wraight Biological Control 43 (2007) 237–256
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Why are the fungi so popular?
• Benign, for
people,
environment
• Easily mass-
produced
• Easily applied,
just like a chemical…
• Invite the
imagination?
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Entomopathogenic Ascomycetes
© Koppert
Isaria Lecanicillium
Metarhizium
Beauveria
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Dispersion
of
Aerial
Conidia Germination &
Penetration thru
Cuticle
Transformation upon
Insect’s Death
Typical Life Cycle
(They act as “contact agents”)
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So how are we using these
microbials?
As inundative
(albeit biological)
catastrophic
density independent
mortality factors
i.e. like chemicals …
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A fact of life and death
• A single spore does not a lethal infection make • LD90 Beauveria GHA 100 spores/mm2 (Whitefly) 28 spores/mm2 (Diamondback Moth)
• 10,000,000,000 mm2 in a hectare 80,000,000,000 mm2 in ha canopy w/ L.A.I. of 8
= 2.24x1012 spores (DBM), 8x1012 spores (WF) / ha
= $10.08/ha $36/ha
One needs a LOT of spores
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AND, Delivery of fungus spores is inefficient often VERY INEFFICIENT
Therefore,
DBM: $50/ha WF: $180/ha Per spray
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How to make fungi better, cheaper?
Deliver them more efficiently with better
spray approaches
ULV oil sprays
High volume orchard sprayer
Motorized mist blower
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How to make fungi better, cheaper?
Creative approaches in application to improve
coverage
Jaronski 2010
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How to make fungi better, cheaper?
Oil-based formulation increases efficacy 8900
1,00E+06
1
10
100
1000
10000
100000
1000000
Oil Water
LD5
0 c
on
idia
/in
sect
Bateman et al . 1993. Annals Appl Biol
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How to make fungi better, cheaper?
Improve persistence via formulations
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BotaniGard 22WP
+ 0.06-0.08% Silwet L77®
Spores penetrated in substantial
numbers into 5-6th petals of unopened
flower
Control much better than 22WP alone
How to make fungi better, cheaper?
Make application more efficient thru formulation
additives
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How to make fungi better, cheaper?
Hydromulch with conidia or esp. microsclerotia applied to tree bark (Taryn Goble, Cornell)
Make application more efficient thru novel
formulations
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How to make fungi better, cheaper?
Nutritive granule
Microsclerotial granule
Sweet whey microfactory on leaf surface
Magnify the rate of fungus by allowing it to
reproduce in the field
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How to make fungi better, cheaper?
Make application more efficient thru novel
formulations
Carnuba wax carrier e.g. Entostat®;
or Candelilla wax powder
Exosect Ltd.
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How to make fungi better, cheaper?
Combine chemical stressors with fungi
to stress insects’ immunity
alter behavior
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Make (get) a ‘better’ microbe
How?
Traditional • Screening for the ‘best’ isolates
– Let Nature provide
• Classical mutation selection – tradeoffs
Novel • Transgenic approaches: virulence factors,
enhanced detox mechanisms
– BUT, regulatory, societal challenges
How to make fungi work better, cheaper?
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How to make fungi better, cheaper?
Bring the insect to the microbe
Alginate beads with Metarhizium or Beauveria spores and CO2-releasing yeast to attract wireworms to a fatal candy Vemmer et al 2016
• Fatty acids attractive to
grasshoppers mixed with
fungus spores sprayed in
strips
Jaronski and Lockwood
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Use the insect to vector the microbe to its
kin
How to make fungi better, cheaper?
• Japanese beetle
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Use another insect to transfer the microbe to
where it’s needed
How to make fungi better, cheaper?
• Honeybees vectoring Beauveria to flowers
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Take advantage of target insect’s behavior to
increase transfer efficiency:
How to make fungi better, cheaper?
• Put the spores where larvae
(e.g. cherry fruit fly) larvae
fall to pupate
• Spray mosquito resting
habitat (resting boxes
in urban area) with
spores
• Spray treebark over which
gypsy moth larvae ,
Asian longhorned beetles
must crawl USDA
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And the newest possibilities
The insect pathogenic fungi,
Beauveria, Metarhizium, Isaria
as plant endophytes
Beauveria are endophytic in maize, cocoa, date palm, coffee, grapes,
tomato, banana, sorghum, medicinal poppy, jute,
broad bean, cassava, cotton, strawberries,
wheat …
Metarhizium are endophytic in rape, beans, switchgrass, yew, rice;
more famously associated with root systems.
Gomez-Vidal et al., 2006
Micron 37 (2006) 624–632
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Beauveria, Metarhizium, Isaria
as endophytes
Can be artificially introduced
in at least some plants
- Foliar application
- Seed treatment
- Root dip
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Courtesy Anant Patel, Bielefeld University
Visualization of the concept by one university-industry group
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How do these endophytic fungi act on an
insect pest?
Direct infection of insects
Indirect effect – secreted metabolites
from Quesada-Moraga et al. 2006
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Induction of Systemic Resistance,
to herbivores, by endophytes
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Induction of Systemic Resistance, to
herbivores
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Endophytic colonization by, or more likely, exposure to Beauveria and Trichoderma affected survival of two leaf miners in beans
Induction of Systemic Resistance, to
herbivores — leaf miners and beans
Akutse et al. 2013 Fungal Ecology 6, 293 – 301
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And more…
Metarhizium anisopliae LHL07-inoculated
soybean plants displayed significantly
- higher shoot length,
- shoot fresh and dry biomass,
- chlorophyll contents,
- transpiration rate,
- photosynthetic rate and leaf area,
under salt-induced salt stress as compared
to non-inoculated control plants.
Metarhizium LHL07
elevated proline and
reduced superoxide
dismutase and
malondialdehyde,
reduced abscisic acid and
elevated jasmonic acid
levels Phytohormone Effects?
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Induction of Systemic Resistance
without endophytism does occur
”
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Induction of Systemic Resistance
PathogenAssocMPs
MicrobialAMPs
DamageAMPs
HerbivoreAMPs
Plants can recognize molecular patterns
Microbes on plant’s cuticle are a molecular pattern …
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Newman et al 2013 Frontiers in Plant Science Vol. 4 Art. 139
There are all sorts of molecular patterns ‘perceived’ by plants
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Beyond just using
entomopathogenic fungi…
Manipulating
the existing crop plant microbiome
for better plant health
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10,000,000 microbes / cm2 leaf surface;
10,000,000,000 microbes, 30,000 species / gram
root associated soil
100s microbe species living in xylem, phloem
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A plant can be a crowded ‘hotel.’
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There is no such thing as a “magic bullet”
(If you think there is,
I would like to sell
you a bridge - cheap).
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Microbials Predators & parasites
Cultural practices
Chemicals
Physical measures
Plant resistance
Pheromones/repellents
Sterile insects
Integrated Pest Management
Many tools used as a system
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Integrated Pest Management (U.S. National IPM Network).
“Integrated Pest Management (IPM) is a
• sustainable approach
• to managing pests
• by combining biological, cultural, physical and chemical tools in a way that
• minimizes economic, health, and environmental risks”.
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INFONET Biovision
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Microbials Predators & parasites
Cultural practices
Chemicals
Physical measures
Plant resistance
Pheromones/repellents
Sterile insects
Integrated Pest Management
Many tools used as a system
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e.g., Integration of fungi with parasitoids
and/or predators
1. Beauveria with Eretmocerous mundus Bemisia tabaci
(Jaronski et al 1995)
2. Beauveria with Aphidius matricariae Aphis glycines (Rashki
et al 2009)
3. Metarhizium with Orius albidipennis Thrips tabaci
(Purian et al 2011)
4. Beauveria with Anthocorous nemorum (Meyling et al. 2006)
5. Beauveria and Metarhizium with Hadrobracon hebetor
Helicoverpa armigera (Mahdavi et al 2013)
6. Beauveria with Diglyphus hesperus and Encarsia formosa
Trialeurodes vaporariorum (Labbe et al 2009)
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Integration of Metarhizium with Cover Crop
(Majumdar et al.)
6.1
4.5
6.7
4.5
5.6 5.5
4.5
5.1
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
Rye
3OBE
Rye
3OBE
Ma G
Ma G Rye
3OBE
Ma S
Ma S UTC Rye
3OBE
Ma G
Ma G
Treatment
Ro
ot
Dam
ag
e I
nd
ex (
0-9
)
p=.06p<.0001p<.0001
2002
Plots had significantly lower root damage with Metarhizium when integrated with rye cover crop. OBE: Oat-Bushel- Equivalents
e.g., Integration with cover crops
(sugar beets)
2003
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Strawberry IPM system — UC Dept of Agriculture & Natural Resources Coop Extension
Another example:
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Week 1: introduce Dacnusa for leafminers
Week 2: early in week, Beauveria for thrips late in week, apply fungicides
Week 4: Apply predaceous mites Apply Beauveria
Week 6-8: Spray Beauveria weekly Apply Bt for lepidoptera, as needed Apply cinnamaldehyde (cinnamon oil) only to mite “hotspots”
Week 8: Introduce Diglyphus for leafminers
IPM with microbial in chrysanthemums
And this program was designed by a farmer
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Resistance/ Tolerance
Disease Biocontrol
Insect / nematode Biocontrol
Cultural Practices
Acquired Systemic Resistance
Chemical Inputs
(limited)
Ultimate goal is
An integrated, bio-based
pest and pathogen management system
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Thank you for your attention