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Cucumber Mosaic Virus Management in Cucumber Mosaic Virus Management in
Snap Beans and PeppersSnap Beans and Peppers
Russell L. GrovesRussell L. Groves
Department of EntomologyDepartment of Entomology
537 Russell Laboratories537 Russell Laboratories
1630 Linden Drive1630 Linden Drive
Madison, WI 53706Madison, WI 53706
[email protected]@entomology.wisc.edu
Central Wisconsin, Processing Crops ConferenceCentral Wisconsin, Processing Crops Conference
March 12, 2008March 12, 2008
New Insect Vector, Soybean aphidNew Insect Vector, Soybean aphid
Lee 2002
Aphis glycines, soybean aphid
2005 Distribution
Seasonal Ecology and DispersalSeasonal Ecology and Dispersal
Soybean aphidSoybean aphid
Overwinters on RhamnusOverwinters on Rhamnus
spp.spp.
Colonizes soybeanColonizes soybean
MidMid--summer dispersal summer dispersal ––
crowding, host qualitycrowding, host quality
Soybean aphids Soybean aphids -- soybeansoybean
Action Threshold onAction Threshold on
soybean: 250 / plantsoybean: 250 / plant
NonNon--circulative (CMV, WMVcirculative (CMV, WMV--2, PRV, ZYMV)2, PRV, ZYMV)
often referred to as “stylet-borne”
NonNon--propagativepropagative
Phloem
Hindgut
ASGPSG
Midgut
FoodCanal
SalivaryCanal
Haemocoel
NonNon--Persistent Virus TransmissionPersistent Virus Transmission
NonNon--Persistent Transmission: Movement Persistent Transmission: Movement
in Insectsin Insects
IngestionIngestion SalivationSalivation
Food IngestionFood Ingestion
Pathogen particles attach
to maxillary lumen
EgestionEgestion
Pathogen particles released
with saliva
Acquisition time Acquisition time -- time required to acquire pathogentime required to acquire pathogen
•• SecondsSeconds
Inoculation time Inoculation time -- time required by infectious insect to inoculate a time required by infectious insect to inoculate a susceptible hostsusceptible host
•• SecondsSeconds
Latent period Latent period -- (minimum time between acquisition of a pathogen and (minimum time between acquisition of a pathogen and ability to transmit) ability to transmit)
•• ZeroZero
Retention time Retention time -- time after acquisition that a time after acquisition that a
vector remains capable of transmitting the virusvector remains capable of transmitting the virus
•• Minutes Minutes –– hour; time to fly to next planthour; time to fly to next plant
Nonpersistent TransmissionNonpersistent Transmission
Chemical controls = no option!!Chemical controls = no option!!
Major Viruses:Major Viruses:
Cucumber mosaic virus (CMV)
Watermelon mosaic virus-2 (WMV-2)
Papaya ringspot virus (PRV - WMV-1)
Zucchini Yellow Mosaic Virus (ZYMV)
Minor Viruses:Minor Viruses:
Alfalfa mosaic virus (AMV)
Clover yellow vein virus (ClYVV)
Beet curly top virus (BCTV)
Beet pseudo-yellows virus (BpYV)
Emerging Virus ComplexEmerging Virus Complex
CMV CMV –– CucumberCucumber
Watermelon mosaic virus Watermelon mosaic virus -- 22
Cucumber Mosaic Virus (CMV)Cucumber Mosaic Virus (CMV)
Widely distributedWidely distributed
NonNon--crop inoculum sourcescrop inoculum sources
Stunting, leaf curlStunting, leaf curl
Elongate, shoestring leafElongate, shoestring leaf
Color breakingColor breaking
CMV CMV -- PeppersPeppers
CMV CMV –– Snap beanSnap bean
CMV CMV –– CucumberCucumber
Emerging bean viruses: Emerging bean viruses:
the problem (2003)the problem (2003)
German et al. (2004)
0
10
20
30
40
50
60
70
80
Wisconsin Snap Bean Survey, 2003Wisconsin Snap Bean Survey, 2003
CentralCentral NewNew SpringSpring Door Co.Door Co. Oconto Co.Oconto Co.
Sands Sands Richmond Richmond GreenGreen
CMVCMV
AMVAMV
BCMVBCMV
BYMVBYMV
ClYVVClYVV
59.1%59.1%
2.9%2.9%
43.3%43.3%
2.8%2.8%0.6%0.6%
78.4%78.4%
22.8%22.8%
53.2%53.2%
22.4%22.4%
18.9%18.9%
0.8%0.8%
20.0%20.0%
0
10
20
30
40
50
60
70
80
90
Wisconsin Snap Bean Survey, 2006Wisconsin Snap Bean Survey, 2006
CentralCentral SpringSpring Manitowoc Co.Manitowoc Co.
Sands Sands GreenGreen
CMVCMV
AMVAMV
BCMVBCMV
BYMVBYMV
ClYVVClYVV
72.0%72.0%
3.9%3.9%
81.9%81.9%
3.1%3.1%
88.4%88.4%
12.8%12.8%
0.9%0.9% 1.2%1.2%
7.2%7.2%
Emerging Bean Viruses: Emerging Bean Viruses:
The problem (2006)The problem (2006)
Groves (2006)
0
10
20
30
40
50
60
70
80
90
100
Wisconsin Snap Bean Survey, 2007Wisconsin Snap Bean Survey, 2007
CentralCentral NewNew SpringSpring Door Co.Door Co.
Sands Sands Richmond Richmond GreenGreen
CMVCMV
AMVAMV
BCMVBCMV
BYMVBYMV
ClYVVClYVV
93.2%93.2%
1.3%1.3%
78.3%78.3%
11.1%11.1%
80.5%80.5%
90.2%90.2%
0.5%0.5% 2.7%2.7%
Emerging Bean Viruses: Emerging Bean Viruses:
The problem (2007)The problem (2007)
Groves (2007)
Emerging Pepper Viruses: Emerging Pepper Viruses:
Locations and Cultivars, 2007Locations and Cultivars, 2007
CMV CMV –– Jalapeno,Jalapeno,
mature leafmature leaf
CMV CMV –– Bell pepperBell pepper CMV CMV –– JalapenoJalapeno
fruitfruit
LL
Warm, Moist AirWarm, Moist Air
Colder, Dry AirColder, Dry Air
Hypothesis I. Long Distance TransportHypothesis I. Long Distance Transport
Viruliferous Soybean AphidsViruliferous Soybean Aphids
Long Distance TransportLong Distance Transport
Viruliferous Soybean AphidsViruliferous Soybean Aphids
CMVCMV--infected aphidinfected aphid
NonNon--infected aphidinfected aphid
Hypothesis II. Long Distance Aphid Dispersal:Hypothesis II. Long Distance Aphid Dispersal:
Local InoculumLocal Inoculum
CMVCMV--infected aphidinfected aphid
NonNon--infected aphidinfected aphid
Affects a number of importantAffects a number of important
vegetable and ornamental plantsvegetable and ornamental plants
(tomato, cucumber, squash, pepper,(tomato, cucumber, squash, pepper,
snap beans)snap beans)
Is transmitted nonIs transmitted non--persistentlypersistently
by several aphid vectors and by several aphid vectors and
through seed.through seed.
Very broad host range (> 1,000 spp.) Very broad host range (> 1,000 spp.)
and can overwinter in several and can overwinter in several
perennial weed speciesperennial weed species
Epidemiology of CMVEpidemiology of CMV
BurdockBurdock
DandelionDandelion
GoldenrodGoldenrod
NonNon--Crop Weed Crop Weed
Sources of CMV InoculumSources of CMV Inoculum
0 10 20 30 40 50 60 70
lead plant (26)
kura clover (12)
white clover (167)
hairy vetch (22)
alfalfa (209)
milkweed (86)
lambs quarter (16)
alsike (22)
birdsfoot trefoil (42)
sweet clover (26)
silene (55)
eastern nightshade (56)
red clover (181)
Pla
nt
sp
ec
ies
Pla
nt
sp
ec
ies
Incidence (%)Incidence (%)
CMVCMV AMVAMV
Mueller and Gratton, 2007Mueller and Gratton, 2007
Research ObjectivesResearch Objectives
Compare the genetic structure of CMV isolates collected Compare the genetic structure of CMV isolates collected
from (1) infected crops (beans and peppers), (2) dispersing from (1) infected crops (beans and peppers), (2) dispersing
aphids, and (3) potential reservoir hosts.aphids, and (3) potential reservoir hosts.
To identify and characterize the seasonal incidence and To identify and characterize the seasonal incidence and
pathopatho--type profile of CMV isolates of the primary aphid type profile of CMV isolates of the primary aphid
vectorsvectors..
To accurately identify the primary reservoir hosts of CMV To accurately identify the primary reservoir hosts of CMV
in and around selected snap bean fields and to determine in and around selected snap bean fields and to determine
those species, or set of species, which have the greatest those species, or set of species, which have the greatest
epidemiological importance.epidemiological importance.
I. Genetic Structure of CMVI. Genetic Structure of CMV
lambslambs--
quartersquarters
eastern blackeastern black
nightshadenightshade
5`5` 3`3`1a1a
5`5` 3`3`3a3a 3b3b
RNA1 (3.4 Kb)RNA1 (3.4 Kb)
RNA2 (3.1 Kb)RNA2 (3.1 Kb)
RNA3 (2.2 Kb)RNA3 (2.2 Kb)
5`5` 3`3`2a2a
2b2b
CMV is tripartite, + sense RNA, with 5 ORF’sCMV is tripartite, + sense RNA, with 5 ORF’s
RNA2RNA2
milkweedmilkweed
ORF 2b involved in grouping by host associationORF 2b involved in grouping by host association
RNA extracted from CMV symptomatic plants:RNA extracted from CMV symptomatic plants:
RTRT--PCR: (Lin PCR: (Lin et al.et al. 2004) 2004)
-- ORF 2b (370 bp)ORF 2b (370 bp)
-- ORF 3’ NTR (315 bp)ORF 3’ NTR (315 bp)
-- ORF CP (678 bp)ORF CP (678 bp)
PCR fragments cloned, sequenced, PCR fragments cloned, sequenced,
and characterizedand characterized
I. Characterizing CMV host associationI. Characterizing CMV host association
370 bp
1 2 3 41 2 3 4 1 2 3 41 2 3 4 1 2 3 41 2 3 4
ORF: 2b 3’ NTR CPORF: 2b 3’ NTR CP
RNA2 (3.1 Kb)RNA2 (3.1 Kb)
2a2a
2b2b
2a2a
2b2b
2a2a
2b2b
Determine similarity / dissimilarity of CMV isolates within Determine similarity / dissimilarity of CMV isolates within
affected fields.affected fields.
I. Characterizing CMV host associationI. Characterizing CMV host association
CMV-infected
Healthy
2a2a
2b2b
Determine similarity / dissimilarity of CMV isolates withinDetermine similarity / dissimilarity of CMV isolates within
and among affected fields.and among affected fields.
I. Characterizing CMV host associationI. Characterizing CMV host association
CMV-infected
Healthy
Determine similarity / dissimilarity of CMV isolates amongDetermine similarity / dissimilarity of CMV isolates among
affected crops.affected crops.
I. Characterizing CMV host associationI. Characterizing CMV host association
CMV-infected
Healthy
snap beansnap bean pepperpepper
CMVCMV--infected infected
pepper, 2007pepper, 2007
II. Characterizing CMV host II. Characterizing CMV host –– vector(s) vector(s)
associationsassociations
Weekly captures of dispersing aphid species.Weekly captures of dispersing aphid species.
D. Voegtlin, Illinois Natural History SurveyD. Voegtlin, Illinois Natural History Survey
Acyrthosiphon pisumAcyrthosiphon pisum "Pea aphid""Pea aphid"
Aphis craccivoraAphis craccivora "Black legume aphid""Black legume aphid"
Aphis glycines Aphis glycines "Soybean aphid""Soybean aphid"
Aphis gossypiiAphis gossypii "Cotton"Cotton-- melon aphid"melon aphid"
Aphis helianthiAphis helianthi "Sunflower or dogwood aphid""Sunflower or dogwood aphid"
Aphis nasturtiiAphis nasturtii "Buckthorn "Buckthorn -- potato aphid"potato aphid"
Aphis spiraecolaAphis spiraecola "Spiraea aphid""Spiraea aphid"
Brachycaudus helichrysiBrachycaudus helichrysi "Leaf curling plum aphid""Leaf curling plum aphid"
Lipaphis pseudobrassicaeLipaphis pseudobrassicae "Turnip aphid""Turnip aphid"
Macrosiphum euphorbiaeMacrosiphum euphorbiae "Potato aphid""Potato aphid"
Myzus persicaeMyzus persicae "Peach potato aphid""Peach potato aphid"
Rhopalosiphum insertumRhopalosiphum insertum "Apple grass aphid""Apple grass aphid"
Rhopalosiphum maidisRhopalosiphum maidis "Corn leaf aphid""Corn leaf aphid"
Rhopalosiphum padiRhopalosiphum padi "Bird cherry"Bird cherry--oat aphid"oat aphid"
Schizaphis graminumSchizaphis graminum "Greenbug""Greenbug"
Sitobion avenae Sitobion avenae "English grain aphid""English grain aphid"
Therioaphis trifoliiTherioaphis trifolii "Spotted Alfalfa aphid""Spotted Alfalfa aphid"
Acyrthosiphon pisumAcyrthosiphon pisum "Pea aphid""Pea aphid"
Aphis glycines Aphis glycines "Soybean aphid""Soybean aphid"
Rhopalosiphum maidisRhopalosiphum maidis "Corn leaf aphid“"Corn leaf aphid“
Rhopalosiphum padiRhopalosiphum padi "Bird cherry"Bird cherry--oat aphid“oat aphid“
Sitobion avenae Sitobion avenae "English grain aphid“"English grain aphid“
Therioaphis trifoliiTherioaphis trifolii "Spotted Alfalfa aphid""Spotted Alfalfa aphid"
II. Characterizing CMV host II. Characterizing CMV host –– vector(s) vector(s)
associationsassociations
Assess the genotype profile(s) of CMV amplified fromAssess the genotype profile(s) of CMV amplified from
dispersing aphids.dispersing aphids.
snap beansnap bean
Acyrthosiphon pisumAcyrthosiphon pisum "Pea aphid""Pea aphid"
Aphis glycines Aphis glycines "Soybean aphid""Soybean aphid"
Rhopalosiphum maidisRhopalosiphum maidis "Corn leaf aphid“"Corn leaf aphid“
Rhopalosiphum padiRhopalosiphum padi "Bird cherry"Bird cherry--oat aphid“oat aphid“
Sitobion avenae Sitobion avenae "English grain aphid“"English grain aphid“
Therioaphis trifoliiTherioaphis trifolii "Spotted Alfalfa aphid""Spotted Alfalfa aphid"
II. Characterizing CMV host II. Characterizing CMV host –– vector(s) vector(s)
associationsassociations
Assess the genotype profile(s) of CMV amplified fromAssess the genotype profile(s) of CMV amplified from
dispersing aphids.dispersing aphids.
snap beansnap bean
III. Accurate identification of CMV III. Accurate identification of CMV
inoculum sourcesinoculum sources
Field cornField corn
SoybeansSoybeans
AlfalfaAlfalfaCloverClover
Single source of inoculumSingle source of inoculum
III. Accurate identification of CMV III. Accurate identification of CMV
inoculum sourcesinoculum sources
Field cornField corn
SoybeansSoybeans
AlfalfaAlfalfaCloverClover
Multiple sources of inoculumMultiple sources of inoculum
Project Goals / Expected OutcomeProject Goals / Expected Outcome
Improve our understanding of the epidemiology of CMV Improve our understanding of the epidemiology of CMV
Determine which vector, or set of vectors, contribute to Determine which vector, or set of vectors, contribute to
disease cyclesdisease cycles..
To accurately identify the primary reservoir hosts of CMV To accurately identify the primary reservoir hosts of CMV
in and around affected snap bean fields and determine in and around affected snap bean fields and determine
those species, or set of species, which those species, or set of species, which
have the greatest epidemiological importance.have the greatest epidemiological importance.
2a2a
2b2b
QUESTIONS?QUESTIONS?
