Virus Diseases of Small Fruit Crops Raspberry and Blackberry - USDA-ARS-HCRL - Utah State Horticulture Association Utah Berry Growers Annual ...
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Virus Diseases of Small Fruit Crops
Raspberry and Blackberry
Utah State
Horticulture Association
Utah Berry Growers
Annual Winter Meeting
January 23, 2008
Provo, Utah
Bob Martin
USDA-ARS-HCRL
Bob.Martin@ars.usda.gov
Virus vector is primary consideration
in developing control strategies
1. Pollen-borne viruses
2. Nematode transmitted viruses
3. Aphid transmitted viruses
4. Whitefly transmitted viruses
5. Mechanically transmitted viruses
6. Viruses with unknown vectors
Control of Viruses in Utah 1. What viruses are present? A. In commercial plantings B. In native hosts of related species
Raspberry bushy dwarf virus
Tobacco streak virus
1. Pollen-borne to maternal tissue,
spread within a field.
2. Bees assist transmission,
within a field and between fields.
3. Seed transmitted, spread to next generation,
concern to breeders not to growers.
4. Contaminated nursery stocks, can spread anywhere
in the world in a few days.
5. Pollen from native hosts, not sure how important
this is since we do not know the mechanism of
transmission from pollen to maternal tissue.
RBDV-Infected Field Washington
Raspberry bushy dwarf virus
Detection by ELISA
4 3 7 0
Rate of Increase in the Incidence
of RBDV in ‘Meeker’
RBDV Infection Yr. Planted
Northern WA 1996 1998
Field 1 0/72 49/88 1995
Field 2 0/72 15/88 1996
Field 3 2/72 73/88 1994
Oregon/Southern WA 1996 1998
Field 1 4/72 4/72 1984
Field 2 0/72 0/72 1993
Field 3 5/72 4/72 1986
Autumn Bliss
In many cultivars crumbly fruit
is the only symptom of
Raspberry bushy dwarf virusOregon
RBDV in ‘Marion’ Blackberry First observed in field plots in 1997. Yield studies 17/15 paired plants in 1999/2000. Found in growers’ fields 2000. First commercial field taken out due to RBDV in 2004.
Effect of RBDV in ‘Marion’
Blackberry
Drupelet number reduced by >40%.
Fruit size reduced by >30%.
Yield reduced by 40-50%.
No effect on cane number or length.Control Strategies for RBDV 1. Use certified virus-tested planting stocks. 2. Use resistant cultivars if available. 3. Isolation from infected fields or native hosts. 4. Plant large blocks for virus-free raspberries. 5. Transgenic approach, works but not accepted by the public.
Cranberries a symptomless host for BlScV
Tomato ringspot virus
Tobacco ringspot virus
1. Nematode transmitted, short distance
in the field.
2. Moved by equipment in clumps of soil,
movement within and between fields.
3. Infects many broadleaf weeds, inoculum
in a field.
4. Seed-transmitted, move in weed seed by wind,
birds etc, and to next generation.
5. Moved in planting stocks, can move anywhere
in the world in a few days.Stand Decline Caused by ToRSV
ToRSV
The nematode feeds at the root tip causing the root to stop growing Virus attaches to lining of the esophagus. When the nematode feeds, the virus is injected into plant cell. The virus is lost with each molt.
Arabis
mosaic virus
Odontostyle
Odontophore
Challenge is to
detect these.Nematodes on the Move
30 cm
30 cmControl of Nematode-Borne Viruses 1. Certified plants free of these viruses. 2. Immune cultivars. 3. Roguing of diseased bushes, spot treating. 4. Soil fumigation – temporary control. 5. Crop rotation/fallow – forget the nematode/target the virus. 6. Minimize soil movement on equipment – cover crops. 7. Cover crops to reduce weed seed transmission. 8. Transgenic crops for virus resistance.
Crop Rotation
Duration 18 mo.
Raspberry (control)
Raspberry-fumigation
Clean fallow
..
Grass (non-host for ToRSV)
Rape (allelopathic)
Cucumber-ELISA assay
for ToRSV in soil
X. americanum densities
- - - - +
0
Percent change in
- 20
- 40
- 60
- 80
- 100
Clean Canola Grass Raspberry Continuous
fallow (weed- followed by raspberry
free) fumigationRaspberry leaf curl virus
1. Aphid-transmitted, can spread within fields and
between fields.
2. Retained by aphid vector for long periods of
time, usually for its entire life.
3. Some strains are symptomless in red raspberry
and can go undetected if not grafted.
4. Moved in planting stocks, can move anywhere
in the world in a few days.
5. Present on quarantine lists for many countries,
one of the few known raspberry viruses that
we don’t have a lab test for.Aphid-borne Viruses
Raspberry Mosaic Disease
Virus Vector Distribution
Black raspberry necrosis A. agathonica/idaei NA & Europe
Rubus yellow net A. agathonica/idaei NA & Europe
Raspberry mottle A. agathonica/idaei NA & Europe
Raspberry leaf mottle A. idaei Europe
Raspberry leaf spot A. idaei Europe
Raspberry leaf spot (NA) A. agathonica NA4th fruiting year!
Aphid Monitoring Time of Transmission
0
1
2
3
4
5
6
7
10-May
17-May
24-May
31-May
2-Jun
7-Jun
14-Jun
17-Jun
21-Jun
28-Jun
5-Jul
6-Jul
12-Jul
19-Jul
20-Jul
26-Jul
2-Aug
9-Aug
16-Aug
Date
23-Aug 3-Aug 18-Aug
30-Aug
2-Sep
6-Sep
13-Sep
20-Sep
16-Sep
27-Sep
Average Aphids/Trap vs. Positives: 2004
4-Oct
30-Sep
11-Oct
18-Oct
25-Oct
14-Oct 28-Oct
1-Nov
0
2
4
6
8
10
12
14
16
# of BRDaV Positives
# of positives
Average Aphids:RYNV
Aphid-borne Viruses Virus Vector Distribution Raspberry leaf curl Aphis rubicola North America Raspberry vein chlorosis Aphis idaei Europe, NZ Cucumber mosaic Europe Thimbleberry ringspot North America Aphid-borne dsRNA? RLSV-NA North America
Control of Aphid-Borne Viruses
1. Certified plants free of these viruses.
2. Vector resistant cultivars.
3. Virus resistant cultivars.
4. Virus tolerant cultivars.
5. Isolation from virus sources.
6. Use of insecticides to reduce vector
populations at critical times.Control of Viruses in Utah
1. What viruses are present?
A. In commercial plantings
B. In native hosts of related species
2. Viruses are at risk of becoming established.
A. What hosts are being introduced for
for production? Blackberries, Blueberries etc.
B. What vectors of potential viruses are present
in the area?Is There A Virus In That Plant? 1. ELISA tests for usual suspects 2. Sap transmission to herbaceous hosts 3. Electron microscopy 4. DsRNA extraction 5. Cloning, sequencing 6. BLAST search 7. Develop diagnostic RT-PCR test 8. Test other Rubus sources
Virus Identification in Woody Hosts Now we extract dsRNA and do shotgun cloning. Find one virus and we get excited. Find another and we get ecstatic, lab pizza. Find a third it’s time for lab pizza and beer. When we get the fourth it’s, Wine and Hors'dovres, “Good thing I have (had) a good stash of Pinot Noirs” Low concentration virus or viruses with limited amounts of dsRNA likely to be missed with this strategy.
Blackberry Yellow Vein an Emerging
Disease in Southern USAPinwheel Inclusions
9 Kbp
dsRNA
3.0 to
0.8 KbpBLAST Search of Sequences from BYVD
Location Virus(es) Likely Vector
Arkansas
Crinivirus (BYVaV) Whitefly?
Potyvirus (BVY) Aphid?
Flexivirus (BVX) Aphid, unknown
Cripavirus An Insect Virus!!!
Southeast
BPYV, BYVaV Whitefly
BVX
INSV Thrips
TRSV Nematodes
Mississippi
BlVE (Allexi), BVX, BYVaV?Symptoms after Transmission on ‘Chester’
Mite GraftingRT-PCR Detection of BYVaV
and BVY
Agarose gel showing RT-
PCR products for BYVaV
(~500 bp) and BVY (~330
bp)
In ‘Chickasaw’, ‘Chester’
and ‘Navaho’ both viruses
are required for symptom
developmentField Spread of BVY Using
Trap Plants Infected with
BYVaV
Trap April May June July August September
plants
Chickasaw 0/10 0/20 1/20 4/20 4/20 1/20
(2003)*
Chester 0/20 0/20 0/20 4/20 6/20 0/20
(2004)Blackberry Yellow Vein Associated Virus Identified in commercial blackberry from Arkansas, Georgia, Kentucky, North Carolina, Tennessee and South Carolina, West Virginia with symptoms, and from symptomless blackberry from Oregon. Identified in wild blackberry in Arkansas. Symptomless in some cultivars in single infections, maybe in most or all cultivars. Symptomless in red raspberries tested. Symptomless in ‘Munger’ black raspberry.
Is Crumbly Fruit Caused by a Virus
Complex?
• RBDV is becoming more common in
southern WA and OR but crumbly fruit is
not as prevalent as it is in the north.
• Are there other viruses present?
• Extract dsRNA, clone and sequence.
• Two new viruses have been identified.
Both likely are transmitted by aphids.
• Aphid populations in the north are much
higher.BLAST Search Red Raspberry Virus(es) Likely Vector Washington Closterovirus Aphid? dsRNA virus Aphid Scotland Closterovirus Aphid? Flexivirus Aphid? Cripavirus An Insect Virus!!!
Closteroviridae from Rubus Polymerase Heat Shock Protein
Fruit Drop
DsRNA from Blueberry Fruit Drop Plants
RT-PCR detection of Fruit Drop
BFD
dsRNASoil Transmission
Healthy plants plus FD plant
in field soil from site with FD
Healthy plants in field
soil from site with Fruit Drop
Healthy plants in pasteurized field
soil from site with Fruit Drop Healthy plants plus FD plant in
pasteurized soil from site with FDBerkeley 2001, WA
Berkeley 2001
Transmission of BlShV to Trap
Plants by Foraging Honeybees
Trap Plants Persistence
Source Plants Hive in Cage Infected in Hives
BSIV-, Yr. 1 Yes 0/72 0/15
BSIV-, Yr. 1 No 0/74
BSIV+, Yr. 1 Yes 0/34, 28/42 1/45
BSIV+, Yr. 1 No 0/34, 7/42
BSIV-, Yr. 2 Yes 0/71 0/28
BSIV-, Yr. 2 No 0/70
BSIV+, Yr. 2 Yes 16/34, 30/45 2/33
BSIV+, Yr. 2 No 1/34, 3/45Winter Moth?
Pseudomonas Mummy berry
Botrytis Winter Moth
Other Blueberry Scorch Look AlikesCalifornia,
Decline in ‘Totem’ in B.C.
2003 & 2004Viruses From Declining
Strawberries
Virus Watsonville ‘03 PNW ’03 ‘04
SPaV 18/24 4/110
BPYV 9/24 1/110
SMYEV 22/24 79/110
SCV 5/24 84/110
SMoV 0/24 59/110
SVBV 0/24 61/110
Oxnard, over 90% with SPaV and/or BPYV.
Previously, SCV has not been reported in B.C., now very
common!No vector control in 2004
Vector Control B.C. 2005
Isolation
The Workers Karen Keller Yannis Tzanetakis Jennifer Kraus Anne Halgren Nola Mosier Suzannah Taylor Paul Kohnen Lisa Wegener Crystle Chamberlain and numerous undergrads. Collaborators: Bill Wintermantel Rose Gergerich Mark Bolda James Susaimuthu Dan Marcum Zvezdana Pesic Mark Sweeney Gina Fernandez Michael Qian Hugh Barker Helena Mathews Small Fruit Breeders
The Check Writers USDA-ARS ORBC OBC NWCSFR WRRC WBC NASGA BCRIDC BCBC CSC NABGA OWB OSC ARF-OSU WWAGG WSC SBIR FVSGA
So you might ask:
What does Bob do
if he has this big crew?
The Fun Stuff!
Visit Grower’s Fields
Travel and Give Talks
The Not So Fun Stuff:
Write Manuscripts
Get Funding To Pay That Crew,
Deal With Administration,
Worse Than Dealing with a Little Ol’bearAnd of Course the Quality Control
Questions?
Bob Martin
USDA-ARS-HCRL
3420 NW Orchard Ave.
Corvallis, OR 97330
541-738-4041
Bob.Martin@ars.usda.govYou can also read
