berry viruses complexes and what we can do about it
DESCRIPTION
Ioannis E. Tzanetakis [email protected] 479-575-3180TRANSCRIPT
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Berry viruses complexes and what we can do about it
Ioannis E. Tzanetakis
[email protected] 479-575-3180
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Virus transmission 101
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Phylum Arthropoda, Class Insecta, Order Hemiptera
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Phylum Arthropoda, Class Insecta
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Phylum Arthropoda, Class Arachnida, Order Acariformes
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Phylum Nematoda, Class Secernentea, Order Tylenchida
Nematodes
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Kingdom: Fungi
Phylum: Chytridiomycota
Class: Chytridiomycetes
Order: Incertae sedis
Family: Olpidiaceae
Genus: Olpidium
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Kingdom: Rhizaria
Phylum: Cercozoa
Class: Plasmodiophorea
Order: Plasmodiophorida
Family: Plasmodiophoridae
Genus: Polymyxa
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Vectors and plant viruses they transmit
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Four modes of virus transmission
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Strawberry viruses circa 2003 Arabis mosaic – Europe Tomato ringspot Raspberry ringspot – Europe Strawberry latent ringspot-Europe Tomato black ring –Europe Strawberry crinkle Strawberry mild yellow edge Strawberry mottle Strawberry vein banding Fragaria chiloensis latent – Chile Tobacco streak* Strawberry pallidosis Beet pseudo-yellows
Nematodes
Aphids
Pollen
Whiteflies*
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Strawberry decline
A $50M story
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The whitefly viruses
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BPYV-SPaV double infections
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Other strawberry viruses?
Goal: Identify unknown viruses that may contribute to the decline.
Chlorotic fleck Leafroll
Revisited strawberry virus-like diseases.
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* *
* *
*
*
*
* *
Tobacco streak virus Strawberry crinivirus 3 Strawberry crinivirus 4
Identification, characterization and development
of detection techniques for strawberry viruses
* *
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No vector control
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Vector control
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Blackberry yellow vein disease First observed in 2000 in the Carolinas.
Tested for known viruses (RBDV, TRSV etc) – Several viruses were found but none consistently associated with symptoms.
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Tobacco ringspot virus and BYVD
TRSV textbook symptoms Single TRSV-infection
Are symptoms cv. dependent? The majority of plants infected with TRSV are symptomless
BYVD is very similar to what people thought as being TRSV symptoms
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New viruses in Rubus in the last 7 yrs 16 viruses & virus-like agents were known to infect Rubus before we started looking into Rubus complexes – We now have over 40… New Rubus viruses Blackberry yellow vein associated virus Blackberry virus E Blackberry virus X Blackberry virus Y Blackberry virus Z Beet pseudo yellows virus Blackberry yellow mottle virus Blackberry chlorotic ringspot virus Strawberry necrotic shock virus Black raspberry necrosis virus Raspberry leaf mottle virus Rubus canadensis virus -1 Impatiens necrotic spot virus Raspberry latent virus etc…..
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New viruses in Rubus in the last 8 yrs 16 viruses & virus-like agents were known to infect Rubus before we started looking into Rubus complexes – We now have over 40… New Rubus viruses Blackberry yellow vein associated virus Blackberry virus E Blackberry virus X Blackberry virus Y Blackberry virus Z Beet pseudo yellows virus Blackberry yellow mottle virus Blackberry chlorotic ringspot virus Strawberry necrotic shock virus Black raspberry necrosis virus Raspberry leaf mottle virus Rubus canadensis virus -1 Impatiens necrotic spot virus Raspberry latent virus etc…..
Tests are available for all the new viruses
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Same disease-different viruses
AR MS NC
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Arkansas
BYVaV BVY TRSV
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Carolinas BYVaV BVX BPYV INSV TRSV
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Mississippi BYVaV TRSV BVE
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How do we tackle BYVD?
After identification of all (or almost all) viruses that are involved in the disease we need to: A. Make sure that mother plants are being tested for the new
viruses before they are propagated. B. Identify virus combinations that can cause BYVD. C. Identify virus vectors. D. Find alternative hosts of the viruses in the field. E. Minimize or eliminate BYVD by eliminating the weakest link, the
virus vector(s) that is the easiest to control.
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What are the viruses present in your area? The importance of detection
• BYVaV - Multistate sample
collection - 35 isolates
• i
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What are the viruses present in your area? The importance of detection
• BYVaV - Multistate sample
collection - 35 isolates
• i
• Detection
100%
identity
1 2 3 4 5 6 7 8 9
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Virus interactions: The BYVaV/BVY story BVY did not cause symptoms in single infections but together with BYVaV they cause BYVD.
In mixed infections, BVY knocks down concentration of BYVaV to about 0.1% compared to titer in single infections.
In mixed infections, they can cause death
of fruiting canes.
Susaimuthu et al.,2008. Plant Disease 92:1288-1292
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Transmission
Experiment Trialeurodes abutilonea Trialeurodes vaporariorum
Experiment 1 4/7 3/9
Experiment 2 5/8 1/8
Experiment 3 3/10 3/10
Total 12/25 7/27
1
• Both whitefly species transmitted the virus at a rate >30%
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Alternate hosts
Plant species Scientific name Family Number of plants
tested
Garden vetch Vicia sativa Fabaceae 16 Virginia creeper Parthenocissus quinquefolia Vitaceae 16 Red clover Trifolium pretense Fabaceae 16 Wild garlic Allium vineale Amaryllidaceae 16 Creeping woodsorrel Oxalis corniculata Oxalidaceae 16 Carolina geranium Geranium carolinianum Geraniaceae 16 Curly dock Rumex crispus Polygonaceae 16 Dandelion Taraxacum officinale Asteraceae 16 Tall fescue Festuca arundinacea Poaceae 16 Wild wheat Avena fatua Poaceae 16 Grapes Vitis vinifera Vitaceae 16 Peach Prunus persica Rosaceae 16 Blueberry Vaccinium spp. Ericaceae 16 Shepherd’s purse Capsella bursa-pastoris Brassicaceae 16 Nutsedge Cyperus spp. Cyperaceae 16 Horsenettle Solanum carolinense Solanaceae 16 Common ragweed Ambrosia artemisiifolia Asteraceae 16 Tree of heaven Ailanthus altissima Simaroubaceae 16 Apple Malus spp. Rosaceae 200 Rose Rosa multiflora Rosaceae 40 Carpetweed Mollugo verticillata Molluginaceae 16 Amaranthus Amaranthus spp. Amaranthaceae 16 Poor joe Diodia teres Rubiaceae 16 Ground cherry Physalis spp. Solanaceae 16 Sorghum Sorghum spp. Poaceae 16 1
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Vector elimination The BRNV paradigm
New field monitoring Permanent tagged plants
K
A
B J
H
G
L
F
E
D
C
I M
N
O
P
Time of transmission High incidence of virus Potted plants Rotated every month
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Average Aphids/Trap: 2004
0
2
4
6
8
10
12
144-M
ay
11-M
ay
18-M
ay
25-M
ay
1-J
un
8-J
un
15-J
un
22-J
un
29-J
un
6-J
ul
13-J
ul
20-J
ul
27-J
ul
3-A
ug
10-A
ug
17-A
ug
Date
Av
era
ge
Ap
hid
s/
Tra
p
40
50
60
70
80
90
100
110
Max D
aily T
em
p (
oF)
Field 1
Field 2
Field 3
Max Daily Temp
Field 1 Field 2 Field 3
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Field 1 Field 2 Field 3
Average Aphids/Trap: 2005
0
1
2
3
4
5
6
1-M
ar
15-M
ar
29-M
ar
12-A
pr
26-A
pr
10-M
ay
24-M
ay
7-J
un
21-J
un
5-J
ul
19-J
ul
2-A
ug
16-A
ug
30-A
ug
13-S
ep
27-S
ep
11-O
ct
25-O
ct
Date
Ave
rag
e A
ph
ids/
Tra
p
40
50
60
70
80
90
100
Ma
x D
aily T
em
p (
oF)
Field 1
Field 2
Field 3
Maximum Daily Temp
Nearly 100% transmission in three years!
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Average Aphids/Trap vs. Positives: 2004
2-Jun
17-Jun
6-Jul
20-Jul
3-Aug 18-Aug
2-Sep
16-Sep
30-Sep
14-Oct 28-Oct
0
1
2
3
4
5
6
7
10
-Ma
y
17
-Ma
y
24
-Ma
y
31
-Ma
y
7-J
un
14
-Ju
n
21
-Ju
n
28
-Ju
n
5-J
ul
12
-Ju
l
19
-Ju
l
26
-Ju
l
2-A
ug
9-A
ug
16
-Au
g
23
-Au
g
30
-Au
g
6-S
ep
13
-Se
p
20
-Se
p
27
-Se
p
4-O
ct
11
-Oct
18
-Oct
25
-Oct
1-N
ov
Date
Av
era
ge
Ap
hid
s/T
rap
0
2
4
6
8
10
12
14
16
# o
f B
RD
aV
Po
sit
ive
s
# of positives
Average Aphids: 2004
Time of Transmission
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Raspberry crumbly fruit and decline • The Pacific Northwest (PNW) is a primary producer of red
raspberries
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Raspberry crumbly fruit and decline
• Still, in many cases RBDV single infections did not cause symptoms
• ‘Several cultivars are susceptible to crumbly fruit disease (drupelets abortion)
• Raspberry bushy dwarf virus (RBDV), a pollen-borne idaeovirus was considered the causal agent of crumbly fruit
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Another virus complex? Important observations suggested that crumbly fruit
symptoms may be increased by additional viruses:
1. The disorder is more severe in cool areas with high populations of the large raspberry aphid Amphorophora agathonica
2. Two additional viruses found in severely affected fields,
Raspberry leaf mottle (RLMV) and Raspberry latent (RpLV)
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RBDV, RLMV and RpLV interactions
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RLMV qRT-PCR
Amp. Cycle
ΔRn
Threshold
RLMV titer in single and mixed infections over time
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RpLV qRT-PCR RpLV titer in single and mixed infections over time
Threshold
ΔRn
Amp. Cycle
int. control
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RBDV titer enhanced in co-infections with RLMV
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RBDV titer increase verified by conventional methods ELISA RT-PCR (20 cycles)
RBDV titer enhanced in co-infections with RLMV
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• H Control • D RBDV - Dwarf • M RLMV - Mottle • L RpLV - Latent • DM RBDV + RLMV • DL RBDV + RpLV • ML RLMV + RpLV • DML RBDV + RLMV + RpLV
Raspberry virus .... what...?
Mixed virus infections affect on plant growth and fruit crumbliness
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Plant Growth Establishment (2010)
Oct. Height
109.03
44.90
65.43 63.83
26.09
48.93
33.49 32.00
0.00
20.00
40.00
60.00
80.00
100.00
120.00
140.00
H D M L DM DL LM DMLTreatments
cm
a b b cd d bc cd cd
10.07
5.74
7.02
8.18
5.736.30
5.415.94
0.00
2.00
4.00
6.00
8.00
10.00
12.00
H D M L DM DL LM DML
mm
Treatments
Cane diameter
a d bc b cd d cd d
Oct.
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Plant Growth (2011) July
Height
55.57
48.5051.27 51.03
30.20
50.7344.60 42.13
0.00
10.00
20.00
30.00
40.00
50.00
60.00
70.00
H D M L DM DL LM DML
Treatments
cm
c a b b b d b c
Cane diameter
8.06 7.81 7.727.34
6.05
7.18 7.346.99
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
H D M L DM DL LM DML
Treatments
mm
a a b a b c b b
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Plant Growth (2011)
October
Height
2.602.25 2.17 2.25
1.97 2.121.93 1.89
0
0.5
1
1.5
2
2.5
3
H D M L DM DL ML DML
Treatments
m
a b b b cd bc cd d
Cane diameter
12.7911.79 11.52 11.83
10.77 10.82 10.21 10.03
0
2
4
6
8
10
12
14
H D M L DM DL ML DML
Treatments
mm
a b b b bc c c c
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Crumbly Fruit
Weight
3.35
2.432.74 2.87
2.572.08
2.332.05
0.000
1.000
2.000
3.000
4.000
H D M L DM DL LM DML
Treatments
g
a b b bc c cd cd d
No. Drupelets
94
64
89 95
75
59
7666
0
20
40
60
80
100
120
H D M L DM DL LM DML
Treatments
a a a b b c c c
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Crumbly fruit
Firmness
73.54
58.54 61.3865.58
61.83
46.38
61.46
49.15
0.000
10.000
20.000
30.000
40.000
50.000
60.000
70.000
80.000
90.000
H D M L DM DL LM DML
Treatments
g
a b bc bc bc c d d
H DML
D
DML
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Virus Incidence in ‘Meeker’ Fields
Northern Washington Field Age
(years) RLMV
(%) RpLV (%)
1 4 0 1 30 0 1 10 0 2 58 21 2 0 0 2 0 0 2 6 0 2 16 0 3 31 6 3 6 0 3 13 0 3 50 0 4 19 6 4 13 0 5 69 0 5 90 80 5 100 75 5 44 6 5 100 17 6 70 25 6 100 6 6 100 12 7 100 6 8 100 46
Southern Washington/Oregon
Field Age (years)
RLMV (%)
RpLV (%)
1 0 0 5 40 20
6 0 20
7 8 17 8 19 0
8 27 0
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Crumbly Fruit Scouting Crumbly fruit and virus incidence in Washington
Crumbliness
rate 1 Crumbliness
rate 2
Crumbliness
rate 3
Field Age
Crumbliness 0: normal 3: severe
Virus incidence %
0 1 2 3 RBDV RLMV RpLV
4 1 44 25 0
4 2 100 100 0
4 3 93 100 7
5 3 100 100 40
6 3 92 96 40
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Insects in Traps (2011)
Lightle, unpublished data
Empoasca fabae was sporadic
Raspberry aphid A. agathonica
predominant insect
Few numbers of
Macropsis fuscula
0
1
2
3
4
5
6
7
8
1-Apr 16-May 30-Jun 14-Aug 28-Sep
Ave
rag
e #
ap
hid
s / 1
0
lea
ve
s
Harvest clean-up sprays applied
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RLMV spread in the field
Four fields being monitored for virus spread
year 1 year 2 year 3 year 0
Field 4 26 % (24 %)
16 % 32 % Field 1
21 % 33 % Field 2
4 % 44 % Field 3
Age of Field
50 %
(40 %)
(12 %)
(16 %)
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Control Strategies
1. Think long term, identify potential risks of a site
2. Start with clean plants
3. Identify and diagnose problems early
4. Implement control strategies ASAP
5. If a virus complex is involved - identify viruses present and which are the easiest to control
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• Better establishment
The importance of clean plants
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• Better establishment
• Longer life of plantings
The importance of clean plants
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• Better establishment
• Longer life of plantings
• Fewer disease problems/Reduce risk of introducing new viruses to a region or field
The importance of clean plants
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A federally-coordinated effort to secure high quality virus-tested plants for clonally propagated crops.
National Clean Plant Network
NCPN Mission
The NCPN provides high quality asexually propagated plant material free of targeted plant pathogens and pests that cause economic loss to protect the environment and ensure the global competitiveness of specialty crop producers in the United States.
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NCPN Supported Clean Plant Centers
TAMU
USDA at OSU
UC-Davis
UCR
UAZ
WSU
MO State U.
LSU
UAF
FAMU
Auburn Clemson
NCSU
FL-Dept. Ag.
Cornell
UHM
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Berry Clean Plant Centers
USDA at OSU
UAF
NCSU
UC-Davis
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Why care???
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The story:
Propagation from an existing plot
10 ton/acre =$30,000/year
Latent infections with Blueberry scorch
The result?
Removal of infected material
Cumulative loss: ~ 100,000/acre
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Florida 2013
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Start clean!!!!
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The team
The berry virus consortium: 16 individuals from UA, NCSU, USDA-ARS, MSU, UGA Bindu Poudel, Diego Quito, Danielle Lightle, Anne Halgren, James Susaimuthu