Transcript Slide 1

Evaluation of Arkansas Soybean Cultivars to Bean Pod Mottle Virus (BPMV)
Ehsanollah
1
Shakiba ,
Pengyin
1
Chen ,
Rose
2
Gergerich
(1) Dept. of Crop, Soil, and Environmental Science (2) Dept. of Plant Pathology, University of Arkansas, Fayetteville, AR 72701
Introduction
One of the important viral diseases in soybean is caused by Bean pod
mottle virus (BPMV). The virus was found for the first time in Arkansas and
North Carolina in 1958 (Walters, 1970). In the past 40 years, it has spread to
most of the midwestern states of the U.S. (Sundararaman et al., 2000). BPMV
symptoms includes: Chlorotic and leaf rugosity, sustained green stem,
reduction of nodulation, pod formation, seed size, seed weight, seed number,
and seed discoloration (Fig.1) (Ross, 1963 and 1986). BPMV belongs to the
family Comoviridae, genus comovirus and has a bipartite positive-strand RNA
genome consisting of RNA-1 and RNA-2. BPMV isolates are divided into two
subgroups (I and II), which induce moderate and mild symptoms respectively,
and one reasortment group (I/II) inducing severe symptoms in soybean
cultivars. To date, no resistance gene for BPMV has been found in soybean
germplasm (Zheng et al., 2005).
A
each pot was measured in centimeters by a ruler. The statistical analyses
were performed by JMP 6 and SAS 9.1 programs.
To examine the effect of BPMV on plant biomass, 68 cultivars from the
plant height experiment were used. Individual plants, which were 3 plants per
pot (6 plants per cultivar), were collected from the BPMV infected and the
control pots (3 plants per pot from each cultivar) and were kept in paper
bags. The samples included stem and leaves and excluded roots. The
samples were dried in oven at 105oC for 48 hours and weighed in grams. The
statistical analyses were performed by JMP 6 and SAS 9.1 programs.
The results of the screen showed no resistant cultivars to the BPMV mild
isolate (K-Ha1) and severe isolate (K-Ho1) (Fig. 3). All cultivars showed
susceptible reaction to the virus. The results of ELISA and tissue blotting
showed presence of virus in all infected plants. Cultivar Terral TV49R12
showed necrotic symptoms.
A
B
C
Fig.1. A- susceptible symptom in plants (Ozark) infected by BPMV severe
isolate. B- comparison of healthy (control) and BPMV-infected
soybean leaflet in cultivar Manokin.
Objectives
Materials and Methods
I- Plant material and virus inoculation
The experiment was conducted in the greenhouse with appropriate control
of environmental conditions at the Rosen Center of University of Arkansas,
Fayetteville, Arkansas. Three-hundred and three soybean cultivars with different
maturities provided by the Variety Testing Program of the University of Arkansas
were used in the experiment. The cultivars were planted in three separate pots,
one for control and two for inoculation with BPMV. Twelve plants were grown in
each pot. After 10 days, two pots of each cultivar (24 plants) at the unifoliate
stage were inoculated with BPMV. Two isolates of BPMV, K- Hal (subgroup II)
and K-Ho1 (subgroup I/II), were used for inoculation. Inoculum for each isolates
was made from BPMV-infected leaves that were collected and ground in a mortar
and pestle in a potassium phosphate buffer (0.05 M and pH 7.5). Before
inoculation, plants were dusted with silicon oxide (carborundum) an abrasive that
helps the virus penetrate into the plants. Then, plants were inoculated with the
virus by using a cheesecloth pad that had been dipped in the inoculum (Fig.2).
The plants were visually evaluated based on symptom expression for four
weeks after inoculation. Furthermore, the presence of the virus in plant were
detected by two serological methods, Tissue Blotting Immunoassay (TBIM) and
Protein A Enzyme-linked Immunosorbent Assay (ELISA), described by Lin et al.
(1990) and Cooper and Endwards (1981), respectively.
Fig.3. A- Reaction of BPMV mild and severe isolates on cultivar HBK R4724 ; left pot= healthy control, middle pot=infected
with mild isolate, right pot= infected with severe isolate. B- Reaction of BPMV severe isolate in cultivar HBK R4724 caused
significantly reduction of plant height, left pot= healthy control, right= infected pot. C- Reaction of soybean cultivar
USG 540RR to BPMV severe isolate which showed less height reduction. Left= control pot
(uninoculated plants), right pot= BPMV-infected pot (inoculated plants).
The experiment for the effect of the BPMV severe isolate on plant height
showed that the virus caused significant reduction in plant height. This result is
in agreement of reports from previous studies by Giesler et al. (2002). The
mean plant height of the control was 49.52 cm, while that of BPMV infected
plants was 31.88 cm (Fig. 4. A,B).
The results of experiments to determine the effect of BPMV severe
isolate infection on plant biomass showed that the isolate significantly
decreased plant biomass. The mean biomass of the control was 2.03 g
whereas the mean biomass of BPMV-infected plants was 0.99 g (Fig.4.B). The
reason for such a reduction of biomass in BPMV-infected plants was that virus
not only had an effect on plant the stem and caused plant height reduction, but
also reduced leaf weight. These results are supported by observations in
previous studies by Ross (1986) and Giesler (2002). The data from plant
height and biomass followed a normal distribution. However, there was a
range of plant height and biomass reduction by the virus. Therefore, it can be
assumed that such a tolerance to the BPMV severe isolate was genotype
specific and may have impact on virus replication and movement.
A
60
(49.6) †
USG 540NRR, Pioneer 94M30, DGrow 4960RR, PGY 4315, S03-058RR,
Stine S4842-4, PGY 3805, Anand, FFR 5663RR, HBK R5620, HBK R5825,
Morsoy RT5620N, Morsoy RT5773N, Progeny 4949, Delta Grow 3950RR,
Dyna Gro 35B40, Asgrow AG4703, AGS 568RR, Pioneer 94M50, DGrow
4970RR, NK Brand S43-B1, Terral TVX46R213, Asgrow AG4801, PGY
XR5862, RJ00-277, RJ00-100, 47-G7, S03-007RR, Hutcheson, Dyna Gro
37A44, Terral TV48R43, Dyana gro 34J56, N.K. Brand AG5702, DKing 4967,
Garst 4612RR/N, Ozark, DGrow 4860RR, DKing XTJ602, Terral TV45R14, Ax
RR53386, USG 7466nRR, Morsoy RT4480N, SS RT498, DP4933RR,
DPX4919RR, NK Brand S49-Q9, PGY 3905, R00-1940, PGY 4205, Armor GP
530,Armor 53-K3, DGrow 4840RR, Progeny 5650, DP4724RR, Morsoy
RT4485N, 4905nRR, Dyna Gro 36M49 USG 5002T, Southern States
RT4651N, Teejay, HBK R4924, R98-1821, S00-9970-09, SS RT490, SS
RT513, DT 98-7278, Ax RR53776, ARXF47205, 495RC, Croplan RC5555,
Delta Grow 5160RR, Delta Grow 5560RR, Delta King 5066, Delta King 55T6,
Delta King XTJ6025, Delta King XTJ603, Delta King XTJ604, Delta King
XTJ6501, Delta King XTJ652, Delta King XTJ6G510, Delta King XTJ6K54,
Dyna Gro 33B52, Dyna Gro 33X55, Dyna Gro 3535NRR, ES XVT-110RR, ES
XVT-501RR, ES XVT-518RR, Fastart F-50H3RR, Fastart Filler, HBK Filler05,
HBK R5324, HBK R5525, Morsoy RT5553N, MPV 5505nRR, NK Brand S52U3, NK Brand S54-G9, PGY 5005, PGY 5105, PGY 5115, Pioneer 95M30,
Pioneer 95M50, Progeny 5250, R01-3263, R01-4804, S02-3934RR, S03380RR, Southern States RT 5302N, Southern States RT 5540N, Stine
S5142-4, Terral TV52R14, TN05-547RR, USG 7553nRS, DB01-5463, Delta
King 5870, Delta King 5995, Freedom, Progeny 5770, R01-379, R97-1634,
USG 5601T, 586 RC, Amr ARX B57104, AG5605, AG5903, AG5905,
Deltapine 5634RR, DGrow 5960RR, Deltapine 5915RR, DynaGro 3562NRR,
DynaGro 3583NRR, Pioneer 95M80, R01-4834, SS RT 5951N, TV56R45,
TN05-548RR,
Plant Height (cm)
30
(0.99g) ‡
10
0
Control
BPMV
Treatment
Fig. 5. A- Effect of BPMV severe isloate on plant height. †, The mean height
of 273 uninoculated cultivars. ‡, The mean height of 273 inoculated cultivars.
B- Comparison of the effect of BPMV severe islotae on plant biomass,† The
mean biomass of 68 uninoculatedcultivars (3 plants/cultivar). ‡ The mean
biomass of 68 inoculated cultivars (6 plant/cultivar).
Fig. 2. The process of virus inoculation: A, selecting infected leaves; B, grinding
leaves in buffer using a mortar and pestle; C, dusting silicon oxide (carborondum)
on unifoliate leaves; and D, inoculation using cheesecloth pad.
II- Classification of cultivar reaction to BPMV
To examine the effect of virus infection on plant height, 273 cultivars
were grown in a separate experiment at the Rosen Center of University of
Arkansas, Fayetteville, Arkansas. Each cultivar was grown and inoculated as
in the procedure described above. After four weeks, the height of all plants in
Very Low Tolerance
HBK R4724, R01-FILL3, Pioneer 94M80, Armor ARX C53104, Morsoy
RT4914N,Terral TV48R43, Terral TV49R12, Terral TVX49R50, ARXD49104,
Pioneer 94B73, Fastart F-48H8RR, RJ00-277, DKing XTJ6L49, Progeny
5650, SS RT513, Dyna Gro 36Y48, Asgrow AG4902, DKing XTJ650, Terral
TVX43R51, Pioneer 94M70,ARXF47105, Morsoy RTS4955N, Armor ARX
C55105, USG 7434nRR, UA 4805, R00-FILL4, Terral TVX46R223, RJ00FILL, Armor 54-03, DPX4818RR, DKing 4868
Conclusions
(32.0) ‡
20
D
DKing XTJ648, R98-209, S03-393RR, HBK C5894, ES XVT-110RR,
Deltapine DPX5115, Morsoy RT5252N, Terral TV55R15, Terral TVX51R50,
USG 7562nRR, TV59R14, TV57R14, Progeny 5622R00-1194F, S02-2238RR,
USG 747R6, GP470, Morsoy RT4731N, Morsoy RT4993N, Progeny 5660,
Progeny 5822, SS RT 5702N, USG 7582nRR, fastart F- 54H2RR, ESXVT488RR, Deltapine 5414RR, Garst 5212RR/N, FFR 5033RR, S03-383RR,
R01-4752, TN05-548RR, Md 96-5722, Pinoneer 95M60, Terral TV48R14,
USG 7455nRR, USG 7484nRR, USG 7499nRR, Fastart F-48H5RR, Fastart
F-48H5RR, Progeny 4910, R01-1017, RJ00-090, USG 7440nRR, DKing
4866, USG 7494nRR, DKing 4766, S02-390RR, R00-1551, Manokin, RJ00261, DKing XTJ6G51, Ax RR53386, , DKing XTJ6025, DKing 4763, Asgrow
AG4902, GP474 ARXF47205, DKing 4967, DKing 4868, R01-FILL5, PGY
4315, Morsoy RT4665N, DKing XTJ6L49, DP4933RR, Dyna Gro 35Z49,
Garst 4999RR/N, Morsoy RT4731N, Morsoy RT4914N, PGY 4805, RJ00FILL2, S02-166RR, SS RT490, USG7475RR, USG 7494nRR, DT 99-17400,
RJ00-277, S00-9925-10, DB01-080, DB01-4249, HBK C5025,RJ00-090,
Terral TV57R14, Aagrow AG5702, , Croplan RC5955,
Armor ARX
A50104Armor ARX C55105, Armor ARX C56105, Armor GP 513, Ax
RR53116 , Terral TVX46R203, R01-375, Dulaney AV57D7, Armor GP 555,
AsgrowAG5301,
Asgrow
AG5501,USG7440nRR,
ARXD49104,Terral
VX46R213, Delta Grow 5260RR, Delta Grow 5260RR, Delta King 5161,Delta
King 5366, Delta King 5466, Delta King 5567
(2.01g) †
50
B
B
40
C
Progeny 4401, HBK R4623, Delta Pine DPX5914RR, Garst 3960RR/N PGY
4315, Dyna Gro 3481NRR, 476RC, Terral TVX41R50, Morsoy RT4802N,
Progeny 3900, Aagrow AG5702, PGY 4805, Dyna Gro 3463NRR, DKing
XTJ6025, HBK R3824, Pioneer 93M90, Croplan C4955, PGY 4615, Garst
3960RR/N.
Low Tolerance
 To identify cultivars with genetic resistance or tolerance to BPMV.
 To classify the reaction of different cultivars to BPMV.
A
High Tolerance
Moderate Tolerance
Results and Discussion
B
Table 1. Classification of Arkansas soybean cultivars to BPMV severe isolate
By using LSD value 15.10 cm for plant height and 25.0g for plant biomass,
all cultivars were classified in four distinct groups based on plant height and
biomass reduction. The cultivar groups are matched perfectly between the two
classification systems based on plant height and plant biomass. Group 1
includes 19 cultivars with high level of tolerance and with 20% plant height (≤10
cm) and 25% biomass reduction as compared to the uninoculated control.
Group 2 includes 90 cultivars that showed 20-35% plant height reduction (about
10 to 20 cm) and 25 to 50% reduction in biomass of infected plants. Group 3
contain 128 cultivars that showed 35-60% plant height reduction (20 to 30 cm)
and 50-75% reduction in plant biomass. The 33 cultivars in group 4 showed a
greater than 60% plant height reduction (≥ 30 cm) and ≥ 75% biomass
reduction in infected-plants with low tolerance. (Table 1).
 No resistant cultivar to BPMV, but some cultivars exhibited
tolerance.
 BPMV causes reduction of plant height and biomass.
 BPMV tolerance related to less plant height & biomass
 reduction.
 cultivars were classified in four groups: high, moderate, low,
and very low tolerance based on 20, 25- 45, 45-60, and 60%
of plant height reduction and 25, 25-50, 50-75, and 75%
plant biomass reduction, respectively.
References
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hybrid poplars and virus detection by ELISA. Ann. Appl. Biol. 99:53-61.
 Giesler, L.J., S.A. Ghabrial, T.E. Hunt, and J.H. Hill. 2002. Bean pod mottle virus: A
threat to U.S. soybean production. Plant Dis. 86:1280-1289.
 Lin, N.S., Y.H. Hsu, and H.T. Hsu. 1990. Immunological detection of plant viruses
and a mycoplasmalike organism by direct tissue blotting on nitrocellulose
membranes. Phytopatghology 80:824- 828.
 Ross, J.P. 1986. Response of early- and late-planted soybean to natural infection
by Bean pod mottle virus. Plant Dis. 70:222-224.
 Ross, J.P. 1963. Transmission of Bean pod mottle virus in soybean by beetles. Pl.
Dis.
 Walters, H.J. 1970. Bean pod mottle virus disease of soybean. Arkansas. Fm. Res.
19:8.
 United States Department of Agriculture, National Agricultural Statistics Service.
2007. Crop production summary 2006. United States, Department of Agriculture,
Washington.
Zheng, C., P. Chen, T. Hymowitz, S. Wickizer, and R. Gergerich. 2005. Evaluation of
Glycine species for resistance to Bean pod mottle virus. Crop Protection 24:49-56.