Mungbean Yellow Mosaic Virus in the AVRDC Mungbean ...

Workshop on Mungbean 159
Mungbean Yellow Mosaic Virus in the
AVRDC Mungbean Improvement Program
S.K. Green ' , D.H. Kim. ' , B.T. Cl1.iangl, and D. Maxwell 3
1 AVRDC, P.O. Box 42, Shanhua, Tainan, Taiwan
' Asian Regional Center, Kasetsart University, Kamphaeng Saen, Thailand
' University of Wisconsin, Madison, USA
Introduction
Mungbean yellow mosaic virus, a whitefly-transmitted geminivirus, causes one
of the most serious diseases of mungbean in all of South Asia. Disease incidence
as high as 100% in farmers' fields is common in the Indian subcontinent, often
resulting in considerable yield losses (Varma et al. 1992). Soybean, blackgram,
mothbean, cowpea and a few other leguminous species have also been reported
hosts of this virus. For the past several decades research has been conducted on
biological characterization of the virus, virus/vector relationship, epidemiology,
and disease control by chemical means and resistance breeding (Varma et al.
1992, Malik 1992). Molecular characterization has been conducted with an
isolate from Thailand, which caused a severe outbreak of the disease in 1977 but
which has never been detected from 1981 onward.
Despite nearly 25 years of resistance breeding efforts and the release of tolerant
lines, the disease still poses a major problem to economic production of this crop
in the Indian subcontinent. This has been attributed to various factors, including
increase in disease and whitefly populations, and unstable levels of resistance.
AVRDC has therefore accorded high priority to yellow mosaic in its mungbean
improvement program. The following are the activities conducted at AVRDC.
Review of MYMV related Literature
Literature relating to MYMV has been reviewed from 1948 to 1992. An Annotated
Bibliography of Mungbean Yellow Mosaic Virus is now available as AVRDC
Library Bibliography Series 6. It includes 235 citations covering the disease, the
virus, the vector, control methods, and host plant resistance.

16 0 International Consultation
International MYMV Workshop
An international workshop was organized by AVRDC in 1991 which brought
together breeders, entomologists, and virologists from countries where MYMV is
a limiting factor to economic production of mungbean (Green and Kim 1992).
From the various papers presented, the following were drawn:
1. As a result of nearly 25 years of varietal screening and breeding for resistance,
germplasm with resistance/tolerance to the virus or the vector has been
identified (Tables 1 and 2) and the mode of inheritance studied. Resistance/
tolerance has been found to involve one or more recessive genes. Breeding
has been complex and involved crosses with MYMV-tolerant mungbean
germplasm, interspecific crosses with highly MYMV-resistant blackgram, and
the use of gamma ray mungbean mutants.
Table 1. Mungbean germplasm and breeding lines with reported resistance to
MYMV
Resistant/tolerant mungbean line '
Bangladesh 2
AVRDC acc. V 2272, V 3485, V 3486
MB 53, MB 57, MB 58, MB 59, MB 133, MB 246
Mutant NOB-10 (25)
Sita Kundu (MB 55)
YH 25
India 2
A11/99, 11/395, 12/333, 16-303-20-3-8, 16-303-20-3-8-g (16-303), 20-3-8, 24-2, 57-7,
192-1, 525, 1066, 1122, 2779, 6601, 6602, 10866, 11148, 15044, 15127, 15176, 15225,
15227, 15229
Ac. No. 20, 214, 215, 216, 217, 218, 222, 223, 320
ADT 2
AMRIT (122)
AVRDC Acc. V 1269, V 2181, V 2182, V 2228, V 2294, V 2297, V 2230, V 3119, V 3404,
V 3417, V 3466, V 3471, V 3472, V 3486, V 3492, V 3493, V 3498, V 4409, V 4433,
V 4483, V 4542, V 4852
Bulk sample Orrsa
B-2
BR-2
China selection
CO 3,4
EC 2815, 277574, 103127
G 65
Green Mung
H-70-6
Hyb. 4-3, 4-3A, 12-4
Jawahar 45
Jyoti (Hyb 4-3)
Kuberi

Workshop on Mungbean 161
Table L. continued
Resistant/tolerant mungbean line '
L 24-2, L 24-2-1
L 23, 45, 52, 53, 61, 65, 66, 80, 87, 115, 127, 129, 143, 213, 213, 271, 304, 323, 349, 404,
294-1
LGG 407, 450
LM 6, 12, 36, 47, 81, 87, 94, 96, 113, 124, 134, 141, 142, 150, 156, 167, 168, 170, 171, 172,
182, 185, 191, 198, 214, 217, 218, 220, 238, 243, 248, 252, 289, 290, 294-1, 298, 310, 316,
254, 355, 356, 364, 383, 392, 404, 438, 459, 492, 496, 508, 509, 520, 529, 531, 542, 625, 626,
630, 662, 671, 686, 687, 692, 695, 696, 711, 744, 755, 775, 784, 873, 1101, 1114
M13, 21-1, 170
MB 5, 53, 57, 58, 59, 60, 132, 133, 142
Mg-1, Mg 588, Mg 589
MI -I 83-20, 309
ML 1, 3, 4, 5, 6, 7, 9, 15, 17, 24, 25, 35, 37, 38, 65, 73, 74, 79, 94, 101, 102, 104, 105, 109,
113, 116, 124 to 130, 131, 131-1, 132, 133, 134, 154, 157, 158, 161, 162 to 168, 170, 171,
186, 192, 194, 195, 197, 234, 237, 238, 266, 267, 267-1-84, 268, 269, 272, 274, 278, 282,284,
285, 286, 288, 289, 290, 296, 298, 302, 304, 306, 307, 313, 314, 315, 322, 326, 336, 337, 338,
353, 359, 360, 368, 369, 370, 371, 372, 375, 376, 382, 388, 390, 393, 394, 395, 405, 408, 409,
418, 421, 422, 423, 427, 428, 435, 436, 437, 438, 459, 466, 475, 484, 489, 506, 520, 536, 537
ML 26-10-3 (Pant Moong 2)
Mohini (S 8)
Moong no. 54
Mu-1
MUG 122, 124, 125, 126, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 144, 145,
169, 170, 173, 176, 242, 262
MUM 1, 2, 3
N 122
No. 525
NDM 88-14
P 1-67, 20, 40, 43-68, 66-68, 68, 82, 84, 86, 91, 94, 131, 156, 179-68, 217, 242, 290, 292,
293, 325, 332, 353-A, 364, 364-68, 366-68, 367, 369, 441-B-2, 453, 473, 614, 617, 670, 679
Pant-Moong-1, Pant-Moong-2 (ML 26-10-3), Pant Moong-3 (Pant M3) (UPM 79-4-12)
Ph 419
Plm 1066
PAIYUR-1
PDM 11, 14, 73, 84-139, 84-140, 84-143, 84-,146
PIMS 1066
PIMSI
PLN 15
PLS 274
PS 16
PUM 79-4-12
Pusa 101, 105
RATILA
Sabarmati (PIMS 4)
Sona
SML 70
ST 7

16 2 International Consultation
Table 1... continued
Resistant/tolerant mungbean line '
SUJATA (I Iyb. 12-4)
Tarai Local
Tarai Local 12 m,r,
T 44
TPM-1
TV 1132, 1133, 1134, 1135
UI I 80-7
UPM 79-4-12 (Pant Moong 3)
V 4116, 4852
Paksitan 2
BRM-114
E-321
M-13-1, M-20-21, M-22-24, M-28
Mut. 19-19, Mut. 20-21, Mut. 121-25, Mut. 131-37, Mut. 94-73, IVIut. 131-98, Mut. 13-1
NCM-5, NCM-7, NCM-68, NCM-69, NCM-87
NIAB Mung 121-25, NIAB Mung 19-19, NIAB Mung 20-21, NIAB Mung 13-1
NM 13-1, NM 19-19, NM 20-21, NM 28, NM 51, NM 121-25, NM 131-98
NIAB-M 51, NIAB-M 54
Pant Moong No. 1, Pant Mung No. 2, Pant Moong No. 3
Pak 3, Pak 13, Pak 18, Pak 22, Pak 32
Var. RC 71-17, Var. AUM 233
6601, 1429, 3198, 3850, 3854, 3881
Sri Lanka 2
CES 55 x M13-133F
M-30-2
N-4-30-5, N-4-30-11, N-4-30-12, N-4-60-1, N-4-60-3, N-4-60-4, N-4-60-5, N-4-60-6, N-4-
60-12, N-4-60-16, N-4-60-21
TV-79-3, 7704, TV-79-6
(CES 55 x MI-3) - 133F
' The list may contain duplicates, because the same lines may have been assigned a different accession
number/name by different institutions.
' The country refers to that country where the resistance screening has been conducted:
Bangladesh: AVRDC 1981, 1990, Chaudhary et al. 1981, Chhabra et al. 1974, 1979, 1980, 1981, 1988,
Chhabra and Kooner 1980, Dahiya et al. 1985, Dey and Singh 1973, Gill et al. 1975, Grewal 1978,
Khatri et al. 1971, Kooner et al. 1973, 1977, 1979, Legaspi et al. 1978, Mishra et al. 1978, National
Bureau of Plant Genetic Resources 1987, Nene et al. 1972, Pandya et al. 1977, Paroda and Thomas
1988, Ramana 1985, Sandhu 1978, 1980, Sandhu et al. 1985, 1988, Shah et al. 1984, Shukla 1977, Shukla
et al. 1978, Shukla and Pandya 1985, Singh 1982, 1986, 1988, Singh et al. 1986, 1977, 1988, 1991, Singh
and Patel 1975, 1977, Singh and Sharma 1981, 1983, 1984, Sinta et al. 1988, Sivaprakasam et al. 1974,
Varma et al. 1973, Verma 1985, Verma et al. 1973, Verma and Sandhu 1992, Verma and Singh 1988,
Vidhyasekaran 1976, Virmani 1983, Virmani et al. 1976.
Pakistan: Bashir et al. 1988, Bashir and Malik 1988, Chadhary et al. 1981, Dey and Singh 1973, Gill et al.
1975, Haq 1980, Malik 1988, 1991, 1992, Malik et al. 1986, Malik and Sarleem 1988, Malik and Sarvar 1988,
NIAB 1987, Shakoor et al. 1977.
Sri Lanka: Jayasekera and Ariyaraatne 1988, Pathirana et al. 1988, Wijetilaka 1989.

Workshop on Mungbean 163
Table 2. Mungbean germplasm with reported vector resistance
Vector resistant/ tolerant line References
G 65 Chhabra et al. 1979, 1980, 1981,
IM 170 1988, Chhabra and Kooner 1980,
LM 47, 141, 170, 364 Kooner et al. 1973, 1977, 1979,
M 170 Singh 1988
ML 1,3,5,6,7,15,24, 186, 192, 194, 195,
197, 235, 337, 423, 428, 711
P 131, 242, 290, 292, 293, 325, 364
T44
191-1, 10866, 11148, 15127
2. Resistance screening has been conducted in the field, under natural epidemic
conditions, using alternating rows of highly susceptible varieties.
Occasionally field screening has been supported by the release of viruliferous
whiteflies (Varma et al. 1992).
3. A positive correlation exists between whitefly populations and disease
incidence in the field (Patel and Srivastava 1990).
4. A number of agronomically acceptable cultivars have been released with
various levels of resistance. However, due to various disease rating systems
used by breeders, the term "resistance" has been used to refer to absence of
symptoms, low disease incidence, or delayed infection.
5. In most cases resistance has been confirmed in multilocational tests
conducted for several years. However, variation in the incidence between
locations and years has been observed occasionally. The presence of
strains or variants of the virus, or possibly even the involvement of
distinct viruses, has been suggested.
6. In the absence of effective diagnostic tools, it has not been possible to confirm
the presence of strains of this virus.
A set of recommendations has been formulated by the participants of this
workshop:
1. Address the MYMV problem by a cooperative regional network with AVRDC
as the coordinating body.
2. Intensify virus/vector studies on the following aspects:
a. exact geographic distribution of the virus
b. variability of MYMV and relationship of MYMV with known
geminiviruses affecting other leguminous crops
c. role of biotypes of the vector
d. information exchange

164 International Consultation
coordination by AVRDC of a biannual information bulletin/
newsletter on all aspects of MYMV research and breeding
▪ regular regional meetings to evaluate progress of MYMV research
Multilocational Testing of MYMV resistant Materials
The most agronomically promising IMMYMV-resistant mungbean materials
developed by the different national agricultural research systems in South Asia
and by AVRDC have been collected and multiplied. These materials now
undergo multilocational testing in India, Pakistan, Nepal, Sri Lanka, and
Bangladesh in the so-called 'hot spots,' where the disease is endemic.
Preliminary results of this multilocational testing are reported elsewhere.
Development of Molecular Markers for MYMV Resistance
A cooperative project with the University of Minnesota is in progress to develop
restriction fragment length polymorphism (RFLP) markers to locate MYMV
resistance genes in mungbean germplasm. Progress on this project is reported
elsewhere.
Development of MYMV specific DNA Probes
The various recommendations drawn up at the international workshop on
MYMV, particularly on the host range and geographic distribution of MYMV, and
on the variability of MYMV and its relationship with other geminiviruses
affecting legumes, clearly showed the need for improved diagnostic tools for the
detection of MYMV.
Because of the difficulty in isolating and purifying the virus, polyclonal antisera
are either not available or, if available, are nonspecific (Honda and Ikegami
1986, Ikegami and Shimizu 1988, Shimizu et al. 1987, Varma et al. 1992).
Similarly, monoclonal antibodies (MABS) produced against specific
geminiviruses, are known to cross-react with other geminiviruses (Macintosh
et al. 1992, Swanson et al. 1992). Virus-specific DNA probes are, therefore,
considered the most effective for the exact diagnosis of geminiviruses.
A DNA specific probe has been produced against the geminivirus which has
caused yellow mosaic of mungbean in Thailand (Chiemsombat 1992, Morinaga et
al. 1990). This virus is, however, clearly distinct from the one affecting mungbean
in the Indian subcontinent because of its transmissibility by sap (Honda et al.
1983, Honda and Ikegami 1986). The virus only occurred sporadically in Thailand
in 1977-81 (Thongmeearkom et al. 1981) but after 1981 its presence in the country
was never confirmed (Chiemsombat 1992). So far it is not known whether the
Indian MYMV is related to or different from the Thailand MYMV.

Workshop on Mungbean 165
AVRDC, in cooperation with the University of Wisconsin, Madison, USA has
developed a DNA probe against an isolate of MYMV from India. Mungbean leaf
samples were collected in the spring of 1993 in Southern India from plants
showing symptoms typical of MYMV infection.
DNA was extracted from air-dried leaf samples by using a modification of the
method described by Dellaporta et al 1983. DNA was amplified by polymerase
chain reaction (PCR). Three primer pairs were used: two general primer pairs for
whitefly-transmitted geminiviruses, i.e., one pair for the top and one pair for the
bottom half of the DNA-A component and another primer pair specifically
designed from the top half of the DNA-B sequence of the Thailand MYMV. PCRderived
DNA fragments and the plasmid pBluescript II KS (+) were digested with
PST 1 restriction enzyme. The DNA-A component was split into three DNA-A
fragments by Pst 1. DNA and the plasmid pBluescript II KS (+) were litigated and
transformed into competent E. coll. Selection for inserted plasmids was on TY
agarose medium, containing IPTG, X-gal, ampicillin, and tetracycline. Four
MYMV-specific clones, pIM 5, pIM 13, pIM 20, and pIM 9 were selected and
identified by DNA sequencing. The first three clones contained fragments of the
DNA-A, whereas pIM 9 contains a fragment of the DNA-B component of MYMV.
It was shown that MYMV from India had 95% sequence homology with the
MYMV from Thailand. However, restriction enzyme digests made from PCR
fragments of geminivirus-infected Croton sparsiflorns and Acalypha indica showed
that each contained a distinct geminivirus which, however, was not MYMV.
These two hosts have been implied as other natural hosts of MYMV.
The cloned DNA-A and DNA-B were used as templates to synthesize MYMVspecific
DNA probes by labeling with digoxigenin-11-dUTP (Boehringer
Mannheim). These will be used for determining levels of resistance, and for
epidemiological studies, such as host range, geographic distribution, and
presence of strains of MYMV. Positive signals have been obtained from yellow
mosaic diseased mungbean, cowpea, and soybean from various regions in India
and also from mungbean in Pakistan.
With its commitment to intensify work on MYMV, AVRDC hopes to coordinate
efforts of the NARS in the Indian subcontinent to develop high-yielding
mungbean lines with stable MYMV resistance which will lead to increased
acreage and sustainable production of this crop in the region and elsewhere in
Asia.
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16 6 International Consultation
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16S International Consultation
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Workshop on Mungbean 169
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