Documentos de Académico
Documentos de Profesional
Documentos de Cultura
B.K. Choi
a
, J.M. Koo
a
, H.J. Ahn
a
, H.J. Yum
a
, C.W. Choi
a,b,
,
K.H. Ryu
c
, P. Chen
d
, S.A. Tolin
e
a
Department of Biology and Medicinal Science, Pai Chai University, Daejeon 302-735, Republic of Korea
b
Bio-medicinal Research Center (RRC), Pai Chai University, Daejeon 302-735, Republic of Korea
c
Plant Virus GenBank, Division of Life and Environmental Sciences, Seoul Womens University, Seoul 139-774, Republic of Korea
d
Department of Crop, Soil, and Environmental Sciences, University of Arkansas, Fayetteville, AR 72701, USA
e
Department of Plant Pathology, Physiology and Weed Science, Virginia Tech, Blacksburg, VA 24061, USA
Received 14 July 2004; received in revised form 9 March 2005; accepted 9 March 2005
Available online 24 May 2005
Abstract
Twelve Rsv resistance-breaking (RB) isolates of Soybean mosaic virus (SMV) were obtained from eld-grown soybean plants showing
mosaic symptoms and subsequently examined biologically and molecularly. All of these RB isolates were identied as SMV based on
serological and infectivity assays, and the amplication of P1 gene products by reverse transcription-polymerase chain reaction (RT-PCR).
Differential soybean cultivars, lines or accessions Lee 68 (rsv), PI 96983, York, Marshall, Ogden, Kwanggyo, Suweon 97 (Rsv1 alleles),
L29 (Rsv3), and V94-5152 (Rsv4), following inoculation with each RB isolate, showed similar systemic symptoms suggesting that these
RB isolates can overcome Rsv resistance at three loci. To differentiate the 12 RB isolates molecularly, the P1 coding region for each isolate
was amplied, cloned, sequenced and compared to known SMV strains. The P1 region from the RB isolates shared 8690% and 9099%
similarities in amino acid (aa) and nucleotide sequence, respectively, with known SMV strains. Comparison of aa sequences indicated that
these RB isolates are newly emerging isolates capable of breaking Rsv resistance. Phylogenetic analysis further suggested that the RB isolates
can be classied as three major types. However, recombination was not observed within the coding region of P1 protein among the types.
This is the rst report on the emergence of SMV isolates capable of overcoming all of the known resistance alleles at the Rsv1 locus, as well
as distinct resistance genes at Rsv3 and Rsv4.
2005 Elsevier B.V. All rights reserved.
Keywords: Rsv resistance-breaking (RB) isolates; Soybean mosaic virus (SMV); P1 protein; Amino acid sequences; Phylogenetic analysis
1. Introduction
Soybean mosaic virus (SMV), a member of the genus
Potyvirus, is one of the most prevalent viral pathogens of
Corresponding author. Tel.: +82 42 520 5617; fax: +82 42 520 5617.
E-mail address: choicw@mail.pcu.ac.kr (C.W. Choi).
soybean [Glycine max (L.) Merr.] worldwide. The virus
has a positive-sense single-stranded RNA genome of 9588
nucleotides with VPg at the 5
C
for 2 min, and DNA synthesis at 72
5
1
Table 3
Matrix of similarities of P1 gene nucleotide and amino acid sequences between new resistance-breaking (RB) isolates and known strains of Soybean mosaic virus
Similarities (%) to isolates and known strains
a
CN 1 CN 2 CN 3 CN 7 CN 9 CN 10 CN 12 CN 13 CN 15 CN 18 CN 31 CN 36 G1 G2 G3 G5a G5b G6 G7 G7d G7H Aa HH5 HZ N
CN1 94 89 89 96 93 99 95 92 95 89 91 95 90 92 95 96 93 87 93 97 93 96 96 95
CN2 95 89 89 95 94 94 95 91 97 89 90 94 89 91 94 95 92 87 92 95 91 95 94 95
CN3 91 91 98 91 88 90 90 89 89 98 90 91 87 90 90 90 88 86 90 91 90 92 91 92
CN7 92 91 99 91 88 89 90 89 89 99 89 91 87 89 90 90 88 86 90 90 89 91 91 92
CN9 96 96 92 93 94 96 98 93 96 91 91 96 91 92 99 97 94 88 93 97 93 97 96 96
CN10 94 96 90 91 95 93 93 91 94 88 90 93 88 90 93 94 91 86 91 94 91 94 93 93
CN12 99 95 92 92 96 94 96 93 95 90 92 96 91 92 96 97 93 88 93 97 93 97 96 96
CN13 95 95 92 92 99 94 96 93 95 90 91 95 90 92 98 96 94 88 93 97 93 96 96 95
CN15 94 94 92 92 95 93 94 95 92 89 95 94 89 94 92 93 91 88 93 93 97 94 94 94
CN18 96 98 92 92 97 96 96 96 95 89 91 95 90 92 95 96 93 88 93 96 92 96 95 96
CN31 92 91 99 99 93 91 92 92 92 92 90 91 87 90 90 90 88 86 90 91 90 92 91 92
CN36 93 93 92 92 94 92 93 93 96 94 92 92 88 92 90 92 90 87 92 93 95 92 92 93
G1 96 95 93 93 96 94 96 96 95 96 93 95 94 94 95 96 93 89 95 97 94 97 96 99
G2 95 94 92 92 95 93 95 95 94 95 92 94 98 89 90 91 89 93 90 92 89 92 91 94
G3 94 94 92 92 95 93 94 94 95 94 92 94 95 94 92 93 90 89 93 93 94 94 94 94
G5a 96 96 92 92 99 95 96 98 95 96 92 93 96 95 94 96 94 87 93 97 93 96 96 95
G5b 97 95 92 93 97 94 97 96 94 96 93 94 96 95 94 96 94 89 94 99 93 98 97 97
G6 95 95 91 92 97 94 95 96 94 96 92 93 95 94 94 96 95 86 91 94 91 94 93 94
G7 93 93 92 92 94 92 94 94 94 94 92 93 95 94 94 94 94 93 93 89 88 89 89 90
G7d 93 93 92 92 94 92 94 94 94 94 92 93 95 94 94 94 95 93 98 94 93 95 94 95
G7H 97 95 93 93 97 94 97 96 95 96 93 94 96 95 95 96 99 95 95 95 94 98 97 97
Aa 95 94 92 93 96 93 95 95 98 95 93 97 96 95 96 95 95 94 94 94 95 94 94 94
HH5 98 96 93 93 97 95 98 96 95 97 93 94 97 96 95 97 98 96 95 95 98 95 98 97
HZ 97 95 92 92 96 94 97 96 95 96 92 93 96 95 95 96 97 95 94 94 97 95 99 97
N 95 95 93 93 96 94 96 96 95 96 93 95 99 98 95 96 96 95 95 95 96 96 97 96
a
Numbers in bold type at upper right corner refer amino acid sequence similarities and numbers in plain type at lower left corner refer to nucleotide sequence similarities.
B.K. Choi et al. / Virus Research 112 (2005) 4251 49
Fig. 2. Phylogenetic analyses for amino acid (aa) sequences of P1 protein of 12 new resistance-breaking SMV isolates and those of 13 known strains and
isolates obtained from the NCBI database. (A) The phlogenetic tree (unrooted) was constructed by the DNAMAN version 5.2.9. The grouping occurred after
bootstrapping the data (only values >50% are shown) and the numbers on the branches indicate bootstrap percentages based on 1000 resamplings. (B) The
phlogenetic tree (unrooted) was constructed by the neighbor-joining method by CLUSTALW and the numbers at the end of branches indicate genetic distance
corrected by Kimuras method (1983). The P1 aa sequences of ZYMV was used as the outgroup.
differences between SMV-G5b (AY294044) from Korea and
-G5a (AF200564) from USA (or between SMV-G7 and -
G7H), even though they shared similar biological characteris-
tics. Perhaps SMVisolates of the same origin could diversify
due to selection pressure overtime imposed by host cultivars
as well as by aphid and seed transmission. The emergence
of a new variant from the same strain group, SMV-G7d, by
serial inoculation of a culture of SMV-G7 originating from
a cDNA clone in the resistant cultivar PI 96983 possess-
ing Rsv1 (Hajimorad et al., 2003), supports this hypothesis.
Similar evidence was reported from rapid emergence of RB
isolates in Rice yellow mottle Sobemovirus (Fargette et al.,
2002). The emergence of SMV RB isolates in Korean soy-
bean cultivars can most likely be attributed to release and
utilization of resistant germplasm and cultivars, to the use of
a narrow genetic base for SMV resistance, to selection pres-
sure overtime on SMV populations, and to mutation and/or
genetic recombination, all common events in plant virus evo-
lution (Roosinck, 1997; Garca-Arenal et al., 2001; Harrison,
2002).
50 B.K. Choi et al. / Virus Research 112 (2005) 4251
Acknowledgement
This workwas partlysupportedbya grant (R05-2003-000-
10293-0) fromthe Basic Research Programof the Korea Sci-
ence and Engineering Foundation (KOSEF), and by a grant
(R12-1999-002-03003-02003) from KOSEF, MOST and the
city of Daejeon through the Research Center for Biomedici-
nal Resources (RRC). We appreciate the ChungnamAgricul-
tural Research and Extension Services for collecting infected
soybean plants and National Crop Experimental Station for
supplying soybean seeds of differential cultivars.
References
Aaziz, R., Tepfer, M., 1999. Recombination between genomic RNAs of
two cucumoviruses under conditions of minimal selection pressure.
Virology 263, 282289.
Bousalem, D., Douzery, E.J.P., Fargette, D., 2000. High genetic diver-
sity, distant phylogenetic relationships and intraspecies recombination
events among natural populations of Yam mosaic virus: a contribu-
tion to understanding potyvirus evolution. J. Gen. Virol. 81, 243
255.
Buss, G.R., Ma, G., Chen, P., Tolin, S.A., 1997. Registration of V94-5152
soybean germplasm resistant to soybean mosaic potyvirus. Crop Sci.
37, 19871988.
Buss, G.R., Ma, G., Kristipati, S., Chen, P., Tolin, S.A., 1999. A new
allele at the Rsv3 locus for resistance to soybean mosaic virus. In:
H.E. Kauffman (Ed.), Proceedings of the World Soybean Research
Conference VI, Superior Printing, Chicago, IL, p. 490.
Chen, P., Buss, G.R., Roane, C.W., Tolin, S.A., 1991. Allelism among
genes for resistance to soybean mosaic virus in strain-differential soy-
bean cultivars. Crop Sci. 31, 305309.
Chen, P., Buss, G.R., Tolin, S.A., Gunduz, I., Cicek, M., 2002a. A valu-
able gene in Suweon 97 soybean for resistance to soybean mosaic
virus. Crop Sci. 42, 333337.
Chen, Y.-K., Goldbach, R., Prins, M., 2002b. Inter- and intramolecu-
lar recombinations in the Cucumber mosaic virus genome related to
adaptation to Alstroemeria. J. Virol. 76, 41194124.
Chen, J., Zheng, H.-Y., Lin, L., Adams, M.J., Antoniw, J.F., Zhao, M.-F.,
Shang, Y.-F., Chen, J.-P., 2004. A virus related to Soybean mosaic
virus from Pinellia ternate in China and its comparison with local
soybean SMV isolates. Arch. Virol. 149, 349363.
Cho, E.K., 1995. Identication of soybean mosaic virus strains and a
consideration on genetics of soybean for resistance to SMV strains.
Plant Dis. Agr. 1, 2225.
Cho, E.K., Chung, K.W., 1986. Strains of soybean mosaic virus caus-
ing soybean necrotic disease in Korea. Korean J. Breed. 18, 150
153.
Cho, E.K., Goodman, R.M., 1979. Strains of soybean mosaic virus: classi-
cation based on virulence in resistant soybean cultivars. Phytopathol-
ogy 69, 467470.
Cho, E.K., Goodman, R.M., 1982. Evaluation of resistance in soybeans
to soybean mosaic virus strains. Crop Sci. 22, 11331136.
Cho, E.K., Chung, B.J., Lee, S.H., 1977. Studies on identication and
classication of soybean virus disease in Korea. II. Etiology of a
necrotic disease of Glycine max. Plant Dis. Rep. 61 (4), 313317.
Cho, E.K., Choi, S.H., Cho, W.T., 1983. Newly recognized soybean mo-
saic virus mutants and sources of resistance in soybeans. Res. Rept.
ORD (S.P.M.U). 25, 1822.
Chung, B.J., Lee, S.H., Cho, E.K., Park, H.C., 1974. Identication of
soybean viruses and soybean varietal reactions. Annu. Rep. C.I.R.C.,
137145.
Domier, L.L., Latorre, I.J., Steinlage, T.A., McCoppin, N., Hartman,
G.L., 2003. Variability and transmission by Aphis glycines of North
American and Asian Soybean mosaic virus isolates. Arch. Virol. 148,
19251941.
Dougherty, W.G., Semler, B.L., 1993. Expression of virus-encoded pro-
teinases: functional and structural similarities with cellular enzymes.
Microbiol. Rev. 57, 781822.
Eggenberger, A.L., Stark, D.M., Beachy, R.N., 1989. The nucleotide se-
quence of a soybean mosaic virus coat protein-coding region and its
expression in Escherichia coli, Agrobacterium tumefaciens and to-
bacco callus. J. Gen. Virol. 70, 18531860.
Fargette, D., Pinel, A., Traor e, O., Ghesqui` ere, A., Konat e, G., 2002.
Emergence of resistance-breaking isolates of Rice yellow mottle virus
during serial inoculations. Eur. J. Plant Pathol. 108, 585591.
Garca-Arenal, F., Fraile, A., Malpica, J.M., 2001. Variability and ge-
netic structure of plant virus populations. Annu. Rev. Phytopathol.
39, 157186.
Glais, L., Tribodet, M., Kerlan, C., 2002. Genomic variability in Potato
potyvirus Y (PVY): evidence that PVY (N) W and PVY (NTN) vari-
ants are single to multiple recombinants between PVY (O) and PVY
(N) isolates. Arch. Virol. 147, 363378.
Gunduz, I., Buss, G.R., Chen, P., Tolin, S.A., 2002. Characterization of
SMV resistance genes in Tousan 140 and Hourei soybean. Crop Sci.
42, 9095.
Gunduz, I., Buss, G.R., Chen, P., Tolin, S.A., 2004. Genetic and pheno-
typic analysis of Soybean mosaic virus resistance in PI 88788 soy-
bean. Phytopathology 94, 687692.
Hajimorad, M.R., Eggenberger, A.L., Hill, J.H., 2003. Evolution of Soy-
bean mosaic virus-G7 molecularly cloned genome in Rsv1 genotype
soybean results in emergence of a mutant capable of evading Rsv1-
mediated recognition. Virology 314, 487509.
Harrison, B.D., 2002. Virus variation in relation to resistance-breaking in
plants. Euphytica 124, 181192.
Higgins, D., Thompson, J., Gibson, T., Thompson, J.D., Higgins, D.G.,
Ginson, T.J., 1994. CLUSTAL W: Improving the sensitivity of pro-
gressive multiple sequence alignment through sequence weighting,
position-specic gap penalties and weight matrix choice. Nucleic
Acids Res. 22, 46734680.
Hillis, D.M., Bull, J.J., 1993. An empirical test of bootstrapping as a
method of assessing condence in phlogenetic analysis. Syst. Biol.
42, 182192.
Hunst, P.L., Tolin, S.A., 1982. Isolation and comparison of two strains
of soybean mosaic virus. Phytopathology 72, 710713.
Jain, R.K., McKern, N.M., Tolin, S.A., Hill, J.H., Barnett, O.W., Tosic,
M., Ford, R.E., Beachy, R.N., Yu, M.H., Ward, C.W., Shukla, D.D.,
1992. Similarity of coat protein peptide proles of fourteen potyvirus
isolates from soybean conrms that they are strains of the one virus.
Phytopathology 82, 294299.
Jayaram, C., Hill, J.H., Miller, W.A., 1992. Complete nucleotide se-
quences of two soybean mosaic virus strains differentiated by re-
sponse of soybean containing the Rsv resistance gene. J. Gen. Virol.
73, 20672077.
Kiihl, R.A.S., Hartwig, E.E., 1979. Inheritance of reaction to soybean
mosaic virus in soybeans. Crop Sci. 19, 372375.
Kim, Y.H., 2000. Molecular classication of soybean mosaic virus strains
and their interaction with soybeans. Ph.D. Thesis, Kyungpook Natl.
Univ., Daegu, Korea.
Kim, Y.H., Kim, O.S., Lee, B.C., Moon, J.K., Lee, S.C., Lee, J.Y., 2003.
G7H, a new Soybean mosaic virus strain: its virulence and nucleotide
sequence of CI gene. Plant Dis. 87, 13721375.
Kimura, M., 1983. The natural theory of molecular evolution. Cambridge
University Press, Cambridge, England.
Lee, Y.H., Hwang, Y.H., Seong, Y.G., Kim, Y.H., Kim, W.H., 1992. Re-
sistance to disease, good in seed quality, high yielding and widely
adapted new soybean variety Taekwangkong. Res. Rep. RDA 34
(2), 1115.
Lee, K.C., Wong, S.M., 1998. Variability of P1 protein of zucchini yellow
mosaic virus for strain differentiation and phylogenetic analysis with
other potyviruses. DNA Seq. 9, 275293.
B.K. Choi et al. / Virus Research 112 (2005) 4251 51
Lim, W.-S., Kim, Y.-H., Kim, K.-H., 2003. Complete genome sequences
of the genomic RNA of soybean mosaic virus strains G7H and G5.
Plant Pathol. J. 19 (3), 171176.
Lin, S.S., Hou, R.F., Yeh, S.D., 2001. Complete genome sequence and
genetic organization of a Taiwan isolate of Zucchini yellow mosaic
virus. Bot. Bull. Acad. Sin. 42, 243250.
Ma, G., Chen, P., Buss, G.R., Tolin, S.A., 1995. Genetic characteristics
of two genes for resistance to soybean mosaic virus in PI 486355
soybean. Theor. Appl. Genet. 91, 907914.
Ma, G., Chen, P., Buss, G.R., Tolin, S.A., 2003. Genetic study of a lethal
necrosis to soybean mosaic virus in PI507389 soybean. J. Hered. 94,
205211.
Martin, D.P., Rybicki, E.P., 2000. RDP: detection of recombination
amongst aligned sequences. Bioinformatics 16, 562563.
Mavankal, G., Rhoads, R.E., 1991. In vitro cleavage at or near the N-
terminus of the helper component protein in the tobacco vein mottling
virus polyprotein. Virology 185, 721731.
Milbrath, G.M., Soong, M., 1976. A local lesion assay for soybean mo-
saic virus using Phaseolus vulgaris L. Top Crop. Phytopathol. Z. 87,
255259.
Moreno, I.M., Malpica, J.M., Daz-Pend on, J.A., Moriones, E., Fraile, A.,
Garca-Arenal, F., 2004. Variability and genetic structure of the popu-
lation of watermelon mosaic virus infecting melon in Spain. Virology
318, 451460.
Nagy, P.D., Christopher, O., Bujarski, J.J., 1999. Mapping sequences
active in homologous RNA recombination in brome mosaic
virus: prediction of recombination hot spots. Virology 254, 92
104.
Pu, Z., Cao, Q., Fang, D., Xi, B., Fang, C., 1982. Identication of strains
of soybean mosaic virus. Acta Phytophyl. Sin. 9, 1520.
Roosinck, M.J., 1997. Mechanisms of plant virus evolution. Annu. Rev.
Phytopathol. 35, 191209.
Saitou, N., Nei, M., 1987. The neighbor-joining method: a new method
for reconstructing phylogenetic trees. Mol. Biol. Evol. 4, 406425.
Takahashi, K., Tanaka, T., Iida, W., Tsuda, Y., 1980. Studies on virus
diseases and casual viruses of soybean in Japan. Bull. Tohoku Natl.
Agr. Exp. Sta. 62, 1130.
Tomimura, K., Gibbs, A.J., Jenner, C.E., Walsh, J.A., Ohshima, K., 2003.
The phylogeny of Turnip mosaic virus; comparisons of 38 genomic
sequences reveal a Eurasian origin and a recent emergence in Asia.
Mole. Ecol. 12, 20992111.
Vance, V.B., Moore, D., Turpen, T.H., Bracker, A., Hollowell, V.C., 1992.
The complete nucleotide sequence of pepper mottle virus genomic
RNA: comparison of the encoded polyprotein with those of other
sequence potyviruses. Virology 191, 1930.
Verchot, J., Carrington, J.C., 1995. Evidence that the potyvirus P1 pro-
teinase functions in trans as an accessory factor for genome ampli-
cation. J. Virol. 69, 36683674.
Verchot, J., Herndon, K.L., Carrington, J.C., 1992. Mutational analysis
of the tobacco etch potyviral 35-kDa-proteinase: identication of es-
sential residues and requirements for autoproteolysis. Virology 190,
298306.