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Complete Nucleotide Sequences and Genome Organization of Two Pepper Mild Mottle Virus Isolates from Capsicum annuum in South Korea Seung-Kook Choi, Gug-Seoun Choi, Sun-Jung Kwon, Ju-Yeon Yoon Horticultural and Herbal Crop Environment Division, National Institute of Horticultural & Herbal Science, RDA, Wanju, Republic of Korea
The complete genome sequences of pepper mild mottle virus (PMMoV)-P2 and -P3 were determined by the Sanger sequencing method. Although PMMoV-P2 and PMMoV-P3 have different pathogenicity in some pepper cultivars, the complete genome sequences of PMMoV-P2 and -P3 are composed of 6,356 nucleotides (nt). In this study, we report the complete genome sequences and genome organization of PMMoV-P2 and -P3 isolates from pepper species in South Korea. Received 5 April 2016 Accepted 8 April 2016 Published 19 May 2016 Citation Choi S-K, Choi G-S, Kwon S-J, Yoon J-Y. 2016. Complete nucleotide sequences and genome organization of two pepper mild mottle virus isolates from Capsicum annuum in South Korea. Genome Announc 4(3):e00411-16. doi:10.1128/genomeA.00411-16. Copyright © 2016 Choi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. Address correspondence to Ju-Yeon Yoon, [email protected].
P
epper mild mottle virus (PMMoV), which belongs to the genus Tobamovirus in the family Virgaviridae, is one of the major viruses in pepper species (Capsicum spp.) in the world (1). Tobamovirus strains isolated from pepper plants have been classified into four subgroups, P0, P1, P1,2 and P1,2,3, on the basis of their responses with the corresponding resistance genes controlled by L1, L2, L3, and L4, which are considered to be alleles at the locus L (2). We described a resistance-breaking pathotype of PMMoV previously, designated PMMoV-P3, that overcomes the L3-mediated resistance in C. annuum and compared the pathogenicity of PMMoV-P3 in paprika cultivars with that of PMMoV-P2 (which overcomes the L2 resistance) (3, 4). To determine complete nucleotide sequences of PMMoV-P2 and -P3, viral RNAs isolated from purified virus particles using the SDS/Phenol/ Proteinase K method (5) were used as templates for reverse transcription-PCR (RT-PCR) with four pairs of PMMoVuniversial primers. Synthesized PCR products covering the complete genomes of PMMoV-P2 and -P3 were cloned into the pCR4TOPO vector for sequencing (Invitrogen, USA). Although PMMoV-P2 and PMMoV-P3 have different pathogenicities in some pepper cultivars, the complete genome sequences of PMMoV-P2 and -P3 were composed of 6,356 nucleotides (nt). Each viral genome encodes 4 proteins; 126 kDa, 183 kDa, movement protein, and coat protein including 5= and 3= untranslated regions with 69 and 198 nt, as described previously. PMMoV-P2 and -P3 shared high sequence identities of nt and
May/June 2016 Volume 4 Issue 3 e00411-16
amino acids with eight PMMoV strains isolated from geographically different regions. To the best of our knowledge, these are the complete genome sequences and genome organization of PMMoV-P3 isolates from pepper species in South Korea. Nucleotide sequence accession numbers. The complete genome sequences of PMMoV-P2 and -P3 have been deposited in GenBank under the accession numbers LC082099 and LC082100. FUNDING INFORMATION This work, including the efforts of Ju-Yeon Yoon, was funded by Rural Development Administration (RDA) (PJ0108782016). This study was supported by a grant (PJ0108782016) from the Basic Research Program of the National Institute of Horticultural and Herbal Science, RDA, South Korea.
REFERENCES 1. Adams MJ, Antoniw JF, Kreuze J. 2009. Virgaviridae: a new family of rod-shaped plant viruses. Arch Virol 154:1967–1972. 2. Boukema IW. 1984. Resistance to TMV in Capsicum chacoense Hunz is governed by an allele of the L-locus. Capsicum Newslett 3:47– 48. 3. Choi GS, Choi SK, Cho JD, Cho IS. 2013. [A pathotype of pepper mild mottle virus causing necrotic spot symptoms in paprika fruit]. Res Plant Dis 19:124 –127. (In Korean.) 4. Choi GS, Choi SK, Cho IS, Kwon SJ. 2014. [Resistance screening to pepper mild mottle virus pathotypes in paprika cultivars]. Res Plant Dis 20: 299 –302. (In Korean.) 5. Gooding GV, Hebert TT. 1967. A simple technique for purification of tobacco mosaic virus in large quantities. Phytopathology 57:1285.
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Complete Nucleotide Sequences and Genome Organization of Two Pepper Mild Mottle Virus Isolates from Capsicum annuum in South Korea Seung-Kook Choi, Gug-Seoun Choi, Sun-Jung Kwon, Ju-Yeon Yoon Horticultural and Herbal Crop Environment Division, National Institute of Horticultural & Herbal Science, RDA, Wanju, Republic of Korea
The complete genome sequences of pepper mild mottle virus (PMMoV)-P2 and -P3 were determined by the Sanger sequencing method. Although PMMoV-P2 and PMMoV-P3 have different pathogenicity in some pepper cultivars, the complete genome sequences of PMMoV-P2 and -P3 are composed of 6,356 nucleotides (nt). In this study, we report the complete genome sequences and genome organization of PMMoV-P2 and -P3 isolates from pepper species in South Korea. Received 5 April 2016 Accepted 8 April 2016 Published 19 May 2016 Citation Choi S-K, Choi G-S, Kwon S-J, Yoon J-Y. 2016. Complete nucleotide sequences and genome organization of two pepper mild mottle virus isolates from Capsicum annuum in South Korea. Genome Announc 4(3):e00411-16. doi:10.1128/genomeA.00411-16. Copyright © 2016 Choi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. Address correspondence to Ju-Yeon Yoon, [email protected].
P
epper mild mottle virus (PMMoV), which belongs to the genus Tobamovirus in the family Virgaviridae, is one of the major viruses in pepper species (Capsicum spp.) in the world (1). Tobamovirus strains isolated from pepper plants have been classified into four subgroups, P0, P1, P1,2 and P1,2,3, on the basis of their responses with the corresponding resistance genes controlled by L1, L2, L3, and L4, which are considered to be alleles at the locus L (2). We described a resistance-breaking pathotype of PMMoV previously, designated PMMoV-P3, that overcomes the L3-mediated resistance in C. annuum and compared the pathogenicity of PMMoV-P3 in paprika cultivars with that of PMMoV-P2 (which overcomes the L2 resistance) (3, 4). To determine complete nucleotide sequences of PMMoV-P2 and -P3, viral RNAs isolated from purified virus particles using the SDS/Phenol/ Proteinase K method (5) were used as templates for reverse transcription-PCR (RT-PCR) with four pairs of PMMoVuniversial primers. Synthesized PCR products covering the complete genomes of PMMoV-P2 and -P3 were cloned into the pCR4TOPO vector for sequencing (Invitrogen, USA). Although PMMoV-P2 and PMMoV-P3 have different pathogenicities in some pepper cultivars, the complete genome sequences of PMMoV-P2 and -P3 were composed of 6,356 nucleotides (nt). Each viral genome encodes 4 proteins; 126 kDa, 183 kDa, movement protein, and coat protein including 5= and 3= untranslated regions with 69 and 198 nt, as described previously. PMMoV-P2 and -P3 shared high sequence identities of nt and
May/June 2016 Volume 4 Issue 3 e00411-16
amino acids with eight PMMoV strains isolated from geographically different regions. To the best of our knowledge, these are the complete genome sequences and genome organization of PMMoV-P3 isolates from pepper species in South Korea. Nucleotide sequence accession numbers. The complete genome sequences of PMMoV-P2 and -P3 have been deposited in GenBank under the accession numbers LC082099 and LC082100. FUNDING INFORMATION This work, including the efforts of Ju-Yeon Yoon, was funded by Rural Development Administration (RDA) (PJ0108782016). This study was supported by a grant (PJ0108782016) from the Basic Research Program of the National Institute of Horticultural and Herbal Science, RDA, South Korea.
REFERENCES 1. Adams MJ, Antoniw JF, Kreuze J. 2009. Virgaviridae: a new family of rod-shaped plant viruses. Arch Virol 154:1967–1972. 2. Boukema IW. 1984. Resistance to TMV in Capsicum chacoense Hunz is governed by an allele of the L-locus. Capsicum Newslett 3:47– 48. 3. Choi GS, Choi SK, Cho JD, Cho IS. 2013. [A pathotype of pepper mild mottle virus causing necrotic spot symptoms in paprika fruit]. Res Plant Dis 19:124 –127. (In Korean.) 4. Choi GS, Choi SK, Cho IS, Kwon SJ. 2014. [Resistance screening to pepper mild mottle virus pathotypes in paprika cultivars]. Res Plant Dis 20: 299 –302. (In Korean.) 5. Gooding GV, Hebert TT. 1967. A simple technique for purification of tobacco mosaic virus in large quantities. Phytopathology 57:1285.
Genome Announcements
genomea.asm.org 1
