Determination and Analysis of the Complete Nucleotide Sequences of Different Isolates of Tobacco bushy top virus in Yunnan, China
LYU Jing1, LIU Fang1, TAN Guanlin2, WEI Wei1, YIN Chaoxian1, MA Yan1, CHEN Xiaojiao1, HE Lihuai1,3, CHEN Hairu1, LI Fan1
1. Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China, Yunnan Agricultural University, Kunming 650201, China; 2. Modern Education Technology Center, Yunnan Agricultural University, Kunming 650201, China; 3. Agricultural Technology Extension Center of Gucheng District, Lijiang 674199, China
[Purpose] The aim of this study was to explore the genetic differences among different isolates of Tobacco bushy top virus (TBTV) in Yunnan and complete the phylogenetic analysis.[Method] The complete genomes of ten TBTV Yunnan isolates were amplified by RT-PCR.[Result] All of the genomic sequences of the ten TBTV isolates were 4 152 nucleotides (nt) in size, and contained four open reading frames (ORFs). ORF2 expressed the fusion protein through -1 frameshift with ORF1, and the sequence GGAUUUU was suggested to be the signature for the frameshift strategies of ORF1-ORF2 in TBTV. Sequence analysis results revealed the nt identity of the complete sequence of the ten TBTV isolates was ranged from 84.9% to 99.1%, and isolate TBTV-DHo was the most variant compared with the other TBTV isolates. Recombinant events were detected among the TBTV Yunnan isolates. Phylogenetic relationships of these isolates involved in the recombinant, and recombinant was mainly occurred in the variant regions of ORF1-ORF2. ORF1 and ORF1-ORF2 were divergent among the four TBTV ORFs, while ORF4 and 3'UTR were conserved. Phylogenetic analysis showed that TBTV was most closely related to Opium poppy mosaic virus (OPMV) among different umbraviruses with nt identity of 61.1% to 61.6%, while was more distant to Carrot mottle virus (CMoV) and Carrot mottle mimic virus (CMoMV) with nt identity of 54%-55.1% in genus Umbravirus. Among the members in the 13 genera of family Tombusviridae, TBTV shared 25.1%-49.8% nt sequence identity with other viruses except umbraviruses, while umbraviruses were distant with the RNA2 of Dianthoviruses in family Tombusviridae.[Conclusion] Sequence analysis indicated a high genetic diversity among the TBTV Yunnan isolates, and recombinant events occurred among these isolates. TBTV was most closely related to OPMV among different umbraviruses.
LYU Jing, LIU Fang, TAN Guanlin, WEI Wei, YIN Chaoxian, MA Yan, CHEN Xiaojiao, HE Lihuai, CHEN Hairu, LI Fan.Determination and Analysis of the Complete Nucleotide Sequences of Different Isolates of Tobacco bushy top virus in Yunnan, China[J] Journal of Yunnan Agricultural University, 2017,V32(3): 410-420
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