Molecular characterization of a Taiwanese isolate of Sweet potato chlorotic stunt virus

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Title:	Molecular characterization of a Taiwanese isolate of Sweet potato chlorotic stunt virus
Authors:	WANG,YUN-CHI
Contributors:	生物科技學系
Keywords:	Sweet potato chlorotic stunt virus;Crinivirus
Date:	2018-02-09
Issue Date:	2018-02-09 08:36:53 (UTC+0)
Publisher:	ASIA
Abstract:	The whitefly-transmitted sweet potato chlorotic stunt virus (SPCSV), belonging to the genus Crinivirus of the family Closteroviridae, severely affects productions of sweet potatoes worldwide. The current SPCSV isolates can be divided into two strains, West African (WA) and East African (EA), according to the genomic phylogeny. SPCSV was reported in Taiwan since 2012, but the molecular characteristic of the local SPCSV isolates is poorly understood. In this study, the whole genome sequence of a virus isolate collected from Chiayi city in 2016, and denoted CYI6 was determined from cDNA fragments amplified by reverse transcription-polymerase chain reaction (RT-PCR), first using the degenerate primers for viruses of Closteroviridae and followed by degenerate and specific primers designed from available sequences of SPCSV isolates. The nucleotide (nt) sequences of RNA1 and RNA2 of the CYI6 shared high identities of 98.7-98.9% and 98.5-98.8%, respectively, with those of SPCSV isolates of the WA strain, but shared lower identities of 70.0-81.5% for RNA1 and 70.3-70.5% for RNA2 with those of the EA strain. Additionally, the RNA-dependent RNA polymerase (RdRp), heat shock protein 70 homolog (Hsp70h) and major coat protein (CP) of CYI6 shared 96.4-99.8% amino acid (aa) identities with those of SPCSV isolates of the WA strain as well as the phylogenetic results, indicating that CYI6 is a SPCSV isolate belonging to the WA strain. A one-step RT-PCR method using CYI6-specific primers was developed to detect SPCSV in symptomatic sweet potato plants in field. A total of 108 sweet potato samples were collected from areas of Taipei, Taichung, Chiayi, Tainan, Kaohsiung and Pingtung during 2016 to 2017 for field survey. A SPCSV detection rate of 44.1% was obtained from the samples collected from Chiayi City. Our result showed that the distribution of SPCSV is limited in Taiwan and its spread should be more concerned.
Appears in Collections:	[Department of Biotechnology] Theses & dissertations

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