Nucleotide sequence analysis and detection application of the coat protein gene of Sweet potato latent virus isolated from central Taiwan

Li-Yuan Wang; Kuan-Chun Chen; Tsung-Chi Chen; Shyi-Dong Yeh; 王麗媛; 陳冠君; 陳宗祺; 葉錫東; Li-Yuan Wang;Kuan-Chun Chen;Tsung-Chi Chen;Shyi-Dong Yeh; 王麗媛;陳冠君;陳宗祺;葉錫東

doi:10.6649/PPB.200709_16(3).0004

Nucleotide sequence analysis and detection application of the coat protein gene of Sweet potato latent virus isolated from central Taiwan

Other Title

甘藷潛伏病毒中台灣分離株鞘蛋白基因序列分析及於檢測上之應用

Journal

植物病理學會刊

Journal Volume

16

Journal Issue

3

Pages

141-148

Date Issued

2007-09

Date

2007-09

Author(s)

Li-Yuan Wang

Kuan-Chun Chen

Tsung-Chi Chen

Shyi-Dong Yeh

王麗媛

陳冠君

陳宗祺

葉錫東

DOI

10.6649/PPB.200709_16(3).0004

URI

https://scholars.tari.gov.tw/handle/123456789/8822

Abstract

A potyvirus-like agent (isolate CY) was isolated from sweet potato with leaf symptoms of chlorotic spots and vein mottling in Chia-Yi area, Taiwan. A 1.9-kb DNA product was amplified by reverse-transcription polymerase chain reaction (RT-PCR) from infected tissues of C. quinoa using the oligo(dT) and degenerate primers for potyviruses, cloned, and then sequenced. The cDNA fragment reflected 1880 nucleotides (nts) corresponding to the 3'-terminal region of a potyvirus and the sequence comparison indicated that the isolate CY belongs to Sweet potato latent virus (SPLV). The deduced amino acid sequence was determined as 561 residues that contains a part of the 3'-terminal region of NIb gene (268 residues) and the complete sequence of coat protein (CP) gene (293 residues). The 3'-terminal region of the cDNA was determined containing a non-coding region (NCR) of 197 nts. The DAG triplet for aphid transmissibility was found at the 7-9 residues from the N-terminus of CP gene. Multiple sequence alignment of the known sequences of SPLV indicated that the CP gene and the NCR of CY isolate share 96.5 and 100% nucleotide identities, respectively, with those of SPLVTW. The phylogenetic analysis indicated that the CY isolate is closely related to Taiwan and China isolates, but distantly related to a Japan isolate. When a pair of specific primers designed from the CP gene of SPLV-CY was used for detection by RT-PCR, a 675 bp specific DNA fragment was amplified from SPLV-CY infected plants. 於嘉義地區，自呈現黃斑及葉脈斑駁的甘藷葉片可分離到似potyvirus的長絲狀病毒(potyvirus-like isolate CY)。利用對應Potyvirus屬病毒基因體核糖核酸3’端之簡併式引子及寡合腺嘌呤引子，可由呈現局部斑之奎藜總量核糖核酸(total RNA)以反轉錄聚合酣連鎖反應增幅出約1.9-kb核酸片段。此核酸片段經選殖及核酸序列分析，全長含1880個核苷酸序列。由5’端起此序列對應的肢基酸共561個，分別為268個肢基酸於細胞核內含體b (nuclear inclusion b, NIb)、及293個肢基酸之全長度鞘蛋白(coat protein, CP)基因。3’端非轉譯區(3' non-coding region, 3'-NCR)則由197個核酸組成。兩者之間蛋白酶切位落於VHHQ/A之間，且由鞘蛋白N端起第7-9個肢基酸位置其有被蚜蟲傳播能力之DAG序列，不同於SPLV-TW之DTG。與登錄於GenBank之SPLV分離株比對核酸序列，與親緣最近之SPLV-TW病毒其CP及3'-NCR區域之核苷酸序列相同度分別有96.5%及100%。鞘蛋白基因演化關係分析此病毒和台灣及中國人陸分離株較為親近而與日本分離株較為疏遠。根據解得SPLV-CY鞘蛋白基因序列，設計之專一性引子對L166/L841，可由SPLV-CY感染之奎藜、煙草及甘藷等病株檢測出專一性675bP 核酸片段。

Subjects

Sweet potato latent virus

gene cloning

sequence analysis

detection

甘藷潛伏病毒

核酸選殖

序列分析

檢測

Publisher

中華民國植物病理學會

Type

journal article

File(s)

Name

16-3-141-148.pdf

Size

2.27 MB

Format

Adobe PDF

Checksum

(MD5):5c7f3478cdec91eeee5f1e0d7c67cf7f