Characterization of Zucchini Yellow Mosaic Virus (ZYMV) Isolates Collected from Taiwan by Host Reactions, Serology, and RT-PCR
Other Title
以寄主反應、血清學及反轉錄酶連鎖反應分析臺灣矮南瓜黃化嵌紋病毒之特性
Journal
植物保護學會會刊
Journal Volume
40
Journal Issue
2
Pages
163-176
Date Issued
1998-06
Author(s)
Shih-Shun Lin
Roger F. Hou
Chiou-Hsiung Huang
Shyi-Dong Yeh
林詩舜
侯豐男
黃秋雄
葉錫東
Abstract
Zucchini yellow mosaic virus (ZYMV) isolates collected from different areas of Taiwan were characterized by host reactions, sodium dodecyl sulfate (SDS) immunodiffusion tests, and restriction enzyme polymorphism of reverse transcription-polymerase chain reaction (RT-PCR) products. The sympotoms on zucchini squash by the 33 ZYMV isolates were divided into three types: yellow mosaic with leaf distortion and narrowing, yellow mosaic with leaf distortion, and yellow mosaic with vein-banding. There ZYMV isolates induced yellow mosaic sympotoms on melon, sponge gourd and cucumber, except that the isolate CY2 acused necrosis on the leaves of sponge gourd and cucumber. In SDS-immunofiffusion tests, the antiserum prepared against the isolate TN3 reacted with all the 33 ZYMV isolates indistinguishabaly by formation of homogenous precipition lines without spurs. In order to provide a fast and sensitive diagnostic tool, a RT-PCR detection method was developed. A 0.76 kb DNA fragment was amplified from the 33 isolates by RT-PCR using primers specific to the NIb and CP genes of ZYMV. To further analyze the variability of the CP genes of five selected ZYMV isolates, EcoRV, TaqI, and NlaIII digestion patterns of RT-PCR products, which were amplified from the variable N-terminal halves of the CP genes, were compared. The results showed that the isolates TC1, CY2, and TNML1 are closely related, while TN3 and NT1 are more variable.
本研究以寄主反應、免疫擴散反應及反轉錄酵素連鎖反應 （RT-PCR） 產物的限制酵素多型性分析來比較從臺灣不同瓜類產區收集之矮南瓜黃化嵌紋病毒（zucchini yellow mosaic virus, ZYMV) 分離株之異同。所得三十三個病毒分離株在矮南瓜葉片上所造成的病徵可歸納成三類：黃化嵌紋與扭曲窄化、黃化嵌紋與扭曲、黃化嵌紋與脈綠。除了 ZYMV-CY2 分離株在絲瓜及胡瓜上產生壞疽病徵之外，其它分離株在甜瓜、絲瓜及胡瓜上則產生黃化嵌紋病徵。以臺南 ZYMV-TN3 分離株純化所得之鞘蛋白所製備的抗血清，經免疫血清擴散反應證實其與其他分離株在血清反應上是不可區分的。為了提供更快更靈敏的診斷工具，反轉錄酵素連鎖反應之診斷方法被發展出來，以針對 ZYMV NIb 及 CP 基因所設計的專一性核酸引子經由 RT-PCR 對三十三個分離株均可增幅出一個 0.76 kb 的 DNA 片段。為了更進一步分析具代表性的五個 ZYMV 分離株 CP 基因的變化，利用 TagⅠ、EcoRV 及 NlaⅢ 剪切這些 CP 基因 N 端變化區的 RT-PCR 產物以比較其異同。結果顯示 ZYMV-TC1、CY2 及 TNML1 的親緣性較接近，而 ZYMV-TN3 及 NT1 與上述三分離株親緣關係比較疏遠。
Subjects
zucchini yellow mosaic virus
host reaction
Immunodiffusion
RT-PCR
restriction enzyme
矮南瓜黃化嵌紋病毒
寄主反應
免疫血清擴散反應
反轉錄酶連鎖反應
限制酶
Publisher
中華植物保護學會
Type
journal article