通过逆转录聚合酶链反应鉴定西葫芦黄花叶马铃薯Y病毒并分析序列变异性。

Identification of Zucchini yellow mosaic potyvirus by RT-PCR and analysis of sequence variability.

作者信息

Thomson K G, Dietzgen R G, Gibbs A J, Tang Y C, Liesack W, Teakle D S, Stackebrandt E

机构信息

Queensland Department of Primary Industries, Gehrmann Laboratories, University of Queensland, Brisbane, Australia.

出版信息

J Virol Methods. 1995 Sep;55(1):83-96. doi: 10.1016/0166-0934(95)00047-x.

DOI:10.1016/0166-0934(95)00047-x

PMID:8576311

Abstract

A reverse transcription-polymerase chain reaction (RT-PCR) method was used to identify Zucchini yellow mosaic virus (ZYMV) in leaves of infected cucurbits. Oligonucleotide primers which annealed to regions in the nuclear inclusion body (NIb) and the coat protein (CP) genes, generated a 300-bp product from ZYMV and also from the closely related watermelon mosaic virus type 2 (WMV-2). However, no product was obtained from papaya ringspot potyvirus which also infects cucurbits. ZYMV and WMV-2 were differentiated using a third primer which was complementary to a sequence in the 3'-untranslated region; a 1186-bp amplified product was obtained for ZYMV only. Nucleotide sequence analysis of the 300-bp fragments of Australian ZYMV and WMV-2 strains revealed 93.7-100% sequence identity between ZYMV strains. Multiple sequence alignments indicated that the nucleotide sequence which codes for the N-terminus of the CP was 74-100% identical for different isolates of ZYMV. The Australian isolate of WMV-2 was 43-46% identical to all isolates of ZYMV and was 84.6% identical to a Florida isolate of WMV-2.

摘要

采用逆转录-聚合酶链反应（RT-PCR）方法在受感染葫芦科植物的叶片中鉴定小西葫芦黄花叶病毒（ZYMV）。与核内含体（NIb）和外壳蛋白（CP）基因区域退火的寡核苷酸引物，从ZYMV以及密切相关的西瓜花叶病毒2型（WMV-2）中产生了一个300 bp的产物。然而，从同样感染葫芦科植物的番木瓜环斑马铃薯Y病毒中未获得产物。使用与3'-非翻译区序列互补的第三条引物区分ZYMV和WMV-2；仅从ZYMV获得了一个1186 bp的扩增产物。对澳大利亚ZYMV和WMV-2菌株300 bp片段的核苷酸序列分析显示，ZYMV菌株之间的序列同一性为93.7%-100%。多序列比对表明，不同ZYMV分离株中编码CP N端的核苷酸序列同一性为74%-100%。澳大利亚WMV-2分离株与所有ZYMV分离株的同一性为43%-46%，与佛罗里达WMV-2分离株的同一性为84.6%。