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园艺学报 ›› 2005, Vol. 32 ›› Issue (3): 403-407.

• 研究论文 • 上一篇    下一篇

草莓轻型黄边病毒的RT-PCR检测及其3'端序列分析

杨洪一;张志宏;高秀岩;杜国栋;代红艳;李 贺   

  1. 沈阳农业大学园艺学院, 沈阳 110161
  • 收稿日期:2004-10-21 修回日期:2005-03-14 出版日期:2005-06-25 发布日期:2005-06-25

Detection of Strawberry mild yellow edge virus with RT-PCR and Analysis of the Sequences of 3' Terminal Region

Yang Hongyi;Zhang Zhihong;Gao Xiuyan;Du Guodong;Dai Hongyan;Li He

  

  1. College of Horticulture, Shenyang Agricultural University, Shenyang 110161, China
  • Received:2004-10-21 Revised:2005-03-14 Online:2005-06-25 Published:2005-06-25

摘要: 利用RT-PCR技术对草莓植株中草莓轻型黄边病毒( SMYEV) 的外壳蛋白基因片段进行特异扩增, 扩增产物经测序证明为SMYEV的特异片段。建立了稳定的SMYEV的RT2PCR检测体系, 并对19个草莓品种和2种野生草莓进行检测, 其中12个品种和五叶草莓带有SMYEV。从SMYEV 的沈阳分离物SY01上扩增出了932 bp的基因组3'末端区域, 其核酸序列与国外分离物的序列同源性为86% ~96%。系统进化树显示SMYEV分为3个组群, 沈阳分离物SY01与欧美分离物位于同一组群内, 但独立形成1个小的分支。RNA二级结构分析显示SMYEV基因组3'末端形成3个茎环结构, 不同分离物的序列变化并未引起茎环结构的明显变化。

关键词: 草莓, 病毒, 草莓轻型黄边病毒(SMYEV), 序列分析

Abstract: Coat protein gene segment of Strawberry mild yellow edge virus ( SMYEV) was amplified with some special primers by reverse transcription-polymerase chain reaction (RT-PCR) from strawberries plants,and testified by sequencing. A stable system for detection of SMYEV with RT-PCR was established, and among nineteen strawberries cultivars and two wild strawberries species checked, twelve cultivars and Fragaria pentaphylla were infected. The 932 bp segment in 3' terminal region of SMYEV genome was amplified from Shenyang isolate SY01. Comparison showed it shared 86%~96% nucleotide acid identity with the published sequences. Phylogenetic analysis shows that all isolates of SMYEV fell into three clades. The Shenyang isolate SY01 lay in the same clade with the Europe and America isolates, but formed a small separate branch. Analysis of RNA secondary structure showed that three stem-loop structures were formed in 3'terminal region, and sequence differences in all isolates impacted little on the stem-loop structures.

Key words: Strawberry, Virus, Strawberry m ild yellow edge virus ( SMYEV), Sequence analysis

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