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应用多重RT-PCR检测草莓斑驳病毒和草莓轻型黄边病毒
Detection of Strawberry mottle virus and Strawberry mild yellow edge virus by Multiplex RT-PCR
【摘要】 建立了多重RT-PCR同时检测草莓斑驳病毒(SMoV)和草莓轻型黄边病毒(SMYEV)的技术体系,对草莓田间植株和试管苗都可以有效进行检测。多重PCR引物根据引物之间的互补性及引物的Tm值进行筛选。适宜的PCR缓冲液的浓度为2×,退火温度为57℃,延伸温度为66℃。分别利用单一PCR和多重PCR对微茎尖培养获得的草莓品种宝交早生的11个株系进行了脱毒效果鉴定。
【Abstract】 A multiplex reverse transcriptase polymerase chain reaction (RT-PCR) assay was developed for simultaneously detecting Strawberry mottle virus(SMoV)and Strawberry mild yellow edge virus(SMYEV). Both field-grown strawberries and microplants were detected effectively. The primers of multiplex PCR were selected by primer-primer interactions and melting temperature. In multiplex PCR for detecting SMoV and SMYEV, the appropriate concentration of PCR buffer was 2×, and the appropriate temperatures for anneal and extension were 57℃ and 66℃ respectively. Eleven lines of strawberry(Fragaria×ananassa) cultivar‘Hokowase’, which were obtained by mini shoot tip culture in vitro, were screened for virus elimination by standard RT-PCR and multiplex RT-PCR.
- 【文献出处】 园艺学报 ,Acta Horticulturae Sinica , 编辑部邮箱 ,2006年03期
- 【分类号】S436.68
- 【被引频次】30
- 【下载频次】440