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

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。

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.