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

作　　者：张志宏[1] 杨洪一[1] 代红艳[1] 李贺[1] 肖敏[1] 

机构地区：[1]沈阳农业大学园艺学院,辽宁沈阳110161

出　　处：《园艺学报》2006年第3期507-510,共4页Acta Horticulturae Sinica

基　　金：国家自然科学基金资助项目(30200187)

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。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.

关 键 词：多重RT-PCR 病毒检测 草莓 

分 类 号：S668.4[农业科学—果树学] S432.1[农业科学—园艺学]