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草莓叶片不同保存方法对RT-PCR检测 草莓斑驳病毒效果的影响
白莹雪,李杨,祝宁,张家兴,任俊达,尚巧霞
0
(北京农学院农业农村部华北都市农业重点实验室,北京 102206;北京市昌平区十三陵林场,北京102200;北京市昌平区农业技术推广站,北京102200)
摘要:
植物样本保存方法不当易导致样本枯萎和腐烂,严重影响病毒检测结果。为提高PCR法检测草莓斑驳病毒(strawberry mottle virus,SMoV)的效果,明确待检测植物样本的有效保存条件,将经过RT-PCR检测感染SMoV的草莓叶片作为试验材料,设置自然风干保存、硅胶干燥保存和氧化钙(CaO)干燥保存3种不同的干燥保存条件。采用试剂盒法提取各处理草莓叶片总RNA,设计特异性引物进行RT-PCR检测。比较得到最佳干燥保存条件后,再以几种常见的低温、干燥保存为对照,分析草莓叶片不同保存方法对草莓斑驳病毒PCR检测效果的影响。电泳结果显示,含有SMoV的草莓叶片在CaO干燥保存条件下PCR检测效果最好,且操作简单易行。该方法可以为高效、便捷的保存带毒草莓叶片样本提供参考,适用于田间大量样品的采集,提高检测效率。
关键词:  草莓叶片  保存方法  SMoV  病毒检测
DOI:10.13610/j.cnki.1672-352x.20220705.020
基金项目:北京市粮经作物产业创新团队项目(BAIC09-2021), 北京农学院青年教师科研基金(QNKJ202001), 北京市教育委员会科技计划一般项目(KM202110020010)和北京农学院青年教师科研创新能力提升计划(QJKC2022002)共同资助。
Effects of different strawberry leaf preserving conditions on RT-PCR detection of strawberry mottle virus
BAI Yingxue,LI Yang,ZHU Ning,ZHANG Jiaxing,REN Junda,SHANG Qiaoxia
(Key Laboratory for Northern Urban Agriculture of Ministry of Agriculture and Rural Affairs, Beijing University of Agriculture, Beijing 102206;Ming Tombs Forest Farm of Changping District in Beijing, Beijing 102200;Agricultural Technology Extension Station of Changping District in Beijing, Beijing 102200)
Abstract:
Plant samples are easy to wither or rot due to improper preservation methods, exerting great influence on the subsequent virus detection. In order to improve the detection effect of strawberry mottle virus (SMoV) by PCR and clarify the effective storage conditions of plant samples to be detected, strawberry leaves with SMoV infection detected by RT-PCR were used as experimental materials. Three different drying conditions were set up: natural air drying, silica gel drying and calcium oxide (CaO) drying. Total RNA was extracted from strawberry leaves by kit method, and SMoV was detected by RT-PCR with specific primers. After obtaining the optimum dry preserving condition by comparison, several common low temperature and dry treatments were selected as controls to analyze the effects of different preservation methods on the detection of SMoV. The results showed that strawberry leaves containing SMoV had the best PCR detection effect under the condition of CaO drying and preservation with simpler and easier operation. This study provides a reference for efficient and convenient preservation of SMoV in strawberry leaves, and it can be applied to the collection of a large number of samples in the field to improve the detection efficiency.
Key words:  strawberry leaf  preserving method  SMoV  virus detection

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