地被菊CVB病毒基因的RT-PCR检测

发布时间：2018-06-20 22:45

本文选题：地被菊 + 菊花病毒　；参考：《植物保护学报》2016年06期

【摘要】：为建立可靠、灵敏且高效的地被菊中菊花B病毒(Chrysanthemum virus B,CVB)的检测方法,利用特异性CVB基因引物,通过RT-PCR技术进行了特异性、重复性和灵敏度测试,最终利用优化的RT-PCR体系对地被菊植株感染CVB病毒情况进行了检测。结果显示,以染病地被菊植株的总RNA为模板扩增出的特异性片段编码211个氨基酸,与基因数据库中其它来源的CVB基因外壳蛋白同源性为96%~99%,可确定为菊花B病毒外壳蛋白基因;重复性和灵敏度检测中均获得了清晰的目标条带,且1 ng的总RNA即可扩增出特异性片段;对7个地被菊品种共32个植株的检测中,CVB的检出率为56.2%,检测的准确率为68.75%。表明建立的地被菊CVB基因的RT-PCR检测体系可有效用于地被菊植株感染病毒情况的鉴定。
[Abstract]:In order to establish a reliable, sensitive and efficient method for the detection of Chrysanthemum virus CVB in chrysanthemum, the specificity, reproducibility and sensitivity were tested by RT-PCR using specific CVB gene primers. Finally, the infection of CVB virus in ground cover chrysanthemum plants was detected by the optimized RT-PCR system. The results showed that the specific fragment amplified from the total RNA of infected chrysanthemum plants encode 211 amino acids, and the cover-protein homology of CVB gene from other sources in the gene database is 960.99%, which can be identified as the coat protein gene of chrysanthemum B virus. A clear target band was obtained in the detection of repeatability and sensitivity, and a specific fragment could be amplified with a total RNA of 1 ng, and the detection rate of CVB was 56.2% and the detection accuracy was 68.75% in 32 plants of 7 ground cover chrysanthemum cultivars. The results showed that the established RT-PCR detection system of CVB gene could be used to identify the virus infection in ground cover chrysanthemum plants.
【作者单位】：东北林业大学生命科学学院;
【基金】：中央高校基本科研业务费(2572014CA21) 黑龙江省自然科学基金(C201343) 东北林业大学大学生创新创业训练项目(201310225118)
【分类号】：S436.8

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