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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。
To establish a reliable, sensitive and efficient method for the detection of Chrysanthemum virus B (CVB), the specificity, repeatability and sensitivity of RT-PCR were tested using specific CVB gene primers. Finally, By using optimized RT-PCR system, the infection of chrysanthemum with CVB virus was detected. The results showed that the specific fragment amplified from the total RNA of Chrysanthemum morifolium plants encoded 211 amino acids, which shared 96% -99% identity with the coat protein of CVB gene from other sources in the gene database and could be identified as Chrysanthemum B virus coat protein gene; repeatability and sensitivity were detected in the detection of a clear target band, and 1 ng of total RNA can be amplified specific fragments of the seven were chrysanthemum a total of 32 plants were detected The detection rate of CVB was 56.2% and the detection accuracy was 68.75%. The results showed that the established RT-PCR system of CVB gene of Chrysolophus pictus could effectively identify the virus infected by Chrysanthemum morifolium.