摘要
利用实时荧光PCR方法,以18srRNA为内参照,对半夏植株及相应组培苗携带的黄瓜花叶病毒(Cu-cumber Mosaic Virus,CMV)含量进行了相对定量检测.并以DAS-ELISA和RT-PCR方法检测结果相比较.结果表明,相比原始植株,组培苗的病毒含量大大降低.以带病毒半夏母株为外植体培养的组培苗,其病毒含量以叶片中的相对较高,在块茎和愈伤组织中病毒相对较低.而ELISA方法在检测病毒含量低的样品时有假阴性结果;RT-PCR方法灵敏度可靠但不适宜精确量化.
With 18srDNA as as endogenous reference, relative quantification detection of Cucumber Mosaic Virus （CMV） in Pinellia Ternata and its tissue cultured plantlets was carried by using Real-time PCR, and combined with methods of DAS-ELISA and RT-PCR. The results showed that CMV concentration in tissue cultured plantlets was decreased greatly compared with the parent plantlet. CMV quantity in leaf of tissue cultured plantlets which come from explant of CMV positive Pinellia Ternata was relatively higher than that of tubercles and callus. There were fake positive results while using ELISA method to detect samples of low concentration virus. RT-PCR method is sensitive but not quantiffed exactly to detected samples.
出处
《湖南师范大学自然科学学报》
EI
CAS
北大核心
2006年第3期103-106,共4页
Journal of Natural Science of Hunan Normal University
基金
国家'863'资助项目(2002AA24112A)
