茎尖脱毒生姜组培苗病原检测研究

Study on Tip-free and Tissue-culture of Fresh Ginger's Prathogeny Detection

利用PCR技术及扫描电镜技术对茎尖脱毒生姜组培苗进行病原检测。用细菌16SrRNA通用引物对生姜组培种苗根部和固体培养基混合提取的基因组DNA作为模板进行16SrRNA扩增,不能扩增出16S rRNA;Trizol提取生姜组培苗根部和茎段RNA经凝胶电泳检测无明显RNA目的条带;生姜组培苗茎段组织经扫描电镜观察组培苗茎段中无呈球形、杆状、卵圆形、丝状、冠状及蝌蚪形等规则形状的病毒颗粒,只见表皮、微管等组织。研究认为,PCR、RT—PCR和扫描电镜是茎尖脱毒生姜组培苗病原检测的有效手段。

Tip-free and tissue-culture fresh Ginger were used polymerase chain(PCR) reaction technique and scanning electron microscopy technology to detect pathogeny.The admixture genome DNA extracted from tissue-culture fresh Ginger's roots and solid medium were used universal primer of 16s RNA to clone 16s RNA,the results indicated no target pills were detected.RNA which were extracted from tissue-culture fresh Ginger s stems and roots by trizol did not detect by Gel electrophoresis.Tissue-culture fresh Ginger's stems were used scanning electron microscopy(SEM) to observe,the results revealed that there had no global,staff,oval,filamentous,coronal,Tadpole shape and other regulation shape virus particle,only epidermis,microtubule tissue and other structure.The study presume PCR technique and scanning electron microscopy technology were effective way to detect pathogeny of Tip-free and Tissue-culture fresh Ginger.