世界中医药

作者	单位
毛亚平1，马莹1，2，卜俊玲1，曾雯1，郭娟1，陈敏1，黄璐琦1	1 中国中医科学院中药资源中心，北京，100700； 2 北京师范大学中药资源保护与利用北京市重点实验室，北京，100875

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中文摘要:

目的：利用土培植物进行原位侵染获得RNAi型丹参毛状根，并对其干扰效果进行研究。方法：利用转化有目标基因RNAi载体的发根农杆菌对土壤中生长的丹参幼苗根茎部分进行侵染产生毛状根，经荧光显微镜鉴定阳性毛状根，并利用实时荧光定量PCR检测目标基因的表达情况。结果：在土培丹参根茎部分成功侵染产生毛状根，利用荧光显微镜进行检测显示目标基因RNAi型毛状根阳性转化率为72.72%，RNAi空载毛状根阳性转化率为66.67%。实时荧光定量PCR结果表明RNAi型毛状根中目标基因表达受到抑制，表达量下降到空载型对照组的8.61%，证明对土培植物直接进行侵染能够成功诱导RNAi毛状根。结论：采用原位侵染法诱导丹参根茎部位成功产生阳性RNAi型毛状根，侵染过程无需无菌操作，简单快捷，对植株生长无影响，能够用于功能基因的体内功能研究。

English Summary:

To study the jamming effects of RNAi hairy roots of Salvia Miltiorrhiza by in situ infection of soil cultivating plants.Methods:Hairy roots were induced by infection of Agrobacterium rhizogenes transformed with the RNAi vector of target gene into the roots of Salvia Miltiorrhiza which were planted in the soil.Positive hairy roots were identified by fluorescence microscope.The expression of target gene was detected by qRT-PCR analysis.Results:Hairy roots were induced from the roots of Salvia Miltiorrhiza successfully.GFP detection results showed that positive conversion rates of hairy root lines of target gene RNAi and empty vector were 72.72% and 66.7%,respectively.qRT-PCR analysis revealed that the expression of target gene in RNAi hairy roots was significantly decreased to 8.61% compared with empty vector,which indicated that RNAi hairy roots were successfully induced from the soil planted Salvia Miltiorrhiza by in situ infection.Conclusion:Positive target gene-RNAi Salvia Miltiorrhiza hairy roots were successfully induced from the soil planted Salvia Miltiorrhiza by in situ infection.As the infection required no sterile operation,it was not only simple and quick,but also would not affect plant growth and development.It can be applicable to functional characterization of gene in vivo.

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