药用植物中转基因元件筛查策略研究

Strategy research for screening of transgenic elements used in medicinal plants

通过对已有药用植物转基因研究文献进行总结分析,为药用植物和中药材转基因监管提供有效策略。采用关键词检索法对中国知网和SCI两大数据库进行查询,建立48种常用药用植物转基因研究中常用表达元件的数据库,分析转基因元件使用情况,并总结外源转基因的筛查策略。获得1993年4月至2016年5月的转基因研究文献281篇,包括中文文献230篇,英文文献51篇。40.4%的中文文献和54.9%的英文文献研究目的为优化药用植物遗传转化体系。常用启动子包括P-35S、P-Ubi、P-GPD和P-act,其中P-35S的使用频率最高,为68.7%。常用标记基因包括NPTII、HPT、Gent、Bar和aad A,其中NPTII的使用频率最高,为37.4%。常用报告基因包括GUS和GFP,GUS使用频率达到35.2%。常用终止子包括T-NOS、T-35S和T-OCS,其中T-NOS的使用频率最高,为58%。对各元件进行配组分析,发现"P-35S+T-NOS+NPTII+GUS"组合筛选成功率为86.1%,在此基础上加入有一定使用频率的HPT、Bar和GFP,对药用植物转基因元件的筛查率达到91.5%。对于使用同源或内源启动子、标记基因和终止子的研究,可基于T-DNA边界序列进行转基因检测。

With the increase of transgenic research literature in medicinal plants, detection and inspection of transgenic elements in medicinal drugs have been highly concerned. The aim of this study was to provide an approach for the detection of transgenic elements in medicinal materials, so as to provide the effective strategy for the transgenic supervision of medicinal plants and Chinese medicinal materials. The literatures involving transgenic research on 48 medicinal plants was retrieved from the two databases of CNKI and SCI from April 1993 to May 2016, which was used to establish a database of commonly used expression elements in transgenic medicinal plants. Totally 281 papers including 230 Chinese literatures and 51 English literatures were obtained, of which 40.4% of Chinese and 54.9% of English literatures were the researches with aim to establish transformation system. The results showed that commonly used promoter included P-35 S, P-Ubi, P-GPD, and P-act, with P-35 S having the highest frequency of 68.7%. Common marker genes included NPTII, HPT, Gent, Bar, and aad A, with NPTII giving the highest frequency of 37.4%. Common reporter genes were GUS and GFP, with GUS of the highest frequency of 35.2%. Common terminator included T-NOS, T-35 S, and T-OCS, with T-NOS of the highest frequency of 58%. The combination ＂P-35 S ＋ T-NOS ＋ NPTII ＋ GUS＂ increased the screening rate to 86.1% for screening the transgenic elements used in medicinal plants. On this basis, the adding of HPT, Bar and GFP with certain frequency of use contributed to the screening rate of 91.5% in searching for transgenic elements. T-DNA border sequence can be used for the transgenic detection in the studies using homologous or endogenous promoters, marker genes, and terminators.