目的研究野生天麻總DNA對(duì)馬鈴薯的轉(zhuǎn)化,分析馬鈴薯轉(zhuǎn)化植株中野生天麻的藥用成分天麻素.方法采用浸苗法將野生天麻總DNA導(dǎo)入馬鈴薯試管苗,通過(guò)紫外掃描法、PCR擴(kuò)增篩選轉(zhuǎn)化植株,對(duì)轉(zhuǎn)化植株進(jìn)行SDS-聚丙烯酰胺凝膠電泳(SDS-PAGE)蛋白分析,通過(guò)TLC法檢測(cè)天麻素.結(jié)果(1)在200株轉(zhuǎn)化的馬鈴薯中有21株的紫外掃描圖譜與正常對(duì)照組有顯著差異,且在220 nm有明顯吸收峰.(2)5株經(jīng)PCR擴(kuò)增出野生天麻抗真菌蛋白(GAFP)基因.(3)轉(zhuǎn)基因馬鈴薯與正常馬鈴薯的蛋白表達(dá)有明顯差異,并且在轉(zhuǎn)基因馬鈴薯中有-條與GAFP相同的條帶.而正常馬鈴薯中無(wú)此條帶.(4)通過(guò)薄層色譜法檢測(cè)出3株轉(zhuǎn)基因馬鈴薯表達(dá)野生天麻的有效藥用成分天麻素.結(jié)論采用浸苗法進(jìn)行外源總DNA導(dǎo)入是可行的.

Objective Using the soaking seedling method to introduce wildGastrodia elataDNA into potato plantlets and analyze gastrodin of transformed Solanum tuberosum.Methods After the tuber grown up,the solution of gastrodin was extracted from the potatoes which were transformed by wildG.elataDNA.The transformed plants were scaned by ultraviolet.PCR was used to analyze GAFP gene by SDS-PAGE.TLC was used to analyze the gastrodin of transformedS.tuberosum.Results (1)The 21 transformed plants had obvious absorption peak at 220 nm in 200 transformedS.tuberosunr.(2)The fragment of wildG.elataGAFP gene was found by PCR from 5 transformed plants.(3)The protein expressions in transgenicS.tuberosumwere obviously different from the normalS.tuberosum:a band was detected in transgenicS.tuberosumand wildG.elata,but wasn't detected in normalS.tuberosum.(4)The expression of the gastrodin was detected by TLC from three transformed plants.Conclusion It is feasible to use the soaking seedling method to introduce exogenous DNA into plants.

貴州省省長(zhǎng)基金;貴州省科技廳社發(fā)基金(2001112)