【摘要】 发根农杆菌Ar1334工程菌株与商陆叶片共培养,诱导效率不同程度地受到发根农杆菌生理状态、浓度、浸泡时间、共培养时间的影响:采用稀释5倍的处于对数生长期的Ar1334工程菌株,感染商陆无菌苗幼嫩叶圆片5min,在MS培养基上共培养2d,可达最大转化效率82.2%;诱导的根在附加20mg·L-1Kan的MS无激素MS培养基上,呈现旺盛的生长态势和典型的发状根结构特点,经PCR检测扩增出了560bp的rolC基因的目的片断,组织化学检测也证实了GUS基因在发状根中的表达.更多还原

【Abstract】 Hairy roots were induced from Phytolacca acinosa leaves after co-cultured with Agrobacterium rhizogenes Ar1334 harbouring agropin type pRi1855 and binary vector pIG121HM.The root with many root hairs and branches grew vigorously towards all directions on MS medium without any plant hormone.Leaf discs pre-cultured on MS medium for two days were dipped into bacteria solution delusioned 5 times at exponential phase for 5 min,and then transferred onto fresh MS medium supplement with 500 mg·L^-1 carbenicillin to eliminat Ar1334,and 20 mg·L^-1 Kan to selected the transformants.A highest frequency as （82.2%） of root formation was achieved at the optimum transforming conditions.A 560 bp DNA fragment was amplified from the genomic DNA of hairy root by PCR using primes of rol C gene which was one of the four genes related to hairy root induction located on TL-DNA of Ri plasmid.Blue dots were discovered on the hairy root tips after they were dipped into X-gluc solution at 37℃ for one night.These results proved phytolacca acinosa hairy root could be an efficient culture system to express foreign gene and accumulate valuable substance.更多还原