Shi H-P, Kintzios S
College of Life Science, South China Normal University, 510631 Guangzhou, China.
Plant Cell Rep. 2003 Jul;21(11):1103-7. doi: 10.1007/s00299-003-0633-6. Epub 2003 Apr 23.
An efficient transformation system for the medicinal plant Pueraria phaseoloides was established by using agropine-type Agrobacterium rhizogenes ATCC15834. Hairy roots could be obtained directly from the cut edges of petioles of leaf explants or via callus 10 days after inoculation with the bacteria. The highest frequency of explant transformation by A. rhizogenes ATCC15834 was about 70% after infection for 30 days. Hairy roots could grow rapidly on solid, growth regulator-free Murashige and Skoog medium and had characteristics of transformed roots such as fast growth and high lateral branching. Paper electrophoresis revealed that bacteria-free hairy roots of P. phaseoloides could synthesize agropine and mannopine. The polymerase chain reaction amplification of rooting locus genes showed that left-hand transferred DNA of the root inducing plasmid of A. rhizogenes was inserted into the genome of transformed P. phaseoloides hairy roots. The content of puerarin in hairy roots reached a level of 1.190 mg/g dry weight and was 1.067 times the content in the roots of untransformed plants.
利用农杆碱型发根农杆菌ATCC15834建立了药用植物三裂叶野葛的高效转化体系。接种该菌10天后，毛状根可直接从叶片外植体叶柄的切口边缘获得，也可通过愈伤组织获得。发根农杆菌ATCC15834感染30天后，外植体转化的最高频率约为70%。毛状根在不含生长调节剂的固体Murashige和Skoog培养基上能快速生长，具有转化根的特征，如生长迅速和侧枝多。纸电泳显示，无菌的三裂叶野葛毛状根能够合成农杆碱和甘露碱。生根位点基因的聚合酶链反应扩增表明，发根农杆菌根诱导质粒的左手转移DNA插入到了转化的三裂叶野葛毛状根基因组中。毛状根中葛根素的含量达到1.190 mg/g干重，是未转化植株根中含量的1.067倍。
