发根农杆菌诱导栝楼毛状根的研究

Induction hairy root of Trichosanth kirilowii Maxim by Agrobacterium rhizogenes

栝楼是一种重要的药用植物,提取物具有多种活性。以栝楼的无菌苗为材料,用A4、15834和LBA9402等3种发根农杆菌浸染茎段,诱导毛状根生成,并研究毛状根分化不定芽的条件。结果显示,3种农杆菌都能诱导栝楼毛状根产生,其中LBA9402诱导率最高,达到52%,且毛状根在6-BA 2.0 mg.L-1+KT 2.0 mg.L-1+NAA 0.5 mg.L-1的培养基上,不定芽分化率达到了24%。对毛状根进行基因组DNA提取,PCR扩增检测出rolB基因的存在,初步证明R i质粒T-DNA已经整合到栝楼基因组中。

Trichosanthes kirilowii Maxim is one of the medicinal plants. Seedlings of Trichosantks kirilowii Maxim were infected by three kinds ofAgrobacterium rhizogenes （ A4,15834. LBA9402） to induce hairy roots, and the differentiation of adventitious shoots were researched. As a results, three kinds of Agrobacterium rhizogenes could induce hairy roots, but induction rate of LBA9402 was higher than that of others and reached 52% ： Differentiation rates of adventitious shoots could reach 24% with culture medium of 2.0 mg·L^-1 6-BA ＋ 2. 0 mg·L^-1 KT ＋ 0.5 mg·L^-1 NAA. The result of PCR showed that rolB gene in Ri plasmid was integrated into the genome of Trichosanthes kirilowii Maxim.