Li B H, Zhang H M, Xu T F, Ding R X
Department of Pharmacy, Second Military Medical University, Shanghai 200433, China.
Zhongguo Zhong Yao Za Zhi. 2000 Nov;25(11):657-60.
To establish an effective system for genetic transformation of autotetraploid Isatis indigotica by Agrobacterium rhizogenes.
Agrobacterium rhizogenes strains R1601, ATCC15834 and A4 were employed to induce hairy roots from autotetraploid Isatis indigotica, and the obtained hairy roots were developed into regenerated plants on solid MS media with different Kinds of plant growth regulators.
Root induction differed obviously. By three Agrobacterium strains. Hairy roots grew rapidly on solid MS medium without plant growth regulators and showed the typical hairy root phenotype: profuse branching, high density of root hairs and plagiotropism. And the transformation of Ri T-DNA was confirmed by opine analysis. The biomass of hairy roots increased nearly 35 times regulator-free MS after suspended in plant growth two weeks. On solid MS medium with BA, adventitious buds were differentiated directly from hairy roots without callus formation. All of the adventitious buds were rooted on root induction medium and developed into regenerated plants. Opine analysis indicated the integration of Ri T-DNA in the transformed plants.
Agrobacterium rhizogene can induce hairy roots from autotetraploid Isatis indigotica, and the obtained hairy roots can develop into regenerated plants.
建立发根农杆菌介导的同源四倍体菘蓝遗传转化体系。
用发根农杆菌菌株R1601、ATCC15834和A4诱导同源四倍体菘蓝产生毛状根，并将获得的毛状根在添加不同种类植物生长调节剂的固体MS培养基上培养成再生植株。
三种发根农杆菌菌株诱导毛状根的效果差异明显。毛状根在不添加植物生长调节剂的固体MS培养基上生长迅速，呈现典型的毛状根表型：分枝多、根毛密度高且具向地性。通过冠瘿碱分析证实了Ri T-DNA的转化。毛状根在植物生长调节剂中悬浮培养两周后，其生物量在无调节剂的MS培养基上增加了近35倍。在添加BA的固体MS培养基上，不定芽直接从毛状根分化产生，不形成愈伤组织。所有不定芽在生根培养基上生根并发育成再生植株。冠瘿碱分析表明Ri T-DNA已整合到转化植株中。
发根农杆菌能诱导同源四倍体菘蓝产生毛状根，且获得的毛状根能发育成再生植株。
