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本文研究了影响农杆菌介导的木薯基因转化的因素。结果表明,供试的4个菌种中,LBA4404(pBin9GusInt)及LBA4404(pTOK)瞬时表达效果较好。对农杆菌的诱导处理能增强瞬时表达效果,外植体的预处理对瞬时表达无影响,而外植体的预培养显著降低瞬时表达。所有供试的木薯品种都能被农杆菌侵染,但外植体的类型及生理状况对农杆菌的侵染力影响很大,成熟胚状体的子叶(萌发15d)及试管苗完全展开的叶片对农杆菌亲和性最高。四种筛选剂(kanamycin、hygromycin、phosphinothricin及geneticin)均表现出剂量效应且能同步抑制芽器官发生、愈伤生长及芽切段生根。
In this paper, the factors affecting the Agrobacterium-mediated transformation of cassava were studied. The results showed that the transient expression of LBA4404 (pBin9GusInt) and LBA4404 (pTOK) in 4 tested strains was better. The induction of Agrobacterium could enhance the transient expression, the explant pretreatment had no effect on the transient expression, but the explant preincubation significantly reduced the transient expression. All the tested cassava varieties could be infected by Agrobacterium. However, the type and physiological status of explants had a great influence on the infectivity of Agrobacterium. Cotyledons (germination 15 days) and test tube seedlings of mature embryoid bodies were fully expanded Agrobacterium leaves the highest affinity. Four kinds of screening agents (kanamycin, hygromycin, phosphinothricin and geneticin) showed dose-effect and could simultaneously inhibit bud and organ occurrence, callus growth and bud rooting.