影响农杆菌介导的木薯基因转化因素的研究

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。

STUDY ON FACTORS INFLUENCING AGROBACTERIUM-MEDIATED CASSAVA TRANSFORMATION

Factors influencing agrobacterium-mediated cassava transformation were investigated. A-mong the four Agrobacterium strains tested ?LBA 4404 ( pTOK 233 ) and LBA 4404 (pBin9GusInt) gave higher transient expression than C 58 C1 (pIG121Hm) and EHA 105 (pBin9Husint). Pretreatment of explants by bombardment or vaccum had no significant effect on transient expression while preinduction of A-grobacterium with acetosyringone showed better effects, and preculture of explants showed worse effects. All the cultivars tested were susceptible to Agrobacterium infection, while the types of explants and the physiological state of the ex-plants had a strong influence on the transient expression efficiency. The 15-day-old somatic cotyledons and the fully expanded leaves from in vitro plantlets were the most susceptible to Agrobacterium infection. The results also showed that all the four selective reagents (hy-gromycin, geneticin, PPT, and kanamycin) synchronously suppressed the growth of callus, shoot organogenesis and shoot rooting in a dose dependent manner.