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首页> 外文期刊>Propagation of Ornamental Plants >EFFICIENT IN VITRO PLANT REGENERATION AND AGROBACTERIUM-MEDIATED GENETIC TRANSFORMATION OF LISIANTHUS [EUSTOMA GRANDIFLORUM (RAF.) SHINN]
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EFFICIENT IN VITRO PLANT REGENERATION AND AGROBACTERIUM-MEDIATED GENETIC TRANSFORMATION OF LISIANTHUS [EUSTOMA GRANDIFLORUM (RAF.) SHINN]

机译：菊芋的高效体外植株再生和农杆菌介导的遗传转化[EUSTOMA GRANDIFLORUM（SHIN）]。

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In this study, a highly efficient plant regeneration and Agrobacterium-mediated genetic transformation protocols of lisianthus Micky Rose were established. Immature and mature internodal explants were incubated with Agrobacterium tumefaciens strain LBA4404 containing a binary vector pBAL2 and carrying the reporter gene beta-glucuronidase intron (GUS-INT), and the marker gene neomycin phosphotransferase (NPTII). Factors affecting transformation efficiency, such as effect of pre-culture, Agrobacterium concentration, acetosyringone, infection, and co-cultivation time of Agrobacterium, were investigated. After co-cultivation, explants were transferred onto MS medium supplemented with 1.0 mu M a-naphthalene acetic acid (NAA) and 1.5 mu M thidiazuron (TDZ), 100 mg l(-1) kanamycin, and 300 mg l(-1) carbenicillin for callus induction. Regeneration of adventitious shoots from callus was achieved on Murashige and Skoog (MS) medium containing 3.0 mu M TDZ, 0.5 mu M NAA, 100 mg l(-1) kanamycin, and 300 mg l(-1) carbenicillin. Immature internodal explants produced more shoots (30 shoots) than mature internodal explants. The transgenic elongated shoots were rooted (91.3%) on MS medium supplemented with 3.0 mu M indole-3-acetic acid (IAA) and 100 mg l(-1) kanamycin. Three days pre-cultivation, Agrobacterium density, infection time for 30 min, acetosyringone (300 mu M), and four days of co-cultivation proved to be critical factors for greatly improving the transformation efficiency. Molecular studies of transgenic plants and their offspring confirmed the presence of the nptII gene. Transgenic plants were acclimatized (90%) in the greenhouse and transgenic efficiency of 21% was evaluated.|
机译：在这项研究中，建立了桔梗米奇玫瑰的高效植物再生和农杆菌介导的遗传转化方案。将未成熟和成熟的节间外植体与含有二元载体pBAL2，携带报告基因β-葡糖醛酸糖苷酶内含子（GUS-INT）的根癌农杆菌菌株LBA4404和标记基因新霉素磷酸转移酶（NPTII）一起孵育。研究了影响转化效率的因素，如预培养效果，农杆菌浓度，乙酰丁香酮，感染和农杆菌共培养时间。共培养后，将外植体转移到补充有1.0μMα-萘乙酸（NAA）和1.5μM噻唑隆（TDZ），100 mg l（-1）卡那霉素和300 mg l（-1）的MS培养基上羧苄青霉素诱导愈伤组织。在含有3.0μM TDZ，0.5μM NAA，100 mg l（-1）卡那霉素和300 mg l（-1）羧苄青霉素的Murashige和Skoog（MS）培养基上实现了愈伤组织不定芽的再生。未成熟的节间外植体比成熟的节间外植体产生更多的芽（30枝）。将转基因的细长芽生根（91.3％）在补充了3.0μM吲哚-3-乙酸（IAA）和100 mg l（-1）卡那霉素的MS培养基上。预先培养三天，土壤杆菌密度，感染时间30分钟，乙酰丁香酮（300μM）和共培养四天被证明是大大提高转化效率的关键因素。对转基因植物及其后代的分子研究证实了nptII基因的存在。在温室中使转基因植物适应环境（90％），评估转基因效率为21％。

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