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以矮牵牛为试材,通过根癌农杆菌介导ACC脱氨酶基因(ACDS)(分别由CaMV35S、SAG12和CHSA启动子驱动)转化矮牵牛叶盘,探讨了预培养时间、农杆菌菌液浓度、浸染时间、共培养时间、乙酰丁香酮(AS)浓度等因素对转化的影响,比较了ACDS基因在不同启动子驱动下获得转基因植株的效率及转基因植株的生长速率。结果表明:预培养5d的矮牵牛叶盘在含AS 200μmol·L-1、OD600值为0.6的农杆菌菌液中浸染8min后,共培养4d时遗传转化效果最优;不同启动子驱动的ACDS基因转化效率以CHSA启动子效率最高(3%),SAG12启动子效率最低(1.25%)。PCR检测证明外源基因已整合进入矮牵牛基因组中。转ACDS基因矮牵牛植株的获得,为利用基因工程技术培育抗衰老矮牵牛新品种奠定了技术基础。
The petunia were used as experimental material to transform the leaves of Petunia hybrida by Agrobacterium tumefaciens-mediated ACC deaminase gene (ACDS) driven by CaMV35S, SAG12 and CHSA promoters respectively. The effects of pre-culture time, Agrobacterium tumefaciens The effect of ACDS on the efficiency of transgenic plants driven by different promoters and the growth rate of transgenic plants were compared. The results showed that the optimum conditions were as follows: the leaves of Petunia hybrida pre-cultured for 5 days had the highest genetic transformation efficiency after being exposed to Agrobacterium with AS 200 μmol·L-1 and OD600 of 0.6 for 8 min, ACDS gene conversion efficiency with CHSA promoter highest efficiency (3%), SAG12 promoter the lowest efficiency (1.25%). PCR test proved that the foreign gene has been integrated into the petunia genome. ACDS transgenic petunia plant access to gene technology for the development of new varieties of anti-aging petunia laid the technical foundation.