抗旱基因8981转化‘吉粳88’的研究

The study on the transformation of rice 'Jijing 88' with a drought resistant gene 8981

以‘吉粳88’成熟胚为外植体,用农杆菌介导法转入抗旱基因8981,对转化过程中愈伤组织的诱导、转化、筛选等条件进行研究,建立一个稳定、高效的农杆菌介导的遗传转化体系,并对获得的转基因植株进行PCR鉴定、除草剂抗性检测和抗旱性鉴定分析。结果表明:‘吉粳88’在诱导愈伤的过程中,不需要切芽;当菌液浓度OD600=0.2,浸染时间为20min时,转化效率最高且效果最好;在筛选培养前,需要经过恢复培养。PCR检测证明8981基因已整合到‘吉粳88’基因组中,抗性检测发现转基因植株对除草剂和PEG胁迫都有一定的抗性。

In order to build a stable,high-efficient genetic transformation system,the calli induced from rice‘Jijing 88＇mature embryos was used in the experiment,and the gene 8981 was transferred into‘Jijing 88＇via Agrobacterium-mediated transformation.The calli induction,transformation and screening which could influence the transformation efficiency were studied.The obtained transgenic plants were identified by PCR test,herbicide resistance and drought resistance tests.The results showed that‘Jijing 88＇didn＇t need to cut the buds in the process of calli induction.When OD600 of Agrobacterium tumefaciens concentration was 0.2and the infection time was 20 min,the transformation efficiency was the highest and the transformation effect was the best.Before the screening culture process,the recovery training was required.PCR test showed that gene 8981 had been integrated into the genome of‘Jijing 88＇.The test of resistance found that the transgenic plants had a certain resistance to herbicides and PEG stress.