农杆菌介导的水稻‘绥8’转化研究

Study on the Agrobacterium-mediated transformation of 'Sui 8' rice plants

以‘绥8’成熟胚为外植体,研究了愈伤组织的诱导、筛选、再生情况,并对转化植株进行PCR和抗旱性鉴定,以建立高效的愈伤诱导、农杆菌转化和再生体系。结果表明:‘绥8’材料成熟胚易消毒,不易染菌;铺种使用2NBK培养基,可诱导出大量长势良好的愈伤组织,不需切芽;转化后的愈伤组织不需要恢复阶段,在筛选过程中愈伤长势良好但是颜色较浅,区分抗性愈伤较难;分化时绿点率和分化率很高。PCR鉴定证明外源基因成功转化到‘绥8’基因组中,其转化率和阳性率分别为17%和70%。对转基因植株进行抗旱性鉴定,20%PEG胁迫复水后,与野生型相比,表现出良好的抗旱性。

In order to establish an efficient system of calli induction,Agrobacteriumtransformation and regeneration of‘Sui 8＇rice mature embryos,the calli induction,screening and regeneration were studied in this experiment with ‘Sui 8＇explants as materials.Transformed plants were identified by PCR and drought resistance tests.The results showed that the‘Sui8＇embryos were easy to be disinfected,and the contamination rate was low.A large amount of well-grown calli was induced by 2NBK culture medium and there was no need to cut the buds.The calli after induction didn＇t require the recovery phase.During the screening process,the calli grew well but was light in color,leading to the difficulty of distinguishing from the resistant calli.At the differentiation stage,the rates of green dot and differentiation were both high.PCR analysis proved that the target gene had been transformed into‘Sui 8＇genome successfully,and the transformation and positive rates were 17% and 70%,,respectively.The drought resistance tests by 20% PEG and rehydration showed that the transgenetic plants exhibited well drought resistance.