转TaLEA1基因丹参植株的耐盐与耐旱性

摘要

TaLEA1,a gene encoding a late-embryogenesis-abundant protein,was cloned from wheat and was transformed into Salvia Miltiorrhiza Bunge by Agrobacterium-mediated leaf-disk transformation method.Seven transgenic lines were obtained after kanamycin(50 mg/L) screening.Six positive lines were obtained by PCR amplification,and after four additional generations,one stable line was obtained by Southern hybridization.Transgenic plants had better growth states than control plants on mediums containing 1% NaC1 and 8% PEG6000,which demonstrated that TaLEA1 played an important role in increasing the salt and drought tolerance of S.miltiorrhiza.%TaLEA1基因是一个从小麦中克隆得到的编码胚胎晚期丰富蛋白的基因,用农杆菌EHA105(含卡那霉素抗性基因表达载体PBI121-TaLEA1)介导的叶盘转化法将其转入丹参(Salvia miltiorrhiza Bunge)组织培养苗中,在50 mg/L卡那霉素选择压下连续筛选,并扩繁和生根培养,获得了7个丹参转基因株系.经PCR扩增,初步验证得到6个阳性株系,再继代4次后,Southern杂交结果表明已获得了一个能稳定遗传的转基因株系.该株系在含1%NaC1和8%PEG6000培养基中的生长状态优于野生型,表明转入TaLEA1基因可改善丹参组培苗的耐盐和耐旱性.