转AmGS基因红叶石楠的分子检测及抗寒性分析

摘要

对转AmGS基因(从沙冬青中克隆的肌醇半乳糖苷合成酶基因)的红叶石楠植株进行多项分子检测(PCR,Southern,RT-PCR),结果表明AmGS基因已经整合到转基因株系R6和R7的基因组DNA中,并检测到转录水平上的表达.随后,经对R6和R7两个转基因株系进行连续6代芽切扩繁继代株系的PCR检测,发现导入的AmGS基因传递到所有芽切扩繁后代植株中.植株抗寒能力试验结果表明,在相同低温处理条件下,转基因株系均比未转基因植株的存活率要高.相对电导率测定结果表明,随着处理温度的降低,2个转基因株系相对电导率升高的程度明显低于未转基因的对照植株.低温半致死温度(LT50)测定结果表明,2个转基因株系(R6和R7)的LT50明显低于未转基因的对照植株.上述结果说明导入的AmGS基因提高了转基因株系的抗寒性.%A few papers reported the successful transformation and expression of woody plant cold-resistant gene in forest tree. Recently, we have obtained AmGS( galactinol synthase gene of Ammopiptanthus mongolicus ) transgenic Photinia x fraseri ' Red Robin' plants. This paper reports the molecular identification of the transgenic plants by PCR, Southern blot and RT-PCR analyses. Results indicated that the AmGS gene had integrated into the genomic DNA of transgenic plants of R6 and R7 lines and expressed at transcriptional level. After 6 generations of propagation by shoot cuttings, it was found that the transformed AmGS gene was detected in all of the examined plants in all of the 6 generations propagated by cuttage. Results of cold resistance tests pointed out that in all of the low temperature treatments, the transgenic lines exhibited higher survival rates than that of the untransformed control plants. After cold treatment, transgenic plants had less increase in the relative electrical conductivity ( REC ) than untransformed control plants. LT50 of transgenic lines of R6 and R7 was remarkable lower than that of the untransformed control plants. These results indicated that the imported AmGS gene increased the cold resistance of transgenic lines R6 and R7.