转甜菜碱醛脱氢酶(BADH)基因棉花的获得及其耐盐性鉴定

Acquiring Transgenic Cotton Harboring Betaine Aldehyde Dehydrogenase Gene and Identifying Their Salt-tolerance

甜菜碱醛脱氢酶(betaine aldehyde dehydrogenase,BADH)是合成甜菜碱的关键酶,广泛用于植物的耐盐转基因研究。为获得耐盐性提高的棉花植株,本研究通过花粉管通道法将山菠菜甜菜碱醛(BADH)基因转化到陆地棉品种——中棉所35。经卡那霉素田间抗性鉴定、标记基因NPT-Ⅱ和目标基因BADH的PCR检测,以及Southern杂交检测,结果表明BADH已整合到棉花的基因组中,并获得转标记基因NPT-Ⅱ和目标基因BADH棉株5株。通过对T2种子在0.6%Na Cl盐池发芽实验结果表明,转化植株的出苗率高达63.4%,对照材料出苗率2.4%,转化植株耐盐性较对照有明显提高。本研究获得转基因植株可作为抗逆育种的种质材料。

Betaine aldehyde dehydrogenase （BADH）, widely used in studies on salt tolerance of transgenic plants, is a key enzyme in the synthesis of betaine. The BADH gene cloned from Mountain Spinach was transferred into upland cotton--variety ZMS35 by pollen-tube pathway in order to obtain the transgenic cotton plants with increa- sed salt tolerance in this research. Through the kanamycin resistance identification of cotton seedlings in the field, PCR molecular detection of marker gene NPT-Ⅱ and target gene BADH, together with southern hybridization detection, the results proved that BADH had been integrated into the cotton genome. A total of 5 transgenic plants, carrying the marker gene NPT-Ⅱ and the target gene BADH, were obtained. Based on the experiment of T2 seed germination in the 0.6%NaC1 salt pond, the result presented that the salt tolerance of transgenic plants with the seedling emergence rate as high as 63.4% had been improved obviously in comparison with the control, whose seedling emergence rate was only 2.4%. Transgenic plants obtained in this study could be used as germplasm materials for resistant breeding.