盐芥TOR基因的克隆及对盐芥生长发育的影响

Cloning of TOR from Thellungiella halophila and Its Role in Plant Growth

雷帕霉素靶蛋白(TOR)在调节动植物的生长及新陈代谢方面起重要作用.为进一步研究植物中TOR的作用,对盐芥TOR基因进行克隆,采用聚合酶链式反应和cDNA末端快速扩增技术等分子生物学方法获得盐芥TOR基因的cDNA全长及基因组全长,构建了ThTOR基因沉默植物表达载体并成功获得盐芥ThTOR基因沉默的转基因株系.结果表明:ThTOR cDNA全长为7836bp,编码2479个氨基酸,蛋白序列与拟南芥同源性高达97.5%,基因组全长19162bp;盐芥ThTOR基因沉默的转基因株系具有生长缓慢、花及种子的发育受阻的明显表型.

Mammalian target of rapamycin（TOR） plays an important role in the growth regulation and metabolism of animals and plants.In order to reveal the function of TOR in plants,TOR gene of Thellungiella halophila（T.halophila） was cloned,and the full lengths of ThTOR cDNA and genomic DNA were obtained by polymerase chain reaction（PCR） and rapid amplification of cDNA ends（RACE）.Then,RNAi vector ThTOR-RNAi was constructed and the transgenic lines were obtained by transforming ThTOR-RNAi into T.halophila.The results show that the full-length ThTOR cDNA contains 7 836 base pairs and encodes a protein with 2 479 amino acids,that a high degree of homology（97.5%） exists between the proteins encoded by ThTOR and AtTOR,that the full-length ThTOR genomic DNA is of 19 162 base pairs,and that obvious phenotypes of slow growth as well as flowers and seeds abortion occur in T.halophila ThTOR-RNAi transgenic lines.