Abstract
水稻氨基酸转运蛋白OsLHT1与拟南芥AtLHT1的蛋白质序列具有较高的同源性,经比对发现相似度达78%。拟南芥氨基酸转运蛋白AtLHT1突变体(atlht1)相比较野生型生长矮小,在开花后期出现分蘖数和果荚数少、早衰等症状。为了探究水稻OsLHT1在植物生长发育中的分子生物学效应/功能,本研究通过RT-PCR的方法,在水稻中克隆获得了OsLHT1全长ORF片段,构建了35s-OsLHT1超表达载体,异源表达于拟南芥atlht1突变体中,并对OsLHT1在水稻重要的生育时期做了基因表达分析。结果表明,转基因植株(atlht1 + 35s-OsLHT1)能够恢复拟南芥atlht1的生长表型,分枝数与果荚数均比突变体增多,转录水平分析结果表明OsLHT1在水稻重要生育时期都有表达,且在分蘖期的根部和老叶的表达量高,推测OsLHT1在植物生长发育过程中起到了关键作用。 Rice amino acid transporter OsLHT1 is highly homologous to Arabidopsis AtLHT1, with sharing a similarity of 78% at the protein level. The Arabidopsis amino acid transporter AtLHT1 mutant (atlht1), compared with the wild-type, shows a growth in small size, less upper part branches and siliques, as well as early senescence at the late flowering stage. In order to explore the molecular biological effects of OsLHT1 on plant growth and development,we cloned the full length open reading frame (ORF) of OsLHT1 using RT-PCR, constructed a 35s-OsLHT1 overexpression vector, heterologously expressed OsLHT1 in the Arabidopsis atlht1 mutant, and analyzed the expression of OsLHT1 in rice at significant growth stages. The result showed that transgenic plants (atlht1 + 35s-OsLHT1) indeed displayed a restored growth phenotype related to atlht1 mutant, the number of the upper branches and pods was higher than that of mutants; moreover, gene expression study showed that OsLHT1 was expressed at almost all important growth stages of rice, but the highest expression level was observed in roots of the tillering stage and old leaves, thus proposing the critical role for OsLHT1 in plant growth and development.