摘要
采用RT PCR扩增的方法 ,从新疆不同地区的野生植物胡杨中分别克隆获得 1kb和 2 3kb的cDNA片段 ,测序和序列分析表明这两个基因片段均包含NHX基因的部分读码框架 ,分别命名为PtNHX和PwNHX。PtNHX序列同源性分析结果显示胡杨NHX基因与滨藜NHX基因同源性高达 98% ,与碱蓬同源性达到 86 % ,与拟南芥同源性为 84 % ,与水稻同源性为 80 % ,表明它是植物中高度保守的一种基因 ,同时说明野生植物胡杨中也存在与拟南芥相似的植物耐盐相关基因。PwNHX序列分析表明 ,该基因内部含有一种转座酶的读码框 ,大小约 135 0bp ,与已发表的Shigellaflexneri.转座子Tn10基因序列 (AF16 2 2 2 3)同源性为 99%。NHX基因的蛋白产物在植物的耐盐性方面起着重要的作用。推测PwNHX由于插入转座酶读码框可能会导致该基因的功能丧失。胡杨生存于盐碱地 ,其体内可能存在另一些机制使胡杨具有抗盐碱的能力。对于这些机制的研究可能有助于进一步了解植物的抗盐机制。利用PwNHX基因内的转座子作基因标签 ,可进一步研究胡杨的其它基因 ,从而有可能揭示胡杨叶子发育的变态过程、耐盐碱等性状。
The United Nations Environment Program estimates that approximately 20% of agricultural land and 50% of cropland in the world is salt stressed. The gene NHX (Na +/H + exchanger) encodes functional protein that catalyzes the countertransport of Na + and H + across membranes and may play an important role in plant salt tolerance. To clone the NHX from the wild plant Populus euphratica collected in Tarim basin and Xinjiang Wujiaqu district into a T vector, designed primer was used to amplify 1kb NHX cDNA fragment with RT PCR. Total RNA was extracted from Populus euphratica tissue (plant tissue was collected from Tarim basin and Xinjiang Wujiaqu district and stored in liquid nitrogen) according to the Plant RNA Mini Kits of Omega. First cDNAs were synthesized from 1μg total RNA of Populus euphratica seedling. A pair of primers were used to perform RT PCR. The amplified DNA fragment was purified and cloned into pMD18 T vector. However, 1kb and 2 3kb fragment were obtained from Tarim basin and Xinjiang Wujiaqu district and named as PtNHX and PwNHX , respectively. Sequence analysis reveals that the cloned PtNHX fragment of Populus euphratica contains partial NHX coding region with 98%,86%,84% and 80% identity comparing with Atriplex gemelini, Suaeda maritima, Arabidopsis thaliana and Oryza sativa , respectively. This analysis suggests that NHX gene would be highly conserved in terms of evolution in plant; and it also suggests that the NHX gene of Populus euphratica also would have the similarity with that of Arabidopsis . It may be of great importance in improvement of the plant salt tolerance and breed of crop. At the same time, sequence analysis shows that PwNHX gene includes a coding region about 1350bp with 99% identity comparing with transposon Tn10 IS10 left transposase of Shigella flexneri . On the one hand, the NHX gene may lose its function because it was inserted a fragment in coding region. On the other hand, its product may play a important role in salt tolerance. Populus grow in saline soil. It speculates that it may have other salt tolerance mechanism in Populus. The transposon can be used as transposon tagging to clone other genes and it will help us to understand farther the salt tolerance mechanism.
出处
《生物工程学报》
CAS
CSCD
北大核心
2003年第5期628-631,共4页
Chinese Journal of Biotechnology
基金
国家科技攻关西部科技行动项目基金资助 (No .2 0 0 1BA90 1A3 2)~~
关键词
胡杨
NHX基因
细菌转座子
植物耐盐
Populus euphratica, NHX gene, RT PCR, salt tolerance, transposon
