cDNA-AFLP法筛选红树植物盐应答基因  

Differential Analysis of Salt Response Genes by cDNA-AFLP in Mangrove

作　　者：陈银华[1,2] 韩淑梅[2] 沙爱华[3] 朱红林[2] 范吉星[2] 谢俊[1,2] 符秀梅[2] 李小靖[2] 

机构地区：[1]海南大学教育部热带生物资源重点实验室 [2]海南大学农学院,海口570228 [3]中国农业科学院油料作物研究所,武汉430062

出　　处：《中国农业科学》2008年第12期4257-4263,共7页Scientia Agricultura Sinica

基　　金：教育部科技重点项目(207092);海南省自然科学基金(80540;80612);海南省重点学科建设项目-热带植物资源开发与利用

摘　　要：【目的】分析红树植物盐胁迫下基因表达谱,分离识别耐盐相关基因。【方法】分别对红树植物-秋茄进行海水和淡水处理。分时提取总RNA进行等量混合,获得两种不同处理的样品池。采用cDNA-AFLP技术进行表达差异分析。【结果】256对引物组合共筛选了约15 000个cDNA片段,获得差异片段61个。差异基因表达模式分为2类,即海水处理诱导上升和下调表达,其中上升表达的基因片段为36个,下调表达的基因片段为25个;其中38个可视为已知基因。按照其功能分类可分为8大类:基础代谢、跨膜蛋白、信号转导、蛋白质代谢、转录因子、细胞骨架、抗病蛋白、假想蛋白,而其中又以基础代谢相关基因所占比重最大。【结论】构建了红树植物-秋茄在盐胁迫下的基因表达谱,从基因组水平上识别了一批受盐胁迫诱导或抑制表达、与耐盐相关的基因,这些新基因可用于耐盐的分子机理研究。[Objective] This study was conducted to elucidate the gene expression response of mangrove to salt-stess. [Method] Gene expression of Kandelia candel treated with fresh water and sea water was analyzed by the technology of cDNA-AFLP. [ Result] Two hundred and fifty-six primer combinations were used to investigate 15 000 cDNA fragments. Sixty-one differentially expressed cDNAs were found, 36 of which were up-regulated and 25 were down-regulated under the salt stress. The sequence analysis showed that 38 of them were with high homology to the genes in GenBank. The ESTs with significant protein homology were sorted into eight functional categories： basal metabolism, transmenbrane protein, signal transduction, protein metabolism, transcription factor, cytoskeleton, disease resistance protein, hypothetical protein. The rate of the basal metablosim related genes was maximum. [Conclusion] Gene expression profiling in response to salt and differentially expressed genes of mangrove Kandelia candel were revealed and identified via cDNA-AFLP analysis. This work begins to reveal potential molecular metablosim for salt tolerance of mangrove.

关 键 词：红树 耐盐性 基因分离 CDNA-AFLP 

分 类 号：S156.4[农业科学—土壤学]