向日葵盐胁迫相关基因的cDNA-AFLP差异表达  

Differentially Expressed Analysis on the Responsive Genes to Salt Stress in Sunflower by cDNA- AFLP

作　　者：孙瑞芬[1] 张艳芳[1] 郭树春[1] 于海峰[1] 李素萍[1] 乔慧蕾[1] 聂惠[1] 安玉麟[1] 

机构地区：[1]内蒙古农牧业科学院,呼和浩特010031

出　　处：《中国生物工程杂志》2015年第1期34-40,共7页China Biotechnology

基　　金：内蒙古自治区自然科学基金资助项目(2012MS0315)

摘　　要：目的:研究向日葵盐胁迫前后基因表达的变化,分离并鉴定耐盐相关基因。方法:采用c DNA-AFLP技术分析盐胁迫产生的差异表达基因片段。结果:从256对引物组合中筛选到232对有差异表达的引物组合。用其进行选择性扩增,获得差异表达的上调TDFs 845条。经二次PCR扩增及反向Northern blot验证,获得42个阳性TDFs。对其中12个TDFs进行克隆及序列测定,得到10条TDFs核苷酸序列。经Blastx比对及功能分析,10个TDFs均与应答盐胁迫相关,涉及信号转导相关蛋白、胁迫相关功能蛋白、衰老相关蛋白以及与蛋白相互作用有关的蛋白。结论:利用c DNA-AFLP技术鉴定出一批盐胁迫应答基因,为揭示向日葵耐盐分子机制及指导向日葵耐盐分子育种实践奠定基础。Objective： This study was conducted to elucidate the response of genes in sunflower to saltstress,isolate and identify the genes related to salt tolerance. Methods： The differentially expressed gene fragments produced after being treated with salt were analyzed by c DNA-AFLP. Results： 232 primer pairs obtaining differential expressed bands were acquired from 256 primers combinations. Using these primer pairs,a total of 845 differentially expressed up-regulated transcript-derived fragments（ TDFs） were selectively amplified,42 positive TDFs from them were identified after the second PCR amplification and reverse Northern blot. 12 TDFs of which were cloned and sequenced,and 10 nucleotide sequences were obtained. By Blastx analysis,10 TDFs were tightly related to salt tolerence of sunflower,being involved in signal transduction related proteins,abiotic stress related functional proteins,senescene associated proteins and proteins associated with protein interaction. Conclusion： A number of genes that respond to salt stress were identified using c DNA-AFLP,the results will lay a foundation for determining potential molecular mechanisms of salt tolerance and the practice of molecular breeding of sunflower.

关 键 词：向日葵 盐胁迫 CDNA-AFLP 差异表达基因 

分 类 号：Q786[生物学—分子生物学]