论文部分内容阅读
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: To study the changes of gene expression in sunflower before and after salt stress, and to isolate and identify salt-tolerant genes. Methods: The differentially expressed genes fragments induced by salt stress were analyzed by c DNA-AFLP. Results: A total of 232 pairs of differentially expressed primer combinations were screened from 256 pairs of primer combinations. With its selective amplification, differentially expressed 844 TDFs were obtained. After secondary PCR amplification and reverse Northern blot validation, 42 positive TDFs were obtained. Among them, 12 TDFs were cloned and sequenced to obtain 10 TDFs nucleotide sequences. After Blastx alignment and functional analysis, all 10 TDFs were involved in the response to salt stress and involved signal transduction related proteins, stress related functional proteins, senescence related proteins and proteins related to protein interactions. Conclusion: A number of salt stress response genes were identified by c DNA-AFLP technique, which laid the foundation for revealing salt-tolerance molecular mechanism of sunflower and guiding salt-tolerant molecular breeding of sunflower.