摘要
对用固相扣除杂交方法从低温驯化沙冬青克隆得到的AmLEA5基因进行表达模式分析和生物信息学分析,表明该基因编码一种第5族胚胎发育晚期丰富蛋白(LEA),全长693 bp,含有1个297 bp的开放阅读框,编码98个氨基酸,预测Am-LEA5的分子量为10.6 kDa,是一种亲水性蛋白,有多个磷酸化位点。密码子偏好性分析表明该基因略偏好于用A或T结尾的密码子。系统发生分析表明,AmLEA5蛋白与蒺藜苜蓿LEA(ACJ84182.1)亲缘关系最近。qRT-PCR结果显示AmLEA5的表达量在低温、干旱、盐和热胁迫条件下均有上调,尤其在低温胁迫后期富集量最高。亚细胞定位表明,用YFP标记的AmLEA5位于细胞质和细胞核内。一系列试验结果表明AmLEA5基因在沙冬青抵御非生物胁迫,尤其是在抵御低温胁迫机制中发挥重要作用。
Molecular characterization and expression patterns of AmLEA5 （AAW31666.1） under abiotic stresses which cloned from cold-and drought-stressed Ammopiptanthus mongolicus by a modified solid subtractive hybridization method were reported in this study. Bioinformatic analysis showed that the gene encoded a group 5 late embryogenesis abundant protein （LEA）. The overall length of AmLEA5 was 693 bp, and the cDNA contained a 297 bp ORF encoding a polypeptide of 98 amino acids with a calculated molecular mass of 10.6 kDa. AmLEA5 ,with more phosphorylation sites,was a kind of hydrophilic protein. The analysis of codon bias showed that AmLEA5 gene was bias toward the synonymous codons with A and T at the third codon position. Phylogenetic analysis indicated that AmLEA5 protein was most closed to Medicago truncatula LEA （ AcJ84182.1 ） in genetic relationship, qRT-PCR analysis demonstrated that AmLEA5 was up-regulated by cold, drought, salt and hot stresses. Specifically,transcription of AmLEA5 accumulated more at the late stage of cold treatment. Intracellular localization analysis revealed that AmLEA5 proteins tagged with yellow fluorescent proteins （YFP） was more likely localized in the cytoplasm and nucleus. The results suggested that AmLEA5 may played important roles in abiotic stresses responses, especially in cold tolerant mechanism in A. Mongolicus.
出处
《植物遗传资源学报》
CAS
CSCD
北大核心
2013年第3期492-500,共9页
Journal of Plant Genetic Resources
基金
国家自然科学基金(31270737
30671476)
北京市自然科学基金(6112016)
关键词
沙冬青
AmLEA5
生物信息学分析
表达模式
亚细胞定位
Ammopiptanthus mongolicus,AmLEA5, bioinformatic analysis, expression pattern, subcellular local- ization
