基于家族分析的苜蓿逆境应答Ca^(2+) ATPase家族基因筛选与鉴定  

Screening and identification of environmental stress responsive Medicago sativa Ca^(2+) ATPases based on gene family analyses

作　　者：张美萍[1] 杨珺凯 孙明哲[1] 贾博为 孙晓丽[1] ZHANG Mei-Ping YANG Jun-Kai SUN Ming-Zhe JIA Bo-Wei SUN Xiao-Li(Crop Stress Molecular Biology Laboratory, Heilongjiang Bayi Agricultural Universtiy, Daqing, Heilongjiang 163319, China)

机构地区：[1]黑龙江八一农垦大学作物逆境分子生物学实验室,黑龙江大庆163319

出　　处：《植物生理学报》2017年第2期198-208,共11页Plant Physiology Journal

基　　金：国家自然科学基金(31500204);黑龙江省自然科学基金(C2015035)~~

摘　　要：Ca^(2+) ATPase属于P型ATPase超家族,在植物对外界逆境的应答中发挥重要作用,但迄今尚未有苜蓿Ca^(2+) ATPase基因研究的相关报道。为筛选逆境胁迫应答Ca^(2+) ATPases,本研究鉴定获得了16个苜蓿Ca^(2+) ATPase家族成员,分布在7条染色体上,且存在一对旁系同源基因。系统进化分析表明苜蓿Ca^(2+) ATPases分为ACA和ECA亚家族,ACA亚家族分为4组,且组内成员具有相似的内含子-外显子组成模式。此外,苜蓿Ca^(2+) ATPase家族具有高度保守的蛋白序列组成,包括4个功能结构域、10个跨膜结构域和11个保守motif。转录组数据分析发现该家族具有不同的逆境诱导表达模式,且多数苜蓿Ca^(2+) ATPases家族基因受低温和冷冻胁迫诱导表达,其中Mt ACA9和Mt ACA8上调表达最明显。q RT-PCR结果证实Mt ACA9和Mt ACA8表达在低温和冷冻胁迫下显著上调。本研究结果为揭示苜蓿Ca^(2+) ATPase基因在低温冷害应答中的功能提供了重要参考。It is widely accepted that the Ca2＋ ATPase gene family, belonging to the P-type ATPase superfamily, plays an important role in environmental stress responses. However, up to now, no researches reported the function of alfalfa Ca2＋ ATPases. Hence, to screening the stress responsive Ca2＋ ATPases, we used the Hidden Markov Model to search the alfalfa proteome database, and identified a total of 16 Ca2＋ ATPase family genes in alfalfa genome. Chromosomal location and synteny analysis revealed that alfalfa Ca2＋ ATPase family members located on 7 chromosomes, and possessed 1 pairs of duplicated genes. By analyzing the topological structure of the phylogenetic tree, we found that alfalfa Ca2＋ ATPase family was clustered into ACA and ECA subfamilies. The ACA subfamily was further divided into four groups （Group I-IV）, and members in each group displayed high similarity in terms of intron-exon organization pattern. Furthermore, we also suggested that alfalfa Ca2＋ ATPase family protein sequences were highly conserved, including 4 conserved functional domains, 10 transmenbrane domains, and 11 conserved motifs. By using the RNA-seq data, we also investigated the expression profiles of alfalfa Ca2＋ ATPase genes under different environmental stresses, and found that the expression of alfalfa Ca2＋ ATPase genes, especially MtACA8 and MtACA9, was induced by cold and freezing stresses. By using quantitative real-time PCR analysis, we further confirmed that MtACA8 and MtACA9 expression was up-regulated by both cold and freezing stresses. Taken together, results presented in this study will facilitate further research about the biological function of alfalfa Ca2＋ ATPases in plant responses to environmental stress.

关 键 词：苜蓿 Ca2+ATPase 家族分析 逆境应答 表达分析 

分 类 号：Q943.2[生物学—植物学] S541[农业科学—作物学]