陆地棉 CBL基因家族成员鉴定与功能分析
Identification and bioinformatics analysis of CBL family gene in Gossypium hirsutum
钙调磷酸酶B亚基蛋白(calcineurin B-like proteins)在植物非生物逆境应答以及钙信号转导过程发挥重要的作用。为了探究CBL基因在棉花非生物胁迫响应的作用，本研究利用生物信息学的方法进行陆地棉 CBL家族成员的鉴定，利用转录组数据和反转录聚合酶链式反应分析陆地棉 CBL基因在盐胁迫下的表达模式，利用病毒诱导的基因沉默技术对 GhCBL10-1基因进行沉默并进行功能验证。结果表明，在陆地棉中获得25个 CBL基因，该基因家族成员蛋白的理化性质差异不大，CBL蛋白中的氨基酸大部分为酸性。系统进化树分析将陆地棉的CBL蛋白分成了2个组，其中A组包括GhCBL4-1、GhCBL4-2、GhCBL4-3、GhCBL4-4、GhCBL4-5、GhCBL8,其他成员属于B组；通过结构域和保守基序分析发现所有的 CBL基因均含有至少1个 EF-hand结构域。基因结构分析发现同一类群中外显子-内含子结构比较相似，不同组之间的基因结构差异较大。染色体定位分析发现 24个 CBL基因被定位在19条染色体上，其中13个 CBL基因定位在A亚组染色体上，GhCBL1-7不能定位到任何染色体上，剩余的基因定位在D亚组染色体上。陆地棉 CBL家族基因成员启动子区域中均含有多个能够应答逆境和植物激素的顺式作用元件。在盐胁迫下，15个基因在叶和根中有显著的差异表达，且在根和叶中的差异表达模式不同。利用病毒诱导的基因沉默技术成功沉默 GhCBL10-1的棉株相较于对照更加不耐盐。该研究结果有助于了解陆地棉 CBL基因家族的进化与功能，为后续研究其功能提供了一定的理论依据。
Calcineurin B-like(CBL)proteins play an important role in plant abiotic stress response and calcium signal transduction. In order to explore the role of CBL gene in cotton abiotic stress response, this study used bioinformatics methods to identify CBL family members in Gossypium hirsutum, and analyzed the expression pattern of CBL gene in upland cotton under salt stress by using transcriptome data and reverse transcription polymerase chain reaction. The virus induced gene silencing technique was used to silence GhCBL10-1 gene and to verify its function. The results showed that 25 CBL genes were obtained from upland cotton, and there was little difference in physiochemical properties of the member proteins. Most of the amino acids in CBL proteins were acidic. The CBL proteins of upland cotton were divided into two groups by phylogenetic tree analysis. Group A included GhCBL4-1,GhCBL4-2,GhCBL4-3, GhCBL4-4, GhCBL4-5 and GhCBL8, and the other members belonged to group B. Domain and conserved motif analysis showed that all CBL genes contain at least one EF-hand domain. The analysis of gene structure showed that exon-intron structure was similar in the same group, and the gene structure was different among different groups. Chromosome localization analysis showed that 24 CBL genes were located on 19 chromosomes, of which 13 CBL genes were located on subgenome A chromosomes, GhCBL1-7 could not be mapped to any chromosome, and the remaining genes were located on subgenome D chromosomes. The promoter region of CBL family gene members in upland cotton contains multiple cis-acting elements that can respond to stress and plant hormones. Under salt stress, 15 genes were significantly differentially expressed in leaves and roots, and the differentially expressed patterns were different in roots and leaves. Cotton plants successfully silenced with GhCBL10-1 by virus induced gene silencing were more sensitive to salt stress than control plants. The results of this study are helpful to understand the evolution and function of CBL gene family in upland cotton, and provide a theoretical basis for further study of its function.
杨秀;邓艳凤;王涛;肖水平;
江西省经济作物研究所,江西九江332105;江西省经济作物研究所,江西九江332105;江西省经济作物研究所,江西九江332105;江西省经济作物研究所,江西九江332105;
Gossypium hirsutum calcineurin B subunit protein virus induced gene silencing gene function
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