大麦气孔发育相关基因HvSPCH的克隆及初步功能分析  

Cloning and Functional Analysis of Stomatal Development Related Gene HvSPCH in Barley

作　　者：次尔甲玛 王宏鹏 苟筱颖 佟可心 郭瑞超 王俊斌 曹高燚 包曙光 谢晓东 陈小强 CIER Jiama;WANG Hongpeng;GOU Xiaoying;TONG Kexin;GUO Ruichao;WANG Junbin;CAO Gaoyi;BAO Shuguang;XIE Xiaodong;CHEN Xiaoqiang(College of Agronomy and Resources and Environment,Tianjin Agricultural University,Tianjin 300384,China)

机构地区：[1]天津农学院农学与资源环境学院,天津300384

出　　处：《麦类作物学报》2022年第10期1175-1181,共7页Journal of Triticeae Crops

基　　金：国家级大学生创新创业训练计划项目(202010061003);国家级大学生创新创业训练计划项目(202110061037);天津市科技计划项目(21ZYCGSN00130);天津市研究生科研创新项目(2020YJSS131);国家自然科学基金青年基金项目(31801446)。

摘　　要：气孔是植物与外界进行气体交换的通道,对植物的生长发育至关重要。SPCH转录因子与气孔的发育关系密切,为揭示其在大麦中的功能,以大麦G1614为材料,利用PCR方法克隆大麦SPCH(HvSPCH)基因后,进行生物信息学分析,并对该基因在干旱胁迫条件下的表达模式进行分析。结果表明,HvSPCH基因编码区长为1098 bp,可编码365个氨基酸,相对分子量为90504.73 Da,理论等电点为4.96,为疏水的、不稳定的碱性蛋白质。氨基酸序列分析表明,HvSPCH蛋白的氨基酸序列不含有跨膜结构域和信号肽,含有40个磷酸化位点和27个糖基化位点。二级结构分析表明,HvSPCH蛋白包含35.62%的α-螺旋、8.77%的延长链、2.47%的β-转角和53.13%的无规则卷曲。系统进化分析表明,大麦HvSPCH蛋白与水稻、野生二粒小麦、二穗短柄草的SPCH蛋白亲缘关系较近。qRT-PCR分析结果表明,干旱处理4 h和12 h时,HvSPCH基因的相对表达量显著升高,说明其有可能参与大麦的干旱逆境响应过程。Stomata is a gas exchange channel between plants and the outside world,which is very important for plant growth and development.SPCH transcription factor is closely related to stomatal development.In order to reveal its function in barley,HvSPCH gene was cloned by PCR method from barley G1614 for bioinformatics analysis,and the expression pattern of HvSPCH gene under drought stress was analyzed.The results showed that HvSPCH gene contained the open reading frame of 1089 bp and encoded a polypeptide of 365 amino acid residues.HvSPCH protein was a hydrophobic and unstable basic protein with a relative molecular weight of 90504.73 Da and a theoretical isoelectric point of 4.96.Amino acid sequence analysis showed that HvSPCH protein contained 40 phosphorylation sites and 27 glycosylation sites,but without transmembrane domain and signal peptide.Secondary structure analysis showed that HvSPCH protein contained 35.62%α-helix,8.77%extended chain,2.47%β-rotation and 53.13%random crimp.Phylogenetic analysis showed that the HvSPCH protein of barley was closely related to the SPCH protein of rice,wild emmer and Brachypodium distachyon.qRT-PCR analysis showed that the relative expression level of HvSPCH gene was significantly increased at 4 h and 12 h under drought treatment,suggesting that HvSPCH might be involved in the drought stress response process of barley.

关 键 词：大麦 气孔发育 SPCH转录因子 基因克隆 

分 类 号：S512.3[农业科学—作物学] S330