玉米甘油-3-磷酸酰基转移酶基因ZmGPAT13生物信息学及表达特性分析

Bioinformatics and Expression of Glycerol-3-phosphate Acyltransferase 13 Gene of Maize

【目的】对玉米(Zea mays L.)甘油-3-磷酸酰基转移酶(Glycerol-3-phosphate acyltransferase,GPAT)家族成员ZmGPAT13进行生物信息学分析和在不同非生物胁迫处理下的基因表达模式变化检测,为进一步研究ZmGPAT13的生物学功能奠定理论基础。【方法】通过TAIR和MaizeGDB数据库获得拟南芥(Arabidopsis thaliana)和玉米中GPAT家族蛋白序列,利用生物信息学方法进行理化性质、蛋白结构、亚细胞定位、保守基序、系统进化、磷酸化位点和启动子顺式作用元件等分析,并采用荧光定量PCR(qRT-PCR)技术对ZmGPAT13在不同组织部位及甘露醇(Mannitol)、NaCl、脱落酸(ABA)、高温(42℃)、低温(4℃)处理下的相对基因表达量进行分析。【结果】ZmGPAT13是一个定位于线粒体的跨膜蛋白,与部分GPAT家族成员含有排序相同的15个保守基序(motif),且与拟南芥中AtGPAT1的亲缘关系最近;预测分析发现ZmGPAT13含有62个磷酸化位点,可能与玉米中多个GPATs存在相互作用关系;ZmGPAT13基因启动子富含多种与低温和ABA响应等相关的顺式作用元件。通过qRT-PCR技术对ZmGPAT13组织表达模式分析发现ZmGPAT13的相对基因表达量在胚芽鞘(Coleoptile)中最高,根(Root)中次之,叶片(Leaf)中最低;甘露醇(Mannitol)、NaCl、脱落酸(ABA)及高温(42℃)和低温(4℃)处理均能诱导ZmGPAT13的基因表达显著上调。【结论】玉米中ZmGPAT13具有高度的进化保守性,可能通过转录水平的变化参与调控玉米对干旱、盐、高温和低温等非生物胁迫的响应过程。

【Objective】Bioinformatics and expressions under stresses of glycerol-3-phosphate acyltransferase(GPAT)gene family member,ZmGPAT13,in maize were studied.【Method】Sequences of GPAT family members in Arabidopsis thaliana and Zea mays L.were obtained from the TAIR and MaizeGDB databases.Physicochemical properties,protein structure,subcellular localization,conserved motifs,phylogenetic evolution,phosphorylation sites,and cis-acting elements of promoter sequence of the GPATs were analyzed by bioinformatic methods.Expressions of ZmGPAT13 under the stresses of mannitol,NaCl,abscisic acid(ABA),high temperature at 42℃,and low temperature at 4℃were examined by qRT-PCR.【Result】A transmembrane protein,ZmGPAT13 localized in mitochondria,shared the same 15 conserved motifs with other GPAT family members,and had the highest homology with AtGPAT1 in arabidopsis.It contained 62 phosphorylation sites and might interact with multiple GPATs of maize.The promoters of ZmGPAT13 consisted of numerous cis-acting elements associated with the responses to low temperature and ABA stresses.The gene expressed most highly in the coleoptile,followed by the roots and the lowest in the leaves.And,relatively,the expressions were significantly elevated under the stress of mannitol,NaCl,ABA or high or low temperature.【Conclusion】A high evolutionary conservation seemed existed in ZmGPAT13.It was postulated that,at transcription level,ZmGPAT13 might be involved in the responses of maize plants to abiotic stresses such as drought,salt,and extreme temperatures.