玉米丝氨酸羧肽酶基因(ZmSCP)的克隆及表达分析  

Cloning and Expression Analysis of Serine Carboxypeptidases in Maize (Zea mays L.)

作　　者：刘丽[1,2] 王静[2] 张志明[2] 赵茂俊[3] 潘光堂[2] 

机构地区：[1]四川农业大学人事处,四川雅安625014 [2]四川农业大学玉米研究所/西南玉米生物学与遗传育种重点实验室,四川成都611130 [3]四川农业大学生命科学与理学院,四川雅安625014

出　　处：《作物学报》2013年第1期164-171,共8页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(30900901);国家转基因生物新品种培育科技重大专项(2011ZX08003003);四川省科技厅应用基础项目(2006J13-039);四川省教育厅重点项目(07ZA063)资助

摘　　要：丝氨酸羧肽酶(serine carboxypeptidases,SCP)基因在植物生长发育及抗病性方面起着重要作用。本研究在立枯丝核菌胁迫24h后,以RT-PCR技术和RACE技术克隆玉米丝氨酸羧肽酶基因全长cDNA序列,命名为ZmSCP。结果显示,该基因全长为1874bp(GenBank登录号为JF682634),开放阅读框为999bp,编码332个氨基酸,相对分子量为36.505kD,等电点为4.75。ZmSCP基因编码蛋白与其他高等植物中该蛋白具有一定的同源性,同源性比例范围为42%～81%。进化树分析表明ZmSCP基因与水稻、高粱亲缘关系较近,属于同一进化分支。ZmSCP基因编码蛋白序列保守结构域分析显示该基因编码产物具有S10结构域,属于S10超家族。半定量RT-PCR与实时荧光定量PCR结果表明,ZmSCP基因在立枯丝核菌、ABA、JA、低温、高盐胁迫条件下,总体均呈诱导表达的趋势。其中,在立枯丝核菌AG1-IA诱导下,ZmSCP基因表达呈两步诱导趋势,第1次诱导出现在接菌后24h,然后下降,第2次诱导出现在接菌后60h。在ABA、JA、低温和盐胁迫下,ZmSCP基因表达均呈现上调趋势,表达高峰均出现在胁迫后48h。Serine carboxypeptidases play important roles in regulating the growth, development and disease resistance in plants. Reverse transcription-polymerase chain reaction （RT-PCR） and rapid amplification of cDNA ends （RACE） were applied to clone the serine carboxypeptidase gene named ZmSCP using cDNA of the high resistant maize inbred line R15 induced by Rhizoctonia solani. The cDNA full length of serine carboxypeptidase gene is 1874 bp （GenBank accession number： JF682634） containing a 999 bp complete open reading frame, encode 333 amino acids, with the molecular weight of 36.505 kD for the expected encoded proteins, and the isoelectric point of 4.75. The homology analysis indicated that the homology percentage were from 42% to 81% between the deduced amino acids from Zea mays L. and those from other plants. Phylogenetic analysis revealed that ZmSCP showed closer kinship with that of Oryza sativa and sorghum, indicating that they belong to the same evolutionary branch. ZmSCP protein has S10 conserved domain and belongs to S10 superfamily. The ZmSCP mRNA expression was analyzed by semi-quantitative and quantitative methods under different stress conditions. It is showed that the ZmSCP gene expression was basically up-regulated after ABA, JA, low temperature, and salt treatments, and showed two-step trend under induction of Rhizoctonia solani, with the first peak at 24 h after inoculation, and the second peak at 60 h, showing significant differences com- pared with the case of non-inoculated. In addition, the expression level of ZmSCP gene increased under the ABA, JA, low tem- perature, and salt stresses with an expression peak at 48 h.

关 键 词：丝氨酸羧肽酶 基因克隆 表达模式 

分 类 号：S513[农业科学—作物学]