CAJ | 학술논문

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24 h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1874 bp (GenBank登录号为JF682634),开放阅读框为999 bp,编码332个氨基酸,相对分子量为36.505 kD,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24 h,然后下降,第2次诱导出现在接菌后60 h.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48 h.
사안산최태매(serine carboxypeptidases, SCP)기인재식물생장발육급항병성방면기착중요작용.본연구재립고사핵균협박24 h후,이RT-PCR기술화RACE기술극륭옥미사안산최태매기인전장cDNA서렬,명명위ZmSCP.결과현시,해기인전장위1874 bp (GenBank등록호위JF682634),개방열독광위999 bp,편마332개안기산,상대분자량위36.505 kD,등전점위4.75.ZmSCP기인편마단백여기타고등식물중해단백구유일정적동원성,동원성비례범위위42%~81%.진화수분석표명ZmSCP기인여수도、고량친연관계교근,속우동일진화분지.ZmSCP기인편마단백서렬보수결구역분석현시해기인편마산물구유S10결구역,속우S10초가족.반정량RT-PCR여실시형광정량PCR결과표명, ZmSCP기인재립고사핵균、ABA、JA、저온、고염협박조건하,총체균정유도표체적추세.기중,재립고사핵균AG1-IA유도하, ZmSCP기인표체정량보유도추세,제1차유도출현재접균후24 h,연후하강,제2차유도출현재접균후60 h.재ABA、JA、저온화염협박하, ZmSCP기인표체균정현상조추세,표체고봉균출현재협박후48 h.
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.