玉米紫色酸性磷酸酶(PAPs)基因家族的鉴定与低磷响应特征  

Identification of Maize(Zea mays) Purple Acid Phosphatase(PAPs)Genes Family and Their Characterization Responses to Phosphorus Starvation

作　　者：易双[1] 聂治[1] 张啸[1] 罗博文[1] 任志勇[1] 吴玲[1] 刘丹[1] 高世斌[1] 

机构地区：[1]四川农业大学玉米研究所/农业部西南玉米生物学与遗传育种重点实验室,成都611130

出　　处：《农业生物技术学报》2015年第3期281-290,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.31361140364和No.31171566);引进国际先进农业科学技术计划(No.2011-G15-2和No.2013-Z38)

摘　　要：紫色酸性磷酸酶(purple acid phosphatase,PAPs)属于金属磷酸酯酶家族,能催化磷酸酯或酸酐的水解,对于活化植物根际周围的有机态磷及促进植株体内磷素的再循环利用起重要作用。本研究以拟南芥(Arabidopsis thaliana)PAPs基因为基础,在玉米(Zea mays)全基因组水平上对PAPs基因家族进行鉴定,对其基因结构及进化关系进行分析,并运用半定量PCR、实时荧光定量PCR及亚细胞定位对其家族成员进行深入研究。结果表明,从玉米自交系B73全基因组中筛选出24个紫色酸性磷酸酶候选基因,聚类为3个家族和8个亚家族;半定量qRT-PCR(sqRT-PCR)分析的8个家族成员均在低磷胁迫下呈现表达变化,对具有显著表达差异的4个成员(ZmPAP1c、ZmPAP10a、ZmPAP10b和ZmPAP26)进行qRT-PCR分析发现,4个ZmPAPs在不同时间的低磷胁迫处理后,因材料基因型不同及器官不同而呈现出时空特异性及组织特异性,其中ZmPAP1c和ZmPAP10a在维持体内磷素动态平衡中可能发挥重要作用;亚细胞定位结果表明,ZmPAP10a和ZmPAP1c表达产物被定位于细胞膜上;酸性磷酸酶活性分析表明,耐低磷玉米自交系178根系的酸性磷酸酶活性较9782高,并且响应低磷胁迫更灵敏,说明其在遇到低磷环境时通过调节ZmPAPs表达来增强酸性磷酸酶的活性,从而提高磷素利用效率。本研究结果为进一步研究玉米ZmPAPs家族的功能提供了基础资料。Purple acid phosphatases （PAPs）, member of the metallo-phosphoesterase family, are involved in catalyzing the hydrolysis of various phosphate esters or anhydrides and play important role in external phosphorus assimilation and recycling P in plants. In this study, comparative genomics method was used to identify maize（Zea mays） ZmPAPs family in whole-genome scale based on the PAPs orthologs in Arabidopsis thaliana. The gene structure and phylogenetic relationship of ZmPAPs had been analyzed. The methods of sqRT-PCR, qRT-PCR and subcellular localization were performed for further study of ZmPAPs family members. The results showed that 24 ZmPAPs were identified from whole genome sequences of maize inbred line B73 and could be classified into 3 distinct groups including 8 subgroups based on amino acid sequences. Eight ZraPAPs which were analyzed by sqRT-PCR all exhibited differential expression under low phosphorus starvation. The expression profiles of 4 ZmPAPs （ZmPAPlc, ZmPAPIOa, ZmPAPIOb and ZmPAP26） with significant levels of differential expression were assayed by qRT-PCR and indicated that 4 ZmPAPs exhibited different expression profiles in different organs and genotypes under low phosphorus stress, among them ZmPAPlc and ZmPAPIOa played important role in maintaining phosphorus homeostasis. Subcellular location analysis revealed that the products of ZmPAPlc and ZmPAPIOa located in cell membrane. The analysis of acid phosphatase （APase） activity showed that APase activity in maize 178 roots was higher than that of maize 9782 and maize 178 was more sensitive than maize 9782 in responses to phosphorus starvation, indicating that the low-phosphorus tolerant maize enhanced the APase activity through regulating the expression of ZmPAPs to improve P utilization efficiency under the stress of phosphorus deprivation. In conclusion, this work provides basic data for further study on ZmPAPs family.

关 键 词：玉米 紫色酸性磷酸酶 表达分析 亚细胞定位 酸性磷酸酶活性 

分 类 号：S513[农业科学—作物学]