玉米2个光敏色素A基因的克隆、蛋白结构与光诱导表达模式  

Isolation and Primary Analysis of Protein Structures and Expression Patterns Responding to Different Light Treatments of Two Phytochrome A Genes in Maize (Zea mays L.)

作　　者：马燕斌[1,2] 李壮[1,2] 蔡应繁[3] 周朋[2] 肖阳[4] 黄玉碧[1] 付凤玲[1] 潘光堂[1] 杨克诚[1] 杨建平[2,3] 

机构地区：[1]四川农业大学玉米研究所,四川雅安625014 [2]中国农业科学院作物科学研究所,北京100081 [3]重庆邮电大学生物信息学院,重庆400065 [4]中国农业科学院研究生院,北京100081

出　　处：《中国农业科学》2010年第10期1985-1993,共9页Scientia Agricultura Sinica

基　　金：国家"863"高技术研究发展计划(2008AA10Z121);转基因生物新品种培育重大专项(2008ZX08009-003;2008ZX08010-002);重庆市自然科学基金(CSTC2009BA1088);中央级公益性科研院所基本科研业务费专项(2060302-2);教育部归国人员启动费;农业部回国人员择优资助项目

摘　　要：【目的】分析玉米2个光敏色素A基因(ZmPhyA1和ZmPhyA2)的蛋白结构及可能存在的功能区段,明确其响应于不同光线处理的表达模式。【方法】通过RT-PCR方法获得ZmPhyA1和ZmPhyA2编码的完整cDNA序列,推断2个基因编码蛋白的保守结构域,利用酵母双杂交测试其结构域间的物理互作,利用半定量RT-PCR技术分析不同光处理条件下ZmPhyA1和ZmPhyA2的表达模式。【结果】遗传进化树分析表明,ZmPHYA2与高粱光敏色素A的遗传关系要比与ZmPHYA1更近。ZmPHYA1与ZmPHYA2蛋白中含有1个GAF结构域、1个Phytochrome结构域、2个PAS结构域、1个HisKinaseA结构域和1个类似组氨酸激酶的ATP激酶结构。ZmPHYA1与ZmPHYA2自身和相互之间可以通过其C端539个氨基酸区段进行相互作用。ZmPhyA1和ZmPhyA2在黑暗、远红光和蓝光条件下表达量较高,在红光和白光条件下较低,并且前者的表达量均低于后者。【结论】ZmPHYA1与ZmPHYA2蛋白是2个具有功能的光受体蛋白,自身和相互之间可能通过C端539个氨基酸区段互作形成复合体,二者的表达水平受不同光质的调控。【Objective】 The protein structures and domain functions of two phytochrome genes of maize （Zea mays L.） （ZmPhyA1 and ZmPhyA2） were analyzed. The expression patterns of ZmPhyA1 and ZmPhyA2 responding to different light treatments were detected. 【Method】 The full cDNA of ZmPhyA1 and ZmPhyA2 were cloned through RT-PCR from maize inbred line of B73. The putative domains of ZmPHYA1 and ZmPHYA2 proteins were predicted via the NCBI Protein BLAST. The physical interactions between different functional domains of ZmPHYA1 and ZmPHYA2 were confirmed using yeast two hybrid system. Their expression levels were detected using semi-quantitative RT-PCR assay. 【Result】 Phylogenetic tree analysis indicated that ZmPHYA2 was more close with SbPHYA （Sorghum bicolor） than with ZmPHYA1. Both of ZmPHYA1 and ZmPHYA2 were composed of a GAF domain,a phytochrome domain,two PAS domains,a His Kinase A domain and a Histidine kinase-like ATPase domain. ZmPHYA1 and ZmPHYA2 were physically able to interact with themselves and each other through the 539-amino acids in their C terminal domains. Both of ZmPhyA1 and ZmPhyA2 showed higher expression lelvels in darkness,far-red and blue light conditions,but lower in red and white light conditions. 【Conclusion】 The ZmPHYA1and ZmPHYA2 proteins belong to active photo-receptor in maize as PHYA does in model plant Arabidopsis. The expression of ZmPhyA1 and ZmPhyA2 rapidly corresponded to dark,far-red,red and blue light treatments.

关 键 词：玉米 光敏色素A 蛋白互作 表达分析 

分 类 号：S513[农业科学—作物学]