2个玉米ZmCRY1a基因的克隆及其响应光质处理的表达模式  

Molecular Cloning of Two Maize(Zea mays) CRY1a Genes and Their Expression Patterns of in Response to Different Light Treatments

作　　者：闫蕾[1,2] 杨宗举 苏亮[2] 肖阳[3] 郭林[2] 宋梅芳[2,4] 孙蕾[2,3] 孟凡华[2] 白建荣[1,5] 杨建平[2] 

机构地区：[1]山西大学生物工程学院,山西太原030006 [2]中国农业科学院作物科学研究所,北京100081 [3]中国农业科学院研究生院,北京100081 [4]北京市辐射中心,北京100875 [5]山西省农业科学院作物科学研究所,山西太原030031

出　　处：《作物学报》2016年第9期1298-1308,共11页Acta Agronomica Sinica

基　　金：国家重点研发计划试点专项项目(SQ2016ZY03002918);国家转基因生物新品种培育重大专项(2016ZX08010002-003-002);国家自然科学基金项目(3157026);北京市自然科学基金(重点)项目(6151002);中国农业科学院科技创新工程项目资助~~

摘　　要：隐花色素(cryptochrome,CRY)是植物蓝光的主要受体,参与其调节生长发育及生物钟过程。为研究隐花色素在玉米光形态建成及生物钟调控方面的作用,本研究利用同源克隆的方法得到玉米自交系B73的2个ZmCRY1a基因的c DNA序列,分别命名为ZmCRY1a1和ZmCRY1a2。这2个基因的编码区(coding DNA sequence,CDS)序列长度都为2124个核苷酸,编码707个氨基酸。生物信息学分析表明ZmCRY1a1和ZmCRY1a2推测的氨基酸序列均包含DNA photolyase、FAD binding和Crytochrome C结构域;与拟南芥及其他常见作物的CRY比对并构建系统发育树显示,这2个基因与水稻Os CRY1a氨基酸序列一致性最高,而与拟南芥和大豆等双子叶植物的CRY1氨基酸序列一致性相对较低。利用实时荧光定量PCR分析了ZmCRY1a1和ZmCRY1a2在不同器官及响应光质、光质转换及长日照与短日照处理的表达模式。在检测的器官中,ZmCRY1a1的表达丰度均高于ZmCRY1a2;这2个基因在成株期叶片中表达丰度最高,分别是根中ZmCRY1a1的52.1倍和6.2倍。相对于黑暗下,二者在各种持续光质中的表达丰度均较高,尤其在蓝光和远红光条件下。尽管是作为编码蓝光受体的基因,2个ZmCRY1a的表达却能强烈地响应远红光和红光转换处理。同样二者也能响应不同光周期处理,长日照条件下,ZmCRY1a1的转录在一个光周期内共出现5个峰值,而ZmCRY1a2的转录只有4个峰值;短日照条件下,2个ZmCRY1a的表达出现了极其相似的模式,均在进入黑暗后10 h和14 h时出现2个最高峰。由此推测2个ZmCRY1a可能在玉米光形态建成与开花调节中发挥重要作用。Cryptochromes are blue light receptors that regulate the development of growth and circadian clock in plants. To stress the functions of crytochrome 1（CRY1） on photomorphogenesis and flowering regulation in maize（Zea mays L.）, we isolated the c DNA clones of two ZmCRY1 a genes from inbred line B73 by homologous cloning, and designated as ZmCRY1a1 and ZmCRY1a2. The length of both ZmCRY1 a coding DNA sequences were 2124 nucleotides, which encoded 707 amino acid residues. Bioinformatics analyses were employed to predict their function domains and to build a phylogenetic relationship tree among plant CRY1 homologs by the DNAMAN software and the NCBI blast. The two ZmCRY1 a proteins possessed three function domains： DNA photolyase, FAD binding, and crytochrome C domains. Phylogenetic analysis indicated that the two ZmCRY1 a proteins belonged to the same branch with Os CRY1 a, while showing low similarity to other CRY1 proteins from dicotyledonous species, such as A. thaliana and Glycine max. The transcription abundances of two ZmCRY1 a genes in different organs and in response to light treatments were detected using quantitative RT-PCR（q RT-PCR）. q RT-PCR assays indicated that the two ZmCRY1 a genes were highly expressed in leaf with 52.1 or 6.2 times higher than ZmCRY1a1 abundance in root, respectively. The transcription abundances of the both genes were very high under different continuous light conditions, especially in blue and far-red light. Although encoding blue light receptors, they both greatly responded to dark-to-far-red and dark-to-red transitions. In addition, their transcription abundances could also respond to photoperiod treatment（both long-day and short-day conditions）. In long-day condition, The transcription abundances of ZmCRY1a1 and ZmCRY1a2 had five four peaks, respectively. In short-day condition, both ZmCRY1 a genes had two big peaks which happened at 10 h and 14 h after transition into darkness. Our results suggest that both ZmCRY1 a genes may be involved in seedling de-etiol

关 键 词：玉米 隐花色素 光信号转导 基因克隆 表达模式 

分 类 号：S513[农业科学—作物学]