乌塌菜DET1基因的克隆及表达模式分析  

Cloning of Gene DET1 and Analyzing of the Expression Pattern in Wuta-cai

作　　者：张彬[1,2] 周爽[2] 朱明库[2] 尹美强[1] 赵娟[1] 王玉国[1] 

机构地区：[1]山西农业大学农学院,山西太谷030801 [2]重庆大学生物工程学院,重庆400044

出　　处：《山西农业科学》2014年第8期796-799,818,共5页Journal of Shanxi Agricultural Sciences

基　　金：山西省青年科技研究基金项目(2013021024-2);山西农业大学科技创新基金项目(201302);山西农业大学引进人才博士启动基金项目(2012YJ12)

摘　　要：高叶绿素是一种极为优良的农艺性状，有利于提高光合作用效率，增加产量。以芸薹属高叶绿素突变体乌塌菜为试验材料，研究调控叶绿素合成的基因DET1与乌塌菜高叶绿素含量的关系。测序结果显示，乌塌菜DET1基因全长cDNA为1690bp，编码535个氨基酸。与小白菜和羽衣甘蓝相比，乌塌菜DET1基因在第734。739位缺失了6个核苷酸“TGATGA”，导致乌塌菜DET1编码的蛋白质BrDET1w在第245位和第246位氨基酸处缺失了2个甲硫氨酸，其中，第245位的甲硫氨酸位于芸薹属DET1蛋白的保守位点。尽管生物信息学分析显示，乌塌菜BrDET1w的相对分子质量、等电点以及亲水性没有受到明显影响，半定量RT-PCR结果也显示，DET1基因的表达量在小白菜和乌塌菜中差别不大，但DET1基因保守位点的缺失仍可能是由于乌塌菜高叶绿素含量的原因。这一结果为乌塌菜DETI基因的后续研究奠定了基础。The agronomic trait high chlorophyll level is helpful to improve photosynthetic efficiency and boost yield. A Brassica plant, Wuta-cai with high chlorophyll level, was selected as the research material. We identified whether the gene DET1 related to high chlorophyll level caused the mutation of Wuta-cai. The sequencing results showed that the total cDNA length of DET1 was 1 690 bp, encoded 535 amino acids. Compared Wuta-cai with pakchoi and kale, there were 6 nucleotides TGATGA missing on the sites 734-739 of DET1 in Wuta-cai, which led the deletion of two methionines on the 245th and 246th sites of protein BrDETlw. The 245th methionine was located on conserved site of protein DET1 in Brassica plants. The relative molecular mass, isoclectric point and hydrophilia of BrDETlw were not affected significantly according to the bioinformatics analysis results. There was little significant difference of expression quantity of gene DET1 in pakchoi and Wuta-cai based on the results of semiquantitative RT-PCR. However, the deletion of conserved site of gene DET1 would be the reason why there was high chlorophyll level in Wuta-cai. The result laid the foundation for further research on gene DET1 in Wuta-eai.

关 键 词：乌塌菜 DET1 基因克隆 表达模式 

分 类 号：S634.4[农业科学—蔬菜学]