独行菜GRAS转录因子家族分析及LaSCL18基因克隆与冷相关性研究

发布时间：2018-04-24 08:42

本文选题：独行菜 + GRAS转录因子　；参考：《分子植物育种》2017年09期

【摘要】：以耐受低温萌发停滞期的种子为实验组,以低温萌发停滞期的种子为对照组,对独行菜(Lepidium apetalum Willd.)种子转录组进行了高通量测序。基于测序数据库资源,本研究对独行菜GRAS类转录因子进行了分析,除去冗余,发现独行菜种子共有GRAS家族270个成员。其中138个成员相对于低温萌发停滞组,在耐受低温萌发的独行菜种子中上调表达,128个下调表达,4个表达水平无差异。进一步选择了一个极显著上调表达的GRAS基因片段,通过Blastn同源序列比对分析表明,该基因片段与拟南芥GRAS基因家族SCL18全长基因一致性最高,达到96%,故本研究将其命名为La SCL18(Gen Bank登录号KY466159)。结合同源序列克隆法、采用RT-PCR技术克隆获得该GRAS基因c DNA序列,全长1 633 bp,编码一个完整的开放阅读框。生物信息学分析表明,该c DNA序列编码435个氨基酸、具有GRAS转录因子家族特有的保守结构域、亲水性较高、是一个水溶性较好的球状蛋白。编码蛋白分子质量139.883 k D、理论等电点为4.97、不具有跨膜区和DNA结合位点,表明其在调控相关基因转录时可能不直接与DNA序列结合,而是与其它转录因子相互作用后发挥功能。实时荧光定量PCR分析表明,独行菜幼苗在低温胁迫处理后La SCL18基因表达量迅速显著升高,初步推测La SCL18基因表达与独行菜幼苗耐受低温有关。
[Abstract]:The seeds of Lepidium apetalum Willd.were used as experimental group and control group respectively. High throughput sequencing of seed transcriptome was carried out. Based on sequencing database resources, GRAS transcription factors were analyzed in this study. After removing redundancy, 270 members of GRAS family were found. Compared with the cold germinating arrest group, 138 members up-regulated the expression in the seeds of the cold germinating alone, 128 down-regulated expression, and 4 expression levels had no difference. A highly up-regulated GRAS gene fragment was further selected. The results of Blastn homologous sequence alignment analysis showed that the gene fragment had the highest consistency with the full-length SCL18 gene of Arabidopsis GRAS gene family. This study named La SCL18(Gen Bank login number KY466159. Combined with homologous sequence cloning, the GRAS gene c DNA sequence was cloned by RT-PCR technique. The cDNA sequence was 1 633 BP, encoding a complete open reading frame. Bioinformatics analysis showed that the c DNA sequence encodes 435 amino acids, has the conserved domain of GRAS transcription factor family, has high hydrophilicity, and is a better water-soluble globular protein. With a molecular weight of 139.883 KD and a theoretical isoelectric point of 4.97, the encoded protein does not have a transmembrane region or a DNA binding site, indicating that it may not directly bind to the DNA sequence in regulating the transcription of the related genes, but function after interacting with other transcription factors. The results of real-time fluorescence quantitative PCR analysis showed that La SCL18 gene expression increased rapidly after low temperature stress. It was preliminarily assumed that La SCL18 gene expression was related to tolerance to low temperature.
【作者单位】：新疆特殊环境物种保护与调控生物学实验室;北京师范大学生命科学学院抗性基因资源与分子发育北京市重点实验室;
【基金】：国家自然科学基金(3166010371) 新疆维吾尔自治区高等学校科研计划项目(XJEDU2014I027) 新疆维吾尔自治区大学生科研创新项目(201610762095) 新疆师范大学校级研究生科研创新项目(XSY201602003)共同资助
【分类号】：Q943.2

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