紫苏PfLEC1基因的克隆及表达研究
    点此下载全文
引用本文:李 丹,陆俊杏,鲁 庚,唐 鑫,张小梅,张 涛.紫苏PfLEC1基因的克隆及表达研究[J].西北植物学报,2019,39(12):2154~2160
摘要点击次数: 54
全文下载次数: 62
作者单位
李 丹,陆俊杏,鲁 庚,唐 鑫,张小梅,张 涛* (重庆师范大学 生命科学学院重庆 401331) 
基金项目:国家自然科学基金(31171588);
中文摘要:紫苏是目前发现的α 亚麻酸含量最高的药食同源经济作物,其种子油脂中含60%以上的α 亚麻酸。该研究基于紫苏转录组测序结果,从紫苏种子中克隆获得植物种胚发生过程中的关键基因Leafy Cotyledon 1(Pf LEC1),并对其进行相关生物信息学分析、实时荧光定量PCR以及不同时期种子的脂肪酸含量测定,以探讨紫苏α 亚麻酸高效合成积累的分子调控机制。(1)序列分析结果表明,Pf LEC1基因编码区长度为621 bp,可编码206个氨基酸,属于NF YB亚基家族,含有HAP3亚基的保守功能域B区域;在线预测该蛋白为不稳定亲水性蛋白,无信号肽和跨膜区域,定位于细胞质;进化分析表明,该蛋白序列与拟南芥、甘蓝型油菜、水稻和玉米关系较近。(2)实时荧光定量PCR分析表明,PfLEC1基因在紫苏根、茎、叶、花及种子中均有表达,且在种子中表达量最高,根中表达最低;PfLEC1在种子发育的前期表达较高,随着种子发育表达量显著下降。(3)不同阶段紫苏种子脂肪酸含量分析表明,油酸、α 亚麻酸含量随种子发育逐渐增加,而棕榈酸、硬脂酸、亚油酸含量变化则相反,其中变化最为明显的是α 亚麻酸,从种子发育5 d时的33.16%上升至20 d时的65.16%,表明紫苏种子中α 亚麻酸含量是随种子发育快速积累的。研究推测,PfLEC1可能与紫苏种子α 亚麻酸的高含量合成积累密切相关,该研究为今后深入探讨PfLEC1基因的功能奠定了分子基础。
中文关键词:紫苏  LEC1  克隆  序列分析  表达分析
 
Cloning and Expression Analysis of Pf LEC1 Gene from Perilla frutescens
Abstract:Perilla is the medicinal and food homogeneous cash crop with the highest α linolenic acid content. The seed oil contains more than 60% of α linolenic acid. However, the molecular regulation mechanism of efficient synthesis and accumulation of α linolenic acid is still unclear. Based on the sequencing results of perilla transcriptome, this study cloned Leafy Cotyledon 1 (Pf LEC1), a key gene in the process of plant embryogenesis, from Perilla seed, and performed relevant bioinformatics analysis, real time PCR and different determination of fatty acid content of seeds at different stages. (1) Sequence analysis results show that the gene coding region is 621 bp in length and can encode 206 amino acids. It belongs to the NF YB subunit family and contains the conserved functional domain B region of the HAP3 subunit. The protein was predicted online as an unstable,hydrophilic protein with no signal peptide or transmembrane region and localized to the cytoplasm. Evolutionary analysis showed that the protein sequence is closely related to ArabidopsisBrassica napus, rice and corn. (2) Real time quantitative PCR analysis showed that PfLEC1 gene was expressed in roots, stems, leaves, flowers and seeds, with the highest expression in seeds and the lowest expression in roots; PfLEC1 expression was higher in the early stages of seed development, and with seed development, the expression level decreased significantly. (3) Analysis of fatty acid content of perilla seeds at different stages showed that the contents of oleic acid and α linolenic acid gradually increased with the development of the seeds, while the content of palmitic acid,stearic acid,and linoleic acid changed in reverse. It increased from 33.16% in 5 days to 65.16% in 20 days,indicating that the content of α linolenic acid in perilla seeds accumulated rapidly with the development of seeds. It is speculated that PfLEC1 may be closely related to the high level synthesis and accumulation of α linolenic acid in perilla seeds. This study lays the molecular foundation for further exploration of the function of PfLEC1 gene in the future.
keywords:Perilla frutescens  LEC1  cloning  sequence analysis  expression analysis
查看全文  查看/发表评论  下载PDF阅读器
   今日访问:3533    总访问量:10515482

版权所有:《西北植物学报》编辑部

技术支持:北京勤云科技有限公司