柠条锦鸡儿F3H基因克隆及功能分析

发布时间：2018-05-19 10:39

本文选题：柠条锦鸡儿 + CkFH基因　；参考：《广西植物》2017年06期

【摘要】：柠条锦鸡儿为豆科灌木,对各种环境胁迫具有较强的适应能力,类黄酮是天然的抗氧化剂,花青素属类黄酮化合物,逆境胁迫会影响植物体内花青素的合成,而黄烷酮3-羟化酶(F3H)是花青素生物合成所必需的一种关键酶。该研究成功分离克隆了柠条锦鸡儿的F3H基因,命名为CkF3H。CkF3H基因的开放阅读框(ORF)为1095 bp,编码364个氨基酸,推测的蛋白质分子量为41.3 kDa,理论等电点为5.9。生物信息学分析发现,CkF3H基因序列与其它植物F3H有较高的一致性,推测CkF3H蛋白与其它植物F3H蛋白具有相似的功能。利用染色体步移法克隆得到CkF3H起始密码子ATG上游468 bp的启动子序列,PlantCARE软件分析表明,该序列具有启动子的基本元件CAAT-box和TATA-box以及多种与逆境胁迫相关的顺式调控元件。实时荧光定量PCR分析表明,CkF3H在柠条的根、茎和叶中均有表达,没有组织特异性;CkF3H的表达受低温、高盐、干旱和高温胁迫的诱导,并且在低温胁迫下,CkF3H的表达还受到光周期的影响。综上所述,研究结果表明CkF3H基因在柠条锦鸡儿适应低温、高盐、干旱和高温胁迫的过程中发挥作用。
[Abstract]:Caragana korshinskii is a legume shrub. Flavonoids are natural antioxidants and anthocyanins are flavonoids. Stress affects the synthesis of anthocyanins in plants. F3H) is a key enzyme for anthocyanin biosynthesis. In this study, the F3H gene of Caragana korshinskii was successfully isolated and cloned. The open reading frame named CkF3H.CkF3H gene was 1095 BP, encoding 364 amino acids. The deduced molecular weight of protein was 41.3 kDa and the theoretical isoelectric point was 5.9. Bioinformatics analysis showed that the sequence of CkF3H gene was consistent with that of other plants. It was suggested that CkF3H protein had similar function with other plant F3H proteins. A 468bp promoter sequence upstream of CkF3H initiation codon (ATG) was cloned by chromosome step method and analyzed by PlantCARE software. The results showed that the promoter had the basic components of promoter CAAT-box and TATA-box, as well as a variety of cis-regulatory elements related to stress. Real-time fluorescence quantitative PCR analysis showed that CkF3H was expressed in roots, stems and leaves of Caragana korshinskii, and no tissue specific CkF3H expression was induced by low temperature, high salt, drought and high temperature stress. The expression of CkF3H was also affected by photoperiod under low temperature stress. In conclusion, CkF3H gene plays an important role in the adaptation of Caragana korshinskii to low temperature, high salt, drought and high temperature stress.
【作者单位】：西北师范大学生命科学学院;
【基金】：国家自然科学基金(31460099) 西北师范大学青年教师科研能力提升计划项目(NWNU-LKQN-14-23)~~
【分类号】：Q943.2

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