中国中药杂志

2022, v.47(18) 4908-4918

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钩吻GeERF转录因子的鉴定及在低温胁迫下的表达
Identification of GeERF transcription factors in Gelsmium elegans and their expression under low temperature stress

尤垂淮;刘安玉;张婷;赵亚菲;崔天真;谢津津;林海玲;阙友雄;苏亚春;阙万才;
YOU Chui-huai;LIU An-yu;ZHANG Ting;ZHAO Ya-fei;CUI Tian-zhen;XIE Jin-jin;LIN Hai-ling;QUE You-xiong;SU Ya-chun;QUE Wan-cai;School of Life Sciences, Fujian Agriculture and Forestry University;Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture and Rural Affairs, Fujian Agriculture and Forestry University;Department of Pharmacy, Fujian Medical University Union Hospital;

摘要(Abstract):

钩吻Gelsemium elegans因其日益凸显的药用价值遭受人为采挖破坏,导致野生钩吻储量减少。人工栽培钩吻药材成为趋势,但钩吻不耐低温,挖掘低温响应基因对钩吻抗逆育种具有重要意义。该研究基于4℃低温胁迫下的钩吻转录组数据库,挖掘获得29个受低温诱导差异表达的钩吻GeERF基因。对其中21个具有完整编码框的GeERF基因序列进行生物信息学分析,结果显示,GeERF蛋白聚类在DREB亚组和ERF亚组的成员数量分别为12、9个。GeDREB1A-1~GeERF6B-1蛋白相对分子质量为23.78~50.96 kDa,氨基酸长度212~459 aa,推测均为亲水性的核定位蛋白。进一步从钩吻叶片中克隆获得GeERF2B-1基因的cDNA全长序列。亚细胞定位结果显示,GeERF2B-1在细胞核上定位。实时荧光定量PCR(quantitative real-time PCR, qRT-PCR)分析表明,GeERF2B-1基因在钩吻的根、茎和叶中组成型表达,且在根中表达量最高。此外,经4℃处理后,相较于对照,GeERF2B-1基因的表达量在12 h显著升高且达到峰值,表明该基因积极响应低温胁迫。研究结果为钩吻GeERF转录因子的功能研究提供科学依据,并为钩吻品种的抗逆性改良提供潜在的基因资源。
With prominent medicinal value, Gelsemium elegans has been overexploited, resulting in the reduction of the wild resource. As a result, artificial cultivation turns out to be a solution. However, this medicinal species is intolerant to low temperature, and thus genes responding to the low temperature are important for the cultivation of this species. Based on the transcriptome database of G. elegans at 4 ℃, 29 differentially expressed GeERF genes were identified. Bioinformatics analysis of 21 GeERF gene sequences with intact open reading frames showed that 12 and 9 of the GeERF proteins respectively clustered in DREB subgroup and ERF subgroup. GeDREB1 A-1-GeERF6 B-1, with molecular weight of 23.78-50.96 kDa and length of 212-459 aa, were all predicted to be hydrophilic and in nucleus. Furthermore, the full-length cDNA sequence of GeERF2B-1 was cloned from the leaves of G. elegans. Subcellular localization suggested that GeERF2B-1 was located in the nucleus. According to the quantitative reverse-transcription PCR(qRT-PCR), GeERF2B-1 showed constitutive expression in roots, stems, and leaves of G. elegans, and the expression was the highest in roots. In terms of the response to 4 ℃ treatment, the expression of GeERF2B-1 was significantly higher than that in the control and peaked at 12 h, suggesting a positive response to low temperature. This study lays a scientific basis for the functional study of GeERF transcription factors and provides gene resources for the improvement of stress resistance of G. elegans.

关键词(KeyWords): 钩吻;ERF转录因子;序列分析;低温胁迫;表达分析
Gelsmium elegans;ERF transcription factor;sequence analysis;low temperature stress;expression analysis

Abstract:

Keywords:

基金项目(Foundation): 福建省卫生厅中青年骨干人才培养项目(2020GGB024);; 福建省科技厅科技创新联合资金项目(2020Y9066);; 福建省教育厅中青年教师教育科研项目(JAT200140);; 福建农林大学大学生创新创业训练计划项目(202210389289)

作者(Authors): 尤垂淮;刘安玉;张婷;赵亚菲;崔天真;谢津津;林海玲;阙友雄;苏亚春;阙万才;
YOU Chui-huai;LIU An-yu;ZHANG Ting;ZHAO Ya-fei;CUI Tian-zhen;XIE Jin-jin;LIN Hai-ling;QUE You-xiong;SU Ya-chun;QUE Wan-cai;School of Life Sciences, Fujian Agriculture and Forestry University;Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture and Rural Affairs, Fujian Agriculture and Forestry University;Department of Pharmacy, Fujian Medical University Union Hospital;

DOI: 10.19540/j.cnki.cjcmm.20220708.101

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