摘要
NAC转录因子在植物生长发育及盐胁迫响应过程中具有重要的调控作用。芝麻SiNAC77参与盐胁迫响应过程。克隆芝麻SiNAC77并分析其耐盐功能,为芝麻耐盐育种提供分子基础和基因资源。克隆SiNAC77的全长CDS序列,利用生物信息学软件对其基因序列和氨基酸序列特征进行分析。构建SiNAC77过表达载体,利用农杆菌介导法转化拟南芥。对过表达株系的耐盐表型和生理生化指标进行分析。结果表明,成功克隆了长度为1008 bp的SiNAC77 CDS序列,编码335个氨基酸,相对分子量为135.93 kD,预测等电点为4.91,含有33个潜在的磷酸化位点;启动子区域含有MBS、STRE、ARE、ABRE和TCA等多个非生物胁迫及激素响应元件。成功构建了SiNAC77过表达载体,并转化拟南芥,获得12个独立的转基因阳性株系。在盐胁迫下,与野生型拟南芥相比,过表达株系的种子发芽率、初生根长度、鲜重均显著提高,转基因株系中Na~+/K~+和相对MDA含量显著降低,相对SOD和POD抗氧化酶活性则显著增强。过表达芝麻SiNAC77可以增强抗氧化酶活性,减少离子毒害和氧化损伤,提高转基因植株的耐盐性。
NAC transcription factors play an important role in the regulation of plant growth,development,and responses to salt stress.The gene SiNAC77 in Sesamum indicum L.is involved in the response process to salt stress.Here,SiNAC77 was cloned to investigate its salt-tolerant function stress,which may provide a theoretical reference and valuable gene resource for breeding salt-tolerant sesame(Sesamum indicum L.)plants.First,the coding sequence(CDS)of SiNAC77 was cloned from Luzhi No.1,and its gene sequence and amino acid sequence characteristics were analyzed using bioinformatics tools.Subsequently,an SiNAC77-overexpressed vector was constructed and transformed containing SiNAC77 was constructed and the transformation of Arabidopsis was conducted using Agrobacterium tumefaciens.Finally,the salt-tolerant genotypes,physiological and biochemical indexes of the transgenic plants were analyzed under saline stress.As results,the CDS region of SiNAC77,1008 bp in length,was cloned successfully.The SiNAC77 gene encoded a protein containing 335 amino acids,with a molecular weight of 135.93 kD,an isoelectric point of 4.91 and 33 potential phosphorylation sites.The SiNAC77 promoter contained MBS,STRE,ARE,ABRE and TCA elements,functionally known to be involved in abiotic stress and phytohormone responses.The SiNAC77 overexpression vector was constructed and transformed successfully into Arabidopsis,and 12 independent transgenic lines were generated.Compared with wild-type Arabidopsis under salt stress,the transgenic lines had significantly higher seed germination rates,longer primary roots,higher fresh weights,significantly lower Na+/K+ratios and MDA contents and higher SOD and POD antioxidant enzyme activities.In summary,the overexpression of SiNAC77 improved the salt tolerance in transgenic plants by enhancing the activities of antioxidant enzymes and reducing ionic toxicity and oxidative damage.
作者
张玉娟
黎冬华
宫慧慧
崔新晓
高春华
张秀荣
游均
赵军胜
ZHANG Yu-juan;LI Dong-hua;GONG Hui-hui;CUI Xin-xiao;GAO Chun-hua;ZHANG Xiu-rong;YOU Jun;ZHAO Jun-sheng(Institute of Industrial Crops,Shandong Academy of Agricultural Sciences,Jinan 250100;Oil Crops Research Institute,Chinese Academy of Agricultural Sciences,Key Laboratory of Biology and Genetic Improvement of Oil Crops,Ministry of Agriculture and Rural Affairs,Wuhan 430062)
出处
《生物技术通报》
CAS
CSCD
北大核心
2023年第11期308-317,共10页
Biotechnology Bulletin
基金
山东省科技型中小企业创新能力提升工程项目(2021TSGC1209)
山东省农业科学院农业科技创新工程项目(CXGC2023A45,CXGC2022B01,CXGC2022F18)。
关键词
芝麻
盐胁迫
NAC转录因子
耐盐基因
功能分析
sesame
salt stress
NAC transcription factor
salt-tolerant gene
functional analysis
