摘要
【目的】DNA binding with one finger(Dof)基因家族是植物特有的转录因子在植物生长发育和胁迫响应中发挥重要作用。研究Dof转录因子在马铃薯响应逆境胁迫特别是盐胁迫中的功能与机制,为获得抗盐逆境胁迫新品种提供一定的理论和技术基础。【方法】克隆StDof5,并对其进行生物信息学分析,通过同源重组的方法构建StDof5过表达载体,转化马铃薯得到阳性植株,并对其表达进行分析。【结果】StDof5的CDS全长498 bp,编码165个氨基酸,分子式为C800H1236N242O246S9,StDof5原子个数2533个,相对分子量为18468.63 kD,等电点是8.47,脂肪族指数为51.45,不稳定系数为45.16,预测该蛋白为不稳定蛋白。预测得到的跨膜螺旋个数为0,其属于非跨膜蛋白。StDof5蛋白亲水性平均值为-0.876,预测该蛋白属于亲水性蛋白。StDof5蛋白二级结构预测结果显示α螺旋占20%,β转角占0,延长链占9.7%,无规卷曲占66.06%。盐胁迫表型试验表明,过表达植株(OPs)比野生型(DES)的株高、鲜重均有明显程度上升,但根长、根数和叶片数差异不大。【结论】过表达植株(OPs)能够抵御盐胁迫,对盐胁迫呈现脱敏感表型。该基因对盐胁迫响应明显,且存在组织特异性。
【Objective】The DNA binding with one finger(Dof)gene family is a plant-specific transcription factor family that plays important roles in plant growth,development and stress response.Studying the function and mechanism of Dof transcription factors responding to abiotic stress,especially salt stress in potato,provides some theoretical and technical basis for obtaining new varieties resistant to salt stress.【Method】StDof5 was cloned,and it was analyzed via bioinformatics.StDof5 overexpression vector was constructed by homologous recombination,potato was transformed to obtain positive plants lines,and their expressions were analyzed.【Result】CDS of StDof5 is 498 bp,it encodes 165 amino acids,and the molecular formula is C800H1236N242O246S9.The number of StDof5 atoms is 2533,the relative molecular weight is 18468.63 kD,the isoelectric point is 8.47,the aliphatic index is 51.45,and the instability coefficient is 45.16,the protein is predicted to be an unstable protein.The predicted number of StDof5 transmembrane helices is 0,thus it is a non-transmembrane protein.The average value of hydrophilicity of StDof5 protein is-0.876,predicting that it is hydrophilic.The predicted secondary structure of StDof5 protein showed it has 20%α-helices,0β-turn,9.7%extended strands and 66.06%random coils.Salt stress phenotype experiments showed that the plant heights and fresh weights of overexpressing plants(OPs)were significantly higher than those of wild type(DES)plants,but there was no significant difference in root length,root number and leaf number between them.【Conclusion】Overexpressing plants(OPs)can resist salt stress and demonstrate desensitization phenotype to salt stress.StDof5 has significant responses to salt stress and tissue specificity.
作者
梅显军
宋慧洋
李京昊
梅超
宋倩娜
冯瑞云
陈喜明
MEI Xian-jun;SONG Hui-yang;LI Jing-hao;MEI Chao;SONG Qian-na;FENG Rui-yun;CHEN Xi-ming(Maize Research Institute,Shanxi Agricultural University,Xinzhou 034000;College of Agriculture,Shanxi Agricultural University,Taigu 030800;Institute of Crop Science,Shanxi Academy of Agricultural Sciences,Taiyuan 030031)
出处
《生物技术通报》
CAS
CSCD
北大核心
2024年第3期181-192,共12页
Biotechnology Bulletin
基金
山西农业大学杂粮种质创新与分子育种国家实验室(筹)(202204010910001)
山西农业大学博士基金(2021BQ44)
山西农业大学生物育种工程(YZGC069)
山西省博士毕业生来晋工作奖励科研项目(SXBYKY2021084)
山西省农业科学院作物科学研究所博士基金(ZB1102)。
关键词
马铃薯
StDof5
载体构建
遗传转化
potato
StDof5 gene
vector construction
genetic transformation
