Bioinformatics Analysis And Expression Analysis Under Drought Stress Of BHLH Transcription Factor Family In Potato

In this study,the plant transcription factor database PlantTFDB(http://planttfdb.cbi.pku.edu.cn/)was used to obtain the potato and Arabidopsis bHLH gene family proteins and gene sequences,and the biological information of the potato bHLH transcription factor family was systematically studied.The expression pattern of some bHLH TFs under drought stress in potato was studied,the StbHLH1 gene was cloned and analyzed functionally.details as follows:Plant transcription factors obtained using a database 206 potato bHLH transcription factor.The potato bHLH protein consists of an average of 519.8 amino acids,an average GRAVY value of positive.Analysis of gene structure indicated that most StbHLHs genes do not contain introns,of which StbHLH82 contains two introns,and StbHLH29,StbHLH45,StbHLH115,StbHLH129,StbHLH136,StbHLH157,and StbHLH197 contain one intron.In the bHLH domain of the StbHLHs protein,a conserved tryptophan residue repeat region R1,R2 and R3 is included.All genes StbHLHs unevenly distributed on the 12 chromosomes,of which chromosome 3 was the most distributed(27)and chromosome 11 was the least(4).Functional prediction indicated that potato StbHLHs protein was associated with cell growth and development,and 14 StbHLHs genes responding to drought stress were selected based on phylogenetic tree results.14 StbHLHs analyzed gene expression pattern of stress in the potato DM drought and salt by qRT-PCR.Drought stress: The expression level of StbHLH194 in leaf tissue was up-regulated at 12 h and 24 h.In root tissues,StbHLH11 was down-regulated at 12 h and 24 h,but at 24 h,the expression was higher than 12 h.In stem tissues,StbHLH11 was down-regulated at 12 h and 24 h,but at 24 h,it was higher than 12 h.StbHLH54 was down-regulated at 12 h,but up-regulated at 24 h;However,the regulation of stress by the same gene in different tissues is different.The StbHLH1 gene in SWS2 was cloned.The full length of StbHLH1 CDS is 2046 bp,and the number of protein amino acid residues can be 682.Its molecular weight is 75592.74 Da,and the subcellular cells are located in the nucleus.The proteins interacted with String online software were PGSC0003DMT400036281,PGSC0003DMT400021018,P,PGSC0003DMT400008569 and AN11,and PGSC0003DMT400008569 and AN11 also interacted.Construction of expression vector for StbHLH1 gene.First,the CDS sequence of the target gene was ligated into the intermediate vector pDONR207 to obtain pDONR207-StbHLH1.Secondly,the plasmid obtained in the BP reaction was recombined with the expression vector pJAM1502 to obtain the expression vector pJAM1502-StbHLH1.After sequencing,the plasmid was extracted and transferred to Agrobacterium tumefaciens(AGL1)by freeze-thaw method.Obtainment of StbHLH1 transgenic tobacco.the constructed plant expression vector pJAM1502-bHLH1 was transformed into by Agrobacterium infection.As a result,54 transformed seedlings were obtained,Among them,27 positive seedlings and the positive rate was 50%.The positive transgenic StbHLH1 seedlings were further tested,The water loss rate measurement showed that the water retention of the transgenic StbHLH1 tobacco was higher than that of the wild type.