Genome-Wide Identification And Library Construction Of Potato Transcription Factors

Potato is an important food crop,but the average global potato production is much lower than the ideal yield due to a variety of stresses.When the potato suffers from adverse environmental effects,it induces the expression of a large number of transcription factors in the body.Transcription factors are a class of transcriptionally active proteins that can directly bind to cis-acting elements of the target gene promoter region or interact with other proteins.The polymer binds to the promoter region of the target gene,thereby activating or inhibiting the transcription of the target gene,precisely regulating the target gene,thereby forming a transcriptional regulatory network,and reducing the damage of the plant under stress.Therefore,transcription factors play a key role in the regulation of plant growth and environmental adaptability.It is clear that how transcription factors participate in various pathways to regulate potato growth and development under stress,which is of great significance in production.Although there are some scattered reports on the transcription factor research in potato,the potato transcription factor has not been studied and analyzed from the wide-genome level.To this end,a potato genome-wide transcription factor cDNA library was established in this study.Firstly,the potato genome-wide transcription factors were identified and analyzed.The results showed that a total of 1736 genes in the potato encoded 2405 transcription factors,which were divided into 56 families according to their domain sequence characteristics.It can be seen that one gene may encode two or more transcription factors,and only one representative transcription factor is retained per gene in this study.Taking the ARF family as an example,the ARF family has 33 transcription factors,which are encoded by 18 genes.Among them,10 genes encode two or more different splicing transcription factors.After identification,each gene is retained only one representative transcription factor.The ARF family retained 18 representative transcription factors,and analyzed the gene structure and protein sequence of 18 transcription factors.There were significant differences among different genes,indicating that the ARF family genes are many functions that may be performed in potato growth and development and the environment adaptation process.In order to understand the function and expression pattern of potato ARF family members,this study used potato genomic RNA-seq data to analyze the expression specificity and stress expression patterns of potato ARF family members.In order to further explore the expression pattern of potato ARF family members,six tissues of root,stem,leaf,flower,stolon and tuber of potato cultivar diploid AC142 were sampled.Since some transcription factors were not expressed under normal environment,potato AC142 was treated with ABA solution,NaCl solution,high temperature,low temperature and drought stress and sampled at different time points.The expression levels of ARF family members were analyzed by real-time PCR.The results showed that the ARF family members had higher expression in the stolons,and the expression levels in other tissues were lower.Different genes had different tissue expression specificities.They have different expression levels during potato growth and development,and perform different functions.In this study,2,405 transcription factors of 56 families of potato were identified and analyzed,and 1736 representative transcription factors were retained after analysis.The primers were designed according to their CDS sequences,and the 1101 genes of 19 families of 1736 genes in the wide-genome were cloned by using the mixed cDNA of each tissue of potato AC142 as a template.Approximately 63.42% of the whole genome was successfully cloned,548 transcription factors were successfully cloned,and the target fragment was recombined with the pGADT7 vector and transformed into the recipient strain.If the sequencing result was confirmed to be correct,the bacterial solution was preserved.Analysis of 424 transcription factor clone sequences from 15 families,at least206 genes containing different alleles,and at least 21 genes containing alternative splicing patterns.At least one of the 88 genes has the same sequence as the DM1-3 sequence,and at least 14 of the gene sequences have different splicing patterns than the DM1-3reference sequence,and a total of 654 transcripts are obtained.At present,the libraryconstruction work is completed 31.57%.The establishment of this library provides conditions for studying the regulation of transcription factors at the wide-genome level and provides a very important resource for potato research.Large-scale analysis of transcription factors using efficient screening systems provides a very useful tool for studying potato transcriptional regulatory networks.