Identification Of AT-Hook Gene Family And Functional Analysis Of Gene AHL23 In Peanut

With the improvement of the quality of crop products,improving the quality of peanut seed has become the primary task in peanut breeding.In this study,the transcription factor AHL23 was identified by screening the differentially expressed genes in seed development of conventional peanut variety Kainong 70 and high-oleic acid peanut variety Kainong 176,and bioinformatics correlation analysis of this gene and AT-hook gene family was conducted to provide theoretical reference for further research on the function of peanut AT-hook protein.The target genes regulated by AHL23 transcription factor were mined by DAP-Seq and the construction of Arabidopsis thaliana transgenic plants,which laid a foundation for further exploration of AHL23 gene function.The main research results are as follows:1.64 AT-hook genes were identified in the peanut genome database,which were unevenly distributed on chromosomes.The AT-hook genes were classified into eight subgroups based on phylogenetic tree analysis,containing 5’-UTR and 3’-UTR in most genes,which encode proteins with six conserved domains.Most proteins contain RGRP and PPC motifs.2.The transcription factor AHL23 was identified from the difference between high and low oleic acid Gene expression profiling.The expression of AHL23 in high oleic acid cultivars was significantly lower than that in normal cultivars,the expression level was also different in different tissues of peanut plants,and the highest expression level was found in root and nodule.3.The full-length c DNA sequence of AHL23 gene was cloned from peanut leaves.The CDS sequence of AHL23 contains 882 bases and encodes 293 amino acids.The 79-91 amino acid sequence contains the conserved AT-hook domain and the 107-220 amino acid sequence contains the DUF296 domain.4.The subcellular localization of AHL23-GFP was analyzed using the protoplast system of Arabidopsis thaliana.Compared with the empty GFP vector,the green fluorescent protein of AHL23-GFP was mainly localized in the nucleus,these results suggest that AHL23 was a transcription factor for nuclear localization.5.The binding sites of AHL23 gene on peanut genome were mined by DAP-Seq technique,and 5,6159 Peak-related genes were screened out.5 short-motifs and 9long-motifs were detected.Based on DAP-Seq results and annotation table analysis,292 peaks were screened out,which were mainly distributed between 200 and 598 bp in length,corresponding to 237 genes on the genome,and distributed unevenly on 20 chromosomes of peanut.These genes are mainly involved in lipid transfer protein,seed linoleate9S-lipoxygenase,fatty acyl-Co A reductase,linoleate 13S-lipoxygenase and fatty acid desaturase.6.The AHL23-OX transgenic Arabidopsis thaliana was obtained by agrobacterium mediated method.A total of 47 plant hormones were detected by LC-MS/MS,and 5DS was highly expressed in AHL-OX1 and AHL-OX2 Arabidopsis thaliana,however,the presence of this hormone was not detected in wild arabidopsis thaliana,and 5DS,as the precursor of plant hormone aurogenin biosynthesis,may be involved in the regulation of the 5DS pathway by AHL23 gene,thus promoting aurogenin metabolism,thus affected the plant oil anabolism and the fatty acid content.