Cloning And Functional Study Of StAAP1 And StAAP8 Genes In Potato

Amino acid transporters mediate the transport of various amino acids in plants,which are essential for plant growth and development.The amino acid transporters in plants are mainly divided into ATFs(the amino acid transporter family)and APCs(the amino acid polyamine choline transporter).The AAPs(amino acid permease)subfamily belongs to the ATFs family,and so far AAPs(amino acids Permease)is a relatively well-defined amino acid transporter studied in plants.In this study,the bioinformatics method was used to analyze the gene structure,protein structure and expression pattern of potato AAP gene family members,and the AAP family protein evolution tree was constructed.The StAAP1 and StAAP8 genes were cloned from the potato cultivar Desiree.Moreover,we constructed the over-expression vectors of StAAP1 and StAAP8 and transformed into potato leaves mediated by Agrobacterium.In addition,the subcellular localizations of them in the growth and development of potato.The main results are as follows:(1)Eight AAP genes were identified in the potato genome by bioinformatics analysis,and the potato AAP family genes were divided into two subgroups.The first subgroup is the StAAP gene containing six microporous transport channels(Pore-lining),including StAAP1,StAAP2,StAAP3,StAAP4,StAAP7,StAAP8.the second subgroup are the StAAP gene containing seven microporous transport channels,which includes StAAP5 and StAAP6.(2)Based on the transcriptomics data of PGSC,the expression patterns of StAAP gene family members were analyzed.It was concluded that StAAP8 gene may affect the osmotic potential of potato root cells by regulating the transport of amino acids in potato roots,thus improving the tolerance of plants to abiotic stresses like drought.(3)The StAAP1 and StAAP8 genes were cloned by RT-PCR.The StAAP1 gene was1446 bp in length and encoded 481 amino acids.The StAAP8 gene was 1467 bp in length and encoded 488 amino acids.Subcellular localization analysis revealed that both StAAP1 and StAAP8 were localized on the cell membrane.(4)In order to study the biological functions of StAAP1 and StAAP8 genes in potato,the overexpression vectors of StAAP1 and StAAP8 genes were constructed.The plant expressive vectors were transformed into potato mediated by Agrobacterium.Thetransgenic plants were identified by resistance screening and PCR detection.we also detected the expression levels of StAAP1 and StAAP8 genes in transgenic plants by qRT-PCR.(5)Observing the phenotype of the transgenic plants,it was found that there was no significant difference at the plant height among the OXStAAP1,OXStAAP8 transgenic plants and the wild type,however their leaf area was significantly larger.Phenotypic analysis showed that there were significant differences at the microtubers between overexpression StAAP1 transgenic plants and wild-type,however there was no significant difference between overexpression StAAP8 transgenic plants and wild-type.