Functional Analysis Of Three Genes In ABS3 Subfamily MATE Transporter Genes In Arabidopsis

MATE(Multidrug and Toxic Compound Extrusion)family are ubiquitous protein transporters. And for plants, MATE transporters involve in exporting xenobiotics and endogenous cationic, as well as participate in the regulation of the balance of metabolites and ions inside plants, which guarantee plants’ growth and development. Our lab has already studied 5 genes of the ABS3 subfamily in Arabidopsis MATE family. And from that, we noticed the over expression of the genes in ABS3 subfamily could lead to a serial of special phenotype. So, in this research, we are focus on the other 3 genes in ABS3 subfamily and analysis their function. The results are list as follows:(1)By building the phylogenetic tree of Arabidopsis MATE transporter family as well as doing the homologous sequence alignment between ABS3L4, ABS3L5, ABS3L6 and ABS3, we confirmed the ABS3L4, ABS3L5, ABS3L6 belong to the ABS3 subfamily and have highly protein sequence similarities with ABS3.(2)Analysis of the temporal and spatial specificity expression of ABS3L4, ABS3L5 and ABS3L6 genes from two factors. On the one hand, we use semi-quantitative RT-PCRs to test the genes tissue specific expression respectively, and find these genes have distinctive tissue expression but all of them express at flower tissue. On the other hand, we use GUS stainings to test the genes promoter-GUS transgenic plants respectively, and the results demonstrated that ABS3L6 gene express in distinctive organs in Arabidopsis.(3)Successfully build the gene-fluorescent expression vectors and with the use of protoplast transient transformation and co-localization analysis, we notice the ABS3L4, ABS3L5 and ABS3L6 have distinctive subcellular localizations as well as distinctive co-localization with ABS4.(4)Successfully get the over expression mutants of ABS3L4, ABS3L5 and ABS3L6, and observe the similar phenotype as abs3-1, and confirm the genes’ knock on in these mutants respectively.(5)Successfully get the ABS3L4, ABS3L5 and ABS3L6 loss-of-function mutants respectively, and have the 8 genes loss-of-function mutants.