Study On Expressions Of AtGLR Ii Gene Subfamily In Arabidopsis Thaliana

The 20 genes that encode the Arabidopsis thaliana glutamate receptor gene family (AtGLRs) share significant similarity in amino acid coding sequence with animal ionotropic glutamate receptor (iGluR) subunits. The family could be divided into three subfamilies in which the type II subfamily is composed of 9 AtGLR genes. In order to study the expression profiles of the type II subfamily, vectors which fused AtGLR genes promoters with GUS gene have been used in this research. The expressions of those genes in different tissues have been observed in different organs and stages. Meanwhile, two amiRNA interference vectors have been constructed with 9 genes as target-genes. The results are as follows:1. The construction of Promoter::GUS vectors and gene transferWhen the gene transfered plants were screened out, plants growed up for 10 days and 6 weeks were GUS stained respectively. 6 lines of gene transfered Arabidopsis thaliana were obtained, and the AtGLR2.3, AtGLR2.5, AtGLR2.6, AtGLR2.8 and AtGLR2.9 lines could be GUS stained successfully. The results of GUS stain showed that AtGLR2.3,AtGLR2.6,AtGLR2.9and AtGLR2.5 experssed in seedling stage, the first three expressed in different part of seedling leaves each, and the AtGLR2.5 experessed only in seedling root. The expression of AtGLR2.8 were high in seedling leaves, roots and mature plants'trichomes, anthers, siliques. The expression of these genes showed the characteristic of temporal and spatial regulation.2. The construction of amiRNA interference vector and gene silencingamiRNA interference vectors were designed to silence AtGLR genes by transgene. Two amiRNA interference vectors aiming at AtGLR2.1-2.4 and AtGLR2.5-2.9 were designed respectively. As a result, AtGLR2.4, AtGLR2.8 and AtGLR2.9 were silenced, the expression of AtGLR2.3 was down-regulated. The AtGLR gene knock-out mutant lines could be researched later in the physiology and biological experiments.