Identification And Characterization Of Downstream Regulators Of GL2 During Trichome Development In Arabidopsis

Trichome is a special epidermal cell which differentiated from the epidermis of plant aerial tissues.It has important physiological and biological functions.It is not only the good model system of plant cell formation,differentiation and development,but also the vital organ of plant response to abiotic stress.The trichome development is regulated by both the endogenous factors such as genetic factors,horomes and cell cycles and complex environmental factors like water and light.In orde to better understanding the molecular regulation mechanisms on trichome development,we focus on the GLABRA2(GL2)which in HD-Zip family of transcription factors.This factors located at a strategic location of trichomes growth regulation network in Arabidopsis.GL2 is the upstream signal output means,and the regulation of downstream signaling mechanisms remain unclearly.Using modern molecular biology methods and second-generation high-throughput sequencing technology to identify regulatory genes and gene networks of GL2 downstream.To take advantage of ChIP-seq method for screening target genes GL2 direct role in the genome-wide.On the one hand we constructed pGL2::GL2-HA and pGL2::GL2-GFP expression vectors to transform Arabidopsis Col and gl2-3 mutants,on the other hand we prepared the polyclonal antibody of GL2.Construction of the DEX inducible expression vector pGL2::GL2-GR and transformed Arabidopsis Col and gl2-3 mutants for screening genes regulated by GL2 in different stages of trichome development.The main results are as follows:(1)By using prokaryotic expression,affinity chromatography,rabbits immunization,antibody purification and immunoblotting,we obtained the GL2 polyclonal antibody;we alsoevaluated the antibody titers by immunoblot experiments.(2)By using fragment ligation,infusion,we constructed pGL2::GL2-GFP,pGL2::GL2-HA and also the DEX-induced expression vector pGL2::GL2-GR.(3)With agrobacterium mediated transformation,we got the positive transgenic plants and also isolated the lines with higher GL2 expression levels.