Preliminary Study On Function Unknown Gene AtLCR23in Arabidopsis Thaliana

Currently limited researches showed a part of LCR proteins in Arabidopsis are involved in plant defense mechanisms,but the most part of LCR family are function unknown genes.This preliminary study was for function unknown gene AtLCR23in Arabidopsis by Bioinformatics, molecular biology, cell biology method.The main research results are as follows:1.Bioinformatics analysis results showed that AtLCR23Located in the Chromosome4in Arabidopsis, full genomic sequence contain373basic group, including2exons and1introns, encoding77amino acids, from19to77amino acids constitute SLR1-BP domain.2. Screen homozygous mutant Icr23with T-DNA insertion, detect expression of AtLCR23in homozygous mutant by RT-PCR,and result shows Icr23mutant lost AtLCR23gene function.3.Constructing pBI101-pAtLCR23-AtLCR23-GUS fusion expression vector and transforming into Col wide type, GUS histochemical analysis of transformation plant showed that AtLCR23was specific expressed in stigma in Arabidopsis.4.Compare Ws with Icr23of flower organs by paraffin section, found there were no significant differences. Statistical data of siliques between Ws and lcr25,show that the lcr23siliques lack of abortive seeds in slightly more than the Ws.5.Through the RT-qPCR detection related functional genes expression level, like SLR1、 ARC1、PIP5PK.Found the expression of SLR1gene in the Icr23mutant were significantly lowered than wide type, and there no significant difference expression of ARC1、PIP5PK.6AtLCR23-overpressing plant was obtaining by transform vector pCAMBIA1300-35S-AtLCR23into Col,get two homozygous transformation named35S-23-6and35S-23-8respectively.Built RNAi interference vector of pGSA1285-AtLCR23transform into Col and get T2plant.In addition, structure GFP fusion expression vector of pCAMBIA1302-AtLCR23and GUS fusion expression vector with AtLCR21、AtLCR22、AtLCR35gene which sequence similar to AtLCR23. All of these can be used for the next phase of the study.