Preliminaryanalysis Of VvABCG20 In Ovule Denelopment Process And The Key Genes In Antive Gibberellin Metabolize Of Grapevine

In this study, we use the different issues from Piont Noir as our matrial, and we also take the seed of seedlessgrapes and Point Noir grapes in different development stages, in addition,we also use 9 kinds of nuclear grapes and 6 kinds of seedless grapes as our material. Using the RT-PCR and VIGS to verify the VvABCG20 function and analysis the key genes inactive gibberellin synthesis and deactivate of grapevine, in order to find abortion-related key factors in gene level, exploring the impact of the active gibberellin on grapes ovule abortion, and providing a theoretical reference for the seedless grape breeding. The main findings are as follows.1、Using the 9 kinds of nuclear grapes and 6 kinds of seedless grapes as our material, and using RT-PCR to analysis the ovule-speciffic expression gene VvABCG20’s function. The result show that the different sequence between different grapes were small, the rate of amino acid matches are 99.28%, the analysis of SNP show that there are 9 single nucleotide mutation sites, but only one leads to the corresponding amino acid sites, the VvABCG20’s expression in nuclear grapes and seedless grapes have significant differences, and the expression in nuclear grapes are significant higher than the expression in seedless grapes.2、Using the Micro-Tom as material and using the VIGS toverify the VvABCG20 gene’s ortholog gene LeABCG20’s function in tomato. Compared with wild type tomato and TRV2 empty vector, TRV2-LeABCG20-1 infected tomato fruits semi-quantitative strip was significant weaker than the former two, and the seed development picture can also be seen that TRV2-LeABCG20-1 infected tomato fruits on the number of seeds is significant less than the wild type and empty vector infected tomato, this proves LeABCG20 gene plays akey role in tomato seed development, and indirectly confirms VvABCG20 gene has an important regulatory role in grapes seed development.3 、 We use the different developmental stages ovule in Point Noir and Seedless as material, analysising the sequences of the key geneVvGA3 ox in active gibberellin synthesis and detecting it’s function by RT-PCR. The result revealed that VvGA3 ox family memebers containing same domain, indicating the VvGA3 ox is relatively conservative, there are significant differences between Point Noir and Seedless, VvGA3ox1 and VvGA3ox2 haveshown a certain degree of reduction in seedless ovule abortion, and the expression in Point Noir are significant higher than the expression in Seedless, inferred VvGA3 ox family may regulate the grapes ovule through affect activity of gibberellin.4 、 We use the different developmental stages ovule in Point Noir and Seedless as material, analysising the sequences of the key gene VvGA2 ox in active gibberellin deactivate and detecting it’s function by RT-PCR. The result revealed that VvGA2 ox is also relatively conservative, there are significant differences between Point Noir and Seedless, VvGA2 ox presents up-regulated expression in critical period in Seedless ovule, this will conductive to the decomeposition of active GA, and inhibited of embryo development, combining with the expression of VvGA3 ox, taking these two points can decuce the active gibberellin may play akey role in grapes ovule abortion.