Isolation And Identification Of Mutants Related To Plasmodesmata In Rice

In this paper,two methods were screened by fluorescence microscopy and laser scanning confocal microscopy.The 800 mutants of T-DNA inserted from the Chinese Academy of Agricultural Sciences were used.Using the fluorescent dye CFDA,we screened the mutants related to plasmodesmata in rice,and examined the photosynthetic physiological indexes.In order to understand the information of T-DNA insertion site in the mutant plants,the flanking sequences were amplified by TAIL-PCR technique and the flanking sequences were analyzed.Finally,Real-time fluorescence quantitative PCR method was used to detect the expression changes of the upstream and downstream genes of the T-DNA insertion site,so as to find out the change of gene expression.The main results are as follows:(1)The 89 mutants were obtained and 25 mutants were obtained by re-screening.(2)At the same growth stage,the permeability and quantity of plasmodesmata were different.(3)Using TAIL-PCR technique to amplify the flanking sequence of the mutant,the sequence alignment showed that T-DNA of two plants was inserted into chromosome 3and was inserted into the same gene.(4)The agronomic traits of the two mutants obtained by TAIL-PCR were lower than those of wild type,such as plant height,panicle length and 1000-grain weight.May be due to the insertion of T-DNA leading to changes in agronomic traits.(5)Real-time fluorescence quantitative PCR showed that the expression level of the gene near the T-DNA insertion site was lower than that of the wild type,which was not consistent with the related literature.The reason for the experiment was to be explored.