Screening Modified Mutants Of The Trichome Mutant Gl2-3 And Cloning The Genes In Arabidopsis Thaliana

Trichomes widely distribution in overground parts of land plants,a special structure of plant epidermal tissue,and serves many biological functions.Arabidopsis thaliana trichome is a typical,specialized single cell structure,often as a study of plant cell differentiation and molecular mechanisms of the excellent model system.The development of Arabidopsis thaliana trichome is a sophisticated molecular regulation process,has been found in a number of direct control of trichome development of key genes.Among them,GL2 is a key factor in the development of trichome of Arabidopsis thaliana,which is involved in the fate of epidermal hair cells and the formation of trichome morphology.It plays an important role in the upstream signal model-activation inhibition model of trichome But the GL2 downstream regulation mechanism is not clear,but it is certain that GL2 downstream regulatory pathway involves a large number of interaction factors,the interaction between different pathways to form a complex internet of GL2 downstream regulation.In order to explore the new genetic interaction between them,we improved the mechanism of GL2 involved in epidermal hair development.We screened the gl2-3 secondary mutagenesis library obtained by mutagenesis of Ethyl methanesulphonate(EMS)The mutant gene was cloned and the mutant gene was cloned to analyze the interaction between the mutant gene and GL2.The main results achieved are as follows:1.Three large trichomes mutants M30-01 and abt2-1 were screened by observing the phenotypic phenotype of Arabidopsis thaliana gl2-3 secondary mutagenesis.M30-01 is a reversal mutant of gl2-3,phenotype compared with gl2-3,a small amount of trichomes in the middle of the rosette leaves,reversing the gl2-3 trichomes development blocked phenotype,and to provide a basis for the study of GL2 regulation of trichomes growth mechanism.abt2-1 is a phenotypic mutant with a significant increase in epidermal hair branching.Genetic analysis showed that the the abt2-1 was single recessive gene mutation in nuclear.2.Gene mutation and cloning of the trichomes mutant abt2-1 were carried out by Map-based cloning.The results showed that the mutation site was located between F20M13 # 3 and T9A14 # 1 molecular marker.A homologous mutation of guanine(G)to adenine(A)was found in the clone of the gene At4g38600(KAKTUS / KAK)in this interval,which was located at the last base of the 13 th exon,immediately adjacent to the intron.The expression of At4g38600 gene in abt2-1 was detected by primers on both sides of the mutation site.It was found that At4g38600 amplified three different bands in this interval.The sequencing results showed that the three bands were due to the synonymous mutation resulting in different ways of splicing.3.The construction of expression vector of the cell cycle protein-related Gene.The overexpression vectors of At2g17620,At4g11920,At4g22910,At5g04470 and At5g06270 genes were constructed.The overexpressed seeds of T2 transgenic plants were obtained,which provided experimental materials for the subsequent development of cell cycle proteins.