Gene Cloning And Functional Analysis Of PpeTAC1

Branching angle determines the plant type,which is one of the factors that makes the plant kingdom in various shapes,and has a great influence on the agricultural and forestry production.Because of branching angle deciding the structure and morphology of the plant,so it affects the structure of the community's living space,such as rice tillering angle affects the plant's capturing lightability and reproducing ability.In the production and cultivation of peach,the columnar tree's crown and branching angle is smaller than the standard tree's,so the shaping and pruning is relatively easy,which can reduce the artificial input.Related genes research of affecting the formation of plant branching angle mainly concentrated in monocotyledon,including rice,com,wheat,and model plant,such as Arabidopsis thaliana.LAZY1,TAC1,PROGland LPA1havebeen acquiredthat had a regulating effect on plant branch angle.And different genes on the mechanisms of regulating plant branching angle is different.To discovering and verifying gene function of different variation types PpeTACl(Tiller Angle control 1),and clarifying molecular mechanism of PpeTACl gene in regulation branching angle.Firstly,researching expression features of PpeTACl genes was conducted in'okubo'(standard)and‘Sahonglongzhu'(pillar).Thenthe variation types of PpeTACl's in genome level and transcriptional level were analyzed.Furthermore,the overexpression vectors of differentvariation types were constructed and transformed into wild-type Arabidopsisand attaclmutant,respectively.Itwould be conducive to improvethe peach tree by molecular breeding,and create excellent new germplasms.It has great significance to increase yield and fruit quality,reduce labor input and enhance market competitiveness.The main results are as follows:1.During the whole growth period,the angle of the‘Okubo'was larger than that of'Sahonglongzhu'.The angle of the 'Okubo'had a trend of increasing,while the angle of 'Sahonglongzhu' showed a gradual decreasing.The period of maximum variation of the two varieties was different,and the basal angle,waist angle and geotropicangle in the sane of variety were different.2.In the six parts of the whole growth period,the expression of TAClin 'Okubo'was higher than that of 'Sahonglongzhu'.During the whole growth period,the expression ofPpeTAClin 'Okubo'was up to 16.56,on the contrary,expression of PpeTAClin'Sahonglongzhu 'was very low in different parts(under 3.5).The trend of relative expression of the different parts of the PpeTAClin 'Okubo'was different,while bark of branch attachment site(top),and bark of branching attachment site(bottom)hadsimilar trends in expression.3.Correlation analysis of between the expression of PpeTACl gene and branching angle in two kinds of trees.In the period of vigorous growth,PpeTACl gene had a greater effect on the change of the branching angle in 'Okubo',whilethe influence of the environment on branching angle may be greater in the later stage of plant growth.The expression of PpeTACl gene was closely related to the opening degree of the shoot in the growth stage,and would be influenced by the environment in the later stage.The expression of PpeTACl genein leaves,bark of branch attachment site(top),bark of branch attachment site(bottom),lyr old branch(top),lyr old branch(middle),lyr old branch(bottom)had the positive correlation withbasal angle,waist angle and geotropic angle.4.The mutation analysis of PpeTACl gene in the DNA level.Compared with the DNA sequence of'okubo',the variation sites of different pillar types were different.The mutation sites mainly appeared in different areas,including 5 'UTR,3 introns,4 introns and 3' UTR.At different variation sites had similarity,such as the sequence of heterozygous sites appearing in 5 'UTR was the same,and the location is consistent,the second SNP site in 3 introns and 4 introns were the same.5.The mutation analysis of Ppe TACl gene in transcriptional level.Compared with the cDNA sequence of'okubo,the cDNA of other pillartypes showed different types of mutations.'Sahonglongzhu'and '18' belonged to Ppetacl-1(inserted three base pairs in 3 expressed region showedinsertion(TGA)and three SNPS),while'Zhouxingshantao',' Terutebeni','Terutemomo','Teruteshiro' and 'Terutehime'belonged to Ppetacl-2(insertion(TGA)and four SNPS,and lack of 4 expressed region).6.Functional verification of variation of PpeTAClgenes.Three overexpression vector of different variation types of PpeTACl gene were constructed.35S::PpeTAC,35S::Ppetac1-1,35S::Ppetacl-2 and empty vector were transferred into wild-type Arabidopsis and attaclmutant.The expression of PpeTACland the phenotypic characteristics in transgenic plants are ongoing.