Cloning Of Chloroplast Genome And Study Of Regulating Chloroplast Development In Pseudosasa Japonica F.Akebonosuji H.Okamura

Pseudosasa japonica f.Akebonosuji H.Okamura is a precious bamboo species.It natives to Japan,and is a variant of Pseudosasa japonica.It could present diverse leaf color such as green,white,and green and white stripes(the shade,shape,quantity,and location of which are different within different leaves).So it is also a widely used bamboo species of landscaping.In our study,the chloroplast genome sequences of P.japonica f.Akebonosuji H.Okamura were sequenced,then we utilized the chloroplast genomes data to research the phylogenetic position of P.japonica f.Akebonosuji H.Okamura and evolution model of chloroplast genome.Furthermore,the leaf of different developmental stages or different colour was collected to study the relations of expression pattern of chloroplast genes and chloroplast development.The main conclusions of this study were as follows:1.The chloroplast genome of the green leaf of P.japonica f.Akebonosuji H.Okamura was139,641 bp in length.Through online software CPGAVAS analysis,the chloroplast genome of P.japonica f.Akebonosuji H.Okamura composed of 124 genes,including 86 protein-coding genes,7 r RNA genes and 31 tRNA genes.2.The 28 chloroplast genomes and GBSSI sequences were analyzed by using MEGA6.0.Two phylogenetic trees were constructed,which showed that P.japonica f.Akebonosuji H.Okamura belongs to Arundinariatae.3.The bioinformatics analysis of 28 chloroplast genomes and GBSSI showed that the structure of bamboo chloroplast genome was highly conserved.And the polymorphism of non-coding regions is higher than that of coding regions.4.The chloroplast ultrastructure and genes expression pattern of normal green leaf of different development stages were investigated.The result showed that the expression level of chloroplast genes were lower and stable in the early developmental stages,but increased significantly in the later developmental stages.Then the expression level increased with thylakoid stacking.This indicated that chloroplast genes had the relationship of positive correlation with chloroplast development.5.The chloroplast ultrastructure and genes expression pattern of albino leaf and revertible albinoleaf of different development stages were investigated.The result showed that thylakoid structure was deleted in chloroplasts of albino leaf.And thylakoid structure was recoverd in revertible albino leaf,as increasement of the green degree.The expression level of chloroplast gene rrn23 S,psbA,psbZ,psaA,and rbc L of albino leaf and revertible albino leaf had significant difference with green leaf.This indicated that the normal expression of these genes play a crucial role in chloroplast development and thylakoid formation.