Temporospatial Expressions And Function Analysis Of Cellulose Synthase Gene In The Growth Process Of Phyllostachys Edilus

Phyllostachys edilus is major main commercial bamboo specie in China.And cellulosesynthesis is a key to the formation of the bamboo.Cellulose is mainly synthesized by cellulose synthase and deposited in primary cell wall and secondary cell wall.Therefore,Study on structure and function of cellulose synthase of bamboo growth and development has an important guidence.Research material for this article is different periods of five heights(10 cm,30 cm,120 cm,600 cm,1400 cm)during young bamboo to mature.Early period of growth is 10 cm and 30 cm,rise period of growth is 120 cm,peak period of growth is 600 cm,last period of growth is 1400 cm.Expression and function of cellulose synthase are researched through bioinformatics analysis and TEM(Transmission electron microscope)and biological microscope and q RT-PCR(Quantitative Real time Polymerase Chain Reaction)and RNA in situ hybridization and Western Blot and Protein expression in vitro.The conclusions are as follows:(1)Microscopic observation showed that Moso bamboo stem structure growth is divided into four periods,and the first period,cells undifferentiated,there is no clear organizational structure,the cells mainly produces more cells.The second period,the native structure formation,a typical structure of vascular bundle,but density is larger,structure is small,phloem cells differentiation is not obvious,fiber cells and parenchyma cells of boundary is not clear.The third period,mature period,the vascular bundle structure of fiber cells and parenchyma cell boundaries clear,there are two typical metaxylem catheter,phloem structure is obvious.The fourth period,fiber cell lignification period,a layer of dark matter around the fiber cells can be seen.Base of fibrous tissue firstly appeared lignification,this time is rise period,the height of the stem is about 120 cm.(2)Bamboo phloem cell ultrastructure observation shows that growth of bamboo can be divided into three stages: the primary wall formation,secondary wall formation,and the secondary wall thickening.Studies have shown that secondary wall thickened with the height increasing,rise period(120 cm)can be observed in secondary wall of bamboo basal part,peak period(600cm)of bamboo basal parts have secondary wall structure,and top and root parts have only primary wall,secondary wall in end period(14 m)of bamboo basal part is thicker than other stage.(3)Analysis of bioinformation shows that Sixteen members of cellulose synthase family are predicted,the proteins contain a cellulose_synt motif and a zinc finger motif on the N terminus bydomain prediction,Bamboo cellulose synthase gene super family is divided into five family,are CESA,CSLD,CSLE,CSLF,CSLH.Phylogenetic tree displays that Ces A genes is likely to be in single before differentiation of dicotyledonous plants.Originally,bamboo cellulose synthase genes are more than arabidopsis thaliana and rice.After species differentiation,bamboo cellulose synthase genes produce more gene copy number,this may be related to bamboo unique rapid growth characteristics.(4)Combinating the basis of previous study of cellulose content in Moso bamboo with micro-,ultrastructural and q RT-PCR results,for cellulose synthase family genes,eight cellulose synthase genes(PeCesA1 ? PeCesA2 ? PeCesA3 ? PeCesA4 ? PeCesA5 ? PeCesA6 ? PeCesA9 ? and PeCesA13)have a high expression on the parts of secondary cell wall,three cellulose synthase genes(PeCesA6 and PeCesA9 and PeCesA13)have a high expression on the parts of primary cell wall.It may find that PeCesA6?PeCesA9?PeCesA13 are likely to involving in building of both primary cell wall and secondary cell wall and PeCesA1?PeCesA2?PeCesA3?PeCesA4?PeCesA5 are main for secondary cell wall.(5)Protein of conservative domain of bamboo cellulose synthase expression in vitro shows that expression product molecular weight is 33 kda,p I is 7.4.Western blot checking shows that the protein is right,and protein is insoluble.