Cloning And Analyzing Of Cellulose Synthase From Populus Ussuriensis

Cellulose is the primary component of the xylem which is the essence of lumber.The way that cloning the genes relate to cellulose synthase,transforming these genes into plant vivo by genetic method,analyzing the function and interaction way of these genes may establish base to show the mechanism of the cellulose synthesis in xylem and explore one method by the genetic engineering to study cellulose biosynthesis mechanism in forest.The new way may carry on the improvement target-oriented to the forest material quality and then enhance the quality and the output of important economic forest lumber and the wood fiber.Also it may overcome so many shortcomings which are the cross-pollination and the hybrid incompatibility and the long breeding period on traditional ways.It can speed up the forest variation speed and reduce the period on forest breeding.Therefore it has important meaning to improvement of the quality of the xylem(lumber) in the arboreal growth process.The poplar tree is the pattern plant on forest heredity breeding and the molecular biology research.Populus ussuriensis which has the characteristics on growth exuberantly and easy to thrive is the fine paper pulp material source.It is a fine economic forest in our nation.This research was cloned the cellulose synthetase genes from PopuIus ussuriensis.Conclusions from the experimental as follows:(1) The cellulose synthase gene was cloned from young leaf DNA of Populus ussuriensis. The sequential analysis indicated that the length of cdg1 was 1846bp,and the coded sequence from it was 921 bp with 98%identity with cellulose synthase gene from Populus trichocarpa (XM 002305024.1),and shared 96%identity with PtrCesA1 from Populus tremuloides.(2) The two DNA fragments,PusC1 and PusC2,were cloned from cDNA prepared from secondary xylem of Populus ussuriensis by reverses transcription PCR.Sequencing results indicated the length of PusC1 was 237bp with 94%identity with CesA from Populus trichocarpa,and PusC2 was 481bp with 95%identity to it.Sequencing analysis also showed the two fragments were shared 92%and 93%identity with PtrCesA1 from Populus tremuloides. The two proteins which were peculated from PusC1 and PusC2 gene were shared 94%and 93%identity with CesA from Populus trichocarpa respectively,and have 93%and 92% similarity with PtrCesA 1 from Populus tremuloides respectively.(3) Through the deep analyze,a suitable PCR reaction system was established,namely 20μL reaction system containing 2.0 U Taq DNA polymerase,1.5mmol·L^-1 Mg²⁺,0.15mmol·L^-1 dNTP,0.5μmol·L^-1 primer,1μl DNA template.(4) For getting an ideal method of extracting Populus ussuriensis genomic DNA and RNA from secondary xylem,the traditional methods were improved according to the experiences of modemourselves.The experiment compared the improved method with others;the results were showed to obtain high quality DNA and RNA with the improved method.It is optimum and suitable for molecular biology researches.