Isolation, Expression And Single Nucleotide Polymorphisms Analysis Of The Genes Involved In The Wood Formation From Cunninghcunia Lanceolata

Cunninghcunia lanceolata is an important fine economic forest in our nation. The objective of the study was to clone cDNA sequences of cellulose synthese genes and cellulose-like protein genes in xylem and done the single nucleotide polymorphism analysis to the partial of ClCCR gene. Conclusions from the experiments as follows:(1)Two full-length cDNAs were cloned by RT-PCR with 5', 3'RACE methods and the primers were designed according to the homologous sequences of CesA from other sequences. Sequence results indicated the cDNA of CesA1 shared over the 74% similarity with the CesA from Populus tomentosa, Pinus taeda, the CesA2 shared over the 78% identity with CesA from Pinus radiata, Acacia mangiam, Populus tremuloides and the CslD1 shared over 70% with the homologue genes from Populus trichocarpa, Populus tremuloides, Arabidopsis thaliana. The multiple comparison sequence analysis showed that the deduced amino acid sequence of CesA1 shared over 73% identity with the homologue genes from Eucalyptus grandis, Betula platyphylla, Pinus taeda and the CesA2 shared over 81% similarity with the CesA genes from Boehmeria nivea,Acacia mangiam,Populus tremuloides and the protein of CslD1 share over 71% similarity from Oryza sativa,Populus trichocarpa, Populus tremuloides, Arabidopsis thaliana. Further analyses indicated that they contained high conserved CXXC motif, transmembrance domains and conserved QXXRW domains.(2)The expression profile of ClCesA1, ClCesA2 and ClCslD1 were analyzed by using quantitative Real Time PCR analyses. The results showed that three genes expressing in root, stem, leaf and the expression of three genes were relatively higher in the stem than other parts(2.37,3.45,4.32). The expression data presented showed that ClCesA1 was lowly expressed in root(0.56) and the ClCesA2 and ClCslD1 were lower in leaf than other parts. Further analyses indicated that three genes were highly expressed in developed xylem(2.33,2.54,3.22)than bark and they were relatively higher in the xylem than the xylem of dwarf fir.(3)The partial CCR gene of 86 variaties were isolated and sequenced, the results showed that thrity SNPs including twenty-eight occurred in introns, and the remainings were observed in extrons.