Abstract:

The cinnamoyl-CoA reductase gene (CCR) of Pinus massoniana was amplified, cloned and sequenced (GenBank Accession No: EU753854). The nucleotide and amino acid sequences of CCR gene among different species were compared and analyzed by MEGA4.0.2. The results showed that the number of exons and introns of CCR gene of P. massoniana were the same as those in Pinus taeda, Populus balsamifera, Leucaena leucocephala, Betula luminifera, Eucalyptus gunnii, Eucalyptus saligna, and Eucalyptus globulus. They all had 5 exons and 4 introns. The size of coding sequence of the CCR gene of P. massoniana, encoding 324 amino acids, was 975 bp. The junction sequences of exons and introns of CCR gene of P. massoniana were in accordance with the law of gene composition (exon/GTPuAG/exon), except Intron Ⅱ/Exon Ⅲ (GTPuGG/Exon Ⅲ). The nucleotide number of Exon Ⅱ to Ⅳ of P. massoniana was the same as those of Eucalyptus,Populus, Betula, and Pinus. The similarities of nucleotide sequences of the coding region of CCR gene between P. massoniana and P. taeda, B. luminifera, P. balsamifera, L. leucocephala, E. gunnii, E. saligna, and E. globulus were 99.2%, 67.8%, 68.0%, 68.9%, 69.5%, 69.3%, 69.9%, respectively, and the similarities of amino-acid sequences were 99.1%, 73.5%, 72.5%, 74.4%, 75.0%, 74.4%, 75.0%, respectively. Molecular phylogenetic trees of P. massoniana with other 13 species (or sequences) reconstructed with CCR and the inferred protein sequence indicated that the 3 conifer species formed a separate clade and evolved earlier.

Key words: Pinus massoniana, cinnamoyl-CoA reductase gene(CCR), cloning, sequence analysis, phylogenetic tree