Cloning and Expression of Key Enzyme Gene GalAT in Ramie Pectin Biosynthesis

To isolate the cDNA partial sequence of key enzyme gene GalAT for pectin biosynthesis in ramie [Boehmeria nivea (L.) Gaud], and thus to understand the expression of GalAT gene in different tissues of ramie, degenerate primer was designed according to GalAT conserved sequence in other species reported, and the cDNA sequence of GalAT gene from ramie variety Zhongzhu 1 was cloned by RT-PCR method based on the degenerate primer. The cDNA revealed a 986-bp in length which encoded 328 amino acids. The cDNA sequence and putative amino acid sequence of GalAT shared high identity with previously reported Arabidopsis thaliana GAUT4 (GalAT) as 77 and 83%, respectively. Molecular evolution analysis showed that the putative amino acid sequence and Arabidopsis thaliana GAUT4 gathered to a same group. Real-time quantitative PCR analysis showed that GalAT mRNA accumulated most abundantly in root, and GalAT transcripts in all kinds of ramie tissues in turn revealed as follows: root > leaf > bast > or ≈ xylem.