Plant & Animal Genomes XIV Conference

January 14-18, 2006
Town & Country Convention Center
San Diego, CA

Comparison Of Fragments Of Cellulose Synthase (CesA) DNA In Widely Divergent Plant Species Using Primers Derived From Zea mays CesA-3 And Arabidopsis thaliana At4g39350.

Joshua N. Cobb¹ , Joseph M. Papenfuss² , Ammon B. Marshall¹ , Jessica K. Sorensen² , Kevin N. Sorensen² , Mikel R. Stevens¹

¹  Department of Plant and Animal Sciences, Brigham Young University 275 WIDB Provo, UT 84602 USA

²  Department of Life Sciences, Snow College 150 College Ave. Ephraim, UT 84627 USA

PCR primers developed from conserved amino acid sequences in Zea mays CesA-3 and Arabidopsis thaliana of At4g39350 cellulose synthase. These primers were used with Phusion-assisted long chain polymerase to amplify the latter portion of the gene including the QVLRW active site and four introns. Resulting PCR products were cloned and sequenced to verify cellulose synthase gene sequences in 27 of the 52 species: 2/2 flowering; 3/5 gymnosperms; 4/4 eusporangiate ferns; 12/32 leptosporangiate ferns; 4/4 lycopods; 1/2 mosses; 1/2 liverworts, and 0/1 hornworts. There were 136/269 clones that were verified to contain 1.0 to 2.3 kb CesA gene fragments. Of these, 99 contained a 1.3 to 2.3 kb fragment with the expected four introns; 14 contained a 1.1 kb fragment with the first intron, and 21 contained a 1.0 kb fragment with no introns. Sequence analysis revealed that the clones contained between 340 and 360 amino acids of the 1,000 or so in the cellulose synthase polypeptide. Approximately 70 amino acids around the first intron were highly conserved, and from 13 amino acids left of the second intron proceeding right to the end of the fragment were also highly conserved. Intron sizes varied among species, but surprisingly, several different gene constructs were identical across two or more widely divergent species even to intron base sequences.