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Acta Agron Sin ›› 2014, Vol. 40 ›› Issue (11): 1925-1935.doi: 10.3724/SP.J.1006.2014.01925

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

cDNA Cloning and Expression of Two Cellulose Synthase Genes from Boehmeria nivea

LIU Yu-Xiang1,**,CHEN Jiang-Rong2,**,PENG Yan1,HUANG Yu1,ZHAO Yan1,HUANG Li-Hua1,GUO Qing-Quan2,ZHANG Xue-Wen1,*   

  1. 1 College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China; 2 Department of Biotechnology and Environmental Science, Changsha University, Changsha 410003, China?
  • Received:2014-03-10 Revised:2014-09-16 Online:2014-11-12 Published:2014-10-01

Abstract:

 Two potentially high homologous fragments CL789 and Unigene20360 were identified as plant cellulose synthase character sequence from the transcriptome data we obtained previously by Blast aligning and homologous screening. The two pairs of specific primers were then designed based on the CL789 and Unigene 20360 sequences information. The intermediate fragments of two cellulose synthase gene cDNA were cloned from ramie variety Xiangzu 3 by RT-PCR. And the whole cDNA was cloned by followed 5' and 3' RACE. The full length cDNAs were sequenced and their encoded putative proteins were identified as cellulose synthase by the conserved domain analysis. These two cDNA sequences were named as BnCesA2 and BnCesA3 respectively. The full-length coding sequence of BnCesA2 gene is 3240 bp, and encodes a putative 1079 amino acids. The coding sequence of BnCesA3 gene is 3120 bp, and could be translated into a 1039 amino acids protein. We designed the specific primers based on cDNA sequences of the two genes and their expression levels were tested by quantitative real-time PCR (qRT-PCR), indicating that BnCesA2 and BnCesA3 were both actively expressed in phloem and xylem in the four different cultivars of ramie. But the level of expression showed significant difference that the BnCesA2 expressionwas 2 to 5 multiples higher than BnCesA3 in both phloem and xylem. It is speculated that both the BnCesA2 and BnCesA3 participate the primary and secondary cell wall biosynthesis.

Key words: Ramie (Boehmeria nivea L.), Cellulose synthase genes, cDNA cloning, Expression analysis

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