Cloning and endogenous expression of a Eucalyptus grandis UDP-glucose dehydrogenase cDNA

Genetics and Molecular Biology

Mônica T. Veneziano Labate¹  Ana L. Ferreira Bertolo¹  Daniela Defávari Do Nascimento¹  Gunta Gutmanis¹  Alexander De Andrade¹  Maria J. Calderan Rodrigues¹  Eduardo L.o. Camargo¹  Luis Felipe Boaretto¹  David H. Moon¹  Juliano Bragatto¹  Carlos A. Labate¹ 

[1] ,Universidade de São Paulo Escola Superior de Agricultura Luiz de Queiroz” orgdiv2= Piracicaba SP ,Brazil

关键词: UDP-glucose;    UDP-glucuronate;    hemicellulose;    pectin;    cell wall;   

DOI  :  10.1590/S1415-47572010005000078

来源: SciELO

【 摘 要 】

UDP-glucose dehydrogenase (UGDH) catalyzes the oxidation of UDP-glucose (UDP-Glc) to UDP-glucuronate (UDP-GlcA), a key sugar nucleotide involved in the biosynthesis of plant cell wall polysaccharides. A full-length cDNA fragment coding for UGDH was cloned from the cambial region of 6-month-old E. grandis saplings by RT-PCR. The 1443-bp-ORF encodes a protein of 480 amino acids with a predicted molecular weight of 53 kDa. The recombinant protein expressed in Escherichia coli catalyzed the conversion of UDP-Glc to UDP-GlcA, confirming that the cloned cDNA encodes UGDH. The deduced amino acid sequence of the cDNA showed a high degree of identity with UGDH from several plant species. The Southern blot assay indicated that more than one copy of UGDH is present in Eucalyptus. These results were also confirmed by the proteomic analysis of the cambial region of 3- and 22-year-old E. grandis trees by 2-DE and LC-MS/MS, showing that at least two isoforms are present. The cloned gene is mainly expressed in roots, stem and bark of 6-month-old saplings, with a lower expression in leaves. High expression levels were also observed in the cambial region of 3- and 22-year-old trees. The results described in this paper provide a further view of the hemicellulose biosynthesis during wood formation in E. grandis.

【 授权许可】

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