Isolation and Characterization of Expressed Sequence Tags (ESTs) from Cambium Tissue of Birch (Betula platyphylla Suk)

Abstract

Cloning and identification of genes from the cambium of woody plants are important steps in the investigation of wood formation. In order to clone and identify the genes involved in wood formation, a cDNA library was constructed from the cambial tissue of Betula platyphylla Suk (birch). From the cDNA library, 2,878 high-quality ESTs were generated, representing 1,540 tentative unique genes (TUGs). Approximately 70% of the ESTs were matched to proteins in the NR database in GenBank and were grouped into 12 functional categories according to their functional annotation. Fifty TUGs potentially involved in wood formation, including the process of lignin biosynthesis, cell wall structure, cell wall polysaccharides synthesis or development regulation, were identified from the EST collection. The time course expression analysis of 13 wood formation related genes was further studied using RNA gel blot and semi-quantitative RT-PCR. The results showed that these genes were primarily expressed in tissue that was collected during two time periods, spring (26 April to 11 June) and later summer (30 July to 18 Sept). The bimodal expression curve of the genes indicates that both of these time periods are critical in wood formation in birch, and may be associated with cell wall expansion earlier in the year, and cell wall strengthening later in the year.