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CELL BIOLOGY & MOLECULAR GENETICS

Preliminary Analysis of Expressed Sequence Tags for Sugarcane

^a Biotechnology Dep., South African Sugar Association Exp. Stn., Private Bag X02, Mount Edgecombe, 4300, South Africa
^b Institute for Plant Biotechnology, University of Stellenbosch, Private Bag X1, Matieland, 7602, South Africa
^c Institute for Plant Biotechnology, Univ. of Stellenbosch, Private Bag X1, Matieland, 7602, South Africa

fcb{at}land.sun.ac.za

Sugarcane (Saccharum spp. hybrids), with its complex polyploid genome, is not well understood at the genetic level. Partial sequencing of anonymous cDNA clones is a widely used technique for gene identification. These partial cDNA sequences, or Expressed Sequence Tags (ESTs) have potential application for the identification of important genes for genetic manipulation. This study aimed to initiate the preliminary development of an EST database for sugarcane and thereby gain some potentially useful information on sugarcane gene sequences. A nondirectional cDNA library has been constructed from sugarcane leaf roll (meristematic region) tissue. Two hundred fifty clones have been randomly selected, subjected to single-pass sequencing from the 5' end of the vector, and identified by sequence similarity searches against gene sequences in international databases. Of the 250 leaf roll clones, 26% exhibit similarity to known plant genes, 50% to non-plant genes, while 24% represent new gene sequences. Analysis of the identified clones indicated sequence similarity to a broad diversity of genes encoding proteins such as enzymes, structural proteins, and regulatory factors. A significant proportion of genes identified in the leaf roll were involved in processes related to protein synthesis and protein modification, as would be expected in meristematic tissues. These results present a successful application of EST analysis in sugarcane and provide a preliminary indication of gene expression in leaf roll tissue.

Abbreviations: bp, base pair • EST, expressed sequence tag • NCBI, National Centre for Biotechnology Information • SuSy, sucrose synthase • pfu, plaque forming units • PAM, Point Acceptable Mutation

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