Plant & Animal Genome IX Conference

Poster: Sequencing & EST
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EST DEVELOPMENT IN ITALIAN RYEGRASS

¹ Japan Forage Seed Association, Nishinasuno Branch,388-5 Higashiakada Nishinasuno, Tochigi 329-2742, Japan

² National Grassland Research Institute, Department Plant Breeding, 768 Senbonmatsu, Nishinasuno, Tochigi 329-2793, Japan

To develop EST (Expressed Sequence Tag) in Italian ryegrass (Lolium multiflorum Lam.), we started a random sequencing project of Italian ryegrass cDNAs. Several cDNA libraries were constructed from various tissues (leaf, stem and fluorescent tissue) and stress-treated leaf (cold / heat treated and powdery mildew-infected leaf). As an initial work, leaf cDNA library was constructed by using oligo dT primer for unidirectional cloning. Three prime tag EST sequencing have been applied for classifying each clone. Up to now, 300 unique sequences (out of 960) have been identified in Italian ryegrass. For other tissues and stress treated-leaves, the subtracted libraries were constructed by using leaf total RNA as a template of driver cDNA, and random sequencing for each library are currently in progress. The goal of this project is to develop approximately 1,000 ESTs. The systematic screening of RFLP markers and development of CAPS markers also have been carried out based on the ESTs. The latest progress will be presented.