Development of an expressed sequence tag (EST) library for Medicago sativa
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Expressed sequence tag (EST) libraries for alfalfa (Medicago sativa L.) were developed from two cDNA populations derived from full sibs from a cross between 'Riley' and the glandular-haired germplasm, KS224. Sibs where sorted into two populations based on the presence or lack of both long- and short-stalked glandular hairs. Glandular hairs have been associated with resistance to pests in alfalfa. Each cDNA population was enriched using the second population as the driver in cDNA subtraction. The differential expression of the majority of the randomly selected cDNAs was verified via reverse Northern hybridization. However, not all sequenced cDNAs were detectable using reverse Northerns, possibly due to their low relative abundance. Fifty-five percent of the ESTs sequenced from the eglandular-haired pool, versus 40% for the glandular-haired cDNA pool, had homology to earlier identified plant genes. When ESTs were compared with the EST database alone, 64% of the 210 glandular-haired ESTs that had no sequence homology to the non-redundant database, also were unique to all plant ESTs sequenced to date. This was twice the percentage for eglandular-haired ESTs. The systematic analysis of ESTs from this study serves as a tool to identify genes regulating development of glandular trichomes and the resistance to insects associated with them. 2001 Elsevier Science Ireland Ltd.
