Abstract:

To isolate seed-expressed sequences, a cDNA library was constructed using wheat (Triticum aestivum L.) seed tissues at 12 d after pollination. Plasmid DNAs of 10 000 clones randomly picked out from the library were prepared. The preparation of high density filters were made with the Biomek 2000 HDRT system, and then hybridized separately with three probes prepared by reverse transcription of RNA of unpollinated ovary, embryo and endosperm. Based on the hybridization results, 800 clones expressed in embryo and/or endosperm were chosen for further analysis of expressed sequence tags (ESTs). Finally, 216 different genes were identified preliminarily. Of them, 24 (11.5%) were considered identical to known wheat genes, 122 (56%) were identified as putative new plant genes which may be involved in seed storage proteins, biochemical metabolisms, development, and other biological processes of seeds, while 70 (32.5%) sequence identities could not be determined.

Key words: wheat seeds, cDNA array, differential screening, expressed sequence tag