Abstract Tartary buckwheat (Fagopyrum tartaricum) seeds contain abundant protein with balanced amino acids and functional flavonoids. It is one typical crop for food and medical functions. Seed-filling development period is the crucial stage for the accumulation of those functional substances, so revealing the gene expression profile of this period is important for further regulation of functional component and improving the seed quality. For this reason, A full-length cDNA library was constructed by swithching mechanism at 5'end of RNA transcript(SMART) technology from filling stage Tartary buckwheat seeds and partial expressed sequence tags(EST) were analyzed. The results indicated that the titer of primary library and amplified library were 4.86×105 pfu/mL and 6.2×109 pfu/mL, respectively. The blue-white screening showed that it had a recombination rate of 99.58%, and the length of cDNA inserts was range from 500 to 3 500 bp. Some ESTs of the cDNA library (GenBank accession No. GO477569 and GO496285~GO496321) were obtained by sequencing of the random circularized plasmid clones. Bioinformatics analysis indicated that 54% ESTs showed high homology with the functional known genes, 28% ESTs had high homology with unknown protein or hypothetical protein, and 5% ESTs had no-hits. Among all known genes, ESTs of storage protein accounted for 17.93%, including 10, 16 and 22 kD storage protein, 11S and 13S globulin, vicilin and oleosin,etc. In addition, two genesrelated to the secondary metabolism named flavonoid 3'-hydroxylase and phenylalanine ammonia-lyase were obtained. All those results can lay a foundation for further isolation the genes related with the metabolism of seed storage protein and secondary metabolites.
|
Received: 09 September 2011
|
|
|
|