Three genes showing distinct regulatory patterns encode the asparagine synthetase of sunflower (Helianthus annuus).

•  Asparagine metabolism in sunflower (Helianthus annuus) was investigated by cDNA cloning, sequence characterization and expression analysis of three genes encoding different isoforms of asparagine synthetase (AS, EC 6.3.5.4). •  The AS-coding sequences were searched for in leaves, roots and cotyledons by using a methodology based on the simultaneous amplification of different cDNAs. Three distinct AS-coding genes, HAS1, HAS1.1 and HAS2, were identified. •  HAS1 and HAS1.1 are twin genes with closely related sequences that share some regulatory features. By contrast, HAS2 is a singular sequence that encodes an incomplete AS polypeptide and shows an unusual regulation. The functionality of both the complete HAS1 and the truncated HAS2 proteins was demonstrated by complementation assays. Northern analysis revealed that HAS1, HAS1.1 and HAS2 were differentially regulated dependent on the organ, the physiological status, the developmental stage and the light conditions. •  Asparagine synthetase from sunflower is encoded by a small gene family whose members have achieved a significant degree of specialization to cope with the major situations requiring asparagine synthesis.