Phytosulfokine-alpha, an enhancer of in vitro regeneration competence in recalcitrant legumes

Oligopeptides have been recognized as signaling molecules playing an important role in plant cell growth and development. Phytosulfokine-alpha (PSK), a plant-specific disulfated pentapeptide, is involved at nanomolar concentrations in initial steps of cellular dedifferentiation, proliferation, and re-differentiation, with a biological function similar to that of plant hormones. On the other hand, legume crops are generally known for their recalcitrance to in vitro regeneration approaches, which has restrained the exploitation of biotechnological tools for their genetic improvement. Against this background, we added PSK at concentrations of 10(-10) to 10(-6)M, to semisolid MS-based culture media previously shown to permit some regeneration responses with pea (Pisum sativum) and highly recalcitrant faba bean (Vicia faba). Callus, cell suspensions and embryo-derived explants of pea cultivars Frisson and Cameor and a low vicin, zero tannin faba bean genotype (Fevita((c))) were assessed for their embryogenic and organogenic regeneration competence. For all genotypes, PSK reliably and significantly enhanced the regeneration competence producing somatic embryos and organs that yielded regenerated plants of both pea cultivars, and had a major organogenic effect leading to plant regeneration with faba bean where somatic embryos, although produced, failed to convert into viable plants. This is the first report on the use of PSK with legume species.