Efficient in vitro regeneration of fertile plants from corm explants of Hypoxis hemerocallidea landrace Gaza -- the "African Potato".
We present efficient protocols for the regeneration of fertile plants from corm explants of Hypoxis hemerocallidea Fisch. and C. A. Mey. landrace Gaza, either by direct multiple shoot formation or via shoot organogenesis from corm-derived calluses. The regeneration efficiency depended on plant growth regulator concentrations and combinations. Multiple direct shoot formation with high frequency (100% with 5-8 shoots/explant) was obtained on a basal medium (BM) supplemented with 3 mg/l kinetin (BM1). However, efficient indirect regeneration occurred when corm explants were first plated on callus induction medium (BM2) with high kinetin (3 mg/l) and naphthalene acetic acid (NAA 1 mg/l), and then transferred to shoot inducing medium (BM3) containing BA (1.5 mg/l) and NAA (0.5 mg/l). Shoot regeneration frequency was 100% and 30-35 shoots per explant were obtained. The regenerated shoots were rooted on a root inducing medium (BM4) containing NAA (0.1 mg/l). Rooted plantlets were transferred to the greenhouse. The regenerants were morphologically normal and fertile. Flow cytometric analyses and chloroplast counts of guard cells suggested that the regenerants were diploid. Efficient cloning protocols described here, have the potential not only to substantially reduce the pressure on natural populations but also for wider biotechnological applications of Hypoxis hemerocallidea-an endangered medicinal plant.
Ndong
Yves Assoumou
YA
Laboratoire Androgenèse et Biotechnologie EA3900, Université de Picardie Jules Verne, 33. Rue Saint-Leu, 80039 Amiens, France.
Wadouachi
Anne
A
Sangwan-Norreel
Brigitte S
BS
Sangwan
Rajbir S
RS
D015084:2,4-Dichlorophenoxyacetic Acid / Q000494:pharmacology; D001593:Benzyl Compounds; D003470:Culture Media; D027802:Hypoxis / Q000187:drug effects / Q000235:genetics / Q000254:growth & development*; D007701:Kinetin / Q000494:pharmacology; D009280:Naphthaleneacetic Acids / Q000494:pharmacology; D010937:Plant Growth Regulators / Q000494:pharmacology; D018517:Plant Roots / Q000254:growth & development; D018520:Plant Shoots / Q000254:growth & development; D011003:Ploidies; D011687:Purines; D012038:Regeneration; D046509:Tissue Culture Techniques / Q000379:methods*
D001593:Benzyl Compounds; D003470:Culture Media; D009280:Naphthaleneacetic Acids; D010937:Plant Growth Regulators; D011687:Purines; D015084:2,4-Dichlorophenoxyacetic Acid; C480551:benzylaminopurine; D007701:Kinetin