Micropropagation of Lycium barbarum through Single Nodal Explants and Seeds from Mature Plants

Studies were conducted to develop protocols for in vitro regeneration of Lycium barbarum, also known as wolfberry or goji.  Wolfberry, which has received rapidly growing attention because of its antioxidant and nutrient values, has been termed a superfruit.  Cultures were established from 3 cm single nodal explants and seeds extracted from mature fruits.  Cuttings and fruit were rinsed for 5 minutes in sterile distilled water plus Tween 20.  Agitation in 10% Clorox for 30 minutes was followed by 3 rinses in sterile distilled water.  Seeds were then removed from the fruit using aseptic technique.  Explants were cultured on MS medium which was supplemented with each of 5 concentrations of cytokinins and auxins: (1) kinetin 2.0 mg/L plus IAA 2.0 mg/L; (2) kinetin 1.0 mg/L plus IAA 0.1 mg/L; (3) 2iP 30 mg/L plus IAA 0.3 mg/L; (4) BA 2.0 mg/L plus NAA 0.5 mg/L; and (5) BA 0.1 mg/L plus NAA 0.5 mg/L.  There were 8 replications of each of the treatments.  Explants were incubated in a 16-h light photoperiod from cool-white fluorescent lamps at 25 °C.  For both the singal nodal explants and seeds, all media variants gave efficient callus formation after 39 days.  Shoot proliferation was achieved at 100% using the BA 2.0 mg/L plus 0.5 mg/L supplement.  Thirty-five days later, root regeneration occurred at 100% after shoots were transferred to BA 0.1 mg/L plus NAA 0.5 mg/L.  Results show that in vitro regeneration protocols can be used for clonal multiplication and possible genetic transformation studies.