The influence of increasing concentrations of naphthaleneacetic acid and the antibiotics cefotaxime, timentin, kanamycin, and hygromycin on eggplant (Solantun melongena L. cv. Embil) somatic embryogenesis was investigated. Cotyledon explants were excised from 16 to 20 days old in vitro grown seedlings. NAA promoted somatic embryogenesis, although ...its concentrations had no influence on the mean number of embryos. Callusing decreaSed significantly with increasing NAA concentrations. Morphogenesis was stopped with 50 to 100 mg L-1 kanamycin and 7.5 to 15 mg L-1 hygromycin. Although early globular embryos were observed up to 15 mg L-1, further embryo development was inhibited at 10 mg L-1. Interestingly, cefotaxime (250 and 500 mg L-1) promoted a marked effect on enhancing fresh weight of calli, accompanied by decrease in embryo regeneration, whereas timentin concentrations (150 and 300 mg L-1) did not affect embryo differentiation as compared to the control treatment.
Some parameters involved in Agrobacterium-mediated transformation in muskmelon Hales best (HBS) were studied. Cotyledon explants excised from 3.5-day-old seedlings were co-cultivated with Agrobacterium tumefaciens harbouring binary vectors which contained GUS and BAR genes. After co-cultivation on a low pH medium, explants were transferred to sele...ctive medium, with higher pH, containing Claforan and Finale. The medium was changed every two weeks till shoots were induced. All shoots rooted on MS medium supplemented with 0.3 mg/L IBA. These parameters combined as a whole led to successful transformation. The expression of the introduced gene construct was confirmed by GUS staining of shoot segments.
Cotyledon protoplasts were isolated from 16-18-day-old in vitro grown seedlings of 9 Brazilian and 3 Hungarian pepper varieties and hybrids. Large numbers (average 9.59 X 106 protoplasts g 14 fresh weight) of highly viable (average 87.0%) protoplasts were released using a pectocellulolytic enzyme mixture. Protoplasts were cultured in K8p mediuni u...sing an alginate disc embedding method. The osmotic pressure of the medium surrounding the alginate-embedded protoplasts was reduced by replenishing the liquid medium at K8p:K8 ratios of 1:0. 2:1, 1:1 in the first. second, and third week, respectively. Initial plating efficiency (IPE) average was 38.5% and after 21 days protoplasts reached microcolonies (15-20 cells) stages. Microcolonies were transferred after 3-4 weeks to a MS-based medium supplemented with 1.0 mg I-1 zeatin, 3.0% (w/v) sucrose, 0.24% (w/v) phytagel and pH 5.8, whereupon they formed callus. Final plating efficiency (FPE) average was 0.29% at a plating density of 1.0 x 105 protoplasts Protoplast-derived calli were cultured on a range of MS-based media supplemented with either BAP, IAA, TDZ; and zeatin. No morphogenic response was observed in any genotype investigated.
Cotyledonary segments of the casaba type muskmelon variety "Hógolyó" were used to induce organogenesis. Fifty different hormone combinations were applied to enhance the induction of shoot formation on the edge of the segments. The phases of organogenesis were followed with light- and scanning electron microscope. Shoot induction was achieved wit...h high frequency. The shoots were transferred to hormone free media for root induction. The rooted plantlets were planted out to soil.
NAA was feasible and the method can be applied in transformation experiments.