| Anthurium andraeanum, one of the most popular tropical flowers, belongs to Araceae. Nowadays, most Anthurium cultivars were propagated by ways of tissue culture. Yet, thanks to the chiasm of genotypes' in vitro regeneration capabilities, many high valued cultivars have no in vitro regeneration system; and the micropropagation method through indirect shoot-genesis via callus formation gives rise to fading vigor of plantlets and off-types. Thus, investigating the factors affecting in vitro regeneration and multiplication ways systematically is the ultimate solvent.Firstly, the in vitro culture systems of several Anthurium cultivars were established using leaf as explant. Sectioning fully expanded young lamina under sterile water could minimize the browning rate of explant; immersing young lamina cut from potted plant in a solution containing high concentration of 6-BA and 2,4-D could promote dediferentiation of explant; callus should be induced under darkness; according to different cultivars, the dedifferentiation and redifferentiation rate varied and different combinations of auxin and cytokinin should be used; generally, the combinations of 0.5~lmg/L 6-BA and 0.1~0.8mg/L 2,4-D added into MN200 (modified Nitsch basal medium) were suitable to most cultivars to be induced into dedifferentiating; the suitable media for redifferentiation were: MN200 + 0.25~0.5mg/L 6-BA + 0.15mg/L KT + 0.2mg/L TFIBA + 6g/L agar.Some cultivars of Anthurium ('B' and 'B6' for example) could be induced to regenerate directly through adventitious shoots or somatic embryos under suitable culture condition ( 1/2MS+0.25mg/L 6-BA+0.14 mg/L KT+ 0.2 mg/L IAA+0.6 % Agar+3 % Sucrose, pH 5.5 ) , while some cultivars could not ('A0' , 'B1' , 'C3' for example). The calli of cultivar 'C3' could regenerate indirect somatic embryo emergence in one medium (l/2MS+0.25mg/L 6-BA+0.2 mg/L TFIBA+0.2%AC). Polar distributed starch-rich cells were found specifically in somatic embryos; the adventitious shoots regenerated from callus could drive from either outer layers or inner layers of callus; excretory cells containing crystal were found universally in parenchyma and meristem tissues; spiral vessels derived from nodules in the inner callus transporting growth substance and nutrients were oberserved indicating that callus might have primary differentiated structures.Taking Anthurium andraeanum Cultivar. 'B' as testing material, different combinations of plant growth regulators, concentrations of sucrose, illumination and culture types were studied on the influence of propagation rates and the quality of axillary shoots. The results showed that Nitsch basal medium supplemented with 1mg/L KT, 0.5mg/L 6-BA and 30mg/l sucrose was the suitable medium to proliferate axillary shoots in liquid flask-shaking culture for the first 25 days under darkness and then in still liquid culture under illumination for another 25 days. Three cultivars, 'A0' , 'B' , 'B6' having been propagated in the established suitable culture system showed different capacity of axillary buds emergence, but all were well propagated above the rate of 8 in 50 days. The plantlet with multiple axillary shoots could be transferred directly into solid subculture medium and then be cut into separated single shoots after 45 days to save labor cost. Compared with the plantlets regenerated from callus, the axillary emergency rate of the plantlets gained from this propagation system was slightly higher.The culture conditions of in vitro plantlets of Anthurium andraeanum were systematically investigated. The suitable cutiure medium was 1/2MS+0.25mg/L 6-BA+0.2mg/L TFIBA+0.1%AC, solidified by 0.5 ~ 0.55% agar. 3100~4000Lux illumination was suitable for the strengthening of in vitro plantlets. Under this culture condition, three cultivars, AO, B, B6 were successfully subcultured for 2 years without apparent growth vigor fading or off-type.And also, one variegated plantlet resulting from chlorophyl loss of partly parenchyma tissue was gained from the adventitious buds regenerated from callus. Three types of chimeras w... |
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