Establishment Of Cell Suspension Culture And Plantlet Regeneration Of Lily

To establish cell suspension cultures of Lilium spp., this research studied the callus induction and regeneration system firstly. Cell suspension cultures were established with the callus which was in good condition and could regenerate successfully. Five cell suspension cultures of three genotypes were established and different cell suspension cultures had different characters and regeneration ability. Specific research methods and the results and conclusions are as follows:(一) Callus induction and regeneration system(1) Callus formation examination: Callus formation were examined in 8 Lilium genotypes,Lilium regale wils., Lilium longiflorum 'White fox' and 'Snow Queen' , Lilium Oriental hybrids' Siberia' and 'Bernini', Lilium Oriental×Trumpet 'Manissa', Lilium davidii var. unicolor, Lilium Asiatic Hybrids 'Vermeer'. Seeds, bulb scales, and flower organs were placed on a medium containing 2.0 mg?L-1 picloram. The results showed that callus formation ability is Lilium regale wils., Lilium longiflorum> Lilium Oriental×Trumpet, Lilium Oriental hybrids, Lilium Asiatic Hybrids > Lilium davidii var. unicolor, respectively. Bulb scales and filament explants showed great ability to form calluses, whereas seeds had poor ability.(2) Callus induction and regeneration: Bulb scales of Lilium regale wils. and Lilium longiflorum 'White fox' were used as explants to study callus induction and regeneration. In the study of callus induction, the effect of picloram concentrations on Lilium regale wils. and the effect of light, initial materials status and hormone types (picloram and 2,4-D) and oncentrations on Lilium longiflorum 'White fox' were analysed . During the regenerating, the effects of sucrose concentrations and mineral element content in MS basic medium on both genotypes were studied. The results showed that the optimum concentration for callus induction of Lilium regale wils. was picloram 2.0mg?L-1, and the induction rate was about 80%; the optimum concentration for callus induction of Lilium longiflorum 'White fox' was picloram 1.0mg?L-1, and the induction rate was about 100%; the optimum regenerating medium of both genotypes was 1/2MS supplemented with sucrose 15 g?L-1.(3) Callus regeneration: Callus in good condition which were obtained from (1) were inoculated on solid 1/2MS medium with 15 g?L-1 sucrose. Genotypes and types of callus had a significant effect on callus regeneration. The genotypes mainly affected the patterns of regeneration and the types of callus had an effect on regeneration capacity. The regenerating rate of embryogenic callus with high regeneration ability was 100%. The non-embryonic callus with low regeneration ability showed lower regenerating rate.(二) Establishment of cell suspension culture and plantlet regeneration: Callus in good condition obtained from (1) and regenerated in (3) were cultured in modified liquid MS medium (NH4NO3 with half-strength) supplemented with picloram 2.0mg?L-1 and sucrose 30g?L-1 , and shook at 100 rpm under dark condition at 25℃. Culture density were 2g(f.w.) per 40mL liquid medium, and cell suspension cultures were subcultured every 1~2w. Suspension cells regenerated plantlets on solid 1/2MS medium with 15 g?L-1 sucrose. The results showed that the state and regeneration ability of cell suspension cultures were related with the initial callus. Stably growing cell suspension cultures were established with loose callus in 10 to 20 days, which had high single-cells ,but low regenerating rate. The cell suspension cultures composed of cell groups,diameter 1~5mm and high regenerating rate,were established by the granular callus.