Immature Embryos Culture, Callus Induction And Differentiation In Osmanthus Fragrans

Osmamthus fragrans ’Dahua Jin’, Osmanthus fragrans ’Zi Yin’ and Osmanthus fragrans’Zi Dan’ were used as materials in this study. The appropriate conditions for immature embryoculture, callus induction and redifferentiation were studied in a series of experiments, in orderto accumulate some experience for establishing the Regeneration system of Sweet Olive.In immature embryo culture experiments, the right sterilization time, basic medium andthe concentration and proportion of exogenous hormones were studied to get the suitableconditions for immature embryo culture. In the rooting experiment, the root were cut off withthree kinds of lengths,to study roots’s growth process with varying degrees of damage indifferent concentrations of NAA. In transplanting hardening experiment, sand, vermiculiteand peat soil were used as matrix, to study the effect on seedling growth. In callus inductionand redifferentiation experiments, placement method, lighting conditions, the species,concentration and proportion of exogenous hormones were studied to get the suitableconditions for callus induction and redifferentiation. The data of the experimental results isanalyzed by DPS.Results are as follows:1. The suitable sterilization time for immature seeds that picked in mid-March is3minutes to6minutes.2. B5is more suitable for immature embryo culture than MS on germination rate andgrowing. The appropriate formula for Osmamthus fragrans ’Dahua Jin’ is B5+GA(3mg/L)+6-BA(1mg/L).The appropriate formula for Osmanthus fragrans ’Zi Yin’ is B5+GA(2mg/L)+BA(2mg/L).The appropriate formula for Osmanthus fragrans ’Zi Dan’ is B5+GA(1mg/L)+BA(1.5mg/L).3. It is Beneficial for the lateral root germination to cut off the main root and addNAA=2mg/L to the medium.4. It has no significant affect on callus induction that place the young leaves with ventralor back upward.5. It can significant accelerate the callus induction and mitigate browning that culturedin the dark. Illumination has no significant affect on callus appreciation and loose degree. The callus showing more green when it growing with illumination and it always showingyellow-white or light yellow-green when cultured in dark.6. It not conducive to callus induction when6-BA or2,4-D added to the medium solely. Itcan produce callus normally when6-BA and NAA added to the medium together. It canproduce more callus than add6-BA and NAA when6-BA and2,4-D added to the mediumtogether, and the callus looks more looser, and most of them browning when it after50days.The appropriate formula for callus induction of young leaves of Osmanthus fragrans ’Zi Yin’is MS+6-BA(2mg/L)+NAA(0.3mg/L), MS+6-BA(3mg/L)+NAA(0.5mg/L) and MS+6-BA (5mg/L)+NAA(0.5mg/L), and the Induction rate is about90%.7. The matrix moisture should be pay attention in transplanting hardening experiment,and peat soil is better than sand and vermiculite to be used as matrix.