| Notholirion hyacinthinum(wils)stapf, Liliaceae, Notholirion, is the Taibai Mountain Nature Reserve Protection Plant. The study, taking bulb, fibrous roots and scale leaves fertile as explants, were investigated by orthogonal design of different media, hormone concentration and light on callus, adventitious buds and adventitious root induction, and were designed to perfect the research method. Extracts from bulbs, calluses, axillary buds and adventitious buds of Notholirion hyacinthinum were examined through UV-Visible spectral analysis and HPLC analysis to study its active ingredients and the difference of wild species, and to better the service for this endangered valuable medicine. The results showed:1,The scale leaves were inoculated on MS, 1/2MS and modified MS medium. It showed that MS basal medium went with the highest rate of callus induction, modified MS followed, and 1/2MS worst. The highest rate of callus induction medium combination was MS + 2,4-D 1.0 mg / L + NAA1.0 mg / L +KT0.4 mg / L, and the induction rate of 80.3%. Role of media and plant growth regulators on callus induction for Notholirion hyacinthinum(wils)stapf was medium> NAA> KT> 2,4-D, and that higher concentrations of NAA and lower concentrations of 2,4-D inhibited callus induction.2,Bulbs, scales and fibrous root, leaf and fertile segments of Notholirion hyacinthinum(wils)stapf were inoculated to the most suitable scale leaf callus induction medium. The results showed that the leaves and bulb scales of Notholirion hyacinthinum(wils)stapf can be produced callus, while the fertile segments did not result in fibrous callus;after 40d in culture, bulb scales of all external browning, only a few scales on the browning of the callus grew pale yellow, while the scale leaf formed large dense agglomerate callus.3,Notholirion hyacinthinum(wils)stapf's scale leaves inoculated to the most suitable medium for callus induction, half were cultured in light, the other half cultured in the dark for the first 20d and then cultured to light in the culture. Cultured in the dark for a period of time at first, it took on the rapid growth of callus and compact structure of Notholirion hyacinthinum(wils)stapf, induction rate greater than only light culture.4,The size, growth, uniform of callus inoculated in different culture medium and subculture on shoot induction, there is a big difference of the number of adventitious buds under different treatments. Medium combination for the largest number of induced adventitious buds was MS + 2,4-D0.5 mg / L + NAA0.5 mg / L + KT 0.1 mg / L +TDZ0.1 mg / L, with the number of callus induction of adventitious buds up to 8.5. The role of the four growth regulators on adventitious bud induction for Notholirion hyacinthinum(wils)stapf was 2,4-D> KT> TDZ> NAA, and with lower concentration of KT ,2,4-D and TDZ on callus induction, adventitious buds were promoted Role, but NAA concentration level was not significant.5,1/2MS as the basic medium supplemented with KT concentration of 0.1 mg / L, and IAA concentration gradients being 0.5, 1.0, 1.5, 2.0 mg / L, single factor experiments were carried out. The results showed that, the role of 1.0 mg / L of IAA on the rate of adventitious rooting was significantly greater than the other three levels. In the rooting medium of 1/2MS + IAA 1.0mg / L + KT 0.1mg / L, the highest rate of rooting was 44.4%, and the average length of the root was roughly 2.0cm.6,Total contents of flavonoids from calluses axillary buds and adventitious buds that had been obtained by tissue culture were 12.6 mg/g,8.2 mg/g,9.2 mg/g respectively,and calluses presented no significant differences compared with 12.9 mg/g of bulbs,which prove that calluses can replace bulbs for officinal.7,Baicalein was found to exist in extracts, with Rutin, Quercetin and Catechin not found; Differences of the amount of Baicalein were evident,calluses,axillary buds and adventitious buds presented minute differences,but having approximately half the amount of Baicalein from bulbs. |
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