Studies On In Vitro Culture And Callus Extraction And Isolation Of Gardenia Jasminoides Ellis.

Gardenia jasminoides Ellis, a member of the family Rubiaceae, it's widelyused for gardening plant for it's beautiful flowers and evergreen. It is also a Chinesetraditional medicine and can be used as both medicine and food that healthydepartment has firstly promulgated, it contains many different chemical ingredients,Gardenia Yellow is rarely natural water-soluble carotenoid pigment with the featuresof nontoxic, safe, strong pigmentation, and can be widely used in food pigmentation.In this paper, flower induction and tissue culture, callus induction andsecondary metabolite's extraction were studied to exploit utilize the resource of theGardenia jasminoides Ellis. in China.Enhanced axillary branching and flower induction of Gardenia jasminoidesEllis were researched first .Much hormone were used to choose better inductioneffect. The results show that: Multiple shoots induction is obtained throughculturing on MS+6-BA 1.0 mg/L+ IBA 0.2 mg/L, in this medium the number of aexplant reaches 8-10 respectively ,stem apex even can reach to 15-20 respectively.The highest quantity of induction is approve to 50 per explant.Test-tube plantlet effloresce in vitro is studied in this paper too. Branchingaxillary buds were used to flower induction. The results show that: flower buds canform on MS+6-BA 1.0 mg/L +NAA0.1 mg/L +GA₃ 0.1 mg/L, and it can bloom andgrowth at last when suitable medium were used. The flower of test-tube plantlethave the same fragrance and appearance.The better root culture medium is 1/2 MS+IBA 0.5 mg/L+GA₃ 0.5 mg/L, in this medium the ratio of rhizogenesis is 98ï¼…. Thesurvival rate could achieve 95ï¼…after 4 weeks radicate induction.Callus induction of Gardenia jasminoides Ellis's pericarpium, leaf, seeds. Theresults show that: different base medium induction diversity callus. Callus inductionof B5 is green color and pykno-texture, but tissue looseness and light yellow in MS.NAA+KT combination is good for callus induction. The best medium is MS+NAA1.0 mg/L+KT 0.25 mg/L, in this condition, the looseness callus grow quickly andhave light yellow. This medium is used to liquid culture to get a great quantity ofsuspension cells.Callus and suspension cells were used to extraction, abstraction, purification,for the analysis of it's secondary metabolite. Four factors in extraction were studied,i.e. extracting agent(ethanol, acetone , methanol), solid/liquid ratio of extractingsystem, reflux time, callus source. The superior extraction condition was that,concentration of alcohol 70ï¼…or 50ï¼…acetone, solid/liquid ratio of 1:6 and reflux 3repetitions, supersonic wave to assist one one hour. A lot of yellow component wereextracted. Then the total extraction were further isolated by macroporous resin.andthe characteristics of isolate were tested by thin-layer chromatography (TLC), UVscan,and high performance liquid chromatography (HPLC). The results show thatcallus extraction may be jasminoidin and chlorogenic acid.