Tissue Culture And Analysis Of Essential Oil From Plant Tissue Cultures Of Thymus Vulgaris

Based on experiments of plant tissue culture, this research focuses on the ability of several Thymus vulgaris tissue cultures synthetizing essential oil in order to obtaining essential oil in vitro. The main results are as follow:A system for callus cultures was established. Callus could be divided two types by color, one is green, and another is white. MS + 2, 4-D 0.10-2.00mg L^-1 + 6-BA 0.10-2.00 mg L^-1 or NAA 5.00 mg L^-1 + KT 10 mg L^-1 was suitable for inducing leave and stem to form green callus. MS + 2, 4-D 0.50 mg L^-1 + AgNO₃ 5.00 mg L^-1 was suitable for inducing stem to form white callus. MS +NAA 1.00 mg L^-1 + KT 1.00 mg L^-1 was suitable for green callus propagating and MS +2, 4-D 0.10 mg L^-1 + 6-BA 2.00 mg L^-1 was suitable for white callus propagating.The research basis on multiplicating adventitious buds of T. vulgaris, an in vitro culture system was set up. Different concentration of IAA and 6-BA were compared for inducing adventitious buds from axillary buds, and for propagation of adventitious buds. The medium containing IAA 0.25 mg L^-1 and 6-BA 0.25 mg L^-1 was suitable for forming adventitious buds from axillary buds, and also was good for propagating adventitious buds. Adventitious buds' propagating was also influenced by different culture styles. In solid medium, adventitious buds were more efficient than in liquid medium.A cell suspension culture system was established. After successive transfer white callus to solid medium which containing 2, 4-D 0.10 mg L^-1 and 6-BA 2.00 mg L^-1, callus would be more fragile. Then fragile callus was transferred to liquid medium containing 2, 4-D 0.10 and 6-BA 2.00 After several times sub-cultivation, a cell suspension culture system of T. vulgaris could be set up.Inducing T. vulgaris to form Hairy root by four kinds Agrobacterium rhizogenes was studied. R1000,A4, ATCC15834 and R1601 could induced stem of T. vulgaris form Hairy root.Components of essential oil extracting from callus, adventitious buds and cell suspension culture system were analyzed. In adventitious buds, thymol could be synthesized and the essential oil constituent was similar to essential oil from T. vulgaris plant.