| Calycanthus chinensis is a deciduous shrub, belongs to Calycanthus, Calycanthaceae, which has a high ornamental value,and the leaves have medicinal value.Currently, Research of Calyca--nthus chinensis mainly concentrated in conservation biology, community ecology, genetic dive--rsity, reproductive technology and breeding, etc.In this paper, explants to induce differentiation and proliferation of adventitious buds were studied by using in viro seeds,stem tips of Calycanth--us chinensis, conduct rooting induction to get complete plants.At the same time, explants to ind--uce callus, adding different browning inhibitors to solve the callus browning in the transgenerati--onal process.The paper aims to provide technical guidance for vitro rapid propagation of Caly--canthus chinensis, to accelerate the extension and application of it. The main finding were as follows:1.Cultivation directly,in in viro seeds,stem tips of Calycanthus chinensis were selected as explants test:(1)75%C2H5OH 20~30s+0.1%HgCl2 7min disinfection processed with best serilization effe--ct for the seeds.75%C2H5OH 20-30s+0.1%HgCl2 10 min disinfection processed with best seriliz--ation effect for the stem tips, browning rate and pollution rate was relatively low,and germination rate was higher.(2)WPM medium were most suitable for the seed and stem tips cultivation, the plants leaves were pale green, the stem was thick and the growth status were better. Adding 0.2mg/l 6-BA could promote the growth of the vaccine effectively.(3)6-BA,ZT prossessed effect on the germination of stem tips,.The best concentration of 6-BA is 2.0mg/l, induction rate and breeding coefficient were 97.06% and 2.41,and the concentra--ton of ZT can be 2.0mg/l or 4.0mg/l. WPM+2.0mg/l 6-BA+0.05mg/l NAA was more suitable for stem tip original culture, induction rate and breeding coefficient were 89.50% and 2.29.(4)6-BA,ZT and TDZ.adding 0.05mg/l IBA prossessed effect on proliferation of tub seedl--ings. 2.0mg/l 6-BA prossessed effect on the germination of stems, Proliferation rate was 98.15% and Prpliferation coefficient was 3.16. 4.0mg/l 6-BA prossessed effect on the germination of see--dling. And the concentraton of ZT can be 2.0mg/l or 4.0mg/l and TDZ was 0.1mg/l WPM+ 2.0mg/l 6-BA+1.0mg/l KT+0.1mg/l NAA was most suitable for buds multiplication compared to the other combinations, proliferation rate was 100% and prpliferation coefficient was 2.60.(5)Transgenerational 3 times in a row, proliferation rate and multiplication coefficient reached to the highest value, effectively promote adventitious bud multiplication effect.(6)1/2MS medium was the best basic medium for inducing of rooting of Calycanthus chinensis,the rooting rate was 97.76% and the average root number was 5.85, when adding 0.5mg/l IBA. 1/2MS+0.1mg/l IBA+0.1 mg/l NAA+500mg/l AC was more suitable for inducing roots than other disposes.2. Cultivation indirectly, leaves of Calycanthus chinensis were selected as explants test:(1)Improved MS medium is more suitable to be the basic medium to induce callus than MS medium.Adding 0.4mg/l NAA prossessed effect best on inducing callus, the rate of callus was higher,that was 97.00%, and the browning degree is smaller.(2)Adding 500mg/l AC,the browning mitigation effect was best,transgenerational effectively can ease the callus browning degree.(3)There is a most significant positive correlation between callus browning and total pHenolic content,but PPO activity was negatively correlated to total pHenolic content and callus browning;POD activity was positively correlated to total pHenolic content and callus browning significantly. |
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