Study On Asexual Propagation Techniques Of Litsea Cubeba

Litsea cubeba as China's energy species,have high economic value for development.The trial use three different provenances of Litsea cubeba for basic materials to study on asexual propagation techniques of the energy species,through opitimizing the process of tissue culture,establishing the callus system by inducting callus as the starting material and opitimizing the process of cutting propagation.Of great realistic significance to raising seedling quality and shorten the breeding cycle and providing theoretical and practical basis for Litsea Cubeba asexual reproduction technology,and also providing technical information and reference for the Propagation of woody plants.The main findings were as follows:1.Tissue culture of Litsea cubeba1.1 Litsea Cubeba explants disinfectionUsing completely randomized experimental design,0.1%Mercuric chloride+20%Sodium hypochlorite was the best combination of disinfection.0.1%Mercuric chloride+20%Sodium hypochlorite disinfection combination of time of 10 rain +5 rain,have the lowest rate of pollution,was the best disinfection options.1.2 Litsea Cubeba initial medium screeningUsing orthogonal design,taking into the effect of different hormones and the concentration of Litsea Cubeba,Screening the initial medium A5:MS+2.0mg/L BA+1.0mg/L KT+0.4mg/L IBA+ Sucrose 30g/L,A15:MS+2.5mg/L BA+0.4mg/L IBA+ Sucrose 30g/L.In the initial medium A5 and A15 the explants budding fast,green seedlings,larger spacing,and the high rate of proliferation.1.3 Anti-browning treatment of Litsea cubebaFrom explants of handling and add some kinds of inhibitors in medium to have the comparison test.Explants of cold treatment to a certain extent the effect of reducing the rate of Browning, Browning reduced rate 11.67 percent.Cutting in the 1 g / L PVP,explants can also reduce Browning, Browning decreased by 10 percent.Cutting in the Sterile water,Browning reduced by 3.34 percent.Using completely randomized experimental design,add different inhibitors into medium to have the comparison test.AC control the browning was the best of all,in the medium containing 0.5 g / L AC,browning decreased by 18.89 percent,and better able to control Litsea Cubeba explants browning.1.4 Effect of different provenances and the site of branches on proliferation of Litsea cubeba Using completely randomized experimental design,Sanming provenance of the Shoot branches have the highest launch rate(81.02%)and Proliferation rate(3.06).Sanming provenance was the optimal,Jiangle provenance was the second one and Shaxian provenance of the worst train,different branches parts of the explants have the different launch rate and the proliferation rate,the shoot have the highest launch rate and the proliferation,and then from top to bottom,the launch rate and the proliferation rate has decreased.1.5 Effect of contamination rate,launch rate and proliferation on different months of Litsea cubebaCollected in different months,the pollution rate and the rate of proliferation activated different, September and November was a better time in the second half of the training time,March,April s a better time in the first half of the training time,with the higher launch rate and proliferation.1.6 The subculture of Litsea cubebaUsing orthogonal design,and selected the first subculture medium for the best was MS+1.0mg/L BA+0.5mg/L KT+0.4mg/L IBA+30g/L Sucrose.the second subculture medium was MS+1.0mg/L BA+0.1mg/L KT+0.4mg/L IBA+30g/L Sucrose +0.5g/L AC.the third subculture medium was MS+0.5mg/L BA+0.2mg/L IBA+20g/L Sucrose +0.5g/L AC.1.7 The rooting culture and transplanting of Litsea cubebaUsing orthogonal design,taking into the effect of different hormones on the rooting of Litsea Cubeba,selected the best medium of the rooting culture was 1/2MS+0.8mg/L IBA+0.2mg/L IAA+20g/L Sucrose.Transplanting media was the sand in the first,and then transplant to the media (sand:yellow soil:peat soil =1:1:1)later.2.Callus induction,subculture and regeneration of Litsea cubebaUsing three different explants(stems,leaves,petioles)to induce the callus,and established callus system of Litsea Cubeba.Different explants(stems,leaves,petioles)had different difficulty to induce the callus:stems＞petioles＞leaves.stems were the best material of all.The optimum medis of stem callus induction was MS+2.0mg/L 2,4-D+1.0mg/L BA+1.0mg/L KT+30g/L Sucrose.The optimum medis of leave callus induction was MS+1.5mg/L 2,4-D+1.0mg/L BA+0.5mg/L KT+30g/L Sucrose. The optimum medis of petioles callus induction was MS+1.5mg/L 2,4-D+1.0mg/L BA+1.0mg/L KT+30g/L Sucrose.The best media of the callus subculture was MS+1.0mg/L 2,4-D+0.5mg/L BA+0.5mg/L IBA+30g/L Sucrose.The best media of the callus differentiation was MS+0.5mg/L IBA+0.5mg/L BA+0.5mg/L KT+30g/L Sucrose.3.The cutting propagation ofLitsea cubeba Three kinds of provenances,the average survival rate of cutting the level of the order was Shaxian＞Jiangle＞Sanming,the average rooting rate of the order was Shaxian＞Jiangle＞Sanming,the materialof Shaxian provenance was the better for cutting.Sand had the highest survival rate and average rooting rate,the most of the average number of roots,the longest average root length,sand had the best effect for cutting.Dfferent cuttings part,the promotions of rooting,cutting treaments on the cutting of Litsea Cubeba had different survival rate.The best combination was germination of the roots+shoot of the branch+200mg/L Ll ABT Immersed 30rain+ Cutting into oblique incision at the bottom.The best of the ear length was 14 cm.The diameter with 0.3-0.4 cm in cutting had the highest survival rate.May was more suitable for transplanting,after the month,the temperature of the external environment was getting higher,the survival rate also decreased.June can be transplanted,and the best in the rainy days,after transplanting attention should be paid to water management.July and August were not suitable for transplanting.