Research On Tissue Culture And Variation Induction Of Chromosome Of Artemisia Annua L.

In this paper we studied the tissue culture method of Artemisia annua L. and the combination of tissue culture method and variation induction of chromosome of Artemisia annua L.by chemical method,aiming at choosing the best method for tissue culture of Artemisia annua L.,and improving variation frequency of somaclones of Artemisia annua L.by using colchicine solution,enlarging variation spectrum,providing basic data for germplasm resource research in the further,optimizing the method of tissue culture and chromosome observation for Artemisia annua L.,and providing evidence for molecular mechanism and cytology theory of metabolism of artemisinin.The results are as follows:1.0.05%HgCl₂ 10min+3%NaClO10min was the best combination for the sterilization of seeds of Artemisia annua L.,and 0.05%HgCl₂ 5min+ Saturated NaClO15min was the best combinations for the sterilization of explant of Artemisia annua L..2.The result of stem induction experiment indicated that 6-BA,NAA had promoting effect for induction and differentiation of explant.6-BA 0.05mg.L^-1+NAA0.01mg.L^-1was the best medium for induction,with the hightest proliferation coefficient of 4.08.Bud formation from leaf of Artemisia annua L.can be induced in 6-BA0.05mg.L^-1+NAA0.01mg.L^-1medium without callus period.The organ types of explant are important for inducing bud different-tiation.Stem was the best position for induction3.The result of bud subculture experiment indicated that 6-BA1.0mg.L^-1 +NAA0.2mg.L^-1+KT0.5 mg.L^-1was the best medium for subculture,with the average proliferation coefficient of 6.65.It was good for subculture of Artemisia annua L.when the sucrose concentration was 30g.L^-1in the medium.Adding coconut in the medium has little effect on the subculture of Artemisia annua L.The best pH for subculture of Artemisia annua L.was pH 5.8.4.Root formation is easy for the tissue culture seedling of Artemisia annua L.,and IBA0.5mg.L^-1was the best concentration for root formation, with average rooting rate of 100%,root length of 2.75cm,root number of 6.04,and seedling height of 4.44cm.5.There were different effects for test-tube seedling transplanting with different medium,and the best ratio of muck:peat:perlite:riversand for the medium was 1:1:1:1.6.There were different effects for chromosome preparation of Artemisia annua L.with different sampling time and pretreatment methods, and the best combination for chromosome preparation of Artemisia annua L. was drawing materials at 10:28am,pretreatmenting with saturated p- Dichlorobenzene for 4.5-6h or with a-bromonaphthalene at 4℃for 24h, dissociating for 8-9min,and treated with 45%acetic acid after dissociation.7.There were different effects for chromosome preparation of Artemisia annua L.with different treatment methods,materials,and time.For soaking treatment with colchicine solution,the best treatment concentration was 0.1%-0.5%,and the best treating time was 5-7days for seed,and the best treatment concentration was 0.25%and 0.5%,and the best treating time was 4-5 days for stem.For solid medium added with colchicine solution,the best treatment concentration was 60mg.L^-1.Treatment with long time and high concentration was better for the induction of chromosome variation of Artemisia annua L..Chromosome number of Artemisia annua L.is 2n=18, 4n=36.