Research On The Mutation Effects Of The Rhizomes Of Cymbidium Kanran Makino In Vitro After EMS And Colchicine Treatment

Using EMS, colchicine as mutagenic agents, and using the rhizomes of Cymbidium kanran Makino as material, we studied the effects on growth of rhizomes treated with mutagenic agents, chosed the semi-lethal dose (LD50), analyzed the physiological and gene changes of rhizomes treated with mutagenic agents. The results were as follows:1. Two mutagenic agents could resrain the growth of rhizomes, with the increase of concentration and the prolonged time, the brown ratio and death ratio rised, the brown ratio were always less than death ratio.2. Calculating the LD50on rhizomes of mutagenic agents, it related to concentration and time, the results could be the referenced dosage for chemical mutation of Cymbidium kanran Makino.The LD50of1,2,3,4d, respectively,1.05%,0.81%,0.59%,0.38%for colchicine. The LD50of1,2,3,4d, respectively,0.607%,0.237%,0.145%and0.119%for EMS.3. After two mutagenic agents treatment,the soluble sugar contents of rhizomes all increased in various degrees compared with the control group.The soluble sugar contents of rhizomes were significantly different from the control group. With the increase of concentration and the prolonged time of EMS, the soluble protein contents rised frist, and then decreased. The soluble protein contents of rhizomes were significantly different from the control group. With the increase of concentration and the prolonged time of colchicine, the soluble protein contents decreased by7.84%-20.2%,the change was not significant.4. After two mutagenic agents treatment,the MDA contents of rhizomes all increased in various degrees compared with the control,but rised frist and then decreased. With the increase of concentration of EMS and colchicine, the H2O2contents of rhizomes rised. The H2O2contents of rhizomes were significantly different from the control. With the increase of treating time of EMS and colchicine, the H2O2contents of rhizomes rised frist and then decreased.5. After two mutagenic agents treatment,the SOD,POD activity of rhizomes all increased in various degrees compared with the control. With the increase of concentration and the prolonged time of EMS, the SOD,POD activity of rhizomes rised frist and then decreased. With the increase of concentration and the prolonged time of colchicine, the SOD,POD,CAT activity of rhizomes rised gradually.6. The ISSR-PCR amplified genome bands of rhizomes treated with two mutagenic agents were different from normal rhizomes. The polymorphic rate was25%between rhizomes from EMS treatment and control.There were no polymorphism bands between rhizomes from colchicine treatment and control.