The Study On Induction And Identification Of Mutations In Cloned Line Of Cardiocrinum Giganteum

The seedlings were used as subjects in this investigation.Based on the early-days work,some seedlings were exposed by the UV(30W) in the distance of 30 cm for 0,3,6, 12 hours respectively,and the others were soaked in the solution of sodium azide(pH=3) with the concentration of 0,1.0,2.0,3.0,4.0mmol/L for 3 hours.The seedlings were investigated for the morphous and physiological functions.And the RAPD system of Cardiocrinum giganteum was established and optimized,and the mutations of Cardiocrinum giganteum were identified.The purpose of this investigation was to gaining the seedlings of Cardiocrinum giganteum,and also making preparation for the introduction and mutation breeding of Cardiocrinum giganteum.The results are as follows,1.Under the treatment of ultraviolet irradiation,the changes of the seedlings were obvious in the comparison with the check group.But considerable seedlings were dead under the treatment of ultraviolet irradiation for 12 hours.After soaked in the solution of sodium azide,almost 1/3 of the seedlings that soaked in the concentration of 3.0mmol/L were dead,and almost all of the seedlings that soaked in the concentration of 4.0mmol/L were dead.These results are very obvious.2.The Cardiocrinum giganteum was untreated,the leaves were strip and thin,and the seeding was tall.While Cardiocrinum giganteum was under the treatment of ultraviolet irradiation,the leaves were orbicular-ovate,the color was light,the leaf vein could be seen easily,the seeding was short,and the stoma was bigger.The longer the time of the treatment was,the more manifest all the changes were.The leaf lengths and the stoma sizes of the he seedlings of Cardiocrinum giganteum were not changed so much under different concentrations of sodium azide.3.The activities of the CAT,POD were increasing in accordance with the accumulation time of the ultraviolet irradiation.The increasing of POD was sharp,and there was significant difference between this group and the CK group.The increasing of SOD,CAT was obvious,and the activity of SOD was decreasing 6h,12 h after the ultraviolet irradiation.The activities of POD increased in accompany with the increase in concentrations of sodium azide,while the activities of SOD decreased.And the changes in the activities of CAT had no regularity. 4.The RAPD system of Cardiocrinum giganteum was established and optimized,and the ideal factors are as follows,Mg²⁺(2.0mmol/L)1.0μl,dNTPs(0.2mmol/L)1.0μl,primer(0.6μmol/L)1.3μl,DNA(30ng/μl)1.0μl,Taq(1U),10×buffer(2μl).The overall reaction system is 20μl.The amplification procedure is as follows,94℃-5 min,than 35 cycles:94℃-30s,37℃-50s,72℃-1 min,final elongation72℃-1 min.The voltage of electrophoresis was 50V,the time consuming is 1 hour.5.According to the RAPD analysis of treated seedlings of Cardiocrinum giganteum,the PCR bands of two seedlings were different from the others.Based on these results, Inductions have occurred already.6.According to the changes of the seedlings,the changes of the enzymatic activity and the RAPD analysis,building a method of identification of Mutations in Cloned Line of Cardiocrinum giganteum...