The Establishment Of Suspension Cell Lines And Salt-tolerant Selection Of Lilium Regale Wils.

On the condition of alkalescence in soils and water in North China,the cut flower quality of lily decreased seriously and the development of lily was restricted.Therefore,breeding of salt tolerant lily variety was a effective way to solve this question.Lilium regale Wils. was the excellent material for salt-toletant selection in Lilium species.In this paper,the bulbscales of L regale were used as explants to induce callus.Suspension cell lines were established and plants regenerated from the cell lines.By posotive selection methods,the callus were selected to obtain the salt-tolerant lines with the selective pressure of NaCl and HYP.The main results described as follows:1. The bulbscales of L regale were used as explants to induce callus in diffrent hormones.Finally,the results showed that the eligible medium to callus induction was MS+6-BA 2.0mg/L+NAA 0.1mg/L+3%sugar.Light was suitable for growth of callus;It was showed that the optimal medium to callus subculture was MS+6-BA 2.0mg/L+NAA 0.1mg/L+3%sugar in the dark.On the condition of dark, callus growed quickly and the color of callus was yellow.On the condition of light,the color of callus changed to green and brown differently.2. By researching the inoculum densities,sucrose concentrations and hormones,it showed that the optimal conditions for the growth of L regale cell lines were as follows:2.0g/40mL,30-60g/L sucrose,optimal medium was MS+1.0mg/L 6-BA. According to the growth curve of cell,it was better to subculture in seven-day cycles.3. The callus were planted onto the media containing maintained different NaCl and HYP concentrations.By posotive selection methods,the salt-tolerant callus of different salt concentrations were obtained.However,the salt-tolerant callus of highest salt concentations (70mmol/LNaCl and 0.15mmol/LHYP) were only researched.4. Through physiological and biochemical analyse,the contents of proline and soluble sugar of the salt-tolerant callus of 0.15mmol/LHYP were 1.88,1.54 times higher than control respectively,the content of MDA of the salt-tolerant callus was lower than control.It was included that the salt-tolerant callus of 0.15mmol/LHYP had significant salt tolerance,but the salt-tolerant callus of 70mmol/LNaCl did not have.So the salt-tolerant callus of 0.15mmol/LHYP was the true salt-tolerant variants,but the salt-tolerant callus of 70mmol/LNaCl was only the cells adaptability to salt. 5. The salt-tolerant callus of 0.15mmol/LHYP and the control were planted onto the media containing maintained different NaCl concentrations.It showed, in 20 days, that the growth rate of the salt-tolerant callus and the content of K~+,Na~+ were higher than the control and the ratio of K~+/Na~+ was the similar to the control.