Study On Callus Induction And The Cell Suspension Culture Of Elymus Dahuricus × Hordeum Brevisubulatum F₁

In this paper,the cell plant culture technique of Elymus dahuricus×Hordeum brevisubulatum F1 as the explant was used,and used different culture medium to induce callus and increase its induction rate.Different culture medium and selected young spike of different length was set.The optimum medium was used to determine the appropriate explant length.After long-term subculture of callus,due to changes in the level of endogenous hormones in the body,its cell activity is low,the growth rate is significantly slowed down,therefore the concentration of hormone in the culture medium was changed and to restore its growth activity and to realize the regulation role on cell growth by hormone.When the cells were cultured in vitro,16 kinds of culture medium were screened by orthogonal test.Different hormones had different effects on cell growth,and liquid culture medium suitable for suspension culture was determined.The growth process of suspended cells was observed,the growth of the suspended cells was understood,the appropriate subculture time was determined,the changes in the process of cell growth was understood the growth of suspended cells was better grasped.In order to improve the growth efficiency of the whole suspension system,the culture conditions were optimized to determine the appropriate initial inoculation amount,pH value and sucrose content.The main findings were as follows:(1)The induction medium was induced with MS 3 mg/L 2,4-D as the basic medium,and the induction rate was 28.3%,the callus was the better embryogenic callus with loose,dense structure.(2)The length of 2?3cm young spike was more suitable for the length of the explants,the induction rate was 31.6%.(3)MS+2,4-D 3mg/L + KT 0.5mg/L + CH 300mg/L and MS+2,4-D 4mg/L can improve the long-term subculture of poor quality callus of the growth of the situation,so that it returned to normal growth status.(4)MS + 2,4-D 1mg/L + 6-BA 0.5mg/L + KT 0.2mg/L + NAA 1mg/L was the ideal liquid medium,cells grew well in the culture medium,dispersed well and the cells showed pale yellow,culture medium was transparent.After 49 days of suspension culture,a stable and well-grown suspension system was initially established.(5)Through the determination of cell growth characteristics and cell activity it was found that subculture once every seven days was more appropriate,adequate nutrients can be timely to the growth and development of cells to provide energy,but also conducive to cell after several generations of cell viability to maintain a stable and efficient cell suspension system.(6)The optimized design of the suspension culture was carried out.The pH value was 5.8,the sucrose concentration was 30g/L,the initial inoculation amount was 1.5g,which was beneficial to the suspension culture of the cells and the growth rate.