The Anther Culture In Alfalfa For Haploid

Five materials were used for anther culture in alfalfa .The research included influence on anther callus, the methods of increasing anther plant getting rate, subculture and taking roots and transplant of anther plant, the ploidy of anther plant . The aim of researches was to set up high-efficient culture technique system of the anther culture of alfalfa .Results were as follows:1. In callus induce capacity, there was significant difference in each genotype.2. The basic medium and sucrose concentration affected on callus formation in anther culture. NB+2.4-D 0.5 mg/L +NAA 0.3 mg/L +6-BA 0.5 mg/L +KT 3.0 mg/L is suitable medium of alfalfa. The suitable sucrose concentration of different basic medium was different,6% sucrose concentration was the best concentration of NB, in the range of callus can be induce, high concentration sucrose can induce haploid callus and inhibition somatic callus induction.3. A change of temperature treatment affected on callus formation in anther culture. As far as the single factor, high temperature treating 48h and 72h was the best to callus induction, low temperature treating 24h was the best to callus induction. The suitable high temperature treatment of low temperature treatment was different, low temperature treating 48h was the best time of high temperature treating 24h and 72h,low temperature treating 24h Was the best time of high temperature teraitng 48h.4. Through different hormones on the differentiation medium callus differentiation rate of the test, we find that BA and NAA coordination use is easier to induce differentiation , but its concentration ratio should not be too large. When the concentration ratio of BA0.2mg/L,NAA 2mg/L and BA0.2mg/L,NAA3mg/L, differentiation frequency were 40% and 54% ,without any hormones or just BA, almost no callus induced sprouting; When KT and NAA coordination use is easier to induce differentiation also , differentiation frequency was 36%.5. Callus on low-temperature processing can improve differentiation rate and the most appropriate treatment is the 4°C for 24h .6. 1 .2 % agar of medium was the most suitable agar content for microspoer embryo becoming shoots culture of alfalfa7. MS + 3% sucrose + 0.5% agar was the suitable subculture and renewal medium of anther culture plant. 1/2MS+ NAA0.1 mg/L+1%sucrose+ 0.5% agar and MS+NAA0.1mg/L+1% sucrose +0.5% agar were suitable for inducing roots of anther culture plants. 8. The distribution of different genotype material, haploid, diploid and tetraploid was different in proportion, diploid is the most .