Study On Potato Haploid Culture

As an efficient breeding method, haploidy breeding in potato has been attended increasingly. At present, anther culture has been the most effective and widely applied one in all ways of haploid production in potato. However, the effect factors, such as genotype, inoculation period, culture mode, medium etc, restrained the further application. So, the technical system of anther culture in potato remains the major factor restraining haploidy breeding.Twenty genotypes of potato were used in this test to study the effects factors on the the disinfection lasting, pretreatment conditions, particularly on the composition of induction medium and differentiation medium.The conclusions were as follows:1. Considering the disinfection result, the best method of disinfection is that disinfecting buds 8 minutes with 0.1% HgCl₂ supplemented with a drop of tween-80 after pretreating 30 seconds with 70 % alcohol.2. The color and size of potato buds could be used as simple and reliable indexes to judge the developmental stage of microspore. When the major buds were green and the size were 5.0 6.0 millimeter, microspore was in mononucleated stage.3. In the 20 genotypes for the trial, different genotype had different ability for induction. Kang 932-2 ranked in the first place for 4.17% in induction rate, while YA03-5, YA03-7, GaoYuan 7 had no callus or embryoid formation. S21, E potato 5, Kang 932-2, 7692-76 had regenerated the anther plants, while the others had no one.4. Optimization regress analysis called 311-A was firstly applied in potato anther culture. The multiplication coefficients of 11 medium with different level 2,4-D and KT and sucrose were investigated. The regress equation was deduced on 2,4-D and KT and sucrose as variable factor, and the better combination was selected. Induction rate may increase to 5.4797% while MS medium contained 2,4-D 0.5636 mg/L, KT 1.0000mg/L and sucrose 28.2187 g/L.5. Pretreatment with 4℃and pretreatment of anther at 35℃after inoculation were all helpful for anther induction. 48 hours of the two kinds of pretreatments was best.6. Medium added with active carbon could reduce the anther brown state effectively and increase anther induction rate. Different genotype had different sensitive to it. 5% potato extract was favorable to anther culture.7. NAA, 6-BA and KT had important roles in the differentiation of potato callus. With high concentration of NAA, the callus only differentiated root. With high concentration of 6-BA and KT, callus brown seriously. MS + NAA0.1mg / L + 6-BA2.0 mg / L + KT2.0 mg / L +5% potato extract was more suitable for callus differentiation.8. Anther plants regenerated through embryoid-way or direct-way were all haploids and these plants had stability in ploidy. Anther plants regenerated by callus differentiation were haploids or tetraploids.