Anther Culture And Identification Of Regenerated Plants In Potato

As an efficient breeding method, anther culture in potato has been attended increasingly. At present, it has been the most effective and widely applied one in all ways of haploid production in potato. However, many factors, such as genotype, inoculation period, pretreatment conditions, medium etc. retrained the further application. So the establishment of the system of anther culture remains the major factor restraining haploidy breeding.Thirty-two genotypes of potato were used in the experiment. Analyzed the effects of the developmental stage of microspores, genotype, hormone combination, pretreatment, carbon source and medium supplement on anther culture and identified the ploidy about the regenerated plants. The main results are as follows:1. The length of potato buds can be used as reliable criteria for judging the developmental stage of microspores. When microspores were in mononuclear stage most buds were about 5.0～6.5mm. Anthers were about 3.8～5.0mm, the colour was yellow-green.2.Cultivated in the same condition, the percentage of callus induction of thirty-two genotypes was differ greatly. Callus induction rate of Kexin 20, Ke GP2-12, Ne16 and Vangal was higher, over 7%. Others was lower and still others had no callus. Neishu 7, Ne16, Bo C and Chunshu 4 had regenerated the anther plants, while others had no one.3.Different concentration and combination of NAA, 2,4-D, KT play an important role in anther culture. Of all the five mediums, D6 is perfect: MS+NAA0.5mg·L-1+2,4-D0.3mg·L-1+KT 0.5 mg·L-1. callus induction rate and regenerated rate were all high.4. Pretreament with lower-temperature and higher-temperature can obviously impove callus induction rate. The result showed that: pretreatment with 4℃for 2d and then pretreatment with 33℃for 3d after inoculation was perfect.5.In sucrose experiment, when the concentration of sucrose was 60g·L-1, callus formation was best.When used the same concentration of sucrose and maltose, callus induction rate was higher with maltose than that with sucrose.6.Medium added with right quantity of AgNO3(20mg·L-1), active carbon(0.50mg·L-1) and potato extract(50mg·L-1)can improve callus induction rate; Right quantity of AgNO3(20mg·L-1) and active carbon(0.50mg·L-1) can also reduce browning rate.7.With callus of Ne16, Neishu 7, Bo C and Chunshu 4 as the material, four kinds of medium and genotypes were analyzed with variance and the difference between medium and genotypes were analyzed respectively. The result as follow: medium﹥genotypes﹥medium x genotypes, medium was the major factor.8.Identifed the ploidy of the regenerated plants by the number of stoma chloroplast in guard cell. The result showed as follow: the number of stoma chloroplast is controlled by ploidy of plant; the number of stoma chloroplast in plants of doubled haploid and tetraploid is different. The trend is that the number of stoma chloroplast in tetraploid plant is higher than that in doubled haploid plant.9.Identified 23 regenerated plants by counting chromosome in root tip The result showed that double haploid plants (2n = 24) 15 ,the rate of induction was 65.2%.