Study On The Technology Of Isolated Unpollinated Ovary Culture In Cabbage

The vegetable crop of unfertilized ovary or ovule culture in vitro can induce artificial gynogenesis.Accrossing this approach,thus effectively shorten the breeding period and expedite breeding course,can also provide beneficial condition for the expression of the recessiveness gene.At the same time,the DH group is also the fine material of molecule marker and genetic map reserch,can improve the accuracy of gene location and plotting to a large extent.Cabbage(Brassica oleracea L.)is an important vegetable crop in our country, so in vitro inducment of unpollinated ovules or ovaries becomes the emphasis of this kind of vegetables haploid research.This research focuses on the major factors affecting isolated culture of unfertilized ovary of cabbage.The major factors were included genotypes,different growth period,concentration of the phytohormones,different heat activation,concentration of Na Cl O of the donor plants.To analyses the effect of various processing conditions about the induction rate,screening out a system in vitro of unfertilized ovary that can induce the plant through single factor and multiple factors,clearing the growth way that cabbage culture in vitro of unfertilized ovary. Ploidy identification of the regeneration plant,Clearing the haploid proportion of the plant,and establish the foundation of the regeneration system on haploid and doubled haploid plants.The main results were as followed:1)The study found that the RM for the induced effect is the best genotypes.It has the highest induction rate of bud points,induction rate of callus and planting percent,the numerical is 3.57, 19.04% and 58.33% respectively,and it has a larger number of regeneration plant.2)The induced effect is the best at the early stage of flower.The average of bud points is 3.15,at the same time,the induction rate of bud points were 86%,these two valu es are the highest,when it formed green callus and embryoid in this period of explant,most of those can be developed into a complete plant with a lower mortality.3)For the unpollinated ovary of cabbage in vitro culture,the first step is to compare differe nt concentrations of NAA on in vitro induced effects in the inducement culture process. The results showed that 0.1 mg/L, 0.3 mg/L,0.5 mg/L effectively stimulated in vitro,the growth of the female nucleus bud points into bud number and embryoid induction rate reached the highest when the concentration of NAA is 0.1 mg/L,it has eight regeneration plants, and the later value is 6%. With a higher concentration of NAA,embryoid induction rate instead of a downward trend.4)Define growth path of in vitro inducment of unpollinated ovaries in cabbage,and establelisthe foundation of the regeneration system of unpollinated ovaries in ca bbage:Inducement culture medium:MS+6-BA(0.5mg/L)+2,4-D(2.0mg/L)+NAA(0.1mg/L)+Suc(30g/L)+agar(8g/L),p H5.8~6.0;Polarization culture medium:MS+6-BA(0.2mg/L)+TDZ(0.05mg/L)+NAA(0.1mg/L)+Suc(30g/L)+agar(8g/L),p H5.8~6.0;Radication culture medium:1/2MS+NAA(0.05 mg/L)+Suc(30g/L)+agar(8g/L),p H5.8~6.0.5)The study found by the single factor experiment,if only add a kind of hormone into the medium,no matter the 6 – BA,2,4-D or IAA,IBA, only can induce kale unfertilized ovary bud points or callus, will not be able to make its further differentiation,and cannot develop into a complete plants. In addition,tadd the IAA or IBA in the culture medium for the growth of cabbage unfertilized ovary have no significant effect.6)In vitro of unfertilized female nucleus needs darkness heat shock pretreatment,the suitable temperature for the unfertilized ovary culture of cabbage is 33℃,and treat 1day in this case,the average of bud points,induction rate of bud points and the bud were 2.85,100% and 13,respectively.7)Ploidy identification results show that the haploid plants accounted for 5.6% and the diploid plants accounted for 93% and triploid plants accounted for 1.4% of the tested plants.