Research On The Anther Culture Techniques For Several Vegetable Crops In Brassica Campestris

Six kinds of subspecies or varieties of vegetable crops in Brassica campestris were used for anther culture.Effects of medium components and culture condition on embryoid inducing, development and plantlet regeneration were probed.Methods of ploidy identification for microspores-derived plants were carried out.Horticultural characters of the double haploid plants were identified.F₁ combinations between the double haploid strains were made.The aim of research is to establish an efficient technique system for anther culture in Brassica campestris and accelerate the application of anther culture in breeding practice.Results are as follows.1.Genotype is one of the key factors affecting the embryoid induction of Brassica campestris.There was significant difference in each genotype in the capacity of microspore embryogenesis.Ten genotypes among nine get embryoid successfully in per research of cabbage and Chinese cabbage.Seven genotypes among two get embryoid successfully in greens komatsuma;one genotypes get embryoid successfully in Milk Chinese Cabbage and one genotypes get embryoid successfully in bird rape,but wuta-cai can not get embryoid.2.The stage of late uninucleate was optimal for anther culture in Brassica campestris.the bud length was 2.0～3.5mm in cabbage,2.0～3.0mm in chinese cabbage,2.5～3.5mm in green komatsuma,1.8～2.2mm in milk Chinese cabbage,2.3～3.4mm in bird rape,1.6～2.4mm in wuta-cai.3.Change temperature pretreatment can increase the frequency of microspore embryogenesis.The optimal treatment time was 24 h 4℃low temperature and 48h 33℃high temperature.Another function of low-temperature pretreatment is keeping materials fresh for anther culture temporarily.4.The ability of embryoid induction of double organic matter medium and two half one major element in MS,B5,changeB5 medium were commonly better than basic medium,but the effect of embryoid induction was different among basic mediums in different materials.5.Basic medium appending proper concentration of L-Glutamine could effectively speed embryoid induction,but superfluous L-Glutamine could restrain embryoid induction.The most suitable concentration was 0.3g.L^-1.6.The optimal medium for plantlet regeneration of microspore-derived embryos was "MS+3%sucrose+1.0%agar+100 mg·L^-1active carbon" and the embryos was normal by anther culture,so the plant formation was easy.7."MS+3%sucrose+0.8%agar" is the optimal medium for subculture and renewal of Chinese cabbage,"1/2MS+NAA0.1 mg·L^-1+3%sucrose+0.8%agar" and "MS+NAA 0.1 mg·L^-1+3%sucrose+0.8%agar" are suitable mediums for rooting.but the cabbage is resisent, but Chinese cabbage and greens komatsuna is strict to oxygen,so it must be to ventilative oxygen paper film.8.The methods of ploidy identification of microspore-derived plant include direct identification with chromosome numbers and indirect identifications by comparing the characters of organs,tissues and cells such as stoma size,mature pollen microspore size and so on.Finally,the result can be validated by self-fruitfulness ability of microspore-derived plant.9.Most of the DH plants obtained in the experiment have conspicuous self-incompatibility and live up to the standard of self-incompatible line.10.In the cross combinations from DH strain,two of them were higher in yield than the others in Pactroy,and five of them were better than the others in combining ability and disease resistant in Chinese cabbage.