利用离体小孢子培养技术培育叶用芥菜耐抽薹新品系研究

A Study on Using in Vitro Microspore Culture Technique to Breed New Strains of Leaf Mustard with High Resistance to Bolting

为了快速选育叶用芥菜耐抽薹新品种,以贵州本地资源威宁青菜和修文青菜2个基因型为供体材料进行离体小孢子培养。结果表明,2个基因型均诱导出胚,产胚量分别为29.5胚/蕾和15.6胚/蕾;选取子叶形胚接种到再生培养基,进行诱导培养形成再生植株,成苗率达88.33%。2个基因型诱导出的单倍体在秋水仙素质量浓度为100.0 mg/L时加倍效率最佳,分别为52.00%、67.40%;对260株再生株进行倍性鉴定,检测到单倍体、双单倍体、非整倍体、单倍体+二倍体嵌合体、二倍体+四倍体嵌合体5种类型,其中,威宁青菜单倍体占87.50%,双单倍体占8.75%,其他倍性占3.75%;修文青菜单倍体占77.50%、双单倍体占17.50%,其他倍性占5.00%。不同倍性材料田间表现,单倍体无花粉,不结实,植株矮小;单倍体+二倍体嵌合体正常开花结实,植株长势强,但有部分空荚;非整倍体虽有花粉,但是结荚小,种子较小、不饱满,植株长势较弱;双单倍体、二倍体+四倍体嵌合体长势较好,有花粉,并且种子饱满,量多。因此,可先根据植株田间表现判断倍性,再选择性进行倍性检测。利用小孢子培养技术获得了威宁青菜和修文青菜优良叶用芥菜耐抽薹新品系,新品系均耐抽薹性强,分蘖性弱,纯合度100.00%,解决了常规种经常出现纯合度不够、田间表现不整齐、影响大规模生产的难题。

In order to rapidly select new bolting resistant varieties of leaf mustard,two genotypes of Guizhou local resources Weining leaf mustard and Xiuwen leaf mustard were used as donor materials for microspore culture in vitro.The results indicated that,embryos were induced in both genotypes,29.5 embryos/bud and 15.6 embryos/bud,respectively;Cotyledonous embryos were selected and inoculated into regeneration medium for induction culture to form regenerated plants,and the seedling rate reached 88.33%.The haploid induced by two genotypes showed the best doubling effect at a mass concentration of 100.0 mg/L of colchicine,with 52.00%and 67.40%,respectively.Five types of haploid,double haploid,aneuploid,haploid+diploid chimera,diploid+tetraploid chimera were detected in 260 regenerated plants.Among them,Weining leaf mustard had 87.50%haploid,8.75%double haploid and 3.75%other ploidy;Haploid,double haploid and other ploidy of Xiuwen leaf mustard accounted for 77.50%,17.50%and 5.00%respectively.In terms of field performance,haploid had no pollen,no seed,and the plant grew short;Haploid+diploid chimera showed normal flowering and fruiting,plant growth was strong and partial empty pods were in the chimera;Aneuploidy had pollen,but the pods were small,the seeds were small and not full,and the plant growth was weak;Double haploid,diploid+tetraploid chimera had good growth potential,with pollen,and full and abundant seeds.Therefore,ploidy can be determined based on the field performance of plants,and then selective ploidy detection can be performed.By using microspore culture technology,new excellent strains of leaf mustard with strong bolting resistance and weak tillering ability are obtained from Weining leaf mustard and Xiuwen leaf mustard.The new strains have a purity of 100.00%,solving the problems of insufficient purity and uneven field performance in conventional varieties,which affect large-scale production.