结球甘蓝游离小孢子培养及胚发育成苗研究

Seedling Regeneration From Isolated Microspore-derived Embryo in Cabbage

以20种不同基因型结球甘蓝为试材,采用游离小孢子离体培养方法,研究了结球甘蓝小孢子胚发生和植株再生的影响因素,并对再生植株倍性进行鉴定,以期建立高效的游离小孢子培养体系,为后期快速创新结球甘蓝种质材料提供参考依据。结果表明:基因型‘D3’小孢子出胚率最高,为42.33胚·蕾^(-1),是最低出胚材料‘D9’的17倍,表明基因型是影响小孢子胚发生的重要因素。花瓣与花药长度比值为0.51~0.75时小孢子出胚率最高。0.10~1.00 mg·L^(-1)TDZ能显著促进小孢子胚发生,获得的子叶胚成苗能力强且周期短。采用流式细胞仪对303株再生植株进行倍性鉴定,单倍体占49.18%,双单倍体占43.89%,嵌合体占6.93%。双单倍体和嵌合体植株均能正常结荚,而单倍体植株蕾期授粉后不能结荚。

Twenty different genotypes of cabbage were used as test materials for isolated microspore culture to study the influencing factors of microspore embryogenesis and plant regeneration,and to identify the ploidy and seed setting of regenerated plants,in order to establish an efficient isolated microspore culture system and provide reference for rapid innovation of germplasm materials in the later stage.The results showed that the genotype had a significant impact on the embryoid induction.The highest embryo yield was 42.33 embryo·bud^(-1)in inbred line‘D3’.Additionally,the embryo yield of this variety was 17 folds that the lowest embryo yield of variety‘D9’.The yield of microspore embryo was the highest when the ratio of petal length and anther length ranged from 0.51 to 0.75.The addition of 0.10-1.00 mg·L^(-1)TDZ significantly increased the yields of all type embryo,and the obtained cotyledon embryos had strong seedling capacity.Flow cytometry detection of 303 regenerated plant revealed that 49.18%were haploid,43.89%were doubled haploid,and chimera only accounted for the remaining 6.93%.Double haploid and chimeric plants could set pods normally after pollination at the bud stage,while the haploid plants could not produce pods.