摘要
对3个甘蓝类植物(甘蓝、青花菜、羽衣甘蓝)19个品种小孢子胚胎发生和植株再生的影响因素进行了研究,结果表明,基因型不仅影响供试材料的小孢子胚状体发生频率,也影响胚状体的质量;4℃低温预处理0～2d结合32.5℃热激1d,能显著提高供试基因型小孢子出胚率。胚状体先在NLN-13液体培养基中培养25d,有利于胚状体分化成苗;后在B5固体分化培养基中添加1%～1.2%琼脂,可有效促进胚状体萌发和植株再生。小孢子再生植株先使用流式细胞仪在生长初期鉴定倍性,后对单倍体植株采用200mg·L-1秋水仙碱浸根处理20h,并用流式细胞仪鉴定,此方法可快速、有效地获得双单倍体植株。
Abstract：The factors that affect microspore embryogenesis and plant regeneration of 19 cuhivars of Brassica plants （cabbage, broccoli and kale） were investigated in the present experiment. The results indicated that the genotype affected not only the frequency of microspore embryogenesis, but also the quality of embryos induced. Cold temperature treatment at 4℃ for 0 - 2 d combining with heat shock treatment at 32.5℃ for 1 d could improve embryogenesis efficiency significantly. Culturing embryos in NLN-13 liquid medium for approximately 25 d was best for embryo germination. Addition of 1% -1.2% concentration of agar to the B5 solid differentiation medium could significantly improve microspore-derived embryo germination and plantlet regeneration. Moreover, the method that first identified the ploidy level of regenerated plantlets at early growth with flow cytometry, then soaked roots of haploids for 20 h with 200mg· L^-1 colchicine solution, and finally identified with flow cytometry, could obtain doubled haploid plants quickly and efficiently.
出处
《核农学报》
CAS
CSCD
北大核心
2013年第7期929-937,共9页
Journal of Nuclear Agricultural Sciences
基金
江苏丘陵地区镇江农科所与浙江大学合作项目(zjs2009001)
国家自然科学基金(31000678)
江苏省自主创新项目(cx(10)463)
江苏省科技支撑计划(BE2012348)
江苏省农业自主创新项目(CX(12)2004)
中国博士后基金(2012T50555)
关键词
甘蓝类植物
小孢子培养
胚胎发生
植株再生
倍性鉴定
Brassica plants
Microspore culture
Embryogenesis
Plant regeneration
Ploidy identification
