甘蓝型油菜DH植株创建及其倍性的流式鉴定

以甘蓝型油菜大粒品系和小粒品系的小孢子为试验材料,研究取样时期、花蕾大小、培养基类型及添加成分对小孢子产胚率的影响,利用流式细胞仪对小孢子再生植株及加倍植株进行了准确、快速的倍性分析。结果表明,始花期为最佳取蕾时期,花蕾大小在2.5~3.5mm为宜,培养基以液体培养基最好。培养基中加入0.1mg/L6-BA可以促进小孢子胚的发生,而活性炭对小孢子培养没有明显的促进作用。流式细胞仪倍性检测显示小孢子再生植株加倍后的双单倍体（doubled haploid,DH）占49%,还存在少量的三倍体（11.3%）、四倍体（1.1%）及嵌合体（27.3%）植株。本研究为油菜材料的快速纯合及其DH系群体的构建等提供了新的经验。

Construction of doubled haploid plants and flow detection of their ploidies in Brassica napus

Large and small seed lines of Brassica napus L.were used for microspore culture and haploid identification by flow cytometry method.The factors effected embryo production rate were investigated.It was found that the optimal bud material of 2.5 to 3.5mm grew at primary flowering period.The liquid medium added with 0.1mg/L 6-BA was able to accelerate the embryogenesis.Activated-carbon has no obvious promotion to the occurrence of embryo.The flow cytometry was found to be quick and accurate on ploidy detection.Results showed that after colchicine doubling,the proportion of doubled haploid（DH） plants were 49%.There were also a few of triploid（11.3%）,tetraploid（1.1%）,and chimera plants（27.3%）.The present study offered new experience to obtain pure materials and to construct doubled haploid lines in B.napus breeding.