Honma Yujiro, Taguchi Kazunori, Hiyama Hajime, Yui-Kurino Rika, Mikami Tetsuo, Kubo Tomohiko
Research Faculty of Agriculture, Hokkaido University, N-9, W-9, Kita-ku, Sapporo, 060-8589, Japan.
Theor Appl Genet. 2014 Dec;127(12):2567-74. doi: 10.1007/s00122-014-2398-4. Epub 2014 Oct 7.
By genetically eliminating the major restorer - of - fertility gene ( Rf ), a weak Rf gene was unveiled. It is an allele of Z , long known as an elusive Rf gene in sugar beet. In the hybrid breeding of sugar beet, maintainer-genotype selection is a laborious process because of the dependence on test crossing, despite the very low occurrence of this genotype. Marker-assisted selection (MAS) of the maintainer genotype is highly desired by sugar beet breeders. The major restorer-of-fertility gene (Rf) was identified as Rf1, and its non-restoring allele (rf1) was discriminated at the DNA level; however, some of the rf1rf1 selections retained an as yet unidentified Rf, another target locus for MAS. The objective of this study was to identify this Rf. An rfrf1 plant was crossed to a cytoplasmic male-sterile sugar beet and then backcrossed to obtain progeny segregating the unidentified Rf. The progeny exhibited partial male-fertility restoration that was unstable in single plants. The segregation ratio of restored vs. non-restored plants suggested the involvement of a single Rf in this male-fertility restoration, designated as Rf2. We confirmed the feasibility of molecular tagging of Rf2 by identifying four shared amplified fragment length polymorphism (AFLP) fragments specific to 17 restored plants. Bulked segregant analysis also was performed to screen the Rf2-linked AFLP markers, which were subsequently converted into 17 sequence-tagged site markers. All the markers, as well two additional chromosome-IV-assigned markers, were linked to each other to form a single linkage map, on which Rf2 was located. Our data suggested that Rf2 is likely an allele of Z, long known as an elusive Rf gene in sugar beet. We also discuss the importance of Rf2 for sugar beet breeding.
通过基因敲除主要育性恢复基因(Rf),一个弱Rf基因得以揭示。它是Z基因的一个等位基因,Z基因长期以来在甜菜中一直是难以捉摸的Rf基因。在甜菜杂交育种中,由于依赖测交,尽管这种基因型出现频率极低,但保持系基因型选择仍是一个费力的过程。甜菜育种者非常希望能进行保持系基因型的分子标记辅助选择(MAS)。主要育性恢复基因(Rf)被鉴定为Rf1,其不育等位基因(rf1)在DNA水平上得以区分;然而,一些rf1rf1选择植株仍保留了一个尚未鉴定的Rf,这是MAS的另一个目标位点。本研究的目的是鉴定这个Rf。将一株rfrf1植株与细胞质雄性不育甜菜杂交,然后回交以获得分离出未鉴定Rf的后代。这些后代表现出部分雄性育性恢复,且单株中不稳定。恢复与未恢复植株的分离比例表明,单个Rf参与了这种雄性育性恢复,命名为Rf2。通过鉴定17株恢复植株特有的四个共享扩增片段长度多态性(AFLP)片段,我们证实了Rf2分子标记的可行性。还进行了混合分离群体分析以筛选与Rf2连锁的AFLP标记,随后将其转化为17个序列标签位点标记。所有这些标记以及另外两个定位于四号染色体的标记相互连锁,形成了一个单一的连锁图谱,Rf2位于该图谱上。我们的数据表明,Rf2可能是Z基因的一个等位基因,Z基因长期以来在甜菜中一直是难以捉摸的Rf基因。我们还讨论了Rf2对甜菜育种的重要性。