春甘蓝型油菜育性恢复基因Rfk1在萝卜1号染色体上的大小和定位
Size and location of radish 1 chromosome regions carrying the fertility restorer Rfk1 gene in spring turnip rape
摘要：
春季芜菁油菜（Brassica rapa L.）spp. Ogu-INRA CMS/Rf系统是最有前途的F1杂种系统。将Ogura恢复基因Rfo的同源基因Kosena育性恢复基因Rfk 1成功地从油菜中转入芜菁油菜中，并使携带Ogura不育系的母本恢复育性。然而，这种特性在后代中并不稳定。采用8 GISH（基因组原位杂交）和BAC-FISH（细菌人工染色体荧光原位杂交）方法对萝卜Rfk 1基因的染色体区域进行了物理定位。以萝卜DNA为基因组探针，与Rfo基因连锁的BAC 64探针进行中期染色体杂交。两种探针均在甘蓝型油菜恢复系4021-2 Rfk的染色体扩散中显示信号，而在阴性对照系4021 B中未显示信号。GISH分析清楚地表明，芜菁油菜恢复系是单体（2n=2x=20+ 1 R）或二体（2n=2x=20+ 2 R）附加系，含有一个或两个来自萝卜的外源染色体。在BAC-FISH分析中，在萝卜染色体臂的亚末端部分检测到双点信号，表明育性恢复基因Rfk 1位于这条额外的萝卜染色体上。检测到的二体附加系在甘蓝型油菜杂交种生产中不稳定。利用BAC-FISH技术，在甘蓝型油菜的两条染色体上有时可见微弱的信号，而在甘蓝型油菜的第9染色体上则有一个Rfk 1的同源区域。通过BLAST分析验证了rapa A基因组。将来，A基因组中的这一同源区域可能被携带Rfk 1基因的萝卜染色体区域所取代。
英文摘要：
In spring turnip rape (Brassica rapa L. spp. oleifera) the most promising F1 hybrid system would be the Ogu-INRA CMS/Rf system. A Kosena fertility restorer gene Rfk1, homologue of the Ogura restorer gene Rfo, was successfully transferred from oilseed rape into turnip rape and that restored the fertility in female lines carrying Ogura cms. The trait was, however, unstable in subsequent generations. The physical localization of the radish chromosomal region carrying the Rfk1 gene was investigated using 8 GISH (genomic in situ hybridization) and BAC-FISH (bacterial artificial chromosome fluorescence in situ hybridization) methods. The metaphase chromosomes were hybridized using radish DNA as the genomic probe and BAC64 probe, which is linked with Rfo gene. Both probes showed a signal in the chromosome spreads of the restorer line 4021-2 Rfk of turnip rape but not in the negative control line 4021B. The GISH analyses clearly showed that the turnip rape restorer plants were either monosomic (2n=2x=20+1R) or disomic (2n=2x=20+2R) addition lines with one or two copies of a single alien chromosome region originating from radish. In the BAC-FISH analysis, double dot signals were detected in sub-terminal parts of the radish chromosome arms showing that the fertility restorer gene Rfk1 was located in this additional radish chromosome. Detected disomic addition lines were found to be unstable for turnip rape hybrid production. Using the BAC-FISH analysis, weak signals were sometimes visible in two chromosomes of turnip rape and a homologous region of Rfk1 in chromosome 9 of the B. rapa A genome was verified with BLAST analysis. In the future this homologous area in A genome could be substituted with radish chromosome area carrying the Rfk1 gene.
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