| Pol CMS is a main pollination controlling system and an important type of rape utilization of heterosis. Hybrid production of Pol CMS system was well-documented and widely used. Because the Pol CMS plays an essential role in heterosis utilization, cloning the Rfp has great significance in both breeding applications and understanding the mechanism of male sterility’s restoraion. In order to map the Rfp for gene cloning, a NIL containing the Rfp gene was established. Based on the previous study, closer linkage markers were developed and candidate clones were analysised by bioinformatics and comparative genomics. The main results of present study are as follows:1. A marker named scar24-2 with the distance of 0.04 cM had been developed successfully by comparing the mark’s sequence homologues and searching cabbage database’s BAC sequence,2. Scpl3, Kbr(lab developed) and scar24-2 markers have4,6and3 exchanges respectively by analysising recombinants derived from NIL about 5000 size population.The Rfp gene was mapped between the markers scar24-2 and Scpl3 with the distance of 0.04cM and 0.05cM.3. Scar24-2, Kbr and Scpl3 can identify homologous in cabbage BAC by BLASTn analysis. A possible PPR had found based on gene prediction of this region.4. Based on the results above, the probe (scar24-2) was used to further scaning the Brassica napus BAC library, and 46 positive BAC clones were obtained.13 BAC clones were selected in the 46 positive BAC clones according to the terminal sequential analysis result of positive clones,.Then 4 BAC positive clones were obtained by PCR analysis5. To map the Rfp, A-F2 and B-F2 populations which agreed with the expected ratio of fertility:sterility were constructed derived from Pol CMS. Scpl3 had the polymorphism in the two F2 populations, while Kbr and scar24-2 had not. The result demonstrated that the marker Scpl3 has better generality and wider application prospect in MAS. |
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