The integrated use of molecular marker-assisted selection (MAS) and anther culture has potential to significantly increase efficiency in plant breeding; however, reports on this kind of practical use are very limited. In the present study, we report the development of cytoplasmic male sterile (CMS) maintainers with aroma, disease resistance and red-brown hulls, as an example of integration of MAS and anther culture in rice breeding. A high-resolution melting (HRM)-based functional molecular marker was developed for the red-brown hull trait caused by a unique mutation (rbh1) in OsCAD2. Functional molecular markers for genes of rice blast resistance (Pi2), aroma (fgr) and red-brown hull (rbh1) were used for precise genotyping of individual plants in the BC₁ and BC₂F₂ populations derived from a cross between CMS maintainers Huaxiang B (pi2–/rbh1–/fgr–) and Rong 3B (Pi2+/RBH1+/Fgr+). A total of 89 doubled haploid (DH) lines were generated from selected BC₂F₂ plants (Pi2+/rbh1–/fgr–) by anther culture. Seven DH lines were subsequently selected as the potential new CMS maintainers based on their overall performance and high resistance to blast. Our study demonstrated that integration of MAS and anther culture significantly accelerated the development of CMS maintainers with multiple stacked genes.

分子标记辅助选择（MAS）和花药培养的综合使用有可能显着提高植物育种的效率；但是，关于这种实际使用的报道非常有限。在本研究中，我们报道了具有香气，抗病性和红棕色外壳的细胞质雄性不育（CMS）维持剂的开发，以MAS和花药培养在水稻育种中的整合为例。针对由OsCAD2中的唯一突变（rbh1）引起的红棕色船体性状，开发了基于高分辨率熔解（HRM）的功能分子标记。稻瘟病抗性（Pi2），香气（fgr）和红棕壳（rbh1）基因的功能分子标记）被用于BC ₁和BC ₂ F ₂种群中单个植物的精确基因分型，这些种群是由CMS维持系花香B（pi2– / rbh1– / fgr–）和荣3B（Pi2 + / RBH1 + / Fgr +）之间的杂交得到的。通过花药培养从选定的BC ₂ F ₂植物（Pi2 + / rbh1-fgr-）产生了总共89个双倍单倍体（DH）系。随后，根据其整体性能和高抗爆性，选择了7条DH系作为潜在的新CMS维持剂。我们的研究表明，MAS和花药培养的整合显着加速了具有多个堆叠基因的CMS保持子的发育。