Stacking of five favorable alleles for amylase content, fragrance and disease resistance into elite lines in rice (Oryza sativa) by using four HRM-based markers and a linked gel-based marker
Stacking of five favorable alleles for amylase content, fragrance and disease resistance into elite lines in rice (Oryza sativa) by using four HRM-based markers and a linked gel-based marker
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DOI：
10.1007/s11032-014-0076-5
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发表时间：
2014-10-01
影响因子：
3.1
通讯作者：
Chen, Zhiqiang
中科院分区：
文献类型：
--
作者：
Luo, Wenlong;Guo, Tao;Chen, Zhiqiang
针对谷物品质和对某些疾病的抗性等质量性状的标记辅助选择（MAS）已被证明是非常有效的。负责各种品质成分和抗病性的多个基因可以同时叠加，以提高性能并延长高产品种的商业寿命。谷物品质基因（fgr 和 Wx）和三种抗病基因（Pita、Pik 和 Xa23）已得到充分表征并用于 MAS 育种。然而，尚未报道将它们全部堆叠在一起形成单一品种。我们在此报道了将这五个基因堆叠到水稻优良品系中的情况。我们通过开发 fgr、Wx 和 Pita 因果突变的功能标记、Pik 的基因靶向标记以及使用 Xa23 连锁标记来实现这一目标。我们采用并优化了高分辨率熔解 (HRM) 分析方法，用作 fgr、Wx、Pik 和 Pita 的基因分型平台。通过结合高通量 DNA 分离、多重和巢式 PCR 方法，我们表明 HRM 可以作为小规模 MAS 项目的经济高效、高度自动化、中等通量和可靠的非凝胶基因分型平台。
Marker-assisted selection (MAS) for qualitative traits such as grain quality and resistance to certain diseases has proven to be highly effective. Multiple genes responsible for various quality components and disease resistances can be simultaneously stacked to boost the performance and to lengthen the commercial lifespan of high-yield varieties. Grain quality genes (fgr and Wx) and three disease resistance genes (Pita, Pik and Xa23) have been well characterized and used in MAS breeding. However, stacking all of them together into a single variety has not been reported. We reported here the stacking of the five genes into elite lines in rice. We achieved this through the development of functional markers from causal mutations at fgr, Wx and Pita, a gene-targeted marker at Pik and the use of a linked marker for Xa23. We employed and optimized the high-resolution melting (HRM) analysis method for use as the genotyping platform of fgr, Wx, Pik and Pita. By combining high-throughput DNA isolation, multiplex and nested-PCR methods, we showed that HRM could serve as cost-effective, highly automated, moderate-throughput and reliable non-gel genotyping platform for a small-scale MAS program.