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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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Abstract

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.

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Acknowledgments

We thank Dr. Kunshen Wu and Thomas H. Tai for their critical reading of the manuscript. We are grateful to Dr. Yaoguang Liu and Mr. Xingliang Ma for their help in the beginning of the HRM experiments. This research was supported by the National High Technology Research and Development Program of China (2012AA101201), the Science & Technology Planning Project of Guangdong Province, China (20110203) and the National Natural Science Foundation of China (31200250).

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Authors and Affiliations

  1. Plant Protection Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, 510640, China

    Qiyun Yang & Xiaoyuan Zhu

  2. National Engineering Research Center of Plant Space Breeding, South China Agricultural University, Guangzhou, 510642, China

    Wenlong Luo, Tao Guo, Hui Wang, Yongzhu Liu & Zhiqiang Chen

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  1. Wenlong Luo
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  2. Tao Guo
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  3. Qiyun Yang
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  4. Hui Wang
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  5. Yongzhu Liu
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  6. Xiaoyuan Zhu
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  7. Zhiqiang Chen
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Correspondence to Zhiqiang Chen.

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Wenlong Luo and Tao Guo have contributed equally to this work.

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Luo, W., Guo, T., Yang, Q. et al. 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. Mol Breeding 34, 805–815 (2014). https://doi.org/10.1007/s11032-014-0076-5

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  • DOI: https://doi.org/10.1007/s11032-014-0076-5

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