Abstract
Thermo-sensitive genic male sterility (TGMS) is an important genetic means of two-line hybrid rice breeding. Pollen fertility in TGMS lines is regulated by a single point mutation in TGMS genes. Based on single nucleotide polymorphisms (SNPs) and insertion-deletion mutations, Kompetitive Allele-Specific PCR (KASP) markers were developed and utilized in rice molecular breeding via high-throughput detection, which saves time and money. In this study, we converted the SNPs on TGMS genes (including p/tms12-1and tms9-1) to functional co-dominant KASP markers and bred the resultant two-line hybrid rice. We differentiated the TGMS lines carrying p/tms12-1 or tms9-1 from other TGMS lines using the KASP assay. Pei’ai64S and Hua201S (containing p/tms12-1) had a homozygous GG genotype generating a blue signal. HengnongS-1 (containing tms9-1) had a homozygous CC genotype generating a red signal. The KASP assay for tms9-1 was identified as a recessive Mendelian trait. Seed purity was tested using the KASP marker for the two-line hybrid varieties of Liangyoupeijiu and Hualiangyou1206, which was consistent with findings by the dCAPS marker. Moreover, new TGMS lines were generated by pyramiding tms12-1 and tms9-1 gene under the same genetic background. Therefore, the KASP marker to detect the TGMS genes developed in this study can be widely used in two-line hybrid rice breeding. It provides a visually convenient toolkit for breeders to select individual target plants using high-throughput screening in two-line rice breeding.
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Acknowledgements
Financial support was provided for this research partly by the Project of the Science and Technology Department of Zhejiang Province, China (Grant Number LGN21C130005, 2016C02050), the Project of the Molecular Designed Japonica Breeding with High Quality and Yield in the Middle-Low Reaches of Yangzi River (Grant Number 2017YFD0100302), and the Talent Training Project of Zhejiang Academy of Agricultural Sciences (Grant Number 2018R16R08E01).We would like to thank Editage (www.editage.cn) for English language editing.
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YQ and JW conceived and designed the study. YQ, LW, JS, and GM performed the study test. YQ, LW, and JS contributed to the KASP assay. YQ and GM prepared the materials and populations. YQ and JW wrote the manuscript.
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Qi, Y., Wang, L., Song, J. et al. Development and utilization of the functional co-dominant KASP marker for thermo-sensitive genic male sterility in rice Oryza sativa L.. Genet Resour Crop Evol 69, 635–643 (2022). https://doi.org/10.1007/s10722-021-01249-7
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DOI: https://doi.org/10.1007/s10722-021-01249-7