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Key message <p>Two <i>Restorer-of-fertility </i>loci, <i>Rf-m1</i> and <i>Rf-m2</i>, co-condition fertility restoration in ZD-type CMS soybean though complex genetic interactions in both gametophytic and sporophytic manners.</p> Abstract <p>In soybean (<i>Glycine max</i>), the genetic mechanisms underlying fertility restoration of ZD-type cytoplasmic male sterility (CMS-ZD) are not fully understood. By crossing the CMS-ZD line W931A with its strong restorer WR016, we elucidated the inheritance patterns of fertility restoration. We discovered that restoration by WR016 is primarily governed by two <i>Restorer-of-fertility</i> (<i>Rf</i>) loci, <i>Rf-m1 and Rf-m2</i>, located within ~ 220&#xa0;kb on chromosome 16 and ~ 117&#xa0;kb on chromosome 9, respectively. They exhibited two distinctive inheritance modes across genotype combinations. In plants with the genotype of <i>-/rf-m1 -/rf-m2</i>, such as F<sub>1</sub> hybrids, gametophytic inheritance mode is predominant. In this case, <i>Rf-m1</i> in haploid tissue is essential for pollen fertility restoration, and <i>Rf-m2</i> is a modifier to enhance the effect. Correspondingly, plants displayed low pollen staining rates (sterile to ~ 50% depending on genotype). Conversely, in plants with one locus present as the homozygous <i>Rf/Rf</i> genotype, sporophytic inheritance mode is predominant. Plants exhibited high pollen staining rates (mostly &gt; 80%), and the segregation of heterozygous loci in the next generation followed the Mendelian ratio. Notably, alleles <i>rf-m1</i> and especially <i>rf-m2</i> appear to suppress fertility. Further experiments revealed the best candidates of <i>Rf-m1</i> and <i>Rf-m2</i> are <i>Glyma.16G139800</i> and <i>Glyma.09G171200</i>, encoding mitochondrial pentatricopeptide repeat (PPR) proteins GmPPR567 and GmPPR497. Haplotype analysis revealed that lines carrying homozygous <i>Rf</i> /<i>Rf</i> or <i>rf</i> /<i>rf</i> at both loci are rare among domesticated soybean germplasms. Using locus-specific markers and in silico haplotype screening, we identified two effective restorers and one maintainer line in elite germplasms, facilitating hybrid breeding in soybean.</p>

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Two nuclear loci Rf-m1 and Rf-m2 interplay to determine the fertility restoring efficacy of ZD-type CMS soybean

  • 期刊论文

  • 发表日期：

    2025年6月9日

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  • Xiaoxia Yu,
  • Yupeng Feng,
  • Lulu Zhou,
  • Hongye Gao,
  • Hongbin Wang,
  • Yaru Chen,
  • Jiamai Liang,
  • Tianyu Meng,
  • Meng Li,
  • Yihan Yuan,
  • Junping Yu,
  • Hao Liu,
  • Aigen Fu,
  • Min Xu

摘要

Key message

Two Restorer-of-fertility loci, Rf-m1 and Rf-m2, co-condition fertility restoration in ZD-type CMS soybean though complex genetic interactions in both gametophytic and sporophytic manners.

Abstract

In soybean (Glycine max), the genetic mechanisms underlying fertility restoration of ZD-type cytoplasmic male sterility (CMS-ZD) are not fully understood. By crossing the CMS-ZD line W931A with its strong restorer WR016, we elucidated the inheritance patterns of fertility restoration. We discovered that restoration by WR016 is primarily governed by two Restorer-of-fertility (Rf) loci, Rf-m1 and Rf-m2, located within ~ 220 kb on chromosome 16 and ~ 117 kb on chromosome 9, respectively. They exhibited two distinctive inheritance modes across genotype combinations. In plants with the genotype of -/rf-m1 -/rf-m2, such as F1 hybrids, gametophytic inheritance mode is predominant. In this case, Rf-m1 in haploid tissue is essential for pollen fertility restoration, and Rf-m2 is a modifier to enhance the effect. Correspondingly, plants displayed low pollen staining rates (sterile to ~ 50% depending on genotype). Conversely, in plants with one locus present as the homozygous Rf/Rf genotype, sporophytic inheritance mode is predominant. Plants exhibited high pollen staining rates (mostly > 80%), and the segregation of heterozygous loci in the next generation followed the Mendelian ratio. Notably, alleles rf-m1 and especially rf-m2 appear to suppress fertility. Further experiments revealed the best candidates of Rf-m1 and Rf-m2 are Glyma.16G139800 and Glyma.09G171200, encoding mitochondrial pentatricopeptide repeat (PPR) proteins GmPPR567 and GmPPR497. Haplotype analysis revealed that lines carrying homozygous Rf /Rf or rf /rf at both loci are rare among domesticated soybean germplasms. Using locus-specific markers and in silico haplotype screening, we identified two effective restorers and one maintainer line in elite germplasms, facilitating hybrid breeding in soybean.