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Background <p>Japanese cedar (<i>Cryptomeria japonica</i> D. Don) is a major plantation species in Japan, but its abundant pollen production is a primary cause of seasonal allergic rhinitis (pollinosis). To mitigate this public health issue, the use of male-sterile cultivars has been promoted. Five types of recessive male-sterile mutations (<i>ms1</i>–<i>ms5</i>) have been identified, and the causal genes and mutations for <i>MS1</i> and <i>MS4</i> have been elucidated. However, the gene responsible for MS2-type male sterility remains unknown.</p> Results <p>We aimed to identify the candidate gene responsible for MS2-type male sterility using a map-based cloning strategy. High-resolution linkage mapping localized the <i>MS2</i> locus to a 1.56&#xa0;cM interval on linkage group 5, corresponding to an 8.64&#xa0;Mb region of the reference genome. Ninety-one genes in this region were subjected to functional annotation, gene expression analysis, and mutation screening. Among these, a single gene, SUGI_0493010, encoding a GDSL-type esterase/lipase protein (GELP), fulfilled all three criteria: it showed homology to pollen development genes in <i>Arabidopsis thaliana</i>, was specifically expressed in male strobili, and carried a deleterious amino acid substitution (S40F) within the predicted catalytic domain in <i>ms2</i> mutant. The same mutation was also detected in a heterozygous individual (<i>Ms2/ms2</i>) from a separate breeding population, whose genotype was confirmed through progeny testing. Structural annotation revealed that the affected serine residue lies within the conserved GDSL motif, suggesting a functional disruption of enzymatic activity.</p> Conclusions <p>Our results strongly suggest that SUGI_0493010 (<i>GELP</i>) is the candidate gene for MS2-type male sterility in <i>C. japonica</i>. This finding enhances our understanding of male sterility mechanisms in conifers and provides a valuable genetic resource for breeding pollen-free trees. The study also demonstrates the effectiveness of combining genetic mapping with transcriptomic and mutational data in forest tree genomics.</p>

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Positional identification of a candidate gene for MALE STERILITY 2 (MS2) by linkage mapping and transcriptomic data in Cryptomeria japonica D. Don

  • 期刊论文

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  • 发表日期：

    2026年5月19日

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  • Saneyoshi Ueno,
  • Yoichi Hasegawa,
  • Tokuko Ujino-Ihara,
  • Momi Tsuruta,
  • Hiroyuki Kakui,
  • Junji Iwai,
  • Satoko Hirayama,
  • Katsushi Yamaguchi,
  • Shuji Shigenobu,
  • Takeshi Fujino,
  • Yutaka Suzuki,
  • Masahiro Kasahara,
  • Yoshinari Moriguchi

摘要

Background

Japanese cedar (Cryptomeria japonica D. Don) is a major plantation species in Japan, but its abundant pollen production is a primary cause of seasonal allergic rhinitis (pollinosis). To mitigate this public health issue, the use of male-sterile cultivars has been promoted. Five types of recessive male-sterile mutations (ms1–ms5) have been identified, and the causal genes and mutations for MS1 and MS4 have been elucidated. However, the gene responsible for MS2-type male sterility remains unknown.

Results

We aimed to identify the candidate gene responsible for MS2-type male sterility using a map-based cloning strategy. High-resolution linkage mapping localized the MS2 locus to a 1.56 cM interval on linkage group 5, corresponding to an 8.64 Mb region of the reference genome. Ninety-one genes in this region were subjected to functional annotation, gene expression analysis, and mutation screening. Among these, a single gene, SUGI_0493010, encoding a GDSL-type esterase/lipase protein (GELP), fulfilled all three criteria: it showed homology to pollen development genes in Arabidopsis thaliana, was specifically expressed in male strobili, and carried a deleterious amino acid substitution (S40F) within the predicted catalytic domain in ms2 mutant. The same mutation was also detected in a heterozygous individual (Ms2/ms2) from a separate breeding population, whose genotype was confirmed through progeny testing. Structural annotation revealed that the affected serine residue lies within the conserved GDSL motif, suggesting a functional disruption of enzymatic activity.

Conclusions

Our results strongly suggest that SUGI_0493010 (GELP) is the candidate gene for MS2-type male sterility in C. japonica. This finding enhances our understanding of male sterility mechanisms in conifers and provides a valuable genetic resource for breeding pollen-free trees. The study also demonstrates the effectiveness of combining genetic mapping with transcriptomic and mutational data in forest tree genomics.