Abstract
Key message
Plant male sterility is a valuable trait in breeding and hybrid seed production. The barley male-sterility gene msg26 was mapped to a 0.02-cM region that anchors to a 506-kb low-quality assembly between two cleaved amplified polymorphic sequence (CAPS) markers, SP1M14 and SP1M49. The barley gene HORVU4Hr1G074840, which encodes a putative cytochrome P450 CYP704B protein, appears to be a strong candidate for the MSG26 trait.
Abstract
Barley (Hordeum vulgare L.) is an important cereal crop worldwide. Traditional breeding in barley is time-consuming and labor-intensive. The use of male-sterile genotypes may significantly improve the efficacy of hybrid breeding and seed production. The barley accession ‘GSHO745’ is a spontaneous male-sterile mutant from the barley variety, ‘Unitan’. The male sterility in ‘GSHO745’ is controlled by the recessive gene, msg26 (originally named as ms-u). We revealed that the barley plants homozygous for msg26 proceeded normally through Meiosis II until the tetrad stage, but became fully defective in the late uninucleate microspores and developed pollen-less anthers. Using seven barley F2 populations, we mapped MSG26 to a 0.02-cM region that anchored to a 506-kb low-quality assembly between two cleaved amplified polymorphic sequence markers, SP1M14 and SP1M49. The HORVU4Hr1G074840 gene that encodes a putative cytochrome P450 protein (CYP704B) was identified as the most plausible candidate for MSG26. First, HORVU4Hr1G074840 is located in a collinear region of the rice CYP704B2 and the maize CYP704B1. Both of these genes are essential for male gamete production. Second, the male-sterile allele of HORVU4Hr1G074840 in GSHO745 contained a 4-bp deletion in the last exon. The resulting frame shift causes a Gly436Gln substitution, scrambles the sequence of the remainder of the protein, and forms a new termination site at the 70th triplet of the shifted reading frame. We thus called the variant protein CYP704B:p.G436Qfs*70. Third, the barley HORVU4Hr1G074840 gene was specifically expressed in anthers. Altogether, HORVU4Hr1G074840 represents a strong candidate for MSG26 in barley.
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This work was supported by the China Research and Development Initiative on Genetically Modified Plants (2016ZX08009003-001-006), the National Key Research and Development Program of China (2016YFD0100604), and by the Hatch project IDA01587 from the USDA National Institute of Food and Agriculture.
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Fig. S1
Pollen grains of the Msg26 heterozygous plant. Pollen grains of an F1 plant (derived from GSHO745/Morex, genotype Msg26 msg26) were colored by Alexander’s stain (Alexander 1969). Scale bar = 50 μm. (JPEG 276 kb)
Fig. S2
MSG26 region collinearity between barley, maize, and rice. a-c) Physical map in rice, barley, and maize, respectively. The collinear regions between HORVU4Hr1G074720 and HORVU4Hr1G074810 are 49.3 kb in rice (a), 457.9 kb in barley (b), and 219.6 kb in maize (c). Due to space limitations, a short name of the barley genes was used on the maps, for example 4Hr1G074790 for HORVU4Hr1G074790. The most promising candidate is highlighted by an asterisk symbol. The following genes are collinear: 1) HORVU4Hr1G074720 (or SP1M14), Os03t0167800, Zm00001d027827; 2) HORVU4Hr1G074790, Os03t0168400, and Zm00001d027835; 3) HORVU4Hr1G074830, Os03t0168550, and Zm00001d027843; 4) HORVU4Hr1G074840, Os03t0168600, and Zm00001d027837; 5) HORVU4Hr1G074850, Os03t0168700, and Zm00001d027838; and 6) HORVU4Hr1G074810 (or SP1M4), Os03t0169100, and Zm00001d027841. (JPEG 234 kb)
Fig. S3
Expression of wheat collinear genes in the MSG26 region. The expression data were extracted from the wheat RNA-Seq databases (Ramírez-González et al. 2018). The X-axis separated different tissues, including roots (at the flag-leaf stage), stems, leaves, and spikes (at the booting stage) and grains (at the milk stage). The Y-axis represented the expression count. Expression counts were normalized using the variance stabilizing transformation in DESeq2 (Love et al. 2014). (JPEG 186 kb)
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Qi, J., Ni, F., Wang, X. et al. The anther-specific CYP704B is potentially responsible for MSG26 male sterility in barley. Theor Appl Genet 132, 2413–2423 (2019). https://doi.org/10.1007/s00122-019-03363-8
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DOI: https://doi.org/10.1007/s00122-019-03363-8