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The characterization and candidate gene isolation for a novel male-sterile mutant ms40 in maize

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Abstract

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A novel genic male-sterile mutant ms40 was obtained from EMS treated RP125. The key candidate gene ZmbHLH51 located on chromosome 4 was identified by map-based cloning. This study further enriched the male sterile gene resources for both production applications and theoretical studies of abortion mechanisms.

Abstract

Maize male-sterile mutant 40 (ms40) was obtained from the progeny of the ethyl methanesulfonate (EMS) treated inbred line RP125. Genetic analysis indicated that the sterility was controlled by a single recessive nuclear gene. Cytological observation of anthers revealed that the cuticles of ms40 anthers were abnormal, and no Ubisch bodies were observed on the inner surface of ms40 anthers through scanning electron microscopy(SEM). Moreover, its tapetum exhibited delayed degradation and then blocked the formation of normal microspores. Using map-based cloning strategy, the ms40 locus was found to locate in a 282-kb interval on chromosome 4, and five annotated genes were predicted within this region. PCR-based sequencing detected a single non-synonymous SNP (G > A) that changed glycine (G) to arginine (A) in the seventh exon of Zm00001d053895, while no sequence difference between ms40 and RP125 was found for the other four genes. Zm00001d053895 encodes the bHLH transcription factor ZmbHLH51 which is localized in the nucleus. Phylogenetic analysis showed that ZmbHLH51 had the highest homology with Sb04g001650, a tapetum degeneration retardation (TDR) bHLH transcription factor in Sorghum bicolor. Co-expression analysis revealed a total of 1192 genes co-expressed with ZmbHLH51 in maize, 647 of which were anther-specific genes. qRT-PCR results suggested the expression levels of some known genes related to anther development were affected in ms40. In summary, these findings revealed the abortion characteristics of ms40 anthers and lay a foundation for further studies on the mechanisms of male fertility.

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Acknowledgements

This research was supported by the Sichuan Science and Technology Program (No. 2021YFYZ0011; No. 2021YFYZ0017), the Crop Variety Improvement and Insect Pest Control by Nuclear Irradiation and the Specific Research Supporting Program for Discipline Construction at Sichuan Agricultural University.

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M.C. and Z.Z. conceived and designed the experiments. X.L., Y.Y., Z.G., Q.H., H.Y.and T.Y. performed the experiments. Z.Z., M.Z. and C.L. analysed the data. X.L. wrote the manuscript. Z.P. directed the study and revised the manuscript. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Moju Cao.

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The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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299_2021_2762_MOESM1_ESM.tif

Supplementary file1 (TIF 182 KB) The expression levels of candidate genes in the fine mapping region between ms40 and RP125. PMC, pollen mother cell stage; T, tetrad stage; Error bars indicate the standard error of mean of three biological replicates. Student’s t-test: *(P < 0.05)

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Liu, X., Yue, Y., Gu, Z. et al. The characterization and candidate gene isolation for a novel male-sterile mutant ms40 in maize. Plant Cell Rep 40, 1957–1970 (2021). https://doi.org/10.1007/s00299-021-02762-w

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  • DOI: https://doi.org/10.1007/s00299-021-02762-w

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