Abstract
Weak seed dormancy (SD) is prone to pre-harvest sprouting (PHS), which reduces cereal yield and quality. Here, through map-based analysis, we identify TaCNGC-2A, encoding a cyclic nucleotide-gated channel protein, as a negative regulator of wheat SD. Knocking out of TaCNGC-2A enhances SD and PHS resistance, with no yield penalty. Two transcription factors, TaMYB-5B and TaMYB-5D, directly bind to the T/A mutation site of TaCNGC-2A promoter to synergistically repress its expression. The calmodulin TaCaM-3A interacts with TaCNGC-2A to jointly modulate SD and PHS resistance through influencing calcium and multiple hormonal signaling pathways. Knocking out of TaCaM-3A not only enhances SD and PHS resistance, but also increases grain weight and per-plant yield. Finally, we identify allele combinations of TaCNGC-2A and other known dormancy genes associated with strong SD. This study uncovers a regulatory mechanism underlying SD and PHS resistance and provides gene targets for breeding wheat varieties with PHS resistance.
Similar content being viewed by others
Data availability
The RNA-seq data generated in this study have been deposited in the NCBI Sequence Read Archive (SRA) under BioProject accession PRJNA1328368. All the supporting data for this study are available in this article and its Supplementary Information files. Source Data are provided with this paper.
References
Ali, A. et al. Unraveling molecular and genetic studies of wheat (Triticum aestivum L.) resistance against factors causing pre-harvest sprouting. Agronomy 9, 117 (2019).
Vetch, J. M., Stougaard, R. N., Martin, J. M. & Giroux, M. J. Review: revealing the genetic mechanisms of pre-harvest sprouting in hexaploid wheat (Triticum aestivum L.). Plant Sci. 281, 180–185 (2019).
Xiao, S. H., Zhang, X. Y., Yan, C. S. & Lin, H. Germplasm improvement for preharvest sprouting resistance in Chinese white-grained wheat: an overview of the current strategy. Euphytica 126, 35–38 (2002).
Zhang, Y. J., Xia, X. C. & He, Z. H. The seed dormancy allele TaSdr‑A1a associated with pre‑harvest sprouting tolerance is mainly present in Chinese wheat landraces. Theor. Appl Genet 130, 81–89 (2017).
Mao, X. X. et al. MKK3 cascade regulates seed dormancy through a negative feedback loop modulating ABA signal in rice. Rice 17, 2 (2024).
Liu, S. B. et al. Cloning and characterization of a critical regulator for preharvest sprouting in wheat. Genetics 195, 263–273 (2013).
Osa, M. et al. Mapping QTLs for seed dormancy and Vp1 homologue on chromosome 3A of wheat. Theor. Appl Genet 106, 1491–1496 (2003).
Chen, C. X., Cai, S. B. & Bai, G. H. A major QTL controlling seed dormancy and pre-harvest sprouting resistance on chromosome 4A in a Chinese wheat landrace. Mol. Breed. 21, 351–358 (2008).
Munkvold, J. D., Tanaka, J., Benscher, D. & Sorrells, M. E. Mapping quantitative trait loci for preharvest sprouting resistance in white wheat. Theor. Appl Genet 119, 1223–1235 (2009).
Zhu, Y. L. et al. Identification of major loci for seed dormancy at different post-ripening stages after harvest and validation of a novel locus on chromosome 2AL in common wheat. Mol. Breed. 36, 1–12 (2016).
Zhu, Y. L. et al. Genome-wide association study of pre-harvest sprouting tolerance using a 90K SNP array in common wheat (Triticum aestivum L.). Theor. Appl Genet 132, 2947–2963 (2019).
Tai, L. et al. Pre-harvest sprouting in cereals: genetic and biochemical mechanisms. J. EXP BOT 72, 2857–2876 (2021).
Nakamura, S. et al. A wheat homolog of MOTHER OF FT AND TFL1 acts in the regulation of germination. Plant Cell 23, 3215–3229 (2011).
Liu, S., Li, L., Wang, W., Xia, G. X. & Liu, S. W. TaSRO1 interacts with TaVP1 to modulate seed dormancy and pre-harvest sprouting resistance in wheat. J. Integr. Plant Biol. 66, 36–53 (2024).
Torada, A. et al. A causal gene for seed dormancy on wheat chromosome 4A encodes a MAP Kinase Kinase. Curr. Biol. 26, 782–787 (2016).
Lang, J. et al. Myb10-D confers PHS-3D resistance to pre-harvest sprouting by regulating NCED in ABA biosynthesis pathway of wheat. N. Phytol. 230, 1940–1952 (2021).
Tai, L. et al. A genome-wide association study uncovers that TaPI4K-2A regulates pre-harvest sprouting in wheat. Plant Commun. 13, 5 (2024).
Sugimoto, K. et al. Molecular cloning of Sdr4, a regulator involved in seed dormancy and domestication of rice. Proc. Natl. Acad. Sci. USA 107, 5792–5797 (2010).
Zhang, Y. J., Miao, X. L., Xia, X. C. & He, Z. H. Cloning of seed dormancy genes (TaSdr) associated with tolerance to pre-harvest sprouting in common wheat and development of a functional marker. Theor. Appl Genet 127, 855–866 (2014).
Zhao, B. et al. Sdr4 dominates pre-harvest sprouting and facilitates adaptation to local climatic condition in Asian cultivated rice. J. Integr. Plant Biol. 64, 1246–1263 (2022).
Bentsink, L., Jowett, J., Hanhart, C. J. & Koornneef, M. Cloning of DOG1, a quantitative trait locus controlling seed dormancy in Arabidopsis. Proc. Natl. Acad. Sci. USA 103, 17042–17047 (2006).
Teng, S., Rognoni, S., Bentsink, L. & Smeekens, S. The Arabidopsis GSQ5/DOG1 Cvi allele is induced by the ABA-mediated sugar signalling pathway, and enhances sugar sensitivity by stimulating ABI4 expression. Plant J. 55, 372–381 (2008).
Ashikawa, I., Mori, M., Nakamura, S. & Abe, F. A transgenic approach to controlling wheat seed dormancy level by using Triticeae DOG1-like genes. Transgenic Res. 23, 621–629 (2014).
Wei, X. N. et al. The GATA transcription factor TaGATA1 recruits demethylase TaELF6-A1 and enhances seed dormancy in wheat by directly regulating TaABI5. J. Integr. Plant Biol. 65, 1262–1276 (2023).
Cao, J. J. et al. Identification and validation of new stable QTLs for grain weight and size by multiple mapping models in common wheat. Front Genet 11, 584859 (2020).
Cheng, X. R. et al. Identification of the wheat C3H gene family and expression analysis of candidates associated with seed dormancy and germination. Plant Physiol. Biochem. 156, 524–537 (2020).
Fischer, C., Kugler, A., Hoth, S. & Dietrich, P. An IQ domain mediates the interaction with calmodulin in a plant cyclic nucleotide-gated channel. Plant Cell Physiol. 54, 573–584 (2013).
DeFalco, T. A. et al. Multiple calmodulin-binding sites positively and negatively regulate Arabidopsis CYCLIC NUCLEOTIDE-GATED CHANNEL12. Plant Cell 28, 1738–1751 (2016).
Niu, W. T. et al. Arabidopsis cyclic nucleotide-gated channel 6 is negatively modulated by multiple calmodulin isoforms during heat shock. J. Exp. Bot. 71, 90–104 (2020).
Ashikawa, I., Abe, F. & Nakamura, S. Ectopic expression of wheat and barley DOG1-like genes promotes seed dormancy in Arabidopsis. Plant Sci. 179, 536–542 (2010).
Barrero, J. M. et al. Transcriptomic analysis of wheat near-isogenic lines identifies PM19-A1 and A2 as candidates for a major dormancy QTL. Genome Biol. 16, 93 (2015).
Wei, W. X. et al. Isolation and characterization of TaQsd1 genes for period of dormancy in common wheat (Triticum aestivum L.). Mol. Breed. 39, 150 (2019).
Finkelstein, R., Reeves, W., Ariizumi, T. & Steber, C. Molecular aspects of seed dormancy. Annu. Rev. Plant Biol. 59, 387–415 (2008).
Shu, K., Liu, X. D., Xie, Q. & He, Z. H. Two faces of one seed: hormonal regulation of dormancy and germination. Mol. Plant 9, 34–45 (2016).
Penfield, S. Seed dormancy and germination. Curr. Biol. 27, 874–878 (2017).
Tognacca, R. S. & Botto, J. F. Post-transcriptional regulation of seed dormancy and germination: current understanding and future directions. Plant Commun. 2, 100169 (2021).
Sajeev, N., Koornneef, M. & Bentsink, L. A commitment for life: decades of unraveling the molecular mechanisms behind seed dormancy and germination. Plant Cell 36, 1358–1376 (2024).
Xu, Y. et al. OsCNGC13 promotes seed-setting rate by facilitating pollen tube growth in stylar tissues. PLoS Genet 13, e1006906 (2017).
Zeb, Q. S. et al. The interaction of CaM7 and CNGC14 regulates root hair growth in Arabidopsis. J. Integr. Plant Biol. 62, 887–896 (2019).
Cui, Y. M. et al. CYCLIC NUCLEOTIDE-GATED ION CHANNELs 14 and 16 promote tolerance to heat and chilling in rice. Plant Physiol. 183, 1794–1808 (2020).
Luan, S. & Wang, C. Calcium signaling mechanisms across kingdoms. Annu. Rev. Cell Dev. Bi 37, 311–340 (2021).
Zhao, C. H. et al. A mis-regulated cyclic nucleotide-gated channel mediates cytosolic calcium elevation and activates immunity in Arabidopsis. N. Phytol. 230, 1078–1094 (2021).
Yang, Y. et al. OPEN STOMATA 1 phosphorylates CYCLIC NUCLEOTIDE-GATED CHANNELs to trigger Ca2+ signaling for acid-induced stomatal closure in Arabidopsis. Plant Cell 36, 2328–2358 (2024).
Peng, Y. et al. Differential phosphorylation of Ca2+-permeable channel CYCLIC NUCLEOTIDE–GATED CHANNEL 20 modulates calcium-mediated freezing tolerance in Arabidopsis. Plant Cell 36, 4356–4371 (2024).
Yu, H. H. et al. Transcription factor OsMYB30 increases trehalose content to inhibit α-amylase and seed germination at low temperature. Plant Physiol. 194, 1815–1833 (2024).
Yuan, Y. P. et al. Abscisic acid-induced transcription factor PsMYB306 negatively regulates tree peony bud dormancy release. Plant Physiol. 194, 2449–2471 (2024).
Li, T. et al. Phosphorylation of MdCYTOKININ RESPONSE FACTOR4 suppresses ethylene biosynthesis during apple fruit ripening. Plant Physiol. 191, 694–714 (2023).
Liu, S. et al. Calmodulins and calmodulin-like proteins-mediated plant organellar calcium signaling networks under abiotic stress. Crop J. 12, 1321–1332 (2024).
Li, M. D. et al. CBL1/CIPK23 phosphorylates tonoplast sugar transporter TST2 to enhance sugar accumulation in sweet orange (Citrus sinensis). J. Integr. Plant Biol. 67, 327–344 (2024).
Wang, B. F. et al. OsCPK12 phosphorylates OsCATA and OsCATC to regulate H2O2 homeostasis and improve oxidative stress tolerance in rice. Plant Commun. 5, 3 (2024).
Yu, Q. H. et al. Phosphorylation-dependent VaMYB4a regulates cold stress in grapevine by inhibiting VaPIF3 and activating VaCBF4. Plant Physiol. 197, 2 (2025).
Ju, L. et al. JAZ proteins modulate seed germination through interaction with ABI5 in bread wheat and Arabidopsis. N. Phytol. 223, 246–260 (2019).
Li, Q. F. et al. Gibberellin recovers seed germination in rice with impaired brassinosteroid signalling. Plant Sci. 293, 110435 (2020).
Acknowledgements
We thank for Dr. Shihe Xiao providing JH-RILs and JW-RILs and Dr. Xianchun Xia in CAAS providing 60 Chinese landraces. This work was supported by grants from National Natural Science Foundation of China (31871608, H.Z.; 32372069, H.Z.; Joint Fund Projects, U20A2033, C.M.; F2010123002, Y.R.), Jiangsu Collaborative Innovation Center for Modern Crop Production (JCIC-MCP, C.M.), Hebei Modern Agricultural Industrial Technology System (HBCT2023010201, Y.Z.), and Henan Province Science and Technology R&D Joint Fund (242301420024, Y.F.) and Innovation fund from Anhui Agricultural University (RC312212, Y.R).
Author information
Authors and Affiliations
Contributions
H.Z., Y.R., C.M., C.Chang., H.S., J.L., C.Chen. and W.G. conceived and guided the experiments, and helped in coordinating the project; H.Z. and Y.R. analyzed the data and write the manuscript. All authors read and approved of the manuscript. B.T., X.C., J.C., W.L., Z.Y., J.C., B.L., H.W., S.L., W.G., L.Z., Y.L. and S.W. performed the experiments, analyzed the data and participated in manuscript writing; Y.F., Y.Z., and C.K. participated in the cultivation and management of transgenic and edited materials in the field.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Peer review
Peer review information
Nature Communications thanks the anonymous reviewers for their contribution to the peer review of this work. A peer review file is available.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary information
Source data
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
About this article
Cite this article
Tian, B., Fang, Y., Zhang, Y. et al. TaCNGC-2A suppresses seed dormancy and activates pre-harvest sprouting through modulating calcium and hormonal signaling pathways. Nat Commun (2026). https://doi.org/10.1038/s41467-026-70894-2
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41467-026-70894-2
