Comparative analysis of conserved miRNAs in maize germination and vegetative development

Autores

  • Alberto J. Donayre-Torres Centro de Investigación en Bioingeniería, Universidad de Ingeniería y Tecnología (UTEC), Lima 15063, Perú. https://orcid.org/0000-0003-1689-8428
  • Pedro Toledo-Garcia Departamento de Bioingeniería e Ingeniería Química, Universidad de Ingeniería y Tecnología (UTEC), Lima 15063, Perú. https://orcid.org/0000-0001-5978-6683
  • Estela Sánchez de Jiménez Departamento de Bioquímica, Facultad de Química, Universidad Nacional Autónoma de México, Ciudad de México 04510, México.

DOI:

https://doi.org/10.17268/sci.agropecu.2026.037

Palavras-chave:

microRNA, maize, seed germination, ribosomal proteins, Northern blot

Resumo

Seed germination represents a metabolically active but transcriptionally limited transition that depends largely on post-transcriptional regulation. MicroRNAs (miRNAs) are key regulators of gene expression through mRNA slicing or translational inhibition, and may play an important role in controlling stored mRNAs during early germination. In this study, we combined bioinformatic analysis and experimental validation to identify conserved miRNAs involved in maize (Zea mays) germination. Mature miRNA sequences from Zea mays and Oryza sativa were retrieved from PMRD and miRBase v22.1, identifying conserved candidates across cereals. From this set, ten miRNAs were selected based on sequence conservation and reported roles in seed biology. Target prediction using thermodynamic modelling (ViennaRNA; MFE ≤ −25 kcal/mol) revealed potential interactions between specific miRNAs and mRNAs encoding ribosomal proteins, suggesting regulation of the translational machinery. Expression analysis by Northern blot across germination stages (0, 6, 12, and 24 h) and vegetative tissues showed distinct patterns. miR528 and miR396a were highly expressed in seeds but absent in vegetative tissues, while miR408 decreased after imbibition. miR414 and miR415 displayed transient expression during germination. Notably, miR160a was strongly expressed during germination, suggesting a role beyond vegetative development. These results support a model in which miRNAs regulate stored mRNAs and translation during germination. This work provides candidate miRNAs for functional studies and potential targets for improving seed vigor and early seedling establishment in maize.

Referências

Bai, B., van der Horst, S., Cordewener, J. H. G., America, T. A. H. P., Hanson, J., & Bentsink, L. (2020). Seed-stored mRNAs that are specifically associated to monosomes are translationally regulated during germination. Plant Physiology, 182(1), 378–392. https://doi.org/10.1104/pp.19.00644

Basbouss-Serhal, I., Pateyron, S., Cochet, F., Leymarie, J., & Bailly, C. (2017). 5' to 3' mRNA decay contributes to the regulation of Arabidopsis seed germination by dormancy. Plant Physiology, 173(3), 1709–1723. https://doi.org/10.1104/pp.16.01933

Cock, P. J. A., Antao, T., Chang, J. T., Chapman, B. A., Cox, C. J., Dalke, A., Friedberg, I., Hamelryck, T., Kauff, F., Wilczynski, B., & de Hoon, M. J. L. (2009). Biopython: Freely available Python tools for computational molecular biology and bioinformatics. Bioinformatics, 25(11), 1422–1423. https://doi.org/10.1093/bioinformatics/btp163

Curaba, J., Singh, M. B., & Bhalla, P. L. (2014). miRNAs in the crosstalk between phytohormone signalling pathways. Journal of Experimental Botany, 65(6), 1425–1438. https://doi.org/10.1093/jxb/eru002

Ding, D., Wang, Y., Han, M., Fu, Z., Li, W., Liu, Z., & et al. (2012). MicroRNA transcriptomic analysis of heterosis during maize seed germination. PLoS ONE, 7(6), e39578. https://doi.org/10.1371/journal.pone.0039578

Ding, N., & Zhang, B. (2023). microRNA production in Arabidopsis. Frontiers in Plant Science, 14, 1096772. https://doi.org/10.3389/fpls.2023.1096772

Donayre-Torres, A. J., Esquivel-Soto, E., Gutiérrez-Xicoténcatl, M. de L., Esquivel-Guadarrama, F. R., & Gómez-Lim, M. A. (2009). Production and purification of immunologically active core protein p24 from HIV-1 fused to ricin toxin B subunit in E. coli. Virology Journal, 6, 17. https://doi.org/10.1186/1743-422X-6-17

Dong, Q., Hu, B., & Zhang, C. (2022). microRNAs and their roles in plant development. Frontiers in Plant Science, 13, 824240. https://doi.org/10.3389/fpls.2022.824240

Gao, Y., Feng, B., Gao, C., Zhang, H., Wen, F., Tao, L., Fu, G., & Xiong, J. (2022). The evolution and functional roles of miR408 and its targets in plants. International Journal of Molecular Sciences, 23(1), 530. https://doi.org/10.3390/ijms23010530

Gran, P., Visscher, T. W., Bai, B., Nijveen, H., Mahboubi, A., Bakermans, L. L., Willems, L. A. J., & Bentsink, L. (2025). Unravelling the dynamics of seed-stored mRNAs during seed priming. New Phytologist, 247(5), 2196–2209. https://doi.org/10.1111/nph.70098

Guo, Z., Kuang, Z., Deng, Y., Li, L., & Yang, X. (2022). Identification of species-specific microRNAs provides insights into dynamic evolution of microRNAs in plants. International Journal of Molecular Sciences, 23(22), 14273. https://doi.org/10.3390/ijms232214273

Gutierrez, L., Mongelard, G., Floková, K., Pacurar, D. I., Novák, O., Staswick, P., Kowalczyk, M., Pacurar, M., Demailly, H., Geiss, G., & Bellini, C. (2012). Auxin controls Arabidopsis adventitious root initiation by regulating jasmonic acid homeostasis. Plant Cell, 24(6), 2515–2527. https://doi.org/10.1105/tpc.112.099119

Hunter, J. D. (2007). Matplotlib: A 2D graphics environment. Computing in Science & Engineering, 9(3), 90–95. https://doi.org/10.1109/MCSE.2007.55

Jiang, A., Guo, Z., Pan, J., Yang, Y., Zhuang, Y., Zuo, D., Hao, C., Gao, Z., Xin, P., Chu, J., Zhong, S., & Li, L. (2021). The PIF1-miR408-PLANTACYANIN repression cascade regulates light-dependent seed germination. Plant Cell, 33(5), 1506–1529. https://doi.org/10.1093/plcell/koab060

Kozomara, A., Birgaoanu, M., & Griffiths-Jones, S. (2019). miRBase: From microRNA sequences to function. Nucleic Acids Research, 47(D1), D155–D162. https://doi.org/10.1093/nar/gky1141

Lanet, E., Delannoy, E., Sormani, R., Floris, M., Brodersen, P., Crété, P., Voinnet, O., & Robaglia, C. (2009). Biochemical evidence for translational repression by Arabidopsis microRNAs. Plant Cell, 21(6), 1762–1768. https://doi.org/10.1105/tpc.108.063412

Li, D., Wang, L., Liu, X., Cui, D., Chen, T., Zhang, H., Jiang, C., Xu, C., Li, P., Li, S., Zhao, L., & Chen, H. (2013). Deep sequencing of maize small RNAs reveals a diverse set of microRNA in dry and imbibed seeds. PLoS ONE, 8(1), e55107. https://doi.org/10.1371/journal.pone.0055107

Li, Z., Yang, J., Cai, X., Zeng, X., Zou, J.-J., & Xing, W. (2024). A systematic review on the role of miRNAs in plant response to stresses under the changing climatic conditions. Plant Stress, 14, 100674. https://doi.org/10.1016/j.stress.2024.100674

Lorenz, R., Bernhart, S. H., Höner zu Siederdissen, C., Tafer, H., Flamm, C., Stadler, P. F., & Hofacker, I. L. (2011). ViennaRNA Package 2.0. Algorithms for Molecular Biology, 6, 26. https://doi.org/10.1186/1748-7188-6-26

Luján-Soto, E., Juárez-González, V. T., Reyes, J. L., & Dinkova, T. D. (2021). MicroRNA Zma-miR528 versatile regulation on target mRNAs during maize somatic embryogenesis. International Journal of Molecular Sciences, 22(10), 5310. https://doi.org/10.3390/ijms22105310

Ma, C., Burd, S., & Lers, A. (2015). miR408 is involved in abiotic stress responses in Arabidopsis. The Plant Journal, 84(1), 169–187. https://doi.org/10.1111/tpj.12999

Martinez-Seidel, F., Beine-Golovchuk, O., Hsieh, Y.-C., & Kopka, J. (2020). Systematic Review of Plant Ribosome Heterogeneity and Specialization. Frontiers in Plant Science, 11. https://doi.org/10.3389/fpls.2020.00948

Motomura, K., Le, Q. T., Kumakura, N., Fukaya, T., Takeda, A., & Watanabe, Y. (2012). The role of decapping proteins in the miRNA accumulation in Arabidopsis thaliana. RNA Biology, 9(5), 644–652. https://doi.org/10.4161/rna.19877

Nieves-Cordones, M., Robles, P., & Quesada, V. (2025). Mutations in the plant-conserved uL1m mitochondrial ribosomal protein significantly affect development, growth and abiotic stress tolerance in Arabidopsis thaliana. Plant Growth Regulation, 105, 429–448. https://doi.org/10.1007/s10725-025-01282-x

Nonogaki, H. (2010). MicroRNA gene regulation cascades during early stages of plant development. Plant and Cell Physiology, 51(11), 1840–1846. https://doi.org/10.1093/pcp/pcq154

Palatnik, J. F., Allen, E., Wu, X., Schommer, C., Schwab, R., Carrington, J. C., & Weigel, D. (2003). Control of leaf morphogenesis by microRNAs. Nature, 425(6955), 257–263. https://doi.org/10.1038/nature01958

Pan, J., Huang, D., Guo, Z., Kuang, Z., Zhang, H., Xie, X., Ma, Z., Gao, S., Lerdau, M. T., Chu, C., & Li, L. (2018). Overexpression of microRNA408 enhances photosynthesis, growth, and seed yield in diverse plants. Journal of Integrative Plant Biology, 60(4), 323–340. https://doi.org/10.1111/jipb.12634

Reyes, J. L., & Chua, N. H. (2007). ABA induction of miR159 controls transcript levels of two MYB factors during Arabidopsis seed germination. Plant Journal, 49(4), 592–606. https://doi.org/10.1111/j.1365-313X.2006.02980.x

Rincon-Guzmán, A., Beltrán-Peña, E., Ortíz-López, A., & Sánchez de Jiménez, E. (1998). Ribonucleoprotein particles of quiescent maize embryonic axes. Plant Molecular Biology, 38(3), 357–364. https://doi.org/10.1023/a:1006020121351

Sano, N., Rajjou, L., & North, H. M. (2020). Lost in translation: Physiological roles of stored mRNAs in seed germination. Plants, 9(3), 347. https://doi.org/10.3390/plants9030347

Sarkar Das, S., Yadav, S., Singh, A., Gautam, V., Sarkar, A. K., Prasad, M., Muthamilarasan, M., & Bhatt, D. (2018). Expression dynamics of miRNAs and their targets in seed germination conditions reveals miRNA–ta-siRNA crosstalk as regulator of seed germination. Scientific Reports, 8, 1233. https://doi.org/10.1038/s41598-017-18823-8

Schneider, C., Rasband, W., & Eliceiri, K. (2012). NIH Image to ImageJ: 25 years of image analysis. Nature Methods, 9, 671–675. https://doi.org/10.1038/nmeth.2089

Song, Z., Zhang, L., Wang, Y., Li, H., Li, S., Zhao, H., & Zhang, H. (2018). Constitutive expression of miR408 improves biomass and seed yield in Arabidopsis. Frontiers in Plant Science, 8, 2114. https://doi.org/10.3389/fpls.2017.02114

Tang, X., Bian, S., Tang, M., Lu, Q., Li, S., Liu, X., Tian, G., Nguyen, V., Edward, W., Wang, A., Rothstein, S., Chen, X., & Cui, Y. (2012). MicroRNA-mediated repression of the seed maturation program during vegetative development in Arabidopsis. PLoS Genetics, 8(2), e1003091. https://doi.org/10.1371/journal.pgen.1003091

Tian, T., Zheng, Y., & Bailey-Serres, J. (2025). Translational regulation of plant stress responses: Mechanisms, pathways, and applications in bioengineering. Annual Review of Phytopathology, 63, 117–146. https://doi.org/10.1146/annurev-phyto-121823-032335

Wang, L., Liu, H., Li, D., & Chen, H. (2011). Identification and characterization of maize microRNAs involved in the very early stage of seed germination. BMC Genomics, 12, 154. https://doi.org/10.1186/1471-2164-12-154

Wang, Q., Wan, J., Dang, K., Meng, S., Hu, D., Lin, Y., Qiu, X., Guo, Z., Fu, Z., Ding, D., & Tang, J. (2023). zma-miR159 targets ZmMYB74 and ZmMYB138 transcription factors to regulate grain size and weight in maize. Plant Physiology, 193(4), 2430–2441. https://doi.org/10.1093/plphys/kiad455

Waskom, M. L. (2021). seaborn: Statistical data visualization. Journal of Open Source Software, 6(60), 3021. https://doi.org/10.21105/joss.03021

Xiong, W., Lan, T., & Mo, B. (2021). Extraribosomal Functions of Cytosolic Ribosomal Proteins in Plants. Frontiers in Plant Science, 12. https://doi.org/10.3389/fpls.2021.607157

Zhang, L., Chia, J.-M., Kumari, S., Stein, J. C., Liu, Z., Narechania, A., Maher, C. A., Guill, K., McMullen, M. D., & Ware, D. (2009). A genome-wide characterization of microRNA genes in maize. PLoS Genetics, 5(11), e1000716. https://doi.org/10.1371/journal.pgen.1000716

Zhang, Y., Yu, Y., Wang, C., Li, Z., Liu, Q., Xu, J., Liao, J., Wang, X., Qu, L., Chen, F., Xin, P., Yan, C., Chu, J., Li, H., & Chen, Y. (2013). Overexpression of microRNA OsmiR397 improves rice yield by increasing grain size and promoting panicle branching. Nature Biotechnology, 31(9), 848–852. https://doi.org/10.1038/nbt.2630

Zhang, Z., Yu, J., Li, D., Liu, F., Zhou, X., Wang, T., Ling, Y., & Su, Z. (2010). PMRD: Plant microRNA database. Nucleic Acids Research, 38(Database issue), D806–D813. https://doi.org/10.1093/nar/gkp818

Zhao, Y., Li, X., & Wang, H. (2025). MicroRNAs in plants development and stress resistance. Plant, Cell & Environment, 48(4), 1–22. https://doi.org/10.1111/pce.15546

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Publicado

2026-05-13

Como Citar

Donayre-Torres, A. J., Toledo-Garcia, P., & Sánchez de Jiménez, E. (2026). Comparative analysis of conserved miRNAs in maize germination and vegetative development. Scientia Agropecuaria, 17(2), 529-539. https://doi.org/10.17268/sci.agropecu.2026.037

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