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The RNA polymerase II subunit NRPB2 is required for indeterminate root development, cell viability, stem cell niche maintenance, and de novo root tip regeneration in Arabidopsis

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Abstract

The RNA polymerase II drives the biogenesis of coding and non-coding RNAs for gene expression. Here, we describe new roles for its second-largest subunit, NRPB2, on root organogenesis and regeneration. Down-regulation of NRPB2 activates a determinate developmental program, which correlated with a reduction in mitotic activity, cell elongation, and size of the root apical meristem. Noteworthy, nrpb2-3 mutants manifest cell death in pro-vascular cells within primary root tips of plants grown in darkness or exposed to light, which triggers the expression of the regeneration gene marker ERF115 in neighbor cells close to damage. Auxin and stem cell niche (SCN) gene expression as well as structural analysis revealed that NRPB2 maintains SCN activity through distribution of PIN transporters in root tissues. Wild-type seedlings regenerated the root tip after excision of the QC and SCN, but nrpb2-3 mutants did not rebuild the missing tissues, and this process could be genotypified using pERF115:GFP, DR5:GFP, and pWOX5:GFP reporter constructs. The levels of reactive oxygen species increased in the mutants four days after germination and strongly decreased at later times, whereas nitric oxide accumulated as the root tip differentiates. These results show the importance of the transcriptional machinery for root organogenesis, cell viability, and regenerative capacity for reconstruction of tissues and organs upon injury.

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Acknowledgements

The authors thank to Prof. Xuemei Chen for sharing the nrpb2-3 mutant, Prof. Lieven de Veylder for providing pERF115:GFP line and Profs. Ben Scheres and Alfredo Cruz Ramirez for donation of Arabidopsis transgenic lines. We thank Dr. Jesús Salvador López Bucio for critical reading and correction of this manuscript. This work was funded by grants from SEP-CONACYT A1-S-34768, and FORDECYT-PRONACES/376120/2020).

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Authors and Affiliations

  1. Facultad de Químico Farmacobiología, Universidad Michoacana de San Nicolás de Hidalgo, Avenida Tzintzuntzan 173, Col. Matamoros, 58240, Morelia, Michoacán, México

    Javier Raya-González

  2. Instituto de Investigaciones Químico Biológicas, Universidad Michoacana de San Nicolás de Hidalgo, Edificio B3, Ciudad Universitaria, 58030, Morelia, Michoacán, México

    Adrián Ávalos-Rangel, León Francisco Ruiz-Herrera & José López-Bucio

  3. Centro Multidisciplinario de Estudios en Biotecnología, Facultad de Medicina Veterinaria y Zootecnia, Universidad Michoacana de San Nicolás de Hidalgo, Morelia, Michoacán, México

    Juan José Valdez-Alarcón

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  1. Javier Raya-González
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  2. Adrián Ávalos-Rangel
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  3. León Francisco Ruiz-Herrera
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  4. Juan José Valdez-Alarcón
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  5. José López-Bucio
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Contributions

J.R.G., A.A.R., J.V.A., and L.F.R.H. performed the experiments. J.R.G., A.A.R., and J.L.B. analyzed the data. J.R.G. and J.L.B. designed the experiments and wrote the manuscript. All authors approved the manuscript.

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Correspondence to Javier Raya-González or José López-Bucio.

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Raya-González, J., Ávalos-Rangel, A., Ruiz-Herrera, L.F. et al. The RNA polymerase II subunit NRPB2 is required for indeterminate root development, cell viability, stem cell niche maintenance, and de novo root tip regeneration in Arabidopsis. Protoplasma 259, 1175–1188 (2022). https://doi.org/10.1007/s00709-021-01732-z

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