Abstract
In plants, asymmetric cell divisions result in distinct cell fates forming large and small daughter cells, adding to the cellular diversity in an organ. SCARECROW (SCR), a GRAS domain-containing transcription factor controls asymmetric periclinal cell divisions in flowering plants by governing radial patterning of ground tissue in roots and cell proliferation in leaves. Though SCR homologs are present across land plant lineages, the current understanding of their role in cellular patterning and leaf development is mostly limited to flowering plants. Our phylogenetic analysis identified three SCR homologs in moss Physcomitrium patens, amongst which PpSCR1 showed highest expression in gametophores and its promoter activity was prominent at the mid-vein and the flanking leaf blade cells pointing towards its role in leaf development. Notably, out of the three SCR homologs, only the ppscr1 knock-out lines developed slender leaves with four times narrower leaf blade and three times thicker mid-vein. Detailed histology studies revealed that slender leaf phenotype is either due to the loss of anticlinal cell divisions or failure of periclinal division suppression in the leaf blade. RNA-Seq analyses revealed that genes responsible for cell division and differentiation are expressed differentially in the mutant. PpSCR1 overexpression lines exhibited significantly wider leaf lamina, further reconfirming the role in leaf development. Together, our data suggests that PpSCR1 is involved in the leaf blade and mid-vein development of moss and that its role in the regulation of cell division and proliferation is ancient and conserved among flowering plants and mosses.
Key message
The GRAS domain containing protein PpSCR1 regulates asymmetric cell divisions and governs leaf blade and mid-vein development in moss Physcomitrium patens.
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Data availability
Sequence data from this article can be found in the GenBank data libraries under accession number(s) SAMN36465315 (scr RNA-Seq Replicate 1), SAMN36465316 (scr RNA-Seq Replicate 2), SAMN36465317 (scr RNA-Seq Replicate 3), SRR22982408 (WT RNA-Seq Replicate 1), SRR22982407 (WT RNA-Seq Replicate 2) and SRR22982406 (WT RNA-Seq Replicate 3). The data supporting the findings and claims of this study is mentioned in the main text and is available with the corresponding author.
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Acknowledgements
We thank Prof. Mitsuyasu Hasebe, NIBB, Japan for sharing knockout vectors (pTN182 and pTN186). We sincerely thank Prof. Magdalena Bezanilla, University of Dartmouth, USA, for the kind gift of Gateway vectors. Technical help received from Mr Vyankatesh Rajmane, Ms Sukanya Jogdand, and Ms Arati Vasav is gratefully acknowledged. The present study was supported by a grant (Grant No. EMR/2016/004852) from the Science and Engineering Research Board (SERB), Government of India to AKB. Core funding and infrastructure were provided by the Indian Institute of Science Education and Research (IISER) Pune, India.
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The financial support received from the Science and Engineering Research Board, Government of India, through Grant No. EMR/2016/004852 is gratefully acknowledged.
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B.M and A.K.B designed the experiments and wrote the manuscript. B.M and S.P constructed the vectors and generated transgenic lines. B.M and A.J.B performed histological analyses. S.P carried out microscopic and phenotypic analyses, western blotting and transcriptomic data analyses. A.J.B conducted gene expression analyses and SCR promoter line characterization. M.P. generated PpSCR2 and PpSCR3 knockout lines. K.R. performed propidium iodide-based staining and image analyses. P.G generated PpSCR1 promoter lines. B.M, S.P, and A.J.B, have contributed to experimental planning, troubleshooting and data interpretation. A.K.B conceived the project, obtained funding and all necessary resources, interpreted data, edited the manuscript, and supervised the entire study. All authors read and approved the final manuscript.
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Mohanasundaram, B., Palit, S., Bhide, A.J. et al. PpSCARECROW1 (PpSCR1) regulates leaf blade and mid-vein development in Physcomitrium patens. Plant Mol Biol 114, 12 (2024). https://doi.org/10.1007/s11103-023-01398-6
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DOI: https://doi.org/10.1007/s11103-023-01398-6