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Functionally redundant LNG3 and LNG4 genes regulate turgor-driven polar cell elongation through activation of XTH17 and XTH24

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In this work, we genetically characterized the function of Arabidopsis thaliana, LONGIFOLIA (LNG1), LNG2, LNG3, LNG4, their contribution to regulate vegetative architecture in plant. We used molecular and biophysical approaches to elucidate a gene function that regulates vegetative architecture, as revealed by the leaf phenotype and later effects on flowering patterns in Arabidopsis loss-of-function mutants. As a result, LNG genes play an important role in polar cell elongation by turgor pressure controlling the activation of XTH17 and XTH24.

Abstract

Plant vegetative architecture is related to important traits that later influence the floral architecture involved in seed production. Leaf morphology is the primary key trait to compose plant vegetative architecture. However, molecular mechanism on leaf shape determination is not fully understood even in the model plant A. thaliana. We previously showed that LONGIFOLIA (LNG1) and LONGIFOLIA2 (LNG2) genes regulate leaf morphology by promoting longitudinal cell elongation in Arabidopsis. In this study, we further characterized two homologs of LNG1, LNG3, and LNG4, using genetic, biophysical, and molecular approaches. Single loss-of-function mutants, lng3 and lng4, do not show any phenotypic difference, but mutants of lng quadruple (lngq), and lng1/2/3 and lng1/2/4 triples, display reduced leaf length, compared to wild type. Using the paradermal analysis, we conclude that the reduced leaf size of lngq is due to decreased cell elongation in the direction of longitudinal leaf growth, and not decreased cell proliferation. This data indicate that LNG1/2/3/4 are functionally redundant, and are involved in polar cell elongation in Arabidopsis leaf. Using a biophysical approach, we show that the LNGs contribute to maintain high turgor pressure, thus regulating turgor pressure-dependent polar cell elongation. In addition, gene expression analysis showed that LNGs positively regulate the expression of the cell wall modifying enzyme encoded by a multi-gene family, xyloglucan endotransglucosylase/hydrolase (XTH). Taking all of these together, we propose that LNG related genes play an important role in polar cell elongation by changing turgor pressure and controlling the activation of XTH17 and XTH24.

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Acknowledgements

This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ01365803)” Rural Development Administration, Republic of Korea and the Next-Generation BioGreen 21 Program (PMBC, PJ011912012016 to S.J.P.)

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Correspondence to Young Koung Lee.

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Supplementary Fig. 1

. Phenotype of WT (A), lng1-1D (B) and lngq (C) grown in the ½ MS media condition for the western blot analysis. Supplementary material 1 (JPG 75 KB)

Supplementary Fig. 2

. Gene expression analysis of expansin genes and endo-1,4-beta-glucanase genes. Expansin genes include EXPANSIN 1, 3, 4, 6, 9, 11, 13, 14, 15, 18, 20, 21 and endo-1,4-betaglucanases include At1g48930, At1g65610, At1g75680 and At5g49720. Supplementary material 2 (JPG 83 KB)

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Supplementary material 4 (DOC 78 KB)

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Lee, Y.K., Rhee, J.Y., Lee, S.H. et al. Functionally redundant LNG3 and LNG4 genes regulate turgor-driven polar cell elongation through activation of XTH17 and XTH24. Plant Mol Biol 97, 23–36 (2018). https://doi.org/10.1007/s11103-018-0722-0

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  • DOI: https://doi.org/10.1007/s11103-018-0722-0

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