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Berenguer E, Carneros E, Pérez-Pérez Y, et al. Small molecule inhibitors of mammalian GSK-3β promote in vitro plant cell reprogramming and somatic embryogenesis in crop and forest species. J Exp Bot. 2021;72(22):7808-7825doi: 10.1093/jxb/erab365.
Berenguer, E., Carneros, E., Pérez-Pérez, Y., Gil, C., Martínez, A., & Testillano, P. S. (2021). Small molecule inhibitors of mammalian GSK-3β promote in vitro plant cell reprogramming and somatic embryogenesis in crop and forest species. Journal of experimental botany, 72(22), 7808-7825. https://doi.org/10.1093/jxb/erab365
Berenguer, Eduardo, et al. "Small molecule inhibitors of mammalian GSK-3β promote in vitro plant cell reprogramming and somatic embryogenesis in crop and forest species." Journal of experimental botany vol. 72,22 (2021): 7808-7825. doi: https://doi.org/10.1093/jxb/erab365
Berenguer E, Carneros E, Pérez-Pérez Y, Gil C, Martínez A, Testillano PS. Small molecule inhibitors of mammalian GSK-3β promote in vitro plant cell reprogramming and somatic embryogenesis in crop and forest species. J Exp Bot. 2021 Dec 04;72(22):7808-7825. doi: 10.1093/jxb/erab365. PMID: 34338766; PMCID: PMC8664590.
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J Exp Bot. 2021 Dec 04;72(22):7808-7825. doi: 10.1093/jxb/erab365.

Small molecule inhibitors of mammalian GSK-3β promote in vitro plant cell reprogramming and somatic embryogenesis in crop and forest species.

Journal of experimental botany

Eduardo Berenguer, Elena Carneros, Yolanda Pérez-Pérez, Carmen Gil, Ana Martínez, Pilar S Testillano

Affiliations

  1. Pollen Biotechnology of Crop Plants group, Margarita Salas Center of Biological Research, CIB-CSIC, Ramiro de Maeztu 9, 28040 Madrid, Spain.
  2. Translational Medicinal and Biological Chemistry group, Margarita Salas Center of Biological Research, CIB-CSIC, Ramiro de Maeztu 9, 28040 Madrid, Spain.

PMID: 34338766 PMCID: PMC8664590 DOI: 10.1093/jxb/erab365

Abstract

Plant in vitro regeneration systems, such as somatic embryogenesis, are essential in breeding; they permit propagation of elite genotypes, production of doubled-haploids, and regeneration of whole plants from gene editing or transformation events. However, in many crop and forest species, somatic embryogenesis is highly inefficient. We report a new strategy to improve in vitro embryogenesis using synthetic small molecule inhibitors of mammalian glycogen synthase kinase 3β (GSK-3β), never used in plants. These inhibitors increased in vitro embryo production in three different systems and species, microspore embryogenesis of Brassica napus and Hordeum vulgare, and somatic embryogenesis of Quercus suber. TDZD-8, a representative compound of the molecules tested, inhibited GSK-3 activity in microspore cultures, and increased expression of embryogenesis genes FUS3, LEC2, and AGL15. Plant GSK-3 kinase BIN2 is a master regulator of brassinosteroid (BR) signalling. During microspore embryogenesis, BR biosynthesis and signalling genes CPD, GSK-3-BIN2, BES1, and BZR1 were up-regulated and the BAS1 catabolic gene was repressed, indicating activation of the BR pathway. TDZD-8 increased expression of BR signalling elements, mimicking BR effects. The findings support that the small molecule inhibitors promoted somatic embryogenesis by activating the BR pathway, opening up the way for new strategies using GSK-3β inhibitors that could be extended to other species.

© The Author(s) 2021. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Keywords: Barley; brassinosteroids; cell reprogramming; cork oak; glycogen synthase kinase; microspore embryogenesis; rapeseed; small molecule inhibitors; somatic embryogenesis

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