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Mitogen-activated protein kinases are developmentally regulated during stress-induced microspore embryogenesis in Brassica napus L

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Abstract

Plant mitogen-activated protein kinase (MAPK) cascades are involved in extracellular stress signalling pathways, leading to different cellular responses. Stress-induced microspore embryogenesis involves the internalization of an extracellular stress signal, generating a number of cellular responses where MAPK cascades might be involved. These responses include a change of the developmental programme, the entry into an early proliferative stage and, subsequently, into differentiation stages during haploid embryogenesis. In this work we studied the expression during microspore embryogenesis of several kinases, to assess their putative role in these events. The known Brassica napus MAP kinase kinase kinases (MAP3Ks BnMAP3Kα1, BnMAP3Kβ1 and BnMAP3Kε, the BnBSKθ kinase and B. napus extracellular signal-regulated kinase (ERK) homologues were analysed by electron microscope (EM) in situ hybridization, immuno-gold labelling, immunofluorescence and western blotting. The differential in situ expression of these kinases suggests a role for them during embryogenesis. Two different expression patterns were observed, indicating a different regulation. BnMAP3Kα1, BnMAP3Kε, and the ERKs showed a pattern consistent with a role mainly in proliferative events. Conversely, BnMAP3Kβ1 and BnBSKθ, presented a pattern that suggested an involvement in differentiation stages. In addition, ERK homologues migrate to the nucleus immediately after induction, being found in a phosphorylated state in a larger amount.

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Acknowledgements

Thanks are due to the EM Service of Institute Cajal, CSIC, Madrid and to Ms. Beryl Ligus-Walker for proofreading the manuscript. J.M.S.S. was the recipient of an FPI grant of the Spanish Ministry of Education. This work was supported by research projects funded by EU BIO4-CT-0275, Spanish DGESIC PB98-0678, MCyT BOS2002-03572, and CM 07G/0026/2003.

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  1. José M. Seguí-Simarro

    Present address: Centro de Conservación y Mejora de la Agrodiversidad Valenciana (COMAV), Universidad Politécnica de Valencia, Valencia, Spain

Authors and Affiliations

  1. Plant Development and Nuclear Organization Unit, Centro de Investigaciones Biológicas—C.S.I.C. C, Ramiro de Maeztu 9, 28040, Madrid, Spain

    José M. Seguí-Simarro, Pilar S. Testillano & Maria C. Risueño

  2. Institut de Biotechnologie des Plantes, Université de Paris-Sud, Orsay, France

    Stefan Jouannic & Yves Henry

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  1. José M. Seguí-Simarro
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  2. Pilar S. Testillano
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  5. Maria C. Risueño
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Correspondence to Maria C. Risueño.

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Seguí-Simarro, J.M., Testillano, P.S., Jouannic, S. et al. Mitogen-activated protein kinases are developmentally regulated during stress-induced microspore embryogenesis in Brassica napus L. Histochem Cell Biol 123, 541–551 (2005). https://doi.org/10.1007/s00418-004-0749-y

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