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JXB Advance Access published online on July 4, 2006
Journal of Experimental Botany, doi:10.1093/jxb/erl042
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© The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org
Received March 3, 2006
Accepted May 9, 2006

RESEARCH PAPER

Hordeins are expressed in microspore-derived embryos and also during male gametophytic and very early stages of seed development

Amada Pulido 1, Alberto Hernando 2, Ferenc Bakos 3, Enrique Méndez 2, Martine Devic 4, Beata Barnabás 3, and Adela Olmedilla 1 *

1 Department of Plant Biochemistry, Cell and Molecular Biology, EEZ (CSIC), Profesor Albareda 1, E-18008 Granada, Spain
2 Unidad de Gluten, Centro Nacional de Biotecnología, Cantoblanco, E- 28049 Madrid, Spain
3 Agricultural Research Institute, Hungarian Academy of Science, H-2462 Martonvasar, Hungary
4 Plant Genome and Development Laboratory, UMR (CNRS) 5096, University of Perpignan, France

* To whom correspondence should be addressed.
Adela Olmedilla, E-mail: adela.olmedilla{at}eez.csic.es


  Abstract

Microspore-derived embryos induced by anther or isolated-microspore culture display certain characteristics of zygotic embryos. Furthermore, the expression of certain endosperm genes has been described in these non-zygotic embryos. The expression of hordein genes encoding the main barley endosperm proteins has been studied using a wide range of methods (RT-PCR, in situ hybridization, ELISA sandwich, western blotting immunocytochemistry, and cytochemistry) to ascertain their presence or absence during the induction and first stages of microspore embryogenesis. Due to the very sensitive techniques used it was possible to detect for the first time hordein expression during microspore embryogenesis. Surprisingly, these hordeins were also detected at different stages of male gametophytic development as well as during the very early stages of seed development, when they have not hitherto been detected. The expression and localization of these storage proteins and their corresponding transcripts provide new information about barley microspore embryogenesis and its relationship to zygotic embryogenesis. Although only small quantities of hordeins are accumulated during microspore embryogenesis they seem to be necessary for the initial development of the microspore-derived embryo. This idea is supported by the changes detected in their concentration throughout this process and is in accordance with previously published data concerning the importance of endosperm proteins for embryo development in both microspore culture and in planta.

Keywords: Barley; hordeins; microspore embryogenesis; pollen development; protein bodies; storage proteins.
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