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JXB Advance Access published online on June 27, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri200
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org
Received March 3, 2005
Accepted April 17, 2005

RESEARCH PAPER

In situ expression of two storage protein genes in relation to histo-differentiation at mid-embryogenesis in Medicago truncatula and Pisum sativum seeds

M. Abirached-Darmency 1*, M. R. Abdel-gawwad 1, G. Conejero 2, J. L. Verdeil 2, and R. Thompson 1

1 INRA-URLEG, Unité de Recherche sur les Légumineuses, BP 86510, F-21065 Dijon Cedex, France
2 CIRAD UMR BEPC, UMRBPMP TA 40/02 Avenue Agropolis, F-34198, Montpellier cedex 5, France

* To whom correspondence should be addressed.
M. Abirached-Darmency, E-mail: mona{at}epoisses.inra.fr


  Abstract

The seed consists of several layers of specialized cell-types that divide and differentiate following a highly regulated programme in time and space. A cytological approach was undertaken in order to study the histo-differentiation at mid-embryogenesis in Medicago truncatula as a model legume, and in Pisum sativum using serial sections of embedded immature seed. Little published information is available about seed development in Medicago species. The observations from this study revealed a number of distinctive features of Medicago seed development and differentiation. Transfer cells, involved in nutrient transfer to the embryo, were clearly identified in the thin-walled parenchyma of the innermost integument. Histological Schiff-naphthol enabled carbohydrate accumulation to be followed in the different seed compartments, and revealed the storage protein bodies. Non-radioactive mRNA in situ hybridization, was carried out using mRNA probes from two highly expressed genes encoding the major vicilin and legumin A storage protein types. The timing of mRNA expression was related to that of the corresponding proteins already identified.

Keywords: In situ hybridization; legumin A mRNA; Medicago truncatula; mid-embryogenesis; Pisum sativum; seed; vicilin mRNA.
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