Plant Physiol.
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Evidence that the Plant Host Synthesizes the Heme Moiety of Leghemoglobin in Root Nodules1

Maria A. Santana2, Kaarina Pihakaski-Maunsbach3, Niels Sandal, Kjeld A. Marcker, and Alison G. Smith*

Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge CB2 3EA, United Kingdom (M.A.S., A.G.S.); and Laboratory of Gene Expression, Department of Molecular Biology, University of Aarhus, Gustav Wieds Vej 10, DK-8000 Aarhus, Denmark (K.P.-M., N.S., K.A.M.)

Although it is well established that the plant host encodes and synthesizes the apoprotein for leghemoglobin in root nodules, the source of the heme moiety has been uncertain. We recently found that the transcript for coproporphyrinogen III oxidase, one of the later enzymes of heme synthesis, is highly elevated in soybean (Glycine max L.) nodules compared with roots. In this study we measured enzyme activity and carried out western-blot analysis and in situ hybridization of mRNA to investigate the levels during nodulation of the plant-specific coproporphyrinogen oxidase and four other enzymes of the pathway in both soybean and pea (Pisum sativum L.). We compared them with the activity found in leaves and uninfected roots. Our results demonstrate that all of these enzymes are elevated in the infected cells of nodules. Because these are the same cells that express apoleghemoglobin, the data strongly support a role for the plant in the synthesis of the heme moiety of leghemoglobin.

1   This work was supported by a studentship from the Venezuelan National Academy of Science (M.A.S.), and by a short-term fellowship from the European Molecular Biology Organization and grants from the Joint Committee of the Nordic Natural Science Research Councils (K.P.-M.).
2   Present address: Instituto Internacional de Estudios Avanzados, Centro de Biociencias, Apartado 17606, Parque Central, Caracas 1015 A, Venezuela.
3   Permanent address: Laboratory of Plant Physiology and Molecular Biology, Department of Biology, University of Turku, FIN-20014 Turku, Finland.
*   Corresponding author; e-mail as25{at}cam.ac.uk; fax 44-1223-333953.

Plant Physiol. (1998) 116: 1259-1269
Copyright Clearance Center:   0032-0889/98/116/1259/11
© 1998 American Society of Plant Physiologists




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