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Plant Physiol, November 2001, Vol. 127, pp. 1287-1298

The Glyoxylate Cycle in an Arbuscular Mycorrhizal Fungus. Carbon Flux and Gene Expression

Peter J. Lammers, Jeongwon Jun, Jehad Abubaker, Raul Arreola, Anjali Gopalan, Berta Bago,1 Cinta Hernandez-Sebastia,2 James W. Allen, David D. Douds, Philip E. Pfeffer, and Yair Shachar-Hill*

Department of Chemistry and Biochemistry, New Mexico State University, Las Cruces, New Mexico 88001 (P.J.L., J.J., J.A., R.A., A.G., C.H.-S., J.W.A., Y.S.-H.); and United States Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center, 600 East Mermaid Lane, Wyndmoor, Pennsylvania 19038 (B.B., D.D.D., P.E.P.)

The arbuscular mycorrhizal (AM) symbiosis is responsible for huge fluxes of photosynthetically fixed carbon from plants to the soil. Lipid, which is the dominant form of stored carbon in the fungal partner and which fuels spore germination, is made by the fungus within the root and is exported to the extraradical mycelium. We tested the hypothesis that the glyoxylate cycle is central to the flow of carbon in the AM symbiosis. The results of 13C labeling of germinating spores and extraradical mycelium with 13C2-acetate and 13C2-glycerol and analysis by nuclear magnetic resonance spectroscopy indicate that there are very substantial fluxes through the glyoxylate cycle in the fungal partner. Full-length sequences obtained by polymerase chain reaction from a cDNA library from germinating spores of the AM fungus Glomus intraradices showed strong homology to gene sequences for isocitrate lyase and malate synthase from plants and other fungal species. Quantitative real-time polymerase chain reaction measurements show that these genes are expressed at significant levels during the symbiosis. Glyoxysome-like bodies were observed by electron microscopy in fungal structures where the glyoxylate cycle is expected to be active, which is consistent with the presence in both enzyme sequences of motifs associated with glyoxysomal targeting. We also identified among several hundred expressed sequence tags several enzymes of primary metabolism whose expression during spore germination is consistent with previous labeling studies and with fluxes into and out of the glyoxylate cycle.

1 Present address: Departamento de Microbiología del Suelo y Sistemas Simbióticos, Estación Experimental del Zaidín, calle Profesor Albareda, 1, 18008-Granada, Spain.

2 Present address: Department of Crop Science and Botany, North Carolina State University, Raleigh, NC 27695-7631.

* Corresponding author; e-mail yairhill{at}nmsu.edu; fax 505-646-2649.

© 2001 American Society of Plant Physiologists


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