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Journal of Histochemistry and Cytochemistry
Volume 52 (2): 227-241, 2004
Copyright ©The Histochemical Society, Inc.

Expression of Lipoxygenase During Organogenic Nodule Formation from Hop Internodes

Ana Margarida Fortes, Maria José Coronado, Pilar S. Testillano, Maria del Carmen Risueño and Maria Salomé Pais

Centro de Biotecnologia Vegetal, Faculdade de Ciências, Universidade Lisboa, Lisboa, Portugal (AMF,MSP), and Plant Development and Nuclear Organization Group, Centro de Investigaciones Biologicas, CSIC, Madrid, Spain (MJC,PST,MCR)

Correspondence to: Ana Margarida Fortes, Centro de Biotecnologia Vegetal, Faculdade de Ciências, Univ. Lisboa, Bloco C2, Piso 1, Campo Grande, 1749-016 Lisboa, Portugal. E-mail: mfortes{at}fc.ul.pt

Study of lipoxygenase expression (LOX; EC 1.13.11.12) during organogenic nodule formation in hop (Humulus lupulus var. Nugget) showed that LOXs are developmentally regulated throughout the process, suggesting their involvement in the response of internodes to wounding, nodule formation, and plantlet regeneration from these nodules. LOX activity and lipid peroxides exhibited a huge increase during the first week of culture, which may indicate a role for LOX and LOX products in response to wounding in hop, as reported for other systems. Western blotting analysis showed a de novo synthesis of LOX isoenzymes in response to wounding and the detection of three different isoenzymes. Confocal analysis of LOX immunofluorescence revealed the presence of the enzyme in cortical cells of induced internodes and in prenodular cells, mostly appearing as cytoplasmic spots. Some of them were identified as lipid bodies by cytochemical and double immunofluorescence assays, suggesting the involvement of a lipid body LOX during nodule formation. Immunogold labeling detected LOX in peroxisomes, lipid bodies, and plastids of nodular cells. Quantification of the labeling density provided statistical significance for the localization of LOX (three different isoenzymes) in the three compartments, which suggested a possible involvement of LOX in metabolic functions of these organelles during organogenic nodule formation and plantlet regeneration. (J Histochem Cytochem 52:227–241, 2004)

Key Words: Humulus lupulus var. Nugget • immunolocalization • lipid peroxides • lipoxygenases • morphogenesis


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