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Plant Physiol Biochem. 2011 Feb;49(2):230-4. doi: 10.1016/j.plaphy.2010.11.012. Epub 2010 Dec 4.

Localization of lipoxygenase activity on the oil bodies and in protoplasts using a novel fluorescence imaging method.

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Laboratory of Plant Physiology, Department of Botany, University of Delhi, Delhi 110007, India.


Lipoxygenase (linoleate:oxygen oxidoreductase; EC; LOX) catalyzes oxygenation of polyenoic fatty acids, which precedes the degradation of storage lipids during seed germination in sunflower. In the present work, it has been confirmed that 2',7'-dichlorodihydrofluorescein diacetate (H(2)DCFDA) produces fluorescence in presence of lipid hydroperoxides (LOX reaction products). This work provides new information on spatial localization of transiently enhanced LOX activity in protoplasts from 5 d old seedling cotyledons of sunflower (Helianthus annuus L. cv. Morden) by exploiting H(2)DCFDA as a probe for fluorescence detection from LOX activity sites. Use of LOX inhibitors [nordihydroguaiaretic acid (NDGA) and propyl gallate (PG)] confirms oil bodies as LOX activity sites. Oil body surface has been shown to possess LOX activity in 5 d old seedling cotyledons.

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