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

Authors :: Yadav MK
Bhatla SC
Source :: Plant physiology and biochemistry : PPB [Plant Physiol Biochem] 2011 Feb; Vol. 49 (2), pp. 230-4. Date of Electronic Publication: 2010 Dec 04.
Publication Year :: 2011
Abstract: Lipoxygenase (linoleate:oxygen oxidoreductase; EC 1.13.11.12; 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.<br /> (Copyright Â© 2010 Elsevier Masson SAS. All rights reserved.)

Subjects :: Cotyledon drug effects
Cotyledon enzymology
Fluoresceins metabolism
Helianthus drug effects
Lipid Peroxides metabolism
Masoprocol pharmacology
Propyl Gallate pharmacology
Protoplasts drug effects
Seedlings drug effects
Seedlings enzymology
Seeds drug effects
Seeds enzymology
Helianthus enzymology
Lipoxygenase metabolism
Protoplasts enzymology

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