August 4, 2006

Lipoxygenases during Brassica napus seed germination

The peroxidation of polyunsaturated fatty acids is mostly catalyzed by members of the lipoxygenase enzyme family. Lipoxygenase products can be metabolized further in the oxylipin pathway and are known as signalling substances that play a role in plant development as well as in plant responses to wounding and pathogen attack. Apart from accumulating data in model plants like Arabidopsis, information on the relevance of lipid peroxide metabolism in the crop plant oilseed rape is scarce. Thus we aimed to analyze lipoxygenases and oxylipin patterns in seedlings of oilseed rape. RNA isolated from 3 day etiolated seedlings contains mRNAs for at least two different lipoxygenases. These have been cloned as cDNAs and named Bn-Lox-lfl and Bn-Lox-2fl. The protein encoded by Bn-Lox-2fl was identified as a 13-lipoxygenase by expression in Escherichia coli. The Bn-Lox-lfl yielded an inactive protein when expressed in E. coli. Based on Bn-Lox-lfl active site determinants and on sequence homology the Bn-Lox-lfl is most likely a 9-lipoxygenase. Both genes are expressed in light-grown and etiolated cotyledons as well as in leaves. Bn-Lox-2fl protein is more abundant in cotyledons of etiolated seedlings than in cotyledons of green seedlings. Both 13- and 9-lipoxygenase-derived hydroperoxides can be detected during germination. Etiolated seedlings contain more lipoxygenase-derived hydroperoxides in non esterified fatty acids than green seedlings. The 13-lipoxygenase derivatives are 6-8-fold more abundant than the 9-derivatives. Lipoxygenase-derived hydroperoxides in esterified lipids are almost not present during germination. These results suggest that 13-lipoxygenases acting on free fatty acids dominate during B. napus seed germination.

Source: Phytochemistry (2006) vol. 67, p. 2030-2040

June 23, 2006

Over-expression of chloroplastic lipoxygenase RCI1 causes PR1 transcript accumulation in transiently transformed rice

The acquired resistance state of rice plants, rendering them resistant to subsequent infections with virulent pathogens, can be triggered by treatments with non-host pathogens, necrotizing pathogens and certain chemicals. The 13-lipoxygenase RCI1 (LOX2:Os:2) is induced specifically following treatment with chemical inducers of resistance. Here, we report that the over-expression of RCI1 in a transient transformation assay leads to PR1 transcript accumulation in rice leaves. In addition, we show that this property is due to the enzymatic activity of the transgenic protein. Furthermore, exogenous application of (13S,92,11E,15Z)-13-hydroxy-9,11,15-octadecatrienoic acid (13-HOT), an oxylipin deriving from the reductase branch of the lipoxygenase pathway, is capable of inducing PR1. These results support a role for RCI1 in the generation of signalling molecules during the establishment of the acquired resistance response in rice.

Source: Physiological and molecular plant pathology (2004) vol. 64, p. 37-43

April 28, 2006

The outcomes of concentration-specific interactions between salicylate and jasmonate signaling include synergy, antagonism, and oxidative...

The outcomes of concentration-specific interactions between salicylate and jasmonate signaling include synergy, antagonism, and oxidative stress leading to cell death

Salicylic acid (SA) has been proposed to antagonize jasmonic acid (JA) biosynthesis and signaling. We report, however, that in salicylate hydroxylase-expressing tobacco (Nicotiana tabacum) plants, where SA levels were reduced, JA levels were not elevated during a hypersensitive response elicited by Pseudomonas syringae pv phaseolicola. The effects of cotreatment with various concentrations of SA and JA were assessed in tobacco and Arabidopsis (Arabidopsis thaliana). These suggested that there was a transient synergistic enhancement in the expression of genes associated with either JA (PDF1.2 [defensin] and Thi1.2 [thionin]) or SA (PR1 [PR1a--glucuronidase in tobacco]) signaling when both signals were applied at low (typically 10–100 µM) concentrations. Antagonism was observed at more prolonged treatment times or at higher concentrations. Similar results were also observed when adding the JA precursor, -linolenic acid with SA. Synergic effects on gene expression and plant stress were NPR1- and COI1-dependent, SA- and JA-signaling components, respectively. Electrolyte leakage and Evans blue staining indicated that application of higher concentrations of SA + JA induced plant stress or death and elicited the generation of apoplastic reactive oxygen species. This was indicated by enhancement of hydrogen peroxide-responsive AoPR10--glucuronidase expression, suppression of plant stress/death using catalase, and direct hydrogen peroxide measurements. Our data suggests that the outcomes of JA-SA interactions could be tailored to pathogen/pest attack by the relative concentration of each hormone.

Source: Plant Physiology (2006) vol. 140, p. 249-262