Cyanobacterial flavodoxin complements ferredoxin deficiency in knocked-down transgenic tobacco plants

División Biología Molecular, Facultad de Ciencias Bioquímicas y Farmacéuticas, Instituto de Biología Molecular y Celular de Rosario (IBR, CONICET), Universidad Nacional de Rosario, S2002LRK Rosario, Argentina.
The Plant Journal (Impact Factor: 6.82). 12/2010; 65(6):922-35. DOI: 10.1111/j.1365-313X.2010.04479.x
Source: PubMed

ABSTRACT Ferredoxins are the main electron shuttles in chloroplasts, accepting electrons from photosystem I and delivering them to essential oxido-reductive pathways in the stroma. Ferredoxin levels decrease under adverse environmental conditions in both plants and photosynthetic micro-organisms. In cyanobacteria and some algae, this decrease is compensated for by induction of flavodoxin, an isofunctional flavoprotein that can replace ferredoxin in many reactions. Flavodoxin is not present in plants, but tobacco lines expressing a plastid-targeted cyanobacterial flavodoxin developed increased tolerance to environmental stress. Chloroplast-located flavodoxin interacts productively with endogenous ferredoxin-dependent pathways, suggesting that its protective role results from replacement of stress-labile ferredoxin. We tested this hypothesis by using RNA antisense and interference techniques to decrease ferredoxin levels in transgenic tobacco. Ferredoxin-deficient lines showed growth arrest, leaf chlorosis and decreased CO(2) assimilation. Chlorophyll fluorescence measurements indicated impaired photochemistry, over-reduction of the photosynthetic electron transport chain and enhanced non-photochemical quenching. Expression of flavodoxin from the nuclear or plastid genome restored growth, pigment contents and photosynthetic capacity, and relieved the electron pressure on the electron transport chain. Tolerance to oxidative stress also recovered. In the absence of flavodoxin, ferredoxin could not be decreased below 45% of physiological content without fatally compromising plant survival, but in its presence, lines with only 12% remaining ferredoxin could grow autotrophically, with almost wild-type phenotypes. The results indicate that the stress tolerance conferred by flavodoxin expression in plants stems largely from functional complementation of endogenous ferredoxin by the cyanobacterial flavoprotein.

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Available from: Michael Melzer, Sep 15, 2014
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    • "Flds can be used as biotechnological tools to engineer tolerance to oxidative stress. Transgenic tobacco plants that constitutively express a cyanobacterial Fld exhibit enhanced tolerance to a broad range of stresses that provoke oxidative damage (Tognetti et al. 2006, 2007a,b; Blanco et al. 2011; Ceccoli et al. 2011), while tolerance of nitrogen-fixing activity to salinity is enhanced in transgenic Medicago truncatula plants expressing Fld (Coba de la Pe~ na et al. 2010). Moreover, overexpression of flavodoxin in the heterotrophic bacterium Ensifer meliloti protects free-living bacteria against cadmium stress (Shvaleva et al. 2010), while expression of a heterologous Fld in symbiotic forms of E. meliloti ameliorates the oxidative balance, thus delaying natural nodule senescence (Redondo et al. 2009) and enhancing the tolerance of alfalfa nodules to cadmium (Shvaleva et al. 2010) and saline stress (Redondo et al. 2012). "
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    ABSTRACT: AIM: To determine whether expression of a cyanobacterial flavodoxin in soil bacteria of agronomic interest confers protection against the widely used herbicides paraquat and atrazine. METHODS AND RESULTS: The model bacterium Escherichia coli, the symbiotic nitrogen-fixing bacterium Ensifer meliloti and the plant growth-promoting rhizobacterium Pseudomonas fluorescens Aur6 were transformed with expression vectors containing the flavodoxin gene of Anabaena variabilis. Expression of the cyanobacterial protein was confirmed by Western blot. Bacterial tolerance to oxidative stress was tested in solid medium supplemented with hydrogen peroxide, paraquat or atrazine. In all three bacterial strains, flavodoxin expression enhanced tolerance to the oxidative stress provoked by hydrogen peroxide and by the ROS-inducing herbicides, witnessed by the enhanced survival of the transformed bacteria in the presence of these oxidizing agents. CONCLUSIONS: Flavodoxin overexpression in beneficial soil bacteria confers tolerance to oxidative stress and improves their survival in the presence of the herbicides paraquat and atrazine. Flavodoxin could be considered as a general antioxidant resource to face oxidative challenges in different microorganisms. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of plant growth-promoting rhizobacteria or nitrogen-fixing bacteria with enhanced tolerance to oxidative stress in contaminated soils is of significant agronomic interest. The enhanced tolerance of flavodoxin-expressing bacteria to atrazine and paraquat points to potential applications in herbicide-treated soils. This article is protected by copyright. All rights reserved.
    Journal of Applied Microbiology 04/2013; DOI:10.1111/jam.12224 · 2.39 Impact Factor
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    • "Flavodoxin can replace ferredoxin in the photosynthetic electron transport from photosystem I to NADP ? and in nitrogenase reduction (Sandmann et al. 1990). Blanco et al. (2011) have shown that cyanobacterial flavodoxin expression can complement ferredoxin deficiency in transgenic ferredoxindeficient tobacco lines. In alfalfa nodules, flavodoxin might increase effectiveness of the nitrogenase complex by compensating the ferredoxin decline that occurs under osmotic stress (Zimmermann et al. 2004; Tognetti et al. 2006). "
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    ABSTRACT: Nitrogen fixation by legumes is very sensitive to salinity stress, which can severely reduce the productivity of legume crops and their soil-enriching capacity. Salinity is known to cause oxidative stress in the nodule by generating reactive oxygen species (ROS). Flavodoxins are involved in the response to oxidative stress in bacteria and cyanobacteria. Prevention of ROS production by flavodoxin overexpression in bacteroids might lead to a protective effect on nodule functioning under salinity stress. Tolerance to salinity stress was evaluated in alfalfa nodules elicited by an Ensifer meliloti strain that overexpressed a cyanobacterial flavodoxin compared with nodules produced by the wild-type bacteria. Nitrogen fixation, antioxidant and carbon metabolism enzyme activities were determined. The decline in nitrogenase activity associated to salinity stress was significantly less in flavodoxin-expressing than in wild-type nodules. We detected small but significant changes in nodule antioxidant metabolism involving the ascorbate-glutathione cycle enzymes and metabolites, as well as differences in activity of the carbon metabolism enzyme sucrose synthase, and an atypical starch accumulation pattern in flavodoxin-containing nodules. Salt-induced structural and ultrastructural alterations were examined in detail in alfalfa wild-type nodules by light and electron microscopy and compared to flavodoxin-containing nodules. Flavodoxin reduced salt-induced structural damage, which primarily affected young infected tissues and not fully differentiated bacteroids. The results indicate that overexpression of flavodoxin in bacteroids has a protective effect on the function and structure of alfalfa nodules subjected to salinity stress conditions. Putative protection mechanisms are discussed.
    Planta 08/2012; 236(6). DOI:10.1007/s00425-012-1725-8 · 3.38 Impact Factor
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    • "The decline in Fd amounts leads to overreduction of the PETC due to shortage of electron acceptors. Under such circumstances the electron surplus could be passed straight to oxygen resulting in ROS generation [39] [40] [41]. Iron–sulfur centers are vulnerable to ROS attack to various extents , depending on solvent exposure and the polypeptide environment surrounding the cluster. "
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    ABSTRACT: Ferredoxins are electron shuttles harboring iron-sulfur clusters which participate in oxido-reductive pathways in organisms displaying very different lifestyles. Ferredoxin levels decline in plants and cyanobacteria exposed to environmental stress and iron starvation. Flavodoxin is an isofunctional flavoprotein present in cyanobacteria and algae (not plants) which is induced and replaces ferredoxin under stress. Expression of a chloroplast-targeted flavodoxin in plants confers tolerance to multiple stresses and iron deficit. We discuss herein the bases for functional equivalence between the two proteins, the reasons for ferredoxin conservation despite its susceptibility to aerobic stress and for the loss of flavodoxin as an adaptive trait in higher eukaryotes. We also propose a mechanism to explain the tolerance conferred by flavodoxin when expressed in plants.
    FEBS letters 07/2012; 586(18):2917-24. DOI:10.1016/j.febslet.2012.07.026 · 3.34 Impact Factor
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