Functional analysis of oxidative stress-activated mitogen-activated protein kinase cascade in plants

Kovtun, Yelena; Chiu, Wan-Ling; Tena, Guillaume; Sheen, Jen

doi:10.1073/pnas.97.6.2940

Cited by 1,397 publications

(1,038 citation statements)

References 75 publications

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“…A signalling role of ROS from the chloroplasts, involving induction of nuclear genes encoding defence related proteins, is well established (reviewed in Nott et al 2006;Pfannschmidt et al 2009), with H 2 O 2 as the prime candidate. Excess H 2 O 2 , produced by stress (or in our case elevated SOD activity) can diffuse from the chloroplast and, probably through mitogen-activated protein kinases (Kovtun et al 2000), trigger expression of nuclear encoded defence-related genes such as cAPX encoding an ascorbate peroxidise (Kimura et al 2001), Cat encoding catalase and Gst1 encoding Glutathione-S-transferase (Polidoros and Scandalios 1999), as well (in the C 3 -CAM transition species Mesembryanthemum crystallinum) as genes encoding Cu/Zn-, Mn-, and Fe-SOD (Slesak et al 2003). The current results suggest that increases in H 2 O 2 levels caused by chloroplastic expression of MnSOD, mimic abiotic and biotic stress responses, leading to a similar MAPK induced expression of nuclear-encoded defence proteins, including several organellar SODs, in these transplastomic tobacco lines.…”

Section: Discussionmentioning

confidence: 83%

Modification of reactive oxygen species scavenging capacity of chloroplasts through plastid transformation

et al. 2011

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Reactive oxygen species (ROS), including superoxide anions, hydrogen peroxide and hydroxyl radicals are generated through normal biochemical processes, but their production is increased by abiotic stresses. The prospects for enhancing ROS scavenging, and hence stress tolerance, by direct gene expression in a vulnerable cell compartment, the chloroplast, have been explored in tobacco. Several plastid transformants were generated which contained either a Nicotiana mitochondrial superoxide dismutase (MnSOD) or an Escherichia coli glutathione reductase (gor) gene. MnSOD lines had a three-fold increase in MnSOD activity, but interestingly a five to nine-fold increase in total chloroplast SOD activity. Gor transgenic lines had up to 6 times higher GR activity and up to 8 times total glutathione levels compared to wild type tobacco. Photosynthetic capacity of transplastomic plants, as measured by chlorophyll content and variable fluorescence of PSII was equivalent to non-transformed plants.The response of these transplastomic lines to several applied stresses was examined. In a number of cases improved stress tolerance was observed. Examples include enhanced methyl viologen (Paraquat)-induced oxidative stress tolerance in Mn-superoxidase dismutase overexpressing plants, improved heavy metal tolerance in glutathione reductase expressing lines, and improved tolerance to UV-B radiation in both sets of plants.

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Section: Discussionmentioning

confidence: 83%

Modification of reactive oxygen species scavenging capacity of chloroplasts through plastid transformation

et al. 2011

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“…Since Arabidopsis became the central model system in plant biology with the availability of its whole genome sequence, we and several other laboratories have established the TEAMP system as a cellbased functional genomics tool 1,5 .…”

Section: Overviewmentioning

confidence: 99%

“…Both abiotic and biotic stress signaling can also be studied in the TEAMP system. For example, the functions of the evolutionarily conserved mitogen-activated protein kinase (MAPK) cascades have been identified and characterized in Arabidopsis mesophyll protoplasts for oxidative stress signaling 5 and innate immune responses 15 . Subsequently, the virulence functions of type III effectors, AvrPto and AvrPtoB from the pathogenic bacterium Pseudomonas syringae, have been shown to be potent suppressors of the MAPK-dependent plant innate immunity 16 .…”

Section: Applicationsmentioning

confidence: 99%

Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis

2007

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The transient gene expression system using Arabidopsis mesophyll protoplasts has proven an important and versatile tool for conducting cell-based experiments using molecular, cellular, biochemical, genetic, genomic and proteomic approaches to analyze the functions of diverse signaling pathways and cellular machineries. A well-established protocol that has been extensively tested and applied in numerous experiments is presented here. The method includes protoplast isolation, PEG-calcium transfection of plasmid DNA and protoplast culture. Physiological responses and high-throughput capability enable facile and cost-effective explorations as well as hypothesis-driven tests. The protoplast isolation and DNA transfection procedures take 6-8 h, and the results can be obtained in 2-24 h. The cell system offers reliable guidelines for further comprehensive analysis of complex regulatory mechanisms in whole-plant physiology, immunity, growth and development. INTRODUCTIONPlant protoplasts without cell walls offer a versatile cell-based experimental system. Macromolecules such as DNA, RNA and proteins can be delivered into protoplasts using various methods, e.g., PEG-calcium fusion, electroporation and microinjection. Signal transduction and metabolic pathways as well as transcription and translation machineries can be transiently manipulated to investigate cell-autonomous regulation and responses. The method is especially powerful in functional genomics analysis when combined with rich genetic resources, such as insertion libraries, available in the model plant Arabidopsis thaliana. We describe here a protocol for a system of transient expression in Arabidopsis mesophyll protoplast (TEAMP) and provide troubleshooting advice and detailed information on useful reporter constructs. Applications, advantages and limitations of the TEAMP assay are also discussed. More general information about plant protoplasts can be found in a review 1 and a live performance of the TEAMP procedure can be seen at the Sheen lab website (http://genetics. mgh.harvard.edu/sheenweb/protocols_reg.html).

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“…The auxindependent activation of cell proliferation in the root meristem is associated with oxidation which is maximal in the quiescent centre cells, suggesting that cell proliferation is influenced by cellular redox state (69,70). Redox controls also play a key part in the regulation of cell cycle progression (87,88). In particular, the G1 and G2 checkpoints that regulate the cell cycle are highly responsive to oxidation (89).…”

Section: Gsh Is Required For the Cell Cycle In Rootsmentioning

confidence: 99%

Glutathione – linking cell proliferation to oxidative stress

Díaz‐Vivancos

Simone

Kiddle³

et al. 2015

Free Radical Biology and Medicine

278

177

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Significance:The multifaceted functions of reduced glutathione (gamma-glutamyl-cysteinyl-glycine; GSH) continue to fascinate plants and animal scientists, not least because of the dynamic relationships between GSH and reactive oxygen species (ROS) that underpin reduction/oxidation (redox) regulation and signalling. Here we consider the respective roles of ROS and GSH in the regulation of plant growth, with a particular focus on regulation of the plant cell cycle. Glutathione is discussed not only as a crucial low molecular weight redox buffer that shields nuclear processes against oxidative challenge but also a flexible regulator of genetic and epigenetic functions. Recent Advances:The intracellular compartmentalization of GSH during the cell cycle is remarkably consistent in plants and animals. Moreover, measurements of in vivo glutathione redox potentials reveal that the cellular environment is much more reducing than predicted from GSH/GSSG ratios measured in tissue extracts. The redox potential of the cytosol and nuclei of non-dividing plant cells is about -300 mV. This relatively low redox potential is maintained even in cells experiencing oxidative stress by a number of mechanisms including vacuolar sequestration of GSSG. We propose that regulated ROS production linked to glutathione-mediated signalling events are the hallmark of viable cells within a changing and challenging environment. Critical Issues:The concept that the cell cycle in animals is subject to redox controls is well established but little is known about how ROS and GSH regulate this process in plants. However, it is increasingly likely that similar redox controls exist in plants, 3 although possibly through different pathways. Moreover, redox-regulated proteins that function in cell cycle checkpoints remain to be identified in plants. While GSHresponsive genes have now been identified, the mechanisms that mediate and regulate protein glutathionylation in plants remain poorly defined.Future Directions: The nuclear GSH pool provides an appropriate redox environment for essential nuclear functions. Future work will function on how this essential thiol interacts with the nuclear thioredoxin system and nitric oxide to regulate genetic and epigenetic mechanisms. The characterization of redox-regulated cell cycle proteins in plants, and the elucidation of mechanisms that facilitate GSH accumulation in the nucleus are keep steps to unravelling the complexities of nuclear redox controls.Diaz 4

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Functional analysis of oxidative stress-activated mitogen-activated protein kinase cascade in plants

Cited by 1,397 publications

References 75 publications

Modification of reactive oxygen species scavenging capacity of chloroplasts through plastid transformation

Modification of reactive oxygen species scavenging capacity of chloroplasts through plastid transformation

Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis

Glutathione – linking cell proliferation to oxidative stress

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