Perception by plants of so-called microbe-associated molecular patterns (MAMPs) such as bacterial flagellin, referred to as pattern-triggered immunity, triggers a rapid transient accumulation of reactive oxygen species (ROS). We previously identified two cell wall peroxidases, PRX33 and PRX34, involved in apoplastic hydrogen peroxide (H 2 O 2 ) production in Arabidopsis (Arabidopsis thaliana). Here, we describe the generation of Arabidopsis tissue culture lines in which the expression of PRX33 and PRX34 is knocked down by antisense expression of a heterologous French bean (Phaseolus vulgaris) peroxidase cDNA construct. Using these tissue culture lines and two inhibitors of ROS generation, azide and diphenylene iodonium, we found that perxoxidases generate about half of the H 2 O 2 that accumulated in response to MAMP treatment and that NADPH oxidases and other sources such as mitochondria account for the remainder of the ROS. Knockdown of PRX33/PRX34 resulted in decreased expression of several MAMP-elicited genes, including MYB51, CYP79B2, and CYP81F2. Similarly, proteomic analysis showed that knockdown of PRX33/PRX34 led to the depletion of various MAMP-elicited defense-related proteins, including the two cysteine-rich peptides PDF2.2 and PDF2.3. Knockdown of PRX33/PRX34 also led to changes in the cell wall proteome, including increases in enzymes involved in cell wall remodeling, which may reflect enhanced cell wall expansion as a consequence of reduced H 2 O 2 -mediated cell wall cross-linking. Comparative metabolite profiling of a CaCl 2 extract of the PRX33/PRX34 knockdown lines showed significant changes in amino acids, aldehydes, and keto acids but not fatty acids and sugars. Overall, these data suggest that PRX33/PRX34-generated ROS production is involved in the orchestration of patterntriggered immunity in tissue culture cells.
1. The stag beetle, Lucanus cervus is Nationally Scarce in the UK, yet no methods exist for monitoring the abundance of adults or presence of the subterranean larvae.2. Here, we describe the design of an aerial flight interception trap that can be used to catch adults. Various lures were tested and ginger root was found to attract both sexes in equal numbers.3. Road transect surveys of adults killed by vehicles were found to produce reliable estimates of the total abundance of both sexes in areas up to about 12 km from the survey. 4. A novel use of radial diffusive samplers is described to infer the presence of larvae. Both larvae and adult females produce longifolene, which is highly attractive to males.5. Larvae produce a characteristic stridulation pattern, which can be recorded and distinguished from sounds produced by other saproxylic beetles that may co-occur with L. cervus.6. We conclude that aerial traps baited with ginger, combined with road transect surveys can be used to monitor population abundance of adults, while detection of longifolene and the characteristic stridulation pattern can be used to reveal larval presence, without destroying their fragile habitat.
It has been proposed that extensive wildfires occurred after the Cretaceous-Tertiary (K-T) impact event. An abundance of soot and pyrosynthetic polycyclic aromatic hydrocarbons (pPAHs) in marine K-T boundary impact rocks (BIRs) have been considered support for this hypothesis. However, nonmarine K-T BIRs, from across North America, contain only rare occurrences of charcoal yet abundant noncharred plant remains. pPAHs and soot can be formed from a variety of sources, including partial combustion of vegetation and hydrocarbons whereby modern pPAH signatures are traceable to their source. We present results from multiple nonmarine K-T boundary sites from North America and reveal that the K-T BIRs have a pPAH signature consistent with the combustion of hydrocarbons and not living plant biomass, providing further evidence against K-T wildfires and compelling evidence that a significant volume of hydrocarbons was combusted during the K-T impact event.
The c-Myc (Myc) oncoprotein is deregulated in a large proportion of diverse human cancers. Considerable effort has therefore been directed at identifying pharmacologic inhibitors as potential anti-neoplastic agents. Three such groups of small molecule inhibitors have been described. The first is comprised of so-called “direct” inhibitors, which perturb Myc's ability to form productive DNA-binding heterodimers in association with its partner, Max. The second group is comprised of indirect inhibitors, which largely function by targeting the BET-domain protein BRD4 to prevent the proper formation of transcriptional complexes that assemble in response to Myc-Max DNA binding. Thirdly, synthetic lethal inhibitors cause the selective apoptosis of Myc over-expressing either by promoting mitotic catastrophe or altering Myc protein stability. We report here a common mechanism by which all Myc inhibitors, irrespective of class, lead to eventual cellular demise. This involves the depletion of ATP stores due to mitochondrial dysfunction and the eventual down-regulation of Myc protein. The accompanying metabolic de-regulation causes neutral lipid accumulation, cell cycle arrest, and an attempt to rectify the ATP deficit by up-regulating AMP-activated protein kinase (AMPK). These responses are ultimately futile due to the lack of functional Myc to support the requisite anabolic response. Finally, the effects of Myc depletion on ATP levels, cell cycle arrest, differentiation and AMPK activation can be mimicked by pharmacologic inhibition of the mitochondrial electron transport chain without affecting Myc levels. Thus, all Myc inhibitors promote a global energy collapse that appears to underlie many of their phenotypic consequences.
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