Fluorescent in vivo imaging of reactive oxygen species and redox potential in plants

Reactive oxygen species (ROS) are by-products of aerobic metabolism, and excessive production can result in oxidative stress and cell damage. In addition, ROS function as cellular messengers, working as redox regulators in a multitude of biological processes. Understanding ROS signalling and stress responses requires methods for precise imaging and quantification to monitor local, subcellular and global ROS dynamics with high selectivity, sensitivity and spatiotemporal resolution. In this review, we summarize the present knowledge for in vivo plant ROS imaging and detection, using both chemical probes and fluorescent protein-based biosensors. Certain characteristics of plant tissues, for example high background autofluorescence in photosynthetic organs and the multitude of endogenous antioxidants, can interfere with ROS and redox potential detection, making imaging extra challenging. Novel methods and techniques to measure in vivo plant ROS and redox changes with better selectivity, accuracy, and spatiotemporal resolution are therefore desirable to fully acknowledge the remarkably complex plant ROS signalling networksThis work was funded by a grant from the Spanish Ministry of Economy and Competitiveness ( AGL2014–53771-R ). Alfonso Blázquez-Castro acknowledges funding under the Marie Skłodowska-Curie Action COFUND 2015 (EU project 713366 – InterTalentum

The Early Oxidative Stress Induced by Mercury and Cadmium Is Modulated by Ethylene in Medicago sativa Seedlings

Cadmium (Cd) and mercury (Hg) are ubiquitous soil pollutants that promote the accumulation of reactive oxygen species, causing oxidative stress. Tolerance depends on signalling processes that activate different defence barriers, such as accumulation of small heat sock proteins (sHSPs), activation of antioxidant enzymes, and the synthesis of phytochelatins (PCs) from the fundamental antioxidant peptide glutathione (GSH), which is probably modulated by ethylene. We studied the early responses of alfalfa seedlings after short exposure (3, 6, and 24 h) to moderate to severe concentration of Cd and Hg (ranging from 3 to 30 μM), to characterize in detail several oxidative stress parameters and biothiol (i.e., GSH and PCs) accumulation, in combination with the ethylene signalling blocker 1-methylcyclopropene (1-MCP). Most changes occurred in roots of alfalfa, with strong induction of cellular oxidative stress, H2O2 generation, and a quick accumulation of sHSPs 17.6 and 17.7. Mercury caused the specific inhibition of glutathione reductase activity, while both metals led to the accumulation of PCs. These responses were attenuated in seedlings incubated with 1-MCP. Interestingly, 1-MCP also decreased the amount of PCs and homophytochelatins generated under metal stress, implying that the overall early response to metals was controlled at least partially by ethylen

Tropical Grass Growth Functions Modeling by Using Nonlinear Mixed Models

Nonlinear Growth curves are used for modeling plant physiological variables. These models are preferable because the polynomial coefficients of the equations have a biological significance. The response variables of the curves occurs commonly with repeated measurements over time and measurements are on different environments. The traditional statistical analysis does not include a repeated measures approach, which can lead to improper estimation of the error terms. It is important to study the growth of tropical grass (Da Silva and Carvalho 2005)

Mediterranean-type diet and brain structural change from 73 to 76 years in a Scottish cohort

STUDY FUNDING The data were collected by a Research into Ageing programme grant; research continues as part of the Age UK–funded Disconnected Mind project. The work was undertaken by The University of Edinburgh Centre for Cognitive Ageing and Cognitive Epidemiology, part of the cross-council Lifelong Health and Wellbeing Initiative (MR/K026992/1), with funding from the BBSRC and Medical Research Council. Imaging and image analysis was performed at the Brain Research Imaging Centre (sbirc.ed.ac.uk/), Edinburgh, supported by the Scottish Funding Council SINAPSE Collaboration. Derivation of mean cortical thickness measures was funded by the Scottish Funding Council’s Postdoctoral and Early Career Researchers Exchange Fund awarded by SINAPSE to David Alexander Dickie. L.C.A.C. acknowledges funding from the Scottish Government's Rural and Environment Science and Analytical Services (RESAS) division.Peer reviewedPublisher PD

Synchrotron Radiation-Fourier transformed infrared microspectroscopy (μSR-FTIR) reveals multiple metabolism alterations in microalgae induced by cadmium and mercury

Toxic metals such as cadmium (Cd) and mercury (Hg) represent a threat to photosynthetic organisms of polluted aquatic ecosystems, and knowledge about mechanisms of toxicity is essential for appropriate assessment of environmental risks. We used Synchrotron Radiation-Fourier Transformed Infrared microspectroscopy (μSR-FTIR) to characterise major changes of biomolecules caused by Cd and Hg in the model green microalga Chlamydomonas reinhardtii. μSR-FTIR showed several metabolic alterations in different biochemical groups such as carbohydrates, proteins, and lipids in a time-dose dependent manner, with the strongest changes occurring at concentrations above 10 μM Cd and 15 μM Hg after short-term (24 h) treatments. This occurred in a context where metals triggered intracellular oxidative stress and chloroplast damage, along with autophagy induction by overexpressing AUTOPHAGY-RELATED PROTEIN 8 (ATG8). Thin layer chromatography analysis confirmed that toxic metals promoted remarkable changes in lipid profile, with higher degree of esterified fatty acid unsaturation as detected by gas chromatography coupled with mass spectrometry. Under Cd stress, there was specifically higher unsaturation of free fatty acids, while Hg led to stronger unsaturation in monogalactosyldiacylglycerol. μSR-FTIR spectroscopy proved as a valuable tool to identify biochemical alterations in microalgae, information that could be exploited to optimise approaches for metal decontaminationWork supported by the Spanish State Research Agency (AEI) (Spain) (projects AGL2014–53771-R and AGL2017–87591-R). The FTIR experiments were performed at MIRAS beamline at ALBA Synchrotron with the collaboration of ALBA staff and supported by travel grants for experiments No 2016091860 and 201702211

Biological activity of Ipomoea pauciflora Martens and Galeotti (Convolvulaceae) extracts and fractions on larvae of Spodoptera frugiperda J. E. Smith (Lepidoptera: Noctuidae)

Hexane, chloroform and methanol extracts of different parts of Ipomoea pauciflora were tested for their effects on the survival and development of Fall Armyworm (Spodoptera frugiperda), a Lepidoptera pest. For seven days, neonatal larvae (grown at 27 ± 2°C with a 16: 8 (L: D) h photoperiod) were exposed to different concentrations of crude I. pauciflora extracts (ranging from 0 to 4 mg/ml) that were incorporated into an artificial diet. Surviving larvae were weighed at days 6, 9 and 13 and were maintained until moths emerged. Eleven of the 18 crude extracts showed more than 30% larval mortality. The highest mortality was produced by hexane and chloroform extracts of seeds at 4 mg/ml(96.9 and 93.8%, respectively), with LC50 values of 1.85 mg/ml and 0.54 mg/ml, respectively. Fractions of both seed extracts were isolated by gravity column chromatography over silica gel and analyzed for their active compounds. Eight fractions of the hexane extract and six fractions of the chloroform extract from I. pauciflora seeds, exhibited larvicidal effects at 1 mg/ml (mortality from 33.3 to 88.9% and from 47.2 to 77%, respectively). Changes in larval weight were observed as compared with the control group. Phytochemical analysis through GC-MS and H1 NMR revealed the presence of fatty acids and aldehydes in the active fractions. These results indicate that the bioactive extracts from the seed of I. pauciflora can induce lethal toxicity in S. frugiperda larvae or affect the weight of the surviving larvae

The effect of nutritional condition on the growth to post-flexion of bluefin tuna larvae under cultured condition

These findings indicate that protein synthesis increase in proportion to the flexion stage which is strongly associated with the development of the digestive system and therefore the increase in the nutritional condition. Ontogenetic differences in the RNA and DNA content will be discussed in more detail. Also, the pattern obtained for Atlantic bluefin tuna larva will be compared with those of other species