It is well established that every living organism spontaneously emits photons referred to as ultra-weak photon emission (synonym biophotons or low-level chemiluminescence) which inherently embodies information about the wellbeing of the source. In recent years, efforts have been made to use this feature as a non-invasive diagnostic tool related to the detection of food quality, agriculture and biomedicine. The current study deals with stress resulting from wounding (mechanical injury) on Arabidopsis thaliana and how it modifies the spontaneous ultra-weak photon emission. The ultra-weak photon emission from control (non-wounded) and stressed (wounded) plants was monitored using different modes of ultra-weak photon emission measurement sensors like charge-coupled device (CCD) cameras and photomultiplier tubes (PMT) and the collected data were analyzed to determine the level of stress generated, photon emission patterns, and underlying biochemical process. It is generally considered that electronically excited species formed during the oxidative metabolic processes are responsible for the ultra-weak photon emission. In the current study, a high-performance cryogenic full-frame CCD camera was employed for two-dimensional in-vivo imaging of ultra-weak photon emission (up to several counts/s) and the spectral analysis was done by using spectral system connected to a PMT. The results show that Arabidopsis subjected to mechanical injury enhances the photon emission and also leads to changes in the spectral pattern of ultra-weak photon emission. Thus, ultra-weak photon emission can be used as a tool for oxidative stress imaging and can pave its way into numerous plant application research.

• Klíčová slova
• Arabidopsis, mechanical injury, oxidative radical reaction, reactive oxygen species, spectral properties, ultra-weak photon emission, wounding,
• Publikační typ
• časopisecké články MeSH

Reactive oxygen species (ROS) have been recognized as important signaling compoundsof major importance in a number of developmental and physiological processes in plants. Theexistence of cellular compartments enables efficient redox compartmentalization and ensuresproper functioning of ROS-dependent signaling pathways. Similar to other organisms, theproduction of individual ROS in plant cells is highly localized and regulated bycompartment-specific enzyme pathways on transcriptional and post-translational level. ROSmetabolism and signaling in specific compartments are greatly affected by their chemicalinteractions with other reactive radical species, ROS scavengers and antioxidant enzymes. Adysregulation of the redox status, as a consequence of induced ROS generation or decreasedcapacity of their removal, occurs in plants exposed to diverse stress conditions. During stresscondition, strong induction of ROS-generating systems or attenuated ROS scavenging can lead tooxidative or nitrosative stress conditions, associated with potential damaging modifications of cellbiomolecules. Here, we present an overview of compartment-specific pathways of ROS productionand degradation and mechanisms of ROS homeostasis control within plant cell compartments.

• Klíčová slova
• cell wall, chloroplasts, cytoplasmic membrane, cytosol, glyoxysomes, mitochondria, peroxisomes, plant cell, reactive oxygen species,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Mechanical injury or wounding in plants can be attributed to abiotic or/and biotic causes. Subsequent defense responses are either local, i.e. within or in the close vicinity of affected tissue, or systemic, i.e. at distant plant organs. Stress stimuli activate a plethora of early and late reactions, from electric signals induced within seconds upon injury, oxidative burst within minutes, and slightly slower changes in hormone levels or expression of defense-related genes, to later cell wall reinforcement by polysaccharides deposition, or accumulation of proteinase inhibitors and hydrolytic enzymes. In the current study, we focused on the production of reactive oxygen species (ROS) in wounded Arabidopsis leaves. Based on fluorescence imaging, we provide experimental evidence that ROS [superoxide anion radical (O2 •-) and singlet oxygen (1O2)] are produced following wounding. As a consequence, oxidation of biomolecules is induced, predominantly of polyunsaturated fatty acid, which leads to the formation of reactive intermediate products and electronically excited species.

• Klíčová slova
• Arabidopsis, confocal microscopy, fluorescent probes, mechanical injury, wounding,
• Publikační typ
• časopisecké články MeSH

Formation of singlet oxygen (1O2) was reported to accompany light stress in plants, contributing to cell signaling or oxidative damage. So far, Singlet Oxygen Sensor Green (SOSG) has been the only commercialized fluorescent probe for 1O2 imaging though it suffers from several limitations (unequal penetration and photosensitization) that need to be carefully considered to avoid misinterpretation of the analysed data. Herein, we present results of a comprehensive study focused on the appropriateness of SOSG for 1O2 imaging in three model photosynthetic organisms, unicellular cyanobacteria Synechocystis sp. PCC 6803, unicellular green alga Chlamydomonas reinhardtii and higher plant Arabidopsis thaliana. Penetration of SOSG differs in both unicellular organisms; while it is rather convenient for Chlamydomonas it is restricted by the presence of mucoid sheath of Synechocystis, which penetrability might be improved by mild heating. In Arabidopsis, SOSG penetration is limited due to tissue complexity which can be increased by pressure infiltration using a shut syringe. Photosensitization of SOSG and SOSG endoperoxide formed by its interaction with 1O2 might be prevented by illumination of samples by a red light. When measured under controlled conditions given above, SOSG might serve as specific probe for detection of intracellular 1O2 formation in photosynthetic organisms.

• MeSH
• Arabidopsis metabolismus   MeSH
• barva MeSH
• Chlamydomonas reinhardtii metabolismus   MeSH
• fluorescenční barviva metabolismus   MeSH
• fotosyntéza fyziologie   MeSH
• kyslík metabolismus   MeSH
• oxidace-redukce MeSH
• singletový kyslík metabolismus   MeSH
• světlo MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• fluorescenční barviva MeSH
• kyslík MeSH
• singletový kyslík MeSH

Developmental transitions and stress reactions in both eukaryotes and prokaryotes are tightly linked with fast and localized modifications in concentrations of reactive oxygen and nitrogen species (ROS and RNS). Fluorescent microscopic analyses are widely applied to detect localized production of ROS and RNS in vivo. In this mini-review we discuss the biological characteristics of studied material (cell wall, extracellular matrix, and tissue complexity) and its handling (concentration of probes, effect of pressure, and higher temperature) which influence results of histochemical staining with "classical" fluorochromes. Future perspectives of ROS and RNS imaging with newly designed probes are briefly outlined.

• Klíčová slova
• cell wall, confocal microscopy, fluorescent probes, reactive nitrogen species, reactive oxygen species,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH