Fallopia japonica and Fallopia × bohemica extracts cause ultrastructural and biochemical changes in root tips of radish seedlings.

Japanese knotweed (Fallopia japonica) and Bohemian knotweed (Fallopia × bohemica) are invasive plants that use allelopathy as an additional mechanism for colonization of the new habitat. Allelochemicals affect the growth of roots of neighboring plants. In the present study, we analyze the early changes associated with the inhibited root growth of radish seedlings exposed to aqueous extracts of knotweed rhizomes for 3 days. Here, we show that cells in the root cap treated with the knotweed extracts exhibited reduced cell length and displayed several ultrastructural changes, including the increased abundance of dilated ER cisternae filled with electron-dense material (ER bodies) and the accumulation of dense inclusions. Moreover, mitochondrial damage was exhibited in the root cap and the meristem zone compared to the non-treated radish seedlings. Furthermore, malfunction of the intracellular redox balance system was detected as the increased total antioxidative capacity. We also detected increased metacaspase-like proteolytic activities and, in the case of 10% extract of F. japonica, increased caspase-like proteolytic activities. These ultrastructural and biochemical effects could be the reason for the more than 60% shorter root length of treated radish seedlings compared to controls.

Oxidative Stress in Roots: Detection of Lipid Peroxidation and Total Antioxidative Capacity.

Various abiotic and biotic agents disturb the fine balance between cellular oxidants and antioxidants. The resulting oxidative stress occurs either due to the increasing levels of reactive oxygen species (ROS) or weak antioxidative system that cannot scavenge ROS burst. In addition to their harmful role, ROS can also act as signaling molecules, and oxidative stress is often the initial step in the programmed cell death. Here we describe two parameters of oxidative stress that can be measured spectrophotometrically: lipid peroxidation via the content of the by-product malondialdehyde, and the amount of all non-enzymatic antioxidants named as total antioxidative capacity. Both methods are presented using young radish (Raphanus sativus) seedlings after treatment with extract from the invasive plant species Japanese knotweed (Fallopia japonica).

Plant cell responses to allelopathy: from oxidative stress to programmed cell death.

Allelopathy is a plant-plant interaction in which one plant releases biologically active compounds that have negative effects on the fitness of the target plant. The most pronounced effects are inhibition of seed germination and growth of neighboring plants. The roots of these plants are in contact with the allelochemicals released into the soil, as the primary target of the allelopathic action. To date, the best documented allelopathic activities relate to some weeds and invasive alien plants that show rapid spread and successful growth. A better understanding of the mechanisms of allelopathy will help to improve crop production and to manage and prevent plant invasions. At the cellular level, allelochemicals induce a burst of reactive oxygen species in the target plants, which leads to oxidative stress, and can promote programmed cell death. Lipid peroxidation and cell membrane changes, protein modifications, and increased protease activities are the early signs of cell damage. When enzymatic and nonenzymatic antioxidants cannot scavenge reactive oxidants, this can result in hydrolytic or necrotic degradation of the protoplast. Cell organelles then lose their integrity and function. In roots, the structure and activity of the apical meristem are changed, which affects root growth and water absorption. Such allelopathically active compounds might thus be applied to control and manage weeds and invasive plants in a more sustainable way, to reduce chemical pollution.

Oxidative potential of ultraviolet-A irradiated or nonirradiated suspensions of titanium dioxide or silicon dioxide nanoparticles on Allium cepa roots.

The effect of ultraviolet-A irradiated or nonirradiated suspensions of agglomerates of titanium dioxide (TiO(2)) or silicon dioxide (SiO(2)) nanoparticles on roots of the onion (Allium cepa) has been studied. The reactive potential of TiO(2) nanoparticles, which have photocatalytic potential, and the nonphotocatalytic SiO(2) nanoparticles with the same size of agglomerates was compared. The authors measured the activity of antioxidant enzymes glutathione reductase, ascorbate peroxidase, guaiacol peroxidase, and catalase as well as lipid peroxidation to assess the oxidative stress in exposed A. cepa roots. A wide range of concentrations of nanoparticles was tested (0.1-1000 µg/mL). The sizes of agglomerates ranged in both cases from 300 nm to 600 nm, and the exposure time was 24 h. Adsorption of SiO(2) nanoparticles on the root surface was minimal but became significant when roots were exposed to TiO(2) agglomerates. No significant biological effects were observed even at high exposure concentrations of SiO(2) and TiO(2) nanoparticles individually. Plants appear to be protected against nanoparticles by the cell wall, which shields the cell membrane from direct contact with the nanoparticles. The authors discuss the need to supplement conventional phytotoxicity and stress end points with measures of plant physiological state when evaluating the safety of nanoparticles.

Oxidative stress in duckweed (Lemna minor L.) caused by short-term cadmium exposure.

The mechanisms of plant defence against cadmium toxicity have been studied by short-term exposure of Lemna minor L. (common duckweed) to concentrations of CdCl2 ranging from 0 to 500microM. High accumulation of cadmium was observed (12,320+/-2155microgg(-1) at 500microM CdCl2), which caused a gradual decrease of plant growth, increased lipid peroxidation, and weakened the entire antioxidative defence. Total glutathione concentration decreased significantly; however, the concentration of oxidized glutathione remained stable. The responses of four antioxidant enzymes showed that catalase was the most inhibited after CdCl2 exposure, ascorbate peroxidase and guaiacol peroxidase moderately, and glutathione reductase least. The total antioxidative potential revealed an induced antioxidative network at 0.1microM CdCl2 (137+/-13.2% of the control) and its reduction to only 47.4+/-4.0% of the control at higher cadmium concentrations. The possible application of the examined biomarkers in ecotoxicological research is discussed.

Antioxidative responses of duckweed (Lemna minor L.) to short-term copper exposure.

GOAL, SCOPE AND BACKGROUND: Elevated concentrations of copper in the environment result in accumulation of the metal in plants and cause an increase in reactive oxidative species (ROS). The first response to elevated amounts of ROS is increased levels of enzymatic and non-enzymatic antioxidants that reduce oxidative stress. The aim of our study was to evaluate the early stages of antioxidative responses to the low copper concentrations usually present in moderately polluted environments. In addition, some other parameters were examined to evaluate the effect of copper on plants. METHODS: Duckweed (Lemna minor L.) was exposed to different concentrations of copper sulphate for up to 24 hours. Glutathione concentration and enzymatic activities of catalase, guaiacol peroxidase and glutathione reductase were measured spectrophotometrically. Additionally, delayed and prompt chlorophyll fluorescence was measured by luminometry and fluorometry, respectively. The accumulation of copper in plants exposed for 24 hours to various concentrations of copper sulphate was measured by flame atomic absorption spectrophotometry. RESULTS: The treatment of plants with copper sulphate resulted in an immediate decrease of the glutathione pool, which was replenished after 24 hours at CuSO4 concentrations lower than 2 microM. Higher CuSO4 concentrations caused a decrease of reduced glutathione. The responses of the antioxidant enzymes glutathione reductase, guaiacol peroxidase and catalase to CuSO4 differed during the first six hours of exposure, but their enzyme activities all increased after 24 hours of exposure. All these enzymes displayed biphasic activity curves with maximum values between 0.5 microM and 1 microM CuSO4. The response of guaiacol peroxidase was the most pronounced and statistically significantly specific and that of catalase the least. Delayed chlorophyll fluorescence decreased after exposure to 1 microM CuSO4, but no significant effect on maximum quantum yield of photosystem II (Fv/Fm) was observed. L. minor accumulated relatively high concentrations of copper. The accumulation rate was higher at lower concentrations of copper in the test medium (up to 2 microM CuSO4) than at concentrations above 2 microM CuSO4. DISCUSSION: One of the most pronounced antioxidative responses to copper exposure was modified levels of oxidized and reduced forms of glutathione. The decrease of the glutathione pool is most probably coupled with induced production of phytochelatins. Antioxidative enzymes showed the biphasic enzyme activity characteristic of stress response. Guaiacol peroxidase exhibited the greatest significant increase of activity, even at higher CuSO4 concentrations at which the activity of catalase and glutathione reductase dropped. The intensity of delayed chlorophyll fluorescence decreased, indicating reduced photosynthesis of plants under stress. All the measured parameters showed that plants respond to even low copper concentrations very soon after exposure. The accumulation rate of copper in duckweed tissues indicates that L. minor is an accumulator species. CONCLUSIONS: The synchronized and prompt inducibility of antioxidants indicates their involvement in a general plant defence strategy for coping with metal-induced oxidative stress. Glutathione concentration and guaiacol peroxidase activity were found to be the most sensitive of the early indicators of exposure to copper concentrations present in polluted water bodies. RECOMMENDATION AND PERSPECTIVES: The experimental design of the present study allowed us to compare the sensitivity of various methods and parameters for detecting plant responses to heavy metal-induced oxidative stress. The level of glutathione and the enzyme activities of guaiacol peroxidase and glutathione reductase could be used as a rapidly determined early warning system in toxicity studies.