Oxidative stress in wild‐derived and cultivated peanut genotypes caused by heat stress at flowering

The elucidation of heat tolerance mechanisms is required to combat the challenges of global warming. This study aimed to determine the antioxidant enzyme responses to heat stress, at the enzymatic activity and gene expression levels, and to investigate the antioxidative alterations associated with heat tolerance in the stems and roots of squashes using three genotypes differing in heat tolerance. Plants of heat-tolerant “C. moschata”, thermolabile “C. maxima” and moderately heat-tolerant interspecific inbred line “Maxchata” genotypes were exposed to moderate (37 °C) and severe (42 °C) heat shocks. “C. moschata” exhibited comparatively little oxidative damage, with the lowest hydrogen peroxide (H2O2), superoxide (O2−) and malondialdehyde (MDA) contents in the roots compared to stems, followed by “Maxchata”. The enzyme activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and peroxidase (POD) were found to be increased with heat stress in tolerant genotypes. The significant inductions of FeSOD, MnSOD, APX2, CAT1 and CAT3 isoforms in tolerant genotypes suggested their participation in heat tolerance. The differential isoform patterns of SOD, APX and CAT between stems and roots also indicated their tissue specificity. Furthermore, despite the sequence similarity of the studied antioxidant genes among “C. maxima” and “Maxchata”, most of these genes were highly induced under heat stress in “Maxchata”, which contributed to its heat tolerance. This phenomenon also indicated the involvement of other unknown genetic and/or epigenetic factors in controlling the expression of these antioxidant genes in squashes, which demands further exploration.

This study leads to advances in the field of heat tolerance among different plant species. We concluded that a coordinated, increased antioxidant defense system enabled white clover to reduce heat-induced oxidative damage. The rise in global ambient temperature has a wide range of effects on plant growth, and, therefore, on the activation of various molecular defenses before the appearance of heat damage. Elevated temperatures result in accelerated generation of reactive oxygen species (ROS), causing an imbalance between ROS production and the ability of scavenging systems to detoxify and remove the reactive intermediates. The aim of this study was to determine the role of antioxidant defense systems in the alleviation of heat stress (HS) consequences in white clover (Trifolium repens L.), which is cultivated worldwide. We evaluated how temperature and time parameters contribute to the thermotolerance of white clover at different growth stages. We revealed HS protection in white clover from 37 to 40°C, with 40°C providing the greatest protection of 3-day-old seedlings and 28-day-old adult plants. Heat-provoked oxidative stress in white clover was confirmed by substantial changes in electrolyte leakage, malondialdehyde (MDA), and chlorophyll content, as well as superoxide anion (O2·-) and hydrogen peroxide (H2O2) production. Furthermore, superoxide dismutase (SOD) and ascorbate peroxidase (APX) as well as a high level of GSH non-enzymatic antioxidant were the most responsive, and were associated with acquired thermotolerance through the regulation of ROS generation.We demonstrated, by studying protoplast transient gene expression, direct genetic evidence of endogenous antioxidant-related genes that confer HS tolerance in white clover. Our present study clearly establishes that oxidative stress ensues from HS, which triggers the induction of antioxidant defense systems for ROS scavenging in white clover.

Photosystem II (PSII) is sensitive to heat stress which can overproduce reactive oxygen species (ROS). To investigate the underlying mechanism of exogenously-supplied trehalose to improve photosynthesis and heat tolerance, in this study, we initially determined the antioxidants to get rid of ROS and examined the effects of trehalose pretreatment on D1 protein. Our results show that heat (40°C) stress decreased PSII activity and D1 protein content. However, trehalose pretreatment enhanced PSII activity and the content of D1 protein during heat stress. In the presence of streptomycin, an inhibitor of D1 protein synthesis, the maximal photochemical efficiency of PSII (Fv/Fm) in trehalose pretreated seedlings was enhanced. Hydrogen peroxide content and malondialdehyde content in trehalose pretreated seedlings were less than those in control plants under heat conditions. Consistently, the ratio of reduced ascorbate to dehydroascorbic acid, the activities of catalase (CAT) and ascorbate peroxidase (APX) and the gene expressions of antioxidant enzymes including superoxide dismutase, CAT, peroxidase and APX were increased in trehalose pretreated plants compared to control seedlings. These results suggest that exogenously applied trehalose may play a key role in the recovery of decreased synthesis of D1 protein and protect it from heat-induced photoinhibition in wheat (Triticum aestivum L.) by decreasing the production of ROS and reducing membrane lipid peroxidation via the change of antioxidants including APX, CAT and reduced ascorbate and by increasing gene transcript level of related antioxidant enzymes during heat stress.

Understanding antioxidant mechanisms for heat stress is important for improving heat tolerance in cool-season plant species. The objective of this study was to identify antioxidant enzymes associated with cultivar variations in heat tolerance in kentucky bluegrass (Poa pratensis) by comparing heat responses of activity and isoforms of antioxidant enzymes in two cultivars contrasting in heat tolerance. Plants of heat-tolerant ‘Eagleton’ and heat-sensitive ‘Brilliant’ were exposed to 20 °C (control) or 40 °C (heat stress) for 28 days in growth chambers. Chlorophyll (Chl) a content remained unchanged and Chl b content increased in ‘Eagleton’, while both of them decreased in ‘Brilliant’, and by 28 days, ‘Eagleton’ had significantly higher Chl a and b content than ‘Brilliant’. The activities of superoxide dismutase (SOD) were significantly higher in ‘Eagleton’ than in ‘Brilliant’ by 28 days of heat stress. An isozyme SOD2 was induced early during heat stress in ‘Eagleton’, while isozyme SOD3 degraded, to a lesser extent in ‘Eagleton’ than in ‘Brilliant’. Catalase (CAT) activity significantly increased in ‘Brilliant’ but remained constant in ‘Eagleton’, and ‘Brilliant’ had a significantly higher CAT activity and isozyme CAT1 than ‘Eagleton’ during heat stress. Significant increases in ascorbate peroxidase (APX) activities occurred under heat stress, to a greater extent in ‘Eagleton’, whereas isozymes did not exhibit difference between cultivars. Guaiacol-peroxidase (POD) activity declined during heat stress in both cultivars. The intensity of POD isozymes in ‘Brilliant’ remained constant, while ‘Eagleton’ showed a transient increases in POD1 at 7 days of heat stress. Our results indicated that antioxidant defense mechanisms for heat tolerance in kentucky bluegrass could be mainly associated with changes in activity and forms of isozymes of SOD for O2 scavenging and APX activity for H2O2 scavenging under heat stress.

Heat stress along with low water availability at reproductive stage (terminal growth phase of wheat crop) is major contributing factor towards less wheat production in tropics and sub-tropics. Flag leaf plays a pivotal role in assimilate partitioning and stress tolerance of wheat during terminal growth phase. However, limited is known about biochemical response of flag leaf to combined and individual heat and drought stress during terminal growth phase. Therefore, current study investigated combined and individual effect of terminal drought and heat stress on water relations, photosynthetic pigments, osmolytes accumulation and antioxidants defense mechanism in flag leaf of bread wheat. Experimental treatments comprised of control, terminal drought stress alone (50% field capacity during reproductive phase), terminal heat stress alone (wheat grown inside plastic tunnel during reproductive phase) and terminal drought stress + terminal heat stress. Individual and combined imposition of drought and heat stresses significantly (p≤0.05) altered water relations, osmolyte contents, soluble proteins and sugars along with activated antioxidant defensive system in terms of superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX). Turgor potential, POD and APX activities were lowest under individual heat stress; however, these were improved when drought stress was combined with heat stress. It is concluded that combined effect of drought and heat stress was more detrimental than individual stresses. The interactive effect of both stresses was hypo-additive in nature, but for some traits (like turgor potential and APX) effect of one stress neutralized the other. To best of our knowledge, this is the first report on physiological and biochemical response of flag leaf of wheat to combine heat and drought stress. These results will help future studies dealing with improved stress tolerance in wheat. However, detailed studies are needed to fully understand the genetic mechanisms behind these physiological and biochemical changes in flag leaf in response to combined heat and drought stress.

Heat stress is a major limiting factor for growth of cool-season perennial grass species, and mechanisms of heat tolerance have not been well understood. This study was designed to investigate antioxidant enzyme and hormone metabolism responses to heat stress in two kentucky bluegrass ( Poa pratensis L.) cultivars contrasting in heat tolerance. The plants were subjected to 20/20 °C [day/night (control)] or 38/30 °C [day/night (heat stress)] for 28 days in growth chambers. Heat stress increased leaf electrolyte leakage (EL) and malondialdehyde (MDA) with heat-tolerant cultivar EverGlade exhibiting lower levels of EL and MDA relative to heat-sensitive cultivar Kenblue under heat stress. Superoxide dismutase (SOD) and catalase (CAT) activity increased and then declined during 28 days of heat stress. Peroxidase (POD) and ascorbate peroxidase (APX) activity declined and then increased during heat stress. ‘EverGlade’ had greater activities of SOD, CAT, POD, and APX relative to ‘Kenblue’ under heat stress. In addition, ‘EverGlade’ had two additional SOD isozymes and three additional POD isozymes relative to ‘Kenblue’ under heat stress. Leaf abscisic acid (ABA) increased in response to heat stress. Leaf indole-3-acetic acid (IAA) increased and then declined during heat stress. ‘OverGlade’ had higher ABA and IAA content relative to ‘Kenblue’. At the end of heat stress, leaf IAA and ABA content were 27.8% and 73% higher in ‘EverGlade’ relative to ‘Kenblue’, respectively. The results indicated that antioxidant enzymes and the hormones (ABA and IAA) were associated with kentucky bluegrass heat tolerance. Selection and use of cultivars with higher IAA and ABA content and greater antioxidant enzyme activities may improve kentucky bluegrass growth and quality under heat stress.

Summer decline is typically characterized by heat and drought stress and is a major concern for fine fescue species (Festuca). The objectives of this study were to examine whether heat or drought stress is more detrimental, and to determine the genotypic variations in heat and drought tolerance for fine fescues. A total of 26 cultivars, including seven hard fescues (Festuca trachyphylla), eight chewings fescues (Festuca rubra ssp. commutate), seven strong creeping red fescues (Festuca rubra ssp. rubra), two sheep fescues (Festuca ovina ssp. hirtula), and two slender creeping red fescues (Festuca rubra ssp. littoralis) were subjected to prolonged heat or drought stress in growth chambers. Several physiological parameters, including turf quality (TQ), electrolyte leakage (EL), photochemical efficiency (Fv/Fm) chlorophyll content (Chl), and relative water content (RWC) were measured in plants exposed to heat or drought stress. The results indicated that heat stress was more detrimental than drought stress for fine fescue species. Based on TQ and major physiological parameters (EL and Fv/Fm) under heat stress, several cultivars with good heat tolerance were selected, including ‘Blue Ray’, ‘Spartan II’, ‘MN-HD1’, ‘Shoreline’, ‘Navigator II’, ‘Azure’, ‘Beacon’, ‘Aurora Gold’, ‘Reliant IV’, ‘Marco Polo’, ‘Garnet’, ‘Wendy Jean’, ‘Razor’, and ‘Cindy Lou’. Based on TQ and major physiological parameters (EL, RWC, and Fv/Fm) under drought stress, several cultivars with good drought tolerance were selected, including ‘Spartan II’, ‘MN-HD1’, ‘Reliant IV’, ‘Garnet’, ‘Azure’, and ‘Aurora Gold’. These cultivars could be used in hot, dry, or both environments and as breeding germplasm for developing heat tolerance, drought tolerance, or both.

Reactive oxygen species (ROS), such as hydrogen peroxide, superoxide and hydroxyl radicals, are by-products of biological redox reactions. ROS can denature enzymes and damage important cellular components. Plants develop antioxidant enzymes, such as superoxide dismutase (SOD) and ascorbate peroxidase (APX) to scavenge ROS and detoxify them. The effect of increased cytosolic ascorbate peroxidase (cAPX) on heat and UV-B stress tolerance was studied using transformed tomato (Lycopersicon esculentum cv. Zhongshu No. 5) plants. This research demonstrates, in either laboratory or field tests, the potential to enhance tolerance to heat, UV-B, and sunscald stress by gene transfer. Overexpression of cAPX in transgenic tomato enhanced resistance to heat (40°C) and UV-B stress compared to wild-type plants. When leaf disks were placed at 40°C for 13 hours, the electrolyte leakage of disks from wild-type were 93%, whereas two tested transgenic lines (A9, A16) exhibited 24% and 52% leakage respectively. When fruits of wild-type and transgenic plants were exposed to UV-B (2.5 mW cm-2) for five days, the extent of browning was 95%, and 33%, and 37%, respectively. In field tests, the detached fruits from field-grown transgenic plants showed more resistance to exposure to direct sunlight than fruits from wild-type plants. APX activity in leaves of cAPX transgenic plants was several folds higher than in leaves of wild-type plants when exposed to heat, UV-B, and drought stresses.

Gama-aminobutyric acid (GABA) is a nonprotein amino acid in plant cells, which responds to changes in environmental factors. The objectives of this study were to evaluate the effects of foliar spray of GABA on drought and heat tolerance in creeping bentgrass (Agrostis stolonifera), and to investigate physiological factors altered by GABA application that contribute to improved drought tolerance and heat tolerance. GABA-treated plants (cv. Penncross) or non-GABA-treated control plants were then subjected to the following three treatments in growth chambers: 1) nonstress control [plants irrigated every 2 days to maintain soil water content at the pot capacity and maintained at 21/19 °C (day/night) for 35 days], 2) heat stress [plants exposed to 35/30 °C (day/night) and well-watered conditions for 35 days], and 3) drought stress [plants unirrigated for 9 days and maintained at 21/19 °C (day/night), and then rewatered for 2 days]. As compared with untreated plants, GABA-treated plants showed 22% to 39% and 8% to 21% significantly lower leaf electrolyte leakage (EL) and 35% to 143% and 21% to 24% significantly higher turf quality (TQ), 8% to 17% and 17% to 24% relative water content (RWC), 22% to 39% and 25% to 27% chlorophyll content, 7% to 11% and 6% to 17% photochemical efficiency, and an 84% to 683% and 57% to 76% osmotic adjustment (OA) exposed to heat or drought stress across days of treatment, respectively. GABA-treated plants accumulated 7% to 10% more water-soluble carbohydrates (WSC) and 11% to 43% more free proline than nontreated plants under heat stress, and 12% to 30% higher accumulation of WSC under drought stress. After 2 days of rewatering, a significantly better recovery also was observed in GABA-treated plants than that in nontreated plants previously exposed to drought stress. The results suggest that foliar application of GABA significantly improved heat and drought tolerance of creeping bentgrass, which was associated with maintenance of cell membrane stability, delaying in leaf senescence, and enhancing OA. The effectiveness of exogenous GABA application was more pronounced under heat stress than under drought stress.

Reactive oxygen species (ROS) and the heat stress response of Daphnia pulex: ROS-mediated activation of hypoxia-inducible factor 1 (HIF-1) and heat shock factor 1 (HSF-1) and the clustered expression of stress genes.

Calcium (Ca2+) may be involved in plant tolerance to heat stress by regulating antioxidant metabolism or/and water relations. This study was designed to examine whether external Ca2+ treatment would improve heat tolerance in two C(3), cool-season grass species, tall fescue (Festuca arundinacea L.) and Kentucky bluegrass (Poa pratensis L.), and to determine the physiological mechanisms of Ca2+ effects on grass tolerance to heat stress. Grasses were treated with CaCl(2) (10 mM) or H(2)O by foliar application and then exposed to heat stress (35/30 degrees C) in growth chambers. Some of the Ca2+ -untreated plants were maintained at 20/15 degrees C as the temperature control. Heat stress reduced grass quality, relative water content (RWC), and chlorophyll (Chl) content of leaves in both species, but Ca2+ treatment increased all three factors under heat stress. The Ca2+ concentration in cell saps increased with heat stress and with external Ca2+ treatment in both species. Osmotic potential increased with heat stress, but external Ca2+ treatment had no effect. Osmotic adjustment increased during short-term heat stress, but then decreased with a prolonged period of stress; it was not influenced by Ca2+ treatment. The activity of superoxide dismutase (SOD) in both species increased transiently at 12 d of heat stress and then remained at a level similar to that of the control. External Ca2+ treatment had no effect on SOD activity. The activities of catalase (CAT), ascorbate peroxidase (AP), and glutathione reductase (GR) of both species decreased during heat stress. Plants treated with Ca2+ under heat stress had higher CAT, GR and AP activities than untreated plants. Lesser amounts of malondialdehyde (MDA) accumulated in Ca2+ -treated plants than in untreated plants during extended periods of heat stress. The results suggested that exogenous Ca2+ treatment enhanced heat tolerance in both tall fescue and Kentucky bluegrass. This enhancement was related to the maintenance of antioxidant activities and a decrease in membrane lipid peroxidation, but not to the regulation of osmotic potential and osmotic adjustment.

Calcium (Ca2+) may be involved in plant tolerance to heat stress by regulating antioxidant metabolism or/and water relations. This study was designed to examine whether external Ca2+ treatment would improve heat tolerance in two C3, cool‐season grass species, tall fescue (Festuca arundinacea L.) and Kentucky bluegrass (Poa pratensis L.), and to determine the physiological mechanisms of Ca2+ effects on grass tolerance to heat stress. Grasses were treated with CaCl2 (10 mM) or H2O by foliar application and then exposed to heat stress (35/30 °C) in growth chambers. Some of the Ca2+‐untreated plants were maintained at 20/15 °C as the temperature control. Heat stress reduced grass quality, relative water content (RWC), and chlorophyll (Chl) content of leaves in both species, but Ca2+ treatment increased all three factors under heat stress. The Ca2+ concentration in cell saps increased with heat stress and with external Ca2+ treatment in both species. Osmotic potential increased with heat stress, but external Ca2+ treatment had no effect. Osmotic adjustment increased during short‐term heat stress, but then decreased with a prolonged period of stress; it was not influenced by Ca2+ treatment. The activity of superoxide dismutase (SOD) in both species increased transiently at 12 d of heat stress and then remained at a level similar to that of the control. External Ca2+ treatment had no effect on SOD activity. The activities of catalase (CAT), ascorbate peroxidase (AP), and glutathione reductase (GR) of both species decreased during heat stress. Plants treated with Ca2+ under heat stress had higher CAT, GR and AP activities than untreated plants. Lesser amounts of malondialdehyde (MDA) accumulated in Ca2+‐treated plants than in untreated plants during extended periods of heat stress. The results suggested that exogenous Ca2+ treatment enhanced heat tolerance in both tall fescue and Kentucky bluegrass. This enhancement was related to the maintenance of antioxidant activities and a decrease in membrane lipid peroxidation, but not to the regulation of osmotic potential and osmotic adjustment.

Cancer cells undergo mitosis more frequently than normal cells and thus have increased metabolic needs, which in turn lead to higher than normal reactive oxygen species (ROS) production. Higher ROS production increases cancer cell dependence on ROS scavenging systems to balance the increased ROS. Selectively modulating intracellular ROS in cancers by exploiting cancer dependence on ROS scavenging systems provides a useful therapeutic approach. Essential to developing these therapeutic strategies is to maintain physiologically low ROS levels in normal tissues while inducing ROS in cancer cells. GMX1778 is a specific inhibitor of nicotinamide phosphoribosyltransferase, a rate-limiting enzyme required for the regeneration of NAD(+) from nicotinamide. We show that GMX1778 increases intracellular ROS in cancer cells by elevating the superoxide level while decreasing the intracellular NAD(+) level. Notably, GMX1778 treatment does not induce ROS in normal cells. GMX1778-induced ROS can be diminished by adding nicotinic acid (NA) in a NA phosphoribosyltransferase 1 (NAPRT1)-dependent manner, but NAPRT1 is lost in a high frequency of glioblastomas, neuroblastomas, and sarcomas. In NAPRT1-deficient cancer cells, ROS induced by GMX1778 was not susceptible to treatment with NA. GMX1778-mediated ROS induction is p53-dependent, suggesting that the status of both p53 and NAPRT1 might affect tumor apoptosis, as determined by annexin-V staining. However, as determined by colony formation, GMX1778 long term cytotoxicity in cancer cells was only prevented by the addition of NA to NAPRT1-expressing cells. Exposure to GMX1778 may be a novel way of inducing ROS selectively in NAPRT1-negative tumors without inducing cytotoxic ROS in normal tissue.

Heat stress impacts plant growth at all phases of development, although the particular threshold for heat tolerance varies significantly across different developmental stages. During seed germination, elevated temperatures can either impede or completely halt the process, contingent upon the plant type and the severity of the stress. During advanced stages, high temperatures can have a negative impact on photosynthesis, respiration, water balance, and membrane integrity. Additionally, they can also influence the levels of hormones and primary and secondary metabolites. In addition, during the growth and development of plants, there is an increased expression of various heat shock proteins, as well as other proteins related to stress, and the generation of reactive oxygen species (ROS). These are significant plant responses to heat stress. Plants employ several strategies to deal with heat stress, such as maintaining the stability of their cell membranes, removing harmful reactive oxygen species (ROS), producing antioxidants, accumulating and adjusting compatible solutes, activating mitogen-activated protein kinase (MAPK) and calcium-dependent protein kinase (CDPK) cascades, and, crucially, signaling through chaperones and activating transcription. These molecular-level systems boost the ability of plants to flourish in heat stress. Potential genetic methods to enhance plant heat stress resistance encompass old and modern molecular breeding techniques and transgenic approaches, all of which rely on a comprehensive comprehension of these systems. Although several plants exhibit enhanced heat tolerance through traditional breeding methods, the effectiveness of genetic transformation techniques has been somewhat restricted. The latter results from the current constraints in our understanding and access to genes that have known impacts on plant heat stress tolerance. However, these challenges may be overcome in the future. Besides genetic methods, crops' heat tolerance can be improved through the pre-treatment of plants with various environmental challenges or the external application of osmoprotectants such as glycine betaine and proline. Thermotolerance is achieved through an active process in which plants allocate significant energy to maintain their structure and function to avoid damage induced by heat stress. The practice of nanoparticles has been shown to upgrade both the standard and the quantity of produce when crops are under heat stress. This review provides information on the effects of heat stress on plants and explores the importance of nanoparticles, transgenics, and genomic techniques in reducing the negative consequences of heat stress. Furthermore, it explores how plants might adapt to heat stress by modifying their biochemical, physiological, and molecular reactions.

Heat stress is a major abiotic constraint limiting wheat (Triticum aestivum L.) productivity, primarily by inducing excessive accumulation of reactive oxygen species (ROS) that disrupt cellular homeostasis. Brassinosteroids such as 24-epibrassinolide (24-EBL) are known to enhance plant stress tolerance through modulation of antioxidant defense systems. This field study evaluated the effect of foliar-applied 24-EBL (0.01, 0.02, and 0.03 mM) on antioxidant enzyme activities, ROS accumulation, and grain yield in two contrasting wheat genotypes HUW-510 (heat-tolerant) and HUW-468 (heat-susceptible) grown under normal and late-sown (heat-stressed) conditions. Foliar sprays were applied at pre-flowering (45 DAS) and post-flowering (65 DAS). Antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) and ROS levels were assayed at 65 and 85 DAS. Statistical analyses included ANOVA (LSD, p ≤ 0.05), stepwise multiple linear regression to examine the relationships between biochemical and yield traits. Application of 24-EBL significantly enhanced SOD, CAT, and APX activities by 25–35%, reduced ROS accumulation by 20–30%, and improved grain yield by 80–120% under heat stress, with the 0.02–0.03 mM concentration being most effective. The heat-tolerant genotype HUW-510 exhibited superior antioxidant capacity and yield stability compared with HUW-468. Regression analysis showed strong positive associations between antioxidant enzyme activities and grain yield (R² = 0.78–0.87; p < 0.01), indicating that biochemical fortification through 24-EBL plays a key role in thermotolerance. These findings demonstrate that foliar 24-EBL application can serve as a practical, eco-friendly strategy to mitigate terminal heat stress in wheat.