Heat-sensitive Genotypes Research Articles

A Genome-wide association study identifies candidate genes for heat tolerance in adult cucumber (Cucumis sativus L.) plants

Heat stress causes overgrowth, leaf dryness and fruit malformation, which negatively impacts cucumber quality and yield. Yet, in spite of the devastating consequences of this abiotic stress, few genes for heat tolerance in cucumber have been identified. Here, the heat injury indices of 88 cucumber accessions representing diverse ecotypes were collected in two open-field environments, with naturally occurring high temperatures over two years. Seventeen of the 88 accessions were identified as highly heat-tolerant. Using a genome-wide association study, five loci (gHII3.1, gHII3.2, gHII3.3, gHII4.1 and gHII6.1) on three chromosomes associated with heat tolerance were detected. Pairwise linkage disequilibrium correlation, sequence polymorphisms, and qRT-PCR analyses at these loci, identified five candidate genes predicted to be casual for heat stress response in cucumber. CsaV3_3G04883, CsaV3_4G029050 and CsaV3_6G005370 each had nonsynonymous SNPs, and were significantly up-regulated by heat stress in the heat-tolerant genotypes. CsaV3_3G031890 was also induced by heat stress, but in the heat-sensitive genotypes, and sequence polymorphism was only found in the promoter region. Identifying these candidate genes lays a foundation for understanding cucumber thermotolerance mechanisms. Our study is one of the few to examine heat stress in adult cucumber plants and it therefore fills a critical gap in knowledge. It is also an important first-step towards accelerating the breeding of robust heat-tolerant varieties.

Salicylic acid associated modulation of physiological attributes of heat tolerant and heat sensitive tomato genotypes results in higher yield

Salicylic acid associated modulation of physiological attributes of heat tolerant and heat sensitive tomato genotypes results in higher yield

Understanding the effect of heat stress during seed filling on nutritional composition and seed yield in chickpea (Cicer arietinum L.)

Increasing temperature affects all food crops, thereby reducing their yield potential. Chickpea is a cool-season food legume vital for its nutritive value, but it is sensitive to high temperatures (> 32/20 °C maximum/minimum) during its reproductive and seed-filling stages. This study evaluated the effects of heat stress on yield and qualitative traits of chickpea seeds in a controlled environment. Chickpea genotypes differing in heat sensitivity [two heat-tolerant (HT) and two heat-sensitive (HS)] were raised in pots, initially in an outdoor environment (average 23.5/9.9 °C maximum/minimum), until the beginning of pod set (107–110 days after sowing). At this stage, the plants were moved to a controlled environment in the growth chamber to impose heat stress (32/20 °C) at the seed-filling stage, while maintaining a set of control plants at 25/15 °C. The leaves of heat-stressed plants of the HT and HS genotypes showed considerable membrane damage, altered stomatal conductance, and reduced leaf water content, chlorophyll content, chlorophyll fluorescence, and photosynthetic ability (RuBisCo, sucrose phosphate synthase, and sucrose activities) relative to their corresponding controls. Seed filling duration and seed rate drastically decreased in heat-stressed plants of the HT and HS genotypes, severely reducing seed weight plant–1 and single seed weight, especially in the HS genotypes. Yield-related traits, such as pod number, seed number, and harvest index, noticeably decreased in heat-stressed plants and more so in the HS genotypes. Seed components, such as starch, proteins, fats, minerals (Ca, P, and Fe), and storage proteins (albumin, globulins, glutelin, and prolamins), drastically declined, resulting in poor-quality seeds, particularly in the HS genotypes. These findings revealed that heat stress significantly reduced leaf sucrose production, affecting the accumulation of various seed constituents, and leading to poor nutritional quality. The HT genotypes were less affected than the HS genotypes because of the greater stability of their leaf water status and photosynthetic ability, contributing to better yield and seed quality traits in a heat-stressed environment.

Investigating the impact of terminal heat stress on contrasting wheat cultivars: a comprehensive analysis of phenological, physiological, and biochemical traits.

Terminal heat stress has become one of the major threats due to global climate change which is significantly affecting the production and productivity of wheat crop. Therefore, it is necessary to identify key traits and genotypes to breed heat-tolerant wheat. The present study was undertaken with the objective of comparing the effects of heat stress (HSE) and extended heat stress (EHSE) on phenological-physio-biochemical traits of contrasting heat-tolerant and heat-susceptible genotypes during the reproductive phase. Phenological traits exhibited significant reduction under EHSE compared to HSE. Heat-tolerant genotypes maintained balanced phenological-physio-biochemical traits, while heat-sensitive genotypes showed significant reductions under both stress regimes. Among phenological traits, DM (R2=0.52) and BY (R2=0.44) have shown a positive effect on seed yield, indicating that biomass and crop duration contributed to the yield advantage under stress. During the grain filling stage, both the normalized difference vegetation index (NDVI) and chlorophyll (Chl) exhibited consistently positive impacts on grain yield under both HSE and EHSE conditions. This could be attributed to the enhanced photosynthesis resulting from delayed senescence and improved assimilate remobilization under terminal heat stress. The biochemical activity of superoxide dismutase (SOD), peroxidase (POX), and ascorbate peroxidase (APX) was induced in tolerant genotypes under HSE. The correlation of canopy temperature with phenological-physio-biochemical traits remained static under HSE and EHSE, suggesting CT as the best selection parameter for heat tolerance. The traits showing a positive association with yield and that are less affected under stress could be used for selecting tolerant genotypes under stress environments. These tolerant genotypes can be used to develop mapping populations to decipher the genes conferring tolerance as well as to study the molecular basis of tolerance.

Proteomic and physiological responses of contrasting two different heat-resistant orchardgrass genotypes to heat stress

As an important forage crop worldwide, the growth and productivity of orchardgrass are greatly impacted by high temperatures. However, little information is known about how orchardgrass proteomic changes under heat conditions. Therefore, the present study investigated the proteomics and physiological changes in 667 [AKZ-NRGR667 (heat-tolerant)] and 7602 [PI237602 (heat-sensitive)] under heat stress (40/35 °C). In addition, the responses of translational regulating of heat stress in orchardgrass were analyzed through proteomic changes using the tandem mass tags (TMT) technique. Together, 410 differentially expressed proteins (DEPs) were identified from two orchardgrass genotypes under heat at 24 h. Proteomics analyses indicated that proteins related to substance metabolism, photosynthesis, and heat shock proteins (HSPs) were differentially expressed under heat stress and control conditions. Moreover, a large proportion of HSPs were expressed in the heat-tolerant genotype as compared to the heat-sensitive genotype. In conclusion, genotype 667 has higher adaptability and repairing capability due to stronger heat tolerance capacity that can make it more suited to sustaining its survival and growth than genotype 7602. These findings can provide the basis for genetic improvements in orchardgrass and other crops facing high-temperature stress or heat environment that may lead to heat resistance or tolerance.

Positive response of maize husk traits for improving heat tolerance during flowering by alleviating husk inside temperature

With the increase in high temperature intensity and occurrence frequency, heat stress has become the main threat to the production of majority of crops, including maize (Zea mays L.). Maize husks play protective and energy-providing roles in ear growth; however, the effects of the husk as well as the response of husk traits to heat stress are not fully understood. To address this gap, a series of experiments were conducted to explore the impacts of heat stress during flowering on husk inside temperature and husk functions and identify the difference in husk response between different tolerant genotypes. Husk inside temperature was lower than the air temperature, and the temperature difference between the outside and inside of the husk (Toutside – Tinside) increased with the increased air temperature and reached a maximum (2.5–3.0 °C) at 40/30 °C. The increased stomatal conductance and transpiration rate of husks could contribute to cooling the inside temperature under heat stress. Additionally, a higher husk inside temperature, larger yield losses, and decreased photosynthesis rate were found after manual removal of partial husks, which contributed to reduced husk protection and energy supply effects, especially exposure to heat stress. Under heat stress, heat-tolerant genotypes (i.e., lower yield losses) correspond to lower husk inside temperature, which could be related to the positive response of husk traits to heat stress, including increased/unchanged husk areas, length, width, weight and numbers. In contrast, heat-sensitive genotypes (i.e., higher yield losses) corresponding to higher husk inside temperature and reduction in the above husk parameters. Thus, husk traits that positively respond to heat stress can alleviate the increase in kernel temperature and further decrease yield damage. Considering appropriate husk traits and understanding the potential heat tolerance mechanism have implications for breeding and screening tolerant genotypes under future warmer climates.

Effect of terminal heat stress on osmolyte accumulation and gene expression during grain filling in bread wheat (Triticum aestivum L.).

The grain-filling stage in Triticum aestivum (wheat) is highly vulnerable to increasing temperature as terminal heat stress diminishes grain quality and yield. To examine the mechanism of terminal heat tolerance, we performed the biochemical and gene expression analyses using two heat-tolerant (WH730 and WH1218) and two heat-sensitive (WH711 and WH157) wheat genotypes. We observed a significant increase in total soluble sugar (25%-47%), proline (7%-15%), and glycine betaine (GB) (22%-34%) contents in flag leaf, whereas a decrease in grain-filling duration, 1000-kernel weight (8%-25%), and grain yield per plant (11%-23%) was observed under the late-sown compared to the timely sown. The maximum content of osmolytes, including total soluble sugar, proline, and GB, was observed in heat-tolerant genotypes compared to heat-sensitive genotypes. The expression of 10 heat-responsive genes associated with heat shock proteins (sHsp-1, Hsp17, and HsfA4), flavonoid biosynthesis (F3'-1 and PAL), β-glucan synthesis (CslF6 and CslH), and xyloglucan metabolism (XTH1, XTH2, and XTH5) was studied in flag leaf exposed to different heat treatments (34, 36, 38, and 40°C) at 15 days after anthesis by quantitative real-time polymerase chain reaction. A significant increase in the relative fold expression of these genes with increasing temperature indicated their involvement in providing heat-stress tolerance. The high differential expression of most of the genes in heat-tolerant genotype "WH730" followed by "WH1218" indicates the high adaptability of these genotypes to heat stress compared to heat-sensitive wheat genotypes. Based on the previous results, "WH730" performed better in terms of maximum osmolyte accumulation, grain yield, and gene expression under heat stress.

The interplay of DNA methyltransferases and demethylases with tuberization genes in potato (Solanum tuberosum L.) genotypes under high temperature.

Potato is a temperate crop consumed globally as a staple food. High temperature negatively impacts the tuberization process, eventually affecting crop yield. DNA methylation plays an important role in various developmental and physiological processes in plants. It is a conserved epigenetic mark determined by the dynamic concurrent action of cytosine-5 DNA methyltransferases (C5-MTases) and demethylases (DeMets). However, C5-MTases and DeMets remain unidentified in potato, and their expression patterns are unknown under high temperatures. Here, we performed genome-wide analysis and identified 10 C5-MTases and 8 DeMets in potatoes. Analysis of their conserved motifs, gene structures, and phylogenetic analysis grouped C5-MTases into four subfamilies (StMET, StCMT3, StDRM, and StDNMT2) and DeMets into three subfamilies (StROS, StDML, and StDME). Promoter analysis showed the presence of multiple cis-regulatory elements involved in plant development, hormone, and stress response. Furthermore, expression dynamics of C5-MTases and DeMets were determined in the different tissues (leaf, flower, and stolon) of heat-sensitive (HS) and heat-tolerant (HT) genotypes under high temperature. qPCR results revealed that high temperature resulted in pronounced upregulation of CMT and DRM genes in the HT genotype. Likewise, demethylases showed strong upregulation in HT genotype as compared to HS genotype. Several positive (StSP6A and StBEL5) and negative (StSP5G, StSUT4, and StRAP1) regulators are involved in the potato tuberization. Expression analysis of these genes revealed that high temperature induces the expression of positive regulators in the leaf and stolon samples of HT genotype, possibly through active DNA demethylation and RNA-directed DNA methylation (RdDM) pathway components. Our findings lay a framework for understanding how epigenetic pathways synergistically or antagonistically regulate the tuberization process under high-temperature stress in potatoes. Uncovering such mechanisms will contribute to potato breeding for developing thermotolerant potato varieties.

Exogenously Applied GA3 Enhances Morphological Parameters of Tolerant and Sensitive Cyclamen persicum Genotypes under Ambient Temperature and Heat Stress Conditions.

Cyclamen genus is part of the Primulaceae family consisting of 24 species widely cultivated as ornamental and medicinal plants. They also possess high plasticity in terms of adaptability to alternating environmental conditions. In this regard, the present study investigates the germination and morphological parameters of heat-tolerant and heat-sensitive Cyclamen persicum accessions in the presence of different GA3 solutions (0, 30, 70 and 90 mg/L) under ambient temperature and heat stress conditions. Heat-tolerant genotypes, mainly C3-Smartiz Victoria (6.42%), C15-Merengue magenta (6.47%) and C16-Metis silverleaf (5.12%) had the highest germination rate with 90 mg/L GA3 treatment compared with control. Regarding heat-sensitive genotypes, C11-Verano (5.11%) and C13-Metis Origami (4.28%) had the lowest values in mean germination time, along with the Petticoat genotypes C1 (73.3%) and C2 (80.0%) with a high germination percentage. Heat-tolerant genotypes positively responded to GA3 (70 and 90 mg/L) even under heat stress conditions, by their higher values in plant height, an ascending trend also seen in heat-sensitive genotypes under GA3 treatment (70 and 90 mg/L). According to the hierarchical clustering, several heat-tolerant genotypes showed peculiar behavior under heat stress conditions, namely C3 (Smartiz Victoria), C7 (Halios falbala) and C8 (Latinia pipoca) which proved to be susceptible to heat stress even under GA3 application, compared with the other genotypes which showed tolerance to higher temperatures. In the case of heat-sensitive genotypes, C4 (Smartiz violet fonce), C6 (Metis blank pur), C11 (Verano) and C13 (Metis origami) possessed higher positive or negative values compared with the other heat-sensitive genotypes with increased doses of GA3. These genotypes were shown to be less affected by heat stress, suggesting their positive response to hormone treatment. In conclusion, the above-mentioned genotypes, particularly heat-tolerant C15 and heat-sensitive C2 with the highest germination capacity and development can be selected as heat-resistant genotypes to be deposited in gene banks and used in further amelioration programs under biotic and/or abiotic stresses to develop resistant genotypes.

High Temperature Alters Leaf Lipid Membrane Composition Associated with Photochemistry of PSII and Membrane Thermostability in Rice Seedlings.

Rice cultivated in the tropics is exposed to high temperature (HT) stress which threatens its growth and survival. This study aimed at characterizing the HT response in terms of PSII efficiency and membrane stability, and to identify leaf fatty acid changes that may be associated with HT tolerance or sensitivity of rice genotypes. Twenty-eight-day-old seedlings of two Thai rice cultivars (CN1 and KDML105), a standard heat tolerance (N22), and a heat sensitive (IR64) rice genotype were treated at 42 °C for 7 days. Under HT, N22 showed the highest heat tolerance displaying the lowest increase in electrolyte leakage (EL), no increments in malondialdehyde (MDA) and stable maximum quantum yield of PSII efficiency (Fv/Fm). Compared to KDML105 and IR64, CN1 was more tolerant of HT, showing a lower increase in EL and MDA, and less reduction in Fv/Fm. N22 and CN1 showed a higher percentage reduction of unsaturated fatty acids (C18:2 and C18:3), which are the major components of the thylakoid membrane, rendering the optimum thylakoid membrane fluidity and intactness of PSII complex. Moreover, they exhibited sharp increases in long-chain fatty acids, particularly C22:1, while the heat sensitive IR64 and KDML105 showed significant reductions. Dramatic increases in long-chain fatty acids may lead to cuticular wax synthesis which provides protective roles for heat tolerance. Thus, the reduction in unsaturated fatty acid composition of the thylakoid membrane and dramatic increases in long-chain fatty acids may lead to high photosynthetic performance and an enhanced synthesis of cuticular wax which further provided additional protective roles for heat tolerance ability in rice.

Effects of Elevated Temperature and Salicylic Acid on Heat Shock Response and Growth of Potato Microplants

Potato is a globally important, highly heat-susceptible crop species. We investigated the effects of prolonged exposure to elevated temperatures and exogenous salicylic acid (SA) on microplant growth and heat-shock response (HSR) in three unrelated potato genotypes/cultivars. Long-term exposure to 29 °C (mild heat stress) caused a significant reduction in the number of surviving explants and shoot morphometric parameters in heat-sensitive genotypes, while exposure to 26 °C (warming) caused only a decline in shoot growth. Interestingly, 26 °C-temperature treatment stimulated root growth in some investigated genotypes, indicating a difference between favorable temperatures for potato shoot and root growth. SA showed a protective effect regarding potato shoot growth at 26 °C. At 29 °C, this effect was genotype-dependent. SA did not affect the number of roots and inhibited root elongation at all temperature treatments, indicating the difference between shoot and root responses to applied SA concentration. Although HSR is mainly considered rapid and short-lived, elevated transcript levels of most investigated HSFs and HSPs were detected after three weeks of heat stress. Besides, two StHSFs and StHSP21 showed elevated expression at 26 °C, indicating extreme potato heat-susceptibility and significance of HSR during prolonged warming. SA effects on HSFs and HSPs expression were minor and alterable.

Physiological and molecular response of cotton (Gossypium hirsutum L.) to heat stress at the seedling stage

The ideal temperature range for the optimal growth and development of cotton is 25 °C–32 °C and high temperature adversely affects the metabolic activities of plant cells. This study was aimed to screen heat-tolerant cotton genotypes based on physiological and molecular parameters. Experiments were carried out during 2019–2020 at the MNS-University of Agriculture, Multan, Pakistan. The research comprised two parts. In the first experiment, 30 cotton genotypes were sown in a completely randomized design with three replications under laboratory conditions for the determination of cell membrane thermostability. Principal component analysis was performed, and four genotypes, i.e., two heat-tolerant (‘CRIS-5A’ and ‘VH-338’) and two heat-sensitive (‘FH-242’ and ‘VH-281’) genotypes, were selected. In the second experiment, the screened cotton genotypes were sown in pots in a factorial complete randomized design with three replications and two treatments (normal and heat treatment). Heat stress was applied at the seedling stage, and eight leaf samples (one from each experimental unit) were collected. Two genes were used for molecular analysis and were amplified in all eight cDNA samples. Molecular analysis indicated the presence of HSP70 and HSP26 genes in the cotton genotypes, and the expression of these genes was measured by using ImageJ software. The gene expression level of HSP70 was very high (16.41%) in ‘VH-281’, which is a heat-sensitive genotype under heat stress. The sensitive genotype ‘FH-242’ exhibited the highest gene expression level of HSP26 (20.32%) under normal conditions. A similar sequence of HSP70 gene of Agave sisalana was amplified for the first time in cotton. It is a good indicator for screening heat tolerant cotton genotypes at the molecular level.

Nitric oxide secures reproductive efficiency in heat-stressed lentil (Lens culinaris Medik.) plants by enhancing the photosynthetic ability to improve yield traits.

Rising temperatures, globally and locally, would be detrimental for cool- and summer-season food legumes, such as lentil (Lens culinaris Medik.). Lentil is highly sensitive to supra-optimal temperatures (> 30°C), particularly during reproductive growth, resulting in flower and pod losses. Thus, suitable strategies are needed to introduce heat tolerance in this legume. Here, we evaluated the efficacy of nitric oxide (NO)-applied as foliar treatment of 1mM sodium nitroprusside (SNP), twice (one day before final exposure to high temperature, and again five days later)-on heat-stressed (32/20°C) lentil genotypes, differing in heat sensitivity. As a result of heat stress, endogenous NO increased significantly in heat-tolerant genotypes (46-62% in leaves and 66-68% in anthers, relative to the respective controls), while it decreased in heat-sensitive (HS) genotypes (27-30% in leaves and 28-33% in anthers, relative to the respective controls). Foliar supplementation with SNP markedly increased endogenous NO in leaves and anthers of both the control and heat-treated plants. Heat stress significantly accelerated phenology, damaged membranes, chlorophyll, chlorophyll fluorescence, cellular viability, and decreased leaf water status, carbon fixing and assimilating ability, less so in plants treated with SNP. Heat stress plus SNP significantly improved carbon fixation (as RuBisCo activity) and assimilation ability, (as sucrose concentration (in leaves and anthers), sucrose synthase and vacuolar acid invertase activity, reducing sugars), as well as osmolyte accumulation (proline and glycine betaine) in leaves and anthers. Moreover, SNP-treated plants had significantly less oxidative damage-measured as malondialdehyde and hydrogen peroxide concentrations-in leaves and anthers, relative to the respective control. Reproductive function-assessed as pollen grain germination and viability, stigma receptivity, and ovular viability-decreased markedly in plants exposed to heat stress alone, more so in HS genotypes, but increased significantly with SNP treatment as a consequence of improved leaf and anther function, to significantly increase the pod and seed numbers in heat-stressed lentil plants, relative to heat-stress alone.

Morpho-Physiological Traits and Functional Markers Based Molecular Dissection of Heat-Tolerance in Urdbean.

Urdbean (Vigna mungo L. Hepper) is one of the important pulse crops. Its cultivation is not so popular during summer seasons because this crop is unable to withstand excessive heat stress beside lack of humidity in the atmosphere. Therefore, a panel of 97 urdbean diverse genotypes was assessed for yield under stress and non-stress conditions with an aim to identify heat tolerant genotypes. This study identified 8 highly heat tolerant and 35 highly heat sensitive genotypes based on heat susceptibility index. Further, physiological and biochemical traits-based characterization of a group of six highly heat sensitive and seven highly heat tolerant urdbean genotypes showed genotypic variability for leaf nitrogen balance index (NBI), chlorophyll (SPAD), epidermal flavnols, and anthocyanin contents under 42/25°C max/min temperature. Our results showed higher membrane stability index among heat tolerant genotypes compared to sensitive genotypes. Significant differences among genotypes for ETR at different levels of PAR irradiances and PAR × genotypes interactions indicated high photosynthetic ability of a few genotypes under heat stress. Further, the most highly sensitive genotype PKGU-1 showed a decrease in different fluorescence parameters indicating distortion of PS II. Consequently, reduction in the quantum yield of PS II was observed in a sensitive one as compared to a tolerant genotype. Fluorescence kinetics showed the delayed and fast quenching of Fm in highly heat sensitive (PKGU 1) and tolerant (UPU 85-86) genotypes, respectively. Moreover, tolerant genotype (UPU 85-86) had high antioxidant activities explaining their role for scavenging superoxide radicals (ROS) protecting delicate membranes from oxidative damage. Molecular characterization further pinpointed genetic differences between heat tolerant (UPU 85-86) and heat sensitive genotypes (PKGU 1). These findings will contribute to the breeding toward the development of heat tolerant cultivars in urdbean.

Heat Priming of Lentil (Lens culinaris Medik.) Seeds and Foliar Treatment with γ-Aminobutyric Acid (GABA), Confers Protection to Reproductive Function and Yield Traits under High-Temperature Stress Environments.

Gradually increasing temperatures at global and local scales are causing heat stress for cool and summer-season food legumes, such as lentil (Lens culinaris Medik.), which is highly susceptible to heat stress, especially during its reproductive stages of development. Hence, suitable strategies are needed to develop heat tolerance in this legume. In the present study, we tested the effectiveness of heat priming (HPr; 6 h at 35 °C) the lentil seeds and a foliar treatment of γ-aminobutyric acid (GABA; 1 mM; applied twice at different times), singly or in combination (HPr+GABA), under heat stress (32/20 °C) in two heat-tolerant (HT; IG2507, IG3263) and two heat-sensitive (HS; IG2821, IG2849) genotypes to mitigate heat stress. The three treatments significantly reduced heat injury to leaves and flowers, particularly when applied in combination, including leaf damage assessed as membrane injury, cellular oxidizing ability, leaf water status, and stomatal conductance. The combined HPr+GABA treatment significantly improved the photosynthetic function, measured as photosynthetic efficiency, chlorophyll concentration, and sucrose synthesis; and significantly reduced the oxidative damage, which was associated with a marked up-regulation in the activities of enzymatic antioxidants. The combined treatment also facilitated the synthesis of osmolytes, such as proline and glycine betaine, by upregulating the expression of their biosynthesizing enzymes (pyrroline-5-carboxylate synthase; betaine aldehyde dehydrogenase) under heat stress. The HPr+GABA treatment caused a considerable enhancement in endogenous levels of GABA in leaves, more so in the two heat-sensitive genotypes. The reproductive function, measured as germination and viability of pollen grains, receptivity of stigma, and viability of ovules, was significantly improved with combined treatment, resulting in enhanced pod number (21–23% in HT and 35–38% in HS genotypes, compared to heat stress alone) and seed yield per plant (22–24% in HT and 37–40% in HS genotypes, in comparison to heat stress alone). The combined treatment (HPr+GABA) was more effective and pronounced in heat-sensitive than heat-tolerant genotypes for all the traits tested. This study offers a potential solution for tackling and protecting heat stress injury in lentil plants.

Regulation of Osmotic Balance and Increased Antioxidant Activities under Heat Stress in Abelmoschus esculentus L. Triggered by Exogenous Proline Application

Keeping in view the yield losses instigated by heat stress in several crops, we carried out an experiment to explore the curative effect of exogenous applications of proline on the morpho-physiological, biochemical, and water-related attributes of okra genotypes under high-temperature stress (controlled conditions). Four contrasting genotypes C1, C2, C3, and C4 heat tolerant and heat sensitive genotypes were selected from a diverse panel of okra genotypes (n = 100) to examine plant responses to high-temperature stress and exogenous application of proline. Four-week-old seedlings were subjected to heat stress by gradually increasing the temperature of a growth chamber from 28/22 °C to 45/35 °C (day/night) and sprayed with an optimized proline concentration 2.5 mM. The experiment consisted of a factorial arrangement of treatments in a completely randomized design. The results showed that there were maximum increases in shoot length (32.7%), root length (58.9%), and shoot fresh (85.7%). The quantities of leaves per plant were increased by 52.9%, 123.6%, 82.5%, and 62.2% in C1, C2, C3, and C4 after proline application. On the other hand, only root fresh weight decreased in all genotypes after proline application by 23.1%, 20%, 266.7%, and 280.8% (C1, C2, C3, C4). A lower leaf temperature of 27.72 °C, minimum transpiration of 3.29 mmol m−2 s−1, maximum photosynthesis of 3.91 μmol m−2 s−1, and a maximum water use efficiency of 1.20 μmol CO2 mmol H2O were recorded in the genotypes C2, C1, C3, and C4, respectively. The highest enzymatic activity of superoxide dismutase, peroxidase and catalase were 14.88, 0.31, and 0.15 U mg-protein in C2, C1, and C3, respectively. Maximum leaf proline, glycinebetaine, total free amino acids, and chlorophyll content 3.46 mg g−1, 4.02 mg g−1, 3.46 mg g−1, and 46.89 (in C2), respectively, due to foliar applications of proline. Another important finding was that heat tolerance in okra was highly linked highly linked to genotypes’ genetic potential, having more water use efficiency, enzymatic activities, and physio-biochemical attributes under the foliar applications of proline.

The effect of individual and combined drought and heat stress under elevated CO2 on physiological responses in spring wheat genotypes

Abiotic stress due to climate change with continuous rise of atmospheric CO2 concentration is predicted to cause severe changes to crop productivity. Thus, research into wheat cultivars, capable of maintaining yield under limiting conditions is necessary. The aim of this study was to investigate the physiological responses of spring wheat to individual and combined drought- and heat events and their interaction with CO2 concentration. Two heat sensitive (LM19, KU10) and two heat tolerant (LM62, GN5) genotypes were selected and grown under ambient (400 ppm, aCO2) and elevated (800 ppm, eCO2) CO2 concentrations. At the tillering stage, the wheat plants were subjected to different treatments: control, progressive drought, heat and combined drought and heat stress. Our results showed that eCO2 mitigated the negative impact of the moderate stress in all genotypes. However, no distinctive responses were observed in some of the measured parameters between heat sensitive and tolerant genotypes. All genotypes grown at eCO2 had significantly higher net photosynthetic rates and maintained maximum quantum efficiency of PSII photochemistry under heat and combined stress compared to aCO2. Under heat and combined stress, the chlorophyll a:b ratios decreased only in heat tolerant genotypes at eCO2 compared to the control. Furthermore, the heat tolerant genotypes grown at eCO2 showed an increased glucose and fructose contents and a decreased sucrose content under combined stress compared to aCO2. These findings provide new insights into the underlying mechanisms of different genotypic responses to combined abiotic stresses at eCO2 that differ from the response to individual stresses.

Response of spikelet water status to high temperature and its relationship with heat tolerance in rice

In rice, high-temperature stress (HT) during flowering results in decreased grain yield via a reduction in spikelet fertility; however, the effect of plant water status on spikelet fertility under HT remains unknown. To investigate the relationship between spikelet water status and spikelet fertility under HT, two experiments were performed under temperature-controlled conditions using four genotypes with varying tolerance to HT. Rice plants were exposed to HT for seven consecutive days during the flowering stage under three soil water treatments (soil water potential 0, −20, and −40 kPa), as well as under hydroponic conditions in a separate experiment. HT significantly decreased spikelet fertility, pollen fertility, and anther dehiscence under each of the three water treatments. HT significantly increased the spikelet transpiration rate, and this change was accompanied by a significant decrease in the internal temperature of the spikelets. HT decreased pollen grain diameter in heat-sensitive genotypes. HT had varying effects on the water potential of panicles and anthers but increased anther soluble-sugar concentration. Different aquaporin genes showed different expression profiles under HT, and the expression levels of PIPs for plasma membrane intrinsic proteins and TIPs for tonoplast intrinsic proteins increased in anthers but decreased in glumes. Correlation analyses showed that anther dehiscence and pollen (spikelet) fertility were tightly associated with anther water status, and the expression levels of almost all anther aquaporin genes were significantly correlated with anther dehiscence under HT. In summary, an increased spikelet transpiration rate and decreased internal spikelet temperature were associated with alleviation of the effects of HT in rice genotypes with varying degrees of heat tolerance, and the response of spikelet water status to HT, involving increased total expression of aquaporins and soluble sugar content, thereby improved pollen fertility, anther dehiscence, and spikelet fertility, especially in heat-resistant genotypes. The heat-resistant genotypes N22 and SY63 may adopt different approaches to reduce heat damage.

Impact of elevated CO2 and heat stress on wheat pollen viability and grain production.

Periods of high temperature and an expected increase in atmospheric CO2 concentration as a result of global climate change are major threats to wheat (Triticum aestivum L.) production. Developing heat-tolerant wheat cultivars demands improved understanding of the impacts of high temperature and elevated CO2 on plant growth and development. This research investigated the interactive effects of heat stress and CO2 concentration on pollen viability and its relationship to grain formation and yield of wheat in greenhouse conditions. Nineteen wheat genotypes and a current cultivar, Suntop, were heat stressed at either meiosis or anthesis at ambient (400 µL L-1) or elevated (800 µL L-1) CO2. Elevated CO2 and heat stress at meiosis reduced pollen viability, spikelet number and grain yield per spike; however, increased tillering at the elevated CO2 level helped to minimise yield loss. Both heat-tolerant genotypes (e.g. genotype 1, 2, 10 or 12) and heat-sensitive genotypes (e.g. genotype 6 or 9) were identified and response related to pollen sensitivity and subsequent impacts on grain yield and yield components were characterised. A high-throughput protocol for screening wheat for heat stress response at elevated CO2 was established and meiosis was the most sensitive stage, affecting pollen viability, grain formation and yield.

Screening for heat tolerant genotypes in bread wheat (T. aestivum L.) using stress tolerance indices

Heat is a major stress that severely affects wheat productivity. The objective of the current study was estimating the effect of heat stress during the grain-filling stage for screening heat tolerant wheat genotypes by measuring six stress tolerance indices for grain yield and also measured the % of decline on other important traits. A set of 190 wheat genotypes along with four local checks were evaluated in two separate experiments, one under normal condition (timely sown) and the other under heat stress condition (late sown) in augmented block design at the research farm of Department of Genetics and Plant Breeding, C.C.S. University, Meerut (U.P) during rabi season of 2014-2015. The calculation of % declines revealed that there was a significant decline in the performance of traits under heat stress environment (late sown) in comparison to a normal environment (timely sown). Three traits, namely grain yield/plant, biological yield/plant and grain weight/ spike showed 57.15 %, 53.66 % and 47.73 % decline, respectively under heat stress environment indicating that above mentioned three traits are highly affected by heat stress. To evaluate the susceptible and tolerant genotypes for grain yield under heat stress environment, six stress tolerance indices viz. heat susceptibility index (HSI), mean productivity (MP), tolerance (TOL), stress tolerance index (STI), trait stability index (TSI) and trait index (TI) were calculated based on the grain yield under normal (timely sown) and stress (late sown) environments. The results of correlation between stress tolerance indices and grain yield in normal environment showed a significant positive relationship with MP (0.822**), STI (0.568**), TSI (0.316**), HSI (0.314**), TI (0.188*) and TOL (0.084). However, in stress environment, grain yield showed a significant positive correlation with TI (1.000**), STI (0.907**), MP (0.713**) and TOL (0.129) and negative correlation with HSI (-0.860**) and TSI (-0.808**). Based on the correlation analysis, we have selected four stress indices viz. HSI (heat susceptibility index), MP (mean productivity), STI (stress tolerance index) and TI (trait index) for their use in the selection of heat tolerant wheat genotypes. The results indicated that 9 genotypes (G51, G64, G71, G114, G119, G134, G139, G148 and G150) were heat tolerant and 12 genotypes (G1, G3, G7, G27, G38, G40, G77, G107, G136, G160, G171 and G187) were highly heat susceptible. The genotype G77 was the most heat sensitive genotype while genotype G119 and G139 showed a high yield under heat stress environment. Therefore, genotypes G119 and G139 were identified as a suitable genotype under late sowing environment and can be recommended for heat stress environment. Further, these two genotypes may be also employed in breeding programs aimed for developing heat tolerant varieties of wheat.