The Stress of Suicide: Temporal and Spatial Expression of Putative Heat Shock Protein 70 Protect the Cells from Heat Injury in Wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)

Heat stress adversely affects growth, development, and yield of winter wheat (Triticum aestivum). Plants have, however, evolved mechanisms to adapt to such conditions mainly by the expression of stress-associated chaperones, the heat shock proteins (HSPs), for modulating the tolerance level. Here, we report cloning of cytosolic putative HSP70 of 1678 bp from a thermotolerant cultivar (C306) of wheat (T. aestivum). A BLASTn search showed maximum homology with the predicted HSP70 protein reported from Hordeum vulgare (accession no AK354795.1). In silico characterization showed the presence of a nucleotide-binding domain of the sugar kinase/HSP70/actin superfamily in the sequence. Putative HSP70 showed temporal and spatial variations in the expression under heat stress (HS). We observed the abundance of HSP70 protein, H₂O₂, proline, and guaiacol peroxidase activity during the seed-hardening stage under HS; accumulation was, however, higher in the thermotolerant C306 than in thermosusceptible HD2329 cultivar. A gradual decrease in cell membrane stability (CMS) and an increase in total antioxidant capacity (TAC) were observed in both the cultivars at the different stages of growth. The expression of HSP70 showed a negative correlation with CMS and a positive correlation with TAC under HS; changes were less pronounced in C306 than in HD2329 at all the stages of growth studied. HSP70 seems to play diverse roles associated with thermotolerance, and partially protect wheat from terminal HS. Being the important member of family of the HSPs, HSP70 needs to be studied in detail, to be used for developing climate-smart wheat crops, through genetic engineering/breeding approaches.     

Increased Antioxidant Activity under Elevated Temperatures: A Mechanism of Heat Stress Tolerance in Wheat Genotypes

An experiment was conducted with three wheat (Triticum aestivum L.) genotypes C 306, HD 2285 and HD 2329 (differently susceptible to water and temperature stress) to study the extent of oxidative injury and activities of antioxidant enzymes in relation to heat stress induced by manipulating dates of sowing. Increase in temperature by late sowing significantly decreased leaf relative water content (RWC), ascorbic acid content, and increased H₂O₂ content and lipid peroxidation in all the genotypes at 8 and 23 d after anthesis. Temperature tolerant genotypes C 306, closely followed by HD 2285 were superior to HD 2329 in maintaining high RWC, ascorbic acid content, and lower H₂O₂ content and lipid peroxidation (malondialdehyde content) under high temperature (late sowing) at the two stages. Activities of superoxide dismutase and catalase were highest in HD 2285 followed by C 306 and minimum in HD 2329 while ascorbate peroxidase activity was highest in C 306.     

The influence of exogenous carbohydrate provision and pre-exercise alkalosis on the heat shock protein response to prolonged interval cycling

The aim of this study was to observe the intracellular heat shock protein 72 (HSP72) and heme oxygenase-1 (HSP32) response to prolonged interval cycling following the ingestion of carbohydrates (CHO) and sodium bicarbonate (NaHCO₃). Six recreationally active males (mean ± SD; age 23.2 ± 2.9 years, height 179.5 ± 5.5 cm, body mass 76.5 ± 6.8 kg, and peak power output 315 ± 36 W) volunteered to complete a 90 min interval cycling exercise on four occasions. The trials were completed in a random and blinded manner following ingestion of either: placebo and an artificial sweetener (P–P), NaHCO₃ and sweetener (B–P), placebo and CHO (P–CHO), and NaHCO₃ and CHO (B–CHO). Both HSP72 and HSP32 were significantly increased in monocytes and lymphocytes from 45 min post-exercise (p ≤ 0.039), with strong relationships between both cell types (HSP72, r = 0.83; HSP32, r = 0.89). Exogenous CHO had no influence on either HSP72 or HSP32, but the ingestion of NaHCO₃ significantly attenuated HSP32 in monocytes and lymphocytes (p ≤ 0.042). In conclusion, the intracellular stress protein response to 90 min interval exercise is closely related in monocytes and lymphocytes, and HSP32 appears to be attenuated with a pre-exercise alkalosis.     

Variation in Osmoregulation in Differentially Drought-Sensitive Wheat Genotypes Involves Calcium

Two wheat (Triticum aestivum L.) genotypes differing in their sensitivity to water deficit (stress tolerant - C306 and stress susceptible - HD2329) were subjected to osmotic stress for 7 d using polyethylene glycol (PEG-6000; osmotic potential –1.0 MPa), at initial vegetative growth. The plants were either supplemented with 1 mM CaCl₂ (Ca²⁺) alone or along with verapamil (VP; calcium channel blocker) to investigate the involvement of calcium in governing osmoregulation. Relative elongation rate (RER), dry matter (DM) production, water potential (_w), electrolyte leakage (EL), contents of proline (Pro) and glycine betaine (GB) and activities of -glutamyl kinase (GK) and proline oxidase (PO) in shoots and roots were examined during stress period. C306 showed relatively higher accumulation of Pro while HD2329 accumulated more GB under stress. RER, DM and _w were relatively higher in C306 than HD2329. Roots compared to shoots showed lower content of osmolytes but had faster rate of their accumulation. Presence of Ca²⁺ in the medium increased the activity of GK and decreased that of PO while in the presence of its inhibitor, decrease in activity of both the enzymes was observed. Ca²⁺ appeared to reduce the damaging effect of stress by elevating the content of Pro and GB, improving the water status and growth of seedlings and minimizing the injury to membranes. The protective effect of Ca²⁺ was observed to be more in HD2329 than C306.     

Overexpression of TaHSF3 in Transgenic Arabidopsis Enhances Tolerance to Extreme Temperatures

Heat shock factors (HSFs) in plants regulate heat stress response by mediating expression of a set of heat shock protein (HSP) genes. In the present study, we isolated a novel heat shock gene, TaHSF3, encoding a protein of 315 amino acids in wheat. Phylogenetic analysis showed that TaHSF3 belonged to HSF class B2. Subcellular localization analysis indicated that TaHSF3 localized in nuclei. TaHSF3 was highly expressed in wheat spikes and showed intermediate expression levels in roots, stems, and leaves under normal conditions. It was highly upregulated in wheat seedlings by heat and cold and to a lesser extent by drought and NaCl and ABA treatments. Overexpression of TaHSF3 in Arabidopsis enhanced tolerance to extreme temperatures. Frequency of survival of three TaHSF3 transgenic Arabidopsis lines was 75–91 % after heat treatment and 85–95 % after freezing treatment compared to 25 and 10 %, respectively, in wild-type plants (WT). Leaf chlorophyll contents of the transformants were higher (0.52–0.67 mg/g) than WT (0.35 mg/g) after heat treatment, and the relative electrical conductivities of the transformants after freezing treatment were lower (from 17.56 to 18.6 %) than those of WT (37.5 %). The TaHSF3 gene from wheat therefore confers tolerance to extreme temperatures in transgenic Arabidopsis by activating HSPs, such as HSP70.     

Salegentibacter echinorum sp. nov., isolated from the sea urchin Hemicentrotus pulcherrimus

A novel Gram-negative, strictly aerobic, heterotrophic, non-motile and yellow-pigmented bacterial strain, designated HD4^T, was isolated from the sea urchin Hemicentrotus pulcherrimus collected from the Yellow Sea in China. Optimal growth of the strain was observed at 28–30 °C, pH 6.8–7.3, and in the presence of 3–5 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence revealed that strain HD4^T exhibited high similarity with the members of Salegentibacter (92.3–95.4 %). The DNA G+C content was 37.0 mol%, MK-6 was the main respiratory quinone and summed feature 3 (comprising iso-C_15:0 2-OH/C_16:1ω7c), iso-C_15:0, iso-C_17:0 3-OH and anteiso-C_15:0 were the major cellular fatty acids. The predominant polar lipids in strain HD4^T were phosphatidylethanolamine and two unknown lipids (L2, L4). Based on the phylogenetic, physiological and biochemical characteristics, strain HD4^T should be classified as a novel species within the genus Salegentibacter, for which the name Salegentibacter echinorum sp. nov. is proposed. The type strain is HD4^T (=CICC 10466^T = NRRL B-59666^T).     

Expression analysis of genes encoding translation initiation factor 3 subunit g (TaeIF3g) and vesicle-associated membrane protein-associated protein (TaVAP) in drought tolerant and susceptible cultivars of wheat

The present study deals with the characterization of genes encoding translation initiation factor 3 subunit g (TaeIF3g) and vesicle-associated membrane protein associated-protein (TaVAP), and how their expression is altered during water stress in the drought tolerant (C-306) and susceptible (HD-2329) cultivars of wheat. Bioinformatics analysis revealed that the TaeIF3g gene consists of an open reading frame (ORF) of 870 nucleotides encoding for a protein of 290 amino acid residues, with a likely molecular mass and pI of 31.47 kDa and 6.89, respectively. The TaVAP cDNA consists of an ORF of 714 nucleotides encoding for a protein of 238 amino acid residues having deduced molecular mass and pI of 25.75 kDa and 7.56, respectively. The changes in expression of the two genes in flag leaf and developing grains were studied in response to drought stress at 15 days post anthesis (DPA). The expression of TaeIF3g and TaVAP in the flag leaf, after increasing in response to mild drought stress, decreased under severe stress conditions in C-306, whereas on the contrary, it persisted in cv. HD-2329. Furthermore, the expression of TaeIF3g and TaVAP in response to drought stress was affected in a coordinated manner in leaf of both the cultivars. The effect of drought on expression of TaeIF3g and TaVAP was also different in the grains of the two cultivars thus implying that the adaptive mechanisms operating in the tissues of tolerant and susceptible cultivars are different.     

Tagging of an Aegilops speltoides Derived Leaf Rust Resistance Gene Lr 28 with a Microsatellite Marker in Wheat

Leaf rust resistance gene Lr28 has been transferred form Aegilops speltoides into bread wheat on chromosome 4AL. To identify the molecular markers linked to Lr28 the available microsatellite markers for wheat chromosome arm 4AL were surveyed on near isogenic lines (NILs) of Triticum aestivum cultivars having Lr28 gene, other Lrgenes and susceptible cultivars. A null allele of Xgwm 160 marker was found to be associated with Lr28. Linkage between the marker and the Lr28 resistance gene was confirmed using F₂ mapping population of cross PBW343 and HD2329 + Lr28.     

Bacillus haikouensis sp. nov., a facultatively anaerobic halotolerant bacterium isolated from a paddy soil

A Gram-stain positive, rod-shaped, endospore-forming and facultatively anaerobic halotolerant bacterium, designated as C-89^T, was isolated from a paddy field soil in Haikou, Hainan Province, People’s Republic of China. Optimal growth was observed at 37 °C and pH 7.0 in the presence of 4 % NaCl (w/v). The predominant menaquinone was identified as MK-7, the major cellular fatty acids were identified as anteiso-C_15:0 and iso-C_15:0, and the major cellular polar lipids were identified as phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and two unknown phospholipids. The peptidoglycan type was determined to be based on meso-DAP. Based on 16S rRNA gene sequence similarity, the closest phylogenetic relatives were identified as Bacillus vietnamensis JCM 11124^T (98.8 % sequence similarity), Bacillus aquimaris JCM 11545^T (98.6 %) and Bacillus marisflavi JCM 11544^T (98.5 %). The DNA G+C content of strain C-89^T was determined to be 45.4 mol%. The DNA–DNA relatedness values of strain C-89^T with its closest relatives were below 18 %. Therefore, on the basis of phylogenetic, chemotaxonomic, and phenotypic results, strain C-89^T can be considered to represent a novel species within the genus Bacillus, for which the name Bacillus haikouensis sp. nov., is proposed. The type strain is C-89^T (=KCTC 33545^T = CCTCC AB 2014076^T).     

Characterisation of wheat germplasm for slow rust resistance against Puccinia striiformis

Thirty-seven wheat germplasm were screened under artificial epiphytotic conditions against stripe rust of wheat at University Research Farm, Chatha, during Rabi, 2013–2014 and 2014–2015. On the basis of final rust severity (FRS), AURPC (Area Under Rust Progress Curve) and CI (Coefficient of Infection), 8 genotypes ((DWR 16, HD2281, VL616, K65, UP2121, HD2329, HD2307 and Lal Bahadur) exhibited partial resistance against the disease. PCR profiles revealed co-dominant pattern with distinct fragment of 150 bp in 14 germplasm, showing the presence of Yr18 gene and 229 bp band in 16 germplasm exhibited absence of Yr18.     

Influence of Rhizobial inoculation on yield of wheat (Triticum aestivum L.)

Summary Soil + charcoal (1∶3) carrier based and liquid cultures of Rhizobia were used to inoculate wheat seed cv. HD2329. The plants received 100 kg N in two equal splits and 60 kg P₂O₅ and 40 kg K₂₀ ha⁻¹. Inoculation with rhizobia had little effect on grain yield of wheat. Significant increase in straw yield and N-uptake occurred due to inoculation. A comparison of results of a similar experiment conducted during 1983–84, showed that inoculation with the same strains of rhizobia and application 50 kg N ha⁻¹ as basal dressing, was more effective in increasing yield and N-uptake in wheat cv. HD2329. It appears reasonable to assume occurrence of nitrogen fixation by root nodule bacteria in rhizosphere of wheat.     

Sterol limitation in a pollen-fed omnivorous lady beetle (Coleoptera: Coccinellidae)

Nutritional constraints of non-prey foods for entomophagous arthropods are seldom investigated, yet are crucial to understanding their nutritional ecology and function within natural and managed environments. We investigated whether pollen from five maize hybrids was of variable quality for the lady beetle, Coleomegilla maculata, whether suitability of these pollens was related with their sterol profiles, and how augmenting sterols (β-sitosterol, cholesterol, or ergosterol) affected the fitness and performance of C. maculata. Preimaginal survival, development rates, the duration of the pre-oviposition period, post-mortem adult dry weight, adult hind tibial length, sex ratio, fecundity, cohort generation time (T_c), net replacement rate (R₀) and intrinsic rate of increase (r) were measured. Individual sterols in the pollens were quantified using GC-MS. Pollens were of variable suitability for C. maculata; the development rate was positively correlated with the amount of 24-methylene-cholesterol and r was positively correlated with episterol and 24-methylene-lophenol found in the pollens. Performance of C. maculata was entirely unaffected by augmenting pollen meals with sterols. This research shows that pollens clearly vary in their sterol contents intraspecifically, which affects their suitability for omnivores that rely on pollen. However, sterols appear to be only one of the limiting nutrients in pollens.     

Influence of temperature on the in vitro pollen germination and pollen tube growth of various native Iranian almonds (Prunus L. spp.) species

Pollen germination and pollen tube growth was quantified among various native Iranian wild almonds (P. dulcis (Mill.) D. A. Webb, P. eleaegnifolia Mill., P. orientalis Mill., P. lycioides Spach, P. reuteri Bioss. et Bushe, P. arabica Olivier, P. glauca Browick and P. scoparia Spach in order to identify differences in the tolerance of pollen to temperature variations. Pollen germination and pollen tube growth were observed after incubation in darkness in a germination medium for 24?h at 10?C50°C at 5°C intervals. Maximum pollen germination of the wild almond species and specify that 60% was obtained for P. orientalis pollen and 98% for P. scoparia. Pollen tube length ranged from 860???m was obtained in P. lycioides and 1490???m in P. scoparia. A modified bilinear model best described the response to temperature of pollen germination and pollen tube length. Almond species variation was found for cardinal temperatures (T _min, T _opt and T _max) of pollen germination percentage and pollen tube growth. Mean cardinal temperatures averaged over eight almond species were 14.7, 24.2, and 43.7°C for maximum percentage pollen germination and 14.48, 25.3, and 44.4 °C for maximum pollen tube length. The principal component analysis (PCA) identified maximum percentage pollen germination and pollen tube length of the species, and T _max for the two processes as the most important pollen parameters in describing a species tolerance to high temperature. PCA also classified Prunus L. spp. into four groups according to the tolerance of pollen to temperature variations. The T _min and T _opt for pollen germination and tube growth, rate of pollen tube growth were less predictive in discriminating species for high temperature tolerance.     

Modulation of Antioxidant Machinery and the Methylglyoxal Detoxification System in Selenium-Supplemented Brassica napus Seedlings Confers Tolerance to High Temperature Stress

We investigated the protective role of selenium (Se) in minimizing high temperature-induced damages to rapeseed (Brassica napus L. cv. BINA Sarisha 3) seedlings. Ten-day-old seedlings which had been supplemented with Se (25 μM Na₂SeO₄) or not were grown separately under control temperature (25 °C) or high temperature (38 °C) for a period of 24 or 48 h in nutrient solution. Heat stress caused decrease in chlorophyll and leaf relative water content (RWC) and increased malondialdehyde (MDA), hydrogen peroxide (H₂O₂), proline (Pro), and methylglyoxal (MG) contents. Ascorbate (AsA) content decreased at any duration of heat treatment. The content of reduced glutathione (GSH) increased only at 24 h of stress, while glutathione disulfide (GSSG) markedly increased at both duration of heat exposure with associated decrease in GSH/GSSG ratio. Upon heat treatment the activities of ascorbate peroxidase (APX), glutathione S-transferase (GST) and glyoxalase I (Gly I) were increased, while the activities of monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), and catalase (CAT) were decreased. The activities of glutathione reductase (GR) and glutathione peroxidase (GPX) remained unchanged under heat stress. However, heat-treated seedlings which were supplemented with Se significantly decreased the lipid peroxidation, H₂O₂, and MG content and enhanced the content of chlorophyll, Pro, RWC, AsA, and GSH as well as the GSH/GSSG ratio. Selenium supplemented heat-treated seedlings also showed enhanced activities of MDHAR, DHAR, GR, GPX, CAT, Gly I, and Gly II as compared to heat-treated seedlings without Se supplementation. This study concludes that exogenous Se application confers heat stress tolerance in rapeseed seedlings by upregulating the antioxidant defense mechanism and methylglyoxal detoxification system.     

Response of in vitro pollen germination and pollen tube growth of groundnut (Arachis hypogaea L.) genotypes to temperature

Air temperatures of greater than 35 °C are frequently encountered in groundnut‐growing regions, especially in the semi‐arid tropics. Such extreme temperatures are likely to increase in frequency under future predicted climates. High air temperatures result in failure of peg and pod set due to lower pollen viability. The response of pollen germination and pollen tube growth to temperature was quantified in order to identify differences in pollen tolerance to temperature among 21 groundnut genotypes. Plants were grown from sowing to harvest in a poly‐tunnel under an optimum temperature of 28/22 °C (day/night). Pollen was collected at anther dehiscence and was exposed to temperatures from 10° to 47·5 °C at 2·5 °C intervals. The results showed that a modified bilinear model most accurately described the response to temperature of percentage pollen germination and maximum pollen tube length. Genotypes were found to range from most tolerant to most susceptible based on both pollen characters and membrane thermostability. Mean cardinal temperatures (T_min, T_opt and T_max) averaged over 21 genotypes were 14·1, 30·1 and 43·0 °C for percentage pollen germination and 14·6, 34·4 and 43·4 °C for maximum pollen tube length. The genotypes 55‐437, ICG 1236, TMV 2 and ICGS 11 can be grouped as tolerant to high temperature and genotypes Kadiri 3, ICGV 92116 and ICGV 92118 as susceptible genotypes, based on the cardinal temperatures. The principal component analysis identified maximum percentage pollen germination and pollen tube length of the genotypes, and T_max for the two processes as the most important pollen parameters in describing a genotypic tolerance to high temperature. The T_min and T_opt for pollen germination and tube growth, rate of pollen tube growth were less predictive in discriminating genotypes for high temperature tolerance. Genotypic differences in heat tolerance‐based on pollen response were poorly related (R² = 0·334, P = 0·006) to relative injury as determined by membrane thermostability.