Improvement Of Thermotolerance Research Articles

Thermo-adaptive evolution of Corynebacterium glutamicum reveals the regulatory functions of fasR and hrcA in heat tolerance

BackgroundHigh-temperature fermentation technology is promising in improving fermentation speed and product quality, and thereby widely used in various fields such as food, pharmaceuticals, and biofuels. However, extreme temperature conditions can disrupt cell membrane structures and interfere with the functionality of biological macromolecules (e.g. proteins and RNA), exerting detrimental effects on cellular viability and fermentation capability.ResultsHerein, a microbial thermotolerance improvement strategy was developed based on adaptive laboratory evolution (ALE) for efficient high-temperature fermentation. Employing this strategy, we have successfully obtained Corynebacterium glutamicum strains with superior resistance to high temperatures. Specifically, the genome analysis indicated that the evolved strains harbored 13 missense genetic mutations and 3 same-sense genetic mutations compared to the non-evolved parent strain. Besides, reverse transcription quantitative PCR analysis (RT qPCR) of the hrcA-L119P mutant demonstrated that both groEL genes were upregulated under 42 °C, which enabled the construction of robust strains with improved heat tolerance. Furthermore, a significant increase in FAS-IA and FAS-IB expression of the fasR-L102F strain was proved to play a key role in protecting cells against heat stress.ConclusionsThis work systematically reveals the thermotolerance mechanisms of Corynebacterium glutamicum and opens a new avenue for revolutionizing the design of cell factories to boost fermentation efficiency.

Foliar application of diethyl aminoethyl hexanoate (DA‐6) alleviated summer bentgrass decline and heat damage to creeping bentgrass

AbstractCreeping bentgrass (Agrostis stolonifera) is sensitive to supra‐optimal temperature, which makes summer bentgrass decline (SBD) a main conundrum troubling turf manager. Diethyl aminoethyl hexanoate (DA‐6) is an artificial and bioactive plant growth regulator affecting tolerance to various abiotic stresses. It is commercially available for field application due to multiple advantages such as low cost, environmentally friendly compound, and sustainable effect. However, the positive effect and mechanism of DA‐6‐mediated heat tolerance in cool‐season grass species and SBD are not yet been explored. Objectives of the study were to screen the optimal dose of DA‐6 for improvement in thermotolerance and to investigate roles of DA‐6 in regulating photosynthesis, osmotic balance, and antioxidant homeostasis in two creeping bentgrass cultivars (heat‐tolerant 13 M and heat‐sensitive Seaside II) in controlled heat condition. Summer heat tolerance induced by the DA‐6 was further evaluated for 2 years under field conditions in a subtropical zone. Results demonstrated that the optimal dose of DA‐6 was screened as 0.1 mM based on analyses of chlorophyll content, photochemical efficiency, and cell membrane stability under heat stress. Foliar application of DA‐6 significantly alleviated heat‐induced leaf senescence by improving photosynthetic performance, accumulation of sugars (sucrose, fructose, and glucose) for osmotic homeostasis, and total antioxidant capacity for reactive oxygen species scavenging in the two cultivars. A 2‐year field test was further carried out to demonstrate that turf quality of the two cultivars was significantly improved after being treated by foliar DA‐6 during hot summer months in 2020 and 2021. Current findings suggested that DA‐6‐mediated thermo‐resistant mechanism was associated with photosynthesis, osmotic adjustment, and antioxidant in cool‐season grass species. Foliar application of DA‐6 could be a cheap, effective, and environmentally friendly strategy to alleviate SBD.

Thermotolerance improvement of engineered Saccharomyces cerevisiae ERG5 Delta ERG4 Delta ERG3 Delta, molecular mechanism, and its application in corn ethanol production

BackgroundThe thermotolerant yeast is beneficial in terms of efficiency improvement of processes and reduction of costs, while Saccharomyces cerevisiae does not efficiently grow and ferment at high-temperature conditions. The sterol composition alteration from ergosterol to fecosterol in the cell membrane of S. cerevisiae affects the thermotolerant capability.ResultsIn this study, S. cerevisiae ERG5, ERG4, and ERG3 were knocked out using the CRISPR–Cas9 approach to impact the gene expression involved in ergosterol synthesis. The highest thermotolerant strain was S. cerevisiae ERG5ΔERG4ΔERG3Δ, which produced 22.1 g/L ethanol at 37 °C using the initial glucose concentration of 50 g/L with an increase by 9.4% compared with the wild type (20.2 g/L). The ethanol concentration of 9.4 g/L was produced at 42 ℃, which was 2.85-fold of the wild-type strain (3.3 g/L). The molecular mechanism of engineered S. cerevisiae at the RNA level was analyzed using the transcriptomics method. The simultaneous deletion of S. cerevisiae ERG5, ERG4, and ERG3 caused 278 up-regulated genes and 1892 down-regulated genes in comparison with the wild-type strain. KEGG pathway analysis indicated that the up-regulated genes relevant to ergosterol metabolism were ERG1, ERG11, and ERG5, while the down-regulated genes were ERG9 and ERG26. S. cerevisiae ERG5ΔERG4ΔERG3Δ produced 41.6 g/L of ethanol at 37 °C with 107.7 g/L of corn liquefied glucose as carbon source.ConclusionSimultaneous deletion of ERG5, ERG4, and ERG3 resulted in the thermotolerance improvement of S. cerevisiae ERG5ΔERG4ΔERG3Δ with cell viability improvement by 1.19-fold at 42 °C via modification of steroid metabolic pathway. S. cerevisiae ERG5ΔERG4ΔERG3Δ could effectively produce ethanol at 37 °C using corn liquefied glucose as carbon source. Therefore, S. cerevisiae ERG5ΔERG4ΔERG3Δ had potential in ethanol production at a large scale under supra-optimal temperature.

Alternative Splicing of TaHsfA2-7 Is Involved in the Improvement of Thermotolerance in Wheat.

High temperature has severely affected plant growth and development, resulting in reduced production of crops worldwide, especially wheat. Alternative splicing (AS), a crucial post-transcriptional regulatory mechanism, is involved in the growth and development of eukaryotes and the adaptation to environmental changes. Previous transcriptome data suggested that heat shock transcription factor (Hsf) TaHsfA2-7 may form different transcripts by AS. However, it remains unclear whether this post-transcriptional regulatory mechanism of TaHsfA2-7 is related to thermotolerance in wheat (Triticum aestivum). Here, we identified a novel splice variant, TaHsfA2-7-AS, which was induced by high temperature and played a positive role in thermotolerance regulation in wheat. Moreover, TaHsfA2-7-AS is predicted to encode a small truncated TaHsfA2-7 isoform, retaining only part of the DNA-binding domain (DBD). TaHsfA2-7-AS is constitutively expressed in various tissues of wheat. Notably, the expression level of TaHsfA2-7-AS is significantly up-regulated by heat shock (HS) during flowering and grain-filling stages in wheat. Further studies showed that TaHsfA2-7-AS was localized in the nucleus but lacked transcriptional activation activity. Ectopic expression of TaHsfA2-7-AS in yeast exhibited improved thermotolerance. Compared to non-transgenic plants, overexpression of TaHsfA2-7-AS in Arabidopsis results in enhanced tolerance to heat stress. Simultaneously, we also found that TaHsfA1 is directly involved in the transcriptional regulation of TaHsfA2-7 and TaHsfA2-7-AS. In summary, our findings demonstrate the function of TaHsfA2-7-AS splicing variant in response to heat stress and establish a link between regulatory mechanisms of AS and the improvement of thermotolerance in wheat.

Direct Transfer and Consolidation of Synthetic Yeast Chromosomes by Abortive Mating and Chromosome Elimination.

Large DNA transfer technology has been challenged with the rapid development of large DNA assembly technology. The research and application of synthetic yeast chromosomes have been mostly limited in the assembled host itself. The mutant of KAR1 prevents nuclear fusion during yeast mating, and occasionally single chromosome can be transferred from one parental nucleus to another. Using the kar1 mutant method, four synthetic yeast chromosomes of Sc2.0 (synIII, synV, synX, synXII) were transferred to wild-type yeasts separately. SynIII was also transferred into an industrial strain Y12, resulting in an improvement of thermotolerance. Moreover, by combining abortive mating and chromosome elimination by CRISPR-Cas9, which has been reported in our previous study, we developed a strategy for consolidation of multiple synthetic yeast chromosomes. Compared to the previous pyramidal strategy using endoreduplication backcross, our method is a linear process independent of meiosis, providing a convenient path for accelerating consolidation of Sc2.0 chromosomes. Overall, the method of transfer and consolidation of synthetic yeast chromosomes by abortive mating and chromosome elimination enables a novel route that large DNA was assembled in donor yeast and then in vivo directly transferred to receptor yeasts, enriching the manipulation tools for synthetic genomics.

Genome-Wide Analysis of HSP70s in Hexaploid Wheat: Tandem Duplication, Heat Response, and Regulation.

HSP70s play crucial roles in plant growth and development, as well as in stress response. Knowledge of the distribution and heat response of HSP70s is important to understand heat adaptation and facilitate thermotolerance improvement in wheat. In this study, we comprehensively analyzed the distribution of HSP70s in hexaploid wheat (TaHSP70s) and its relatives, and we found an obvious expansion of TaHSP70s in the D genome of hexaploid wheat. Meanwhile, a large portion of tandem duplication events occurred in hexaploid wheat. Among the 84 identified TaHSP70s, more than 64% were present as homeologs. The expression profiles of TaHSP70s in triads tended to be expressed more in non-stressful and heat stress conditions. Intriguingly, many TaHSP70s were especially heat responsive. Tandem duplicated TaHSP70s also participated in heat response and growth development. Further HSE analysis revealed divergent distribution of HSEs in the promoter regions of TaHSP70 homeologs, which suggested a distinct heat regulatory mechanism. Our results indicated that the heat response of TaHSP70s may experience a different regulation, and this regulation, together with the expression of tandem duplicated TaHSP70s, may help hexaploid wheat to adapt to heat conditions.

Overexpression of wheat transcription factor (TaHsfA6b) provides thermotolerance in barley.

Overexpressing a heat shock factor gene (TaHsfA6bT) from wheat provides thermotolerance in barley by constitutive expression of heat and other abiotic stress-response genes. Temperature is one of the most crucial abiotic factors defining the yield potential of temperate cereal crops, such as barley. The regulators of heat shock response (HSR), heat stress transcription factors (Hsfs), modulate the transcription level of heat-responsive genes to protect the plants from heat stress. In this study, an Hsf from wheat (TaHsfA6b) is overexpressed in barley for providing thermotolerance. Transgenic barley lines overexpressing TaHsfA6b showed improvement in thermotolerance. The constitutive overexpression of a TaHsfA6b gene upregulated the expression of major heat shock proteins and other abiotic stress-responsive genes. RNA-seq and qRT-PCR analysis confirmed the upregulation of Hsps, chaperonins, DNAJ, LEA protein genes, and genes related to anti-oxidative enzymes in transgenic lines. Excessive generation and accumulation of reactive oxygen species (ROS) occurred in wild-type (WT) plants during heat stress; however, the transgenic lines reflected improved ROS homeostasis mechanisms, showing lesser ROS accumulation under high temperature. No negative phenotypic changes were observed in overexpression lines. These results suggest that TaHsfA6b is a regulator of HSR and its overexpression altered the expression patterns of some main stress-related genes and enhanced the thermotolerance of this cereal crop.

New gene functions are involved in the thermotolerance of the wild wheat relative Aegilops umbellulata

Wheat is one of the most important food crops in the world for human consumption, like all plants it is exposed to environmental stresses including high temperatures. The deleterious effect of high temperatures negatively affects plant growth and development, leading to reduced viability and yield. These effects can be reduced by improvement of thermotolerance through innovative breeding strategies, based on the expansion of the genetic pool available, by exploring important genetic functions from wheat wild progenitors. Improving the genetic thermotolerance characteristics of wheat requires greater understanding of genetic bases of thermotolerance, through identification of high temperature stress related genes. A good source of new useful alleles is given by Aegilops species characterized by thermotolerant habits.In this study we have classified as thermotolerant or thermosensitive, on the basis of physiologic tests, some accessions of wheat wild relative species belonging to Aegilops and Triticum genera. A thermotolerant accession of Aegilops umbellulata (AUM5) was selected, subjected to different thermal treatments and analyzed at transcriptional level. By differential display reverse transcriptase polymerase chain reaction (DDRT-PCR), we investigated modulation of gene expression elicited by heat treatments. This approach allowed the identification of various transcript-derived fragments (TDFs) produced by AUM5 in response to different thermal treatments. The functions of the inducible unique genes in the molecular determination of thermotolerance process are discussed.

Combined Proteome and Transcriptome Analysis of Heat-Primed Azalea Reveals New Insights Into Plant Heat Acclimation Memory.

Plants can obtain superinduction of defense against unpredictable challenges based on prior acclimation, but the mechanisms involved in the acclimation memory are little known. The objective of this study was to characterize mechanisms of heat acclimation memory in Rhododendron hainanense, a thermotolerant wild species of azalea. Pretreatment of a 2-d recovery (25/18°C, day/night) after heat acclimation (37°C, 1 h) (AR-pt) did not weaken but enhanced acquired thermotolerance in R. hainanense with less damaged phenotype, net photosynthetic rate, and membrane stability than non-acclimation pretreated (NA-pt) plants. Combined transcriptome and proteome analysis revealed that a lot of heat-responsive genes still maintained high protein abundance rather than transcript level after the 2-d recovery. Photosynthesis-related genes were highly enriched and most decreased under heat stress (HS: 42°C, 1 h) with a less degree in AR-pt plants compared to NA-pt. Sustainably accumulated chloroplast-localized heat shock proteins (HSPs), Rubisco activase 1 (RCA1), beta-subunit of chaperonin-60 (CPN60β), and plastid transcriptionally active chromosome 5 (pTAC5) in the recovery period probably provided equipped protection of AR-pt plants against the subsequent HS, with less damaged photochemical efficiency and chloroplast structure. In addition, significant higher levels of RCA1 transcripts in AR-pt compared to NA-pt plants in early stage of HS showed a more important role of RCA1 than other chaperonins in heat acclimation memory. The novel heat-induced RCA1, rather than constitutively expressed RCA2 and RCA3, showed excellent thermostability after long-term HS (LHS: 42/35°C, 7 d) and maintained balanced Rubisco activation state in photosynthetic acclimation. This study provides new insights into plant heat acclimation memory and indicates candidate genes for genetic modification and molecular breeding in thermotolerance improvement.

Dynamic Transcriptome Analysis of Anther Response to Heat Stress during Anthesis in Thermotolerant Rice (Oryza sativa L.).

High temperature at anthesis is one of the most serious stress factors for rice (Oryza sativa L.) production, causing irreversible yield losses and reduces grain quality. Illustration of thermotolerance mechanism is of great importance to accelerate rice breeding aimed at thermotolerance improvement. Here, we identified a new thermotolerant germplasm, SDWG005. Microscopical analysis found that stable anther structure of SDWG005 under stress may contribute to its thermotolerance. Dynamic transcriptomic analysis totally identified 3559 differentially expressed genes (DEGs) in SDWG005 anthers at anthesis under heat treatments, including 477, 869, 2335, and 2210 for 1, 2, 6, and 12 h, respectively; however, only 131 were regulated across all four-time-points. The DEGs were divided into nine clusters according to their expressions in these heat treatments. Further analysis indicated that some main gene categories involved in heat-response of SDWG005 anthers, such as transcription factors, nucleic acid and protein metabolisms related genes, etc. Comparison with previous studies indicates that a core gene-set may exist for thermotolerance mechanism. Expression and polymorphic analysis of agmatine-coumarin-acyltransferase gene OsACT in different accessions suggested that it may involve in SDWG005 thermotolerance. This study improves our understanding of thermotolerance mechanisms in rice anthers during anthesis, and also lays foundation for breeding thermotolerant varieties via molecular breeding.

Improvement of Thermotolerance of Zymomonas mobilis by Genes for Reactive Oxygen Species-Scavenging Enzymes and Heat Shock Proteins.

Thermotolerant genes, which are essential for survival at a high temperature, have been identified in three mesophilic microbes, including Zymomonas mobilis. Contrary to expectation, they include only a few genes for reactive oxygen species (ROS)-scavenging enzymes and heat shock proteins, which are assumed to play key roles at a critical high temperature (CHT) as an upper limit of survival. We thus examined the effects of increased expression of these genes on the cell growth of Z. mobilis strains at its CHT. When overexpressed, most of the genes increased the CHT by about one degree, and some of them enhanced tolerance against acetic acid. These findings suggest that ROS-damaged molecules or unfolded proteins that prevent cell growth are accumulated in cells at the CHT.

Electrical stimulation boosts seed germination, seedling growth, and thermotolerance improvement in maize (Zea mays L.)

ABSTRACTElectrical signaling, similar to chemical signalings such as calcium (Ca2+), reactive oxygen species (ROS, mainly hydrogen peroxide: H2O2), nitric oxide (NO), and hydrogen sulfide (H2S), regulates many physiological processes. However, the effect of electrical stimulation on seed germination, seedling growth, and thermotolerance improvement in maize was little known. In this study, using maize as materials, the effect of electrical stimulation on seed germination, seedling growth, and thermotolerance improvement in maize was explored. The results suggested that electrical stimulation with optimal intensity boosted germination rate and seedling growth (as indicated in the increase in the length of shoots and roots, as well as fresh weight) under normal germination conditions. In addition, electrical stimulation augmented the survival rate of maize seedlings, mitigated the decrease in the tissue vitality, and reduced the peroxidation of membrane lipids under heat stress. These data suggested that electrical stimulation could boost seed germination, seedling growth, and thermotolerance improvement in maize.

Physiological plasticity to high temperature stress in chickpea: Adaptive responses and variable tolerance

High temperature stress (HTS) is one of the most crucial factors that limits plant growth and development, and reduces crop yields worldwide. Cool-season crops, particularly the legumes, are severely affected by increasing ambient temperature associated with global climate change. We characterized the HTS-induced modulations of morpho-physicochemical traits and gene expression of several chickpea genotypes and the metabolic profile of the tolerant cultivar. Higher water use efficiency and photosynthetic capacity, minimal membrane lipid peroxidation in conjunction with increased abundance of osmolytes and secondary metabolites depicted thermotolerance of ICC 1205. The adaptive responses were accompanied by high transcript abundance of heat shock proteins and antioxidant enzymes. To integrate stress-responsive signalling and metabolic networks, the HTS-induced physicochemical analysis was further extended to metabolite profiling of the thermotolerant cultivar. The screening of the metabolome landscape led to the identification of 49 HTS-responsive metabolites that include polycarboxylic acid, sugar acids, sugar alcohols and amino acids which might confer thermotolerance in chickpea. The present study, to our knowledge, is the most comprehensive of its kind in dissecting cultivar-specific differential adaptive responses to HTS in chickpea, which might potentiate the identification of genetic traits extendible to improvement of thermotolerance of crops.

Improvement of thermotolerance in Lachancea thermotolerans using a bacterial selection pressure

The use of thermotolerant yeast strains is an important attribute for a cost-effective high temperature biofermentation processes. However, the availability of thermotolerant yeast strains remains a major challenge. Isolation of temperature resistant strains from extreme environments or the improvements of current strains are two major strategies known to date. We hypothesised that bacteria are potential “hurdles” in the life cycle of yeasts, which could influence the evolution of extreme phenotypes, such as thermotolerance. We subjected a wild-type yeast, Lachancea thermotolerans to six species of bacteria sequentially for several generations. After coevolution, we observed that three replicate lines of yeasts grown in the presence of bacteria grew up to 37 °C whereas the controls run in parallel without bacteria could only grow poorly at 35 °C retaining the ancestral mesophilic trait. In addition to improvement of thermotolerance, our results show that the fermentative ability was also elevated, making the strains more ideal for the alcoholic fermentation process because the overall productivity and ethanol titers per unit volume of substrate consumed during the fermentation process was increased. Our unique method is attractive for the development of thermotolerant strains or to augment the available strain development approaches for high temperature industrial biofermentation.

Effect of inorganic salt stress on the thermotolerance and ethanol production at high temperature of Pichia kudriavzevii

Application of cross-protection is expected to improve the thermotolerance of yeasts to enhance their ethanol production at high temperature. In this study, the effects of eight kinds of inorganic salts on the thermotolerance and ethanol production at high temperature in Pichia kudriavzevii were investigated. P. kudriavzevii showed strong thermotolerance and the ability to produce ethanol at high temperature, and higher ethanol production of P. kudriavzevii was observed at high temperature (37–42 °C) compared with that at 30 °C. Inorganic salt stresses induced obvious cross-protection of thermotolerance in P. kudriavzevii. The presence of 0.1 mol/L KNO3 or Na2SO4 or 0.2 mol/L NaCl, KCl, NaNO3, K2SO4 or MgCl2 increased the yeast biomass in YEPD medium at 44 °C to 2.72–3.46 g/L, obviously higher than that in the absence of salt stress (2.17 g/L). The addition of NaCl, KCl, NaNO3, KNO3, Na2SO4, K2SO4, CaCl2 and MgCl2 significantly increased the ethanol production of P. kudriavzevii in YEPD fermentation medium at 44 °C by 37–58%. KCl and MgCl2 exhibited the best performance on improving the thermotolerance and ethanol production, respectively, of P. kudriavzevii. A highly significant correlation (P < 0.01) was obtained among ethanol production, biomass and glucose consumption, suggesting the important role of thermotolerance and glucose consumption in enhanced ethanol production. The combination of NaCl, KCl and MgCl2 had a synergistic effect on the improvement of thermotolerance and ethanol production at high temperature in P. kudriavzevii. This study provides some important clues for improving ethanol production of thermotolerant yeasts at high temperature.

Comparative Proteomic Analysis of Flag Leaves Reveals New Insight into Wheat Heat Adaptation.

Hexaploid wheat (Triticum aestivum L.) is an important food crop but it is vulnerable to heat. The heat-responsive proteome of wheat remains to be fully elucidated because of previous technical and genomic limitations, and this has hindered our understanding of the mechanisms of wheat heat adaptation and advances in improving thermotolerance. Here, flag leaves of wheat during grain filling stage were subjected to high daytime temperature stress, and 258 heat-responsive proteins (HRPs) were identified with iTRAQ analysis. Enrichment analysis revealed that chlorophyll synthesis, carbon fixation, protein turnover, and redox regulation were the most remarkable heat-responsive processes. The HRPs involved in chlorophyll synthesis and carbon fixation were significantly decreased, together with severe membrane damage, demonstrating the specific effects of heat on photosynthesis of wheat leaves. In addition, the decrease in chlorophyll content may result from the decrease in HRPs involved in chlorophyll precursor synthesis. Further analysis showed that the accumulated effect of heat stress played a critical role in photosynthesis reduction, suggested that improvement in heat tolerance of photosynthesis, and extending heat tolerant period would be major research targets. The significantly accumulation of GSTs and Trxs in response to heat suggested their important roles in redox regulation, and they could be the promising candidates for improving wheat thermotolerance. In summary, our results provide new insight into wheat heat adaption and provide new perspectives on thermotolerance improvement.

Exogenous Application of Salicylic Acid: Inducing Thermotolerance In Cotton (Gossypium Hirsutum L.) Seedlings

Improvement in thermotolerance of major cash crops by exogenous application of salicylic acid is an important way to promote economic utilization. Salicylic acid is an important plant growth regulator, commercially utilized to enhance tolerance in plants against harsh environments. The present study was designed to investigate the effects of foliar application of different salicylic acid concentrations (0. 0.5, 1.0, 1.5 mM) on thermotolerance of three cotton ( Gossy­pium hirsutum L.) varieties - “Bt 703”, ‘Bt 131”, and “Bt 3701”- at control (28± 2 O C) and heat shocked (HS) (42±2 O C) cotton seedlings. Heat stress damages were quantified by H2O2 production, malondialdehyde content (MDA), osmo­lyte (Proline) accumulation, and total soluble proteins (TSP). Results revealed that foliar application of salicylic acid minimized heat shock damages; however, different concentrations showed varied responses. Heat stressed seedlings with salicylic application showed increased plant dry matter, free proline accumula­tion, and total soluble protein content as compared to minus salicylic acid stressed seedlings. The above mentioned responses were better in “Bt 703” among the cultivars, making its position thermotolerant. The best responses for most of the above parameters were with the application of 1.0 mM SA. These findings suggest that exogenous application of SA could mitigate the deleterious effects of HS on Gossypium hirsutum seedlings and offer an efficient, economic, and simple means to enhance SH tolerance of the cotton.

Molecular regulation and physiological functions of a novel FaHsfA2c cloned from tall fescue conferring plant tolerance to heat stress.

SummaryHeat stress transcription factors (HSFs) compose a large gene family, and different members play differential roles in regulating plant responses to abiotic stress. The objectives of this study were to identify and characterize an A2‐type HSF, FaHsfA2c, in a cool‐season perennial grass tall fescue (Festuca arundinacea Schreb.) for its association with heat tolerance and to determine the underlying physiological functions and regulatory mechanisms of FaHsfA2c imparting plant tolerance to heat stress. FaHsfA2c was localized in nucleus and exhibited a rapid transcriptional increase in leaves and roots during early phase of heat stress. Ectopic expression of FaHsfA2c improved basal and acquired thermotolerance in wild‐type Arabidopsis and also restored heat‐sensitive deficiency of hsfa2 mutant. Overexpression of FaHsfA2c in tall fescue enhanced plant tolerance to heat by triggering transcriptional regulation of heat‐protective gene expression, improving photosynthetic capacity and maintaining plant growth under heat stress. Our results indicated that FaHsfA2c acted as a positive regulator conferring thermotolerance improvement in Arabidopsis and tall fescue, and it could be potentially used as a candidate gene for genetic modification and molecular breeding to develop heat‐tolerant cool‐season grass species.

Association of SSCP variants of HSP genes with physiological and yield traits under heat stress in wheat

Heat stress alters various physiological phenomenons which adversely affect yield components and ultimately lead to a severe reduction in economic yield in wheat (Triticum aestivum L.). To cope with heat stress, plants have army of heat shock proteins (HSPs) which acts as molecular chaperones stabilizing the polypeptides and membranes. The thermo-tolerance ability of different genotypes of a plant species resides in their genetic diversity of HSP genes. In this study, association of HSPs with physiological and yield traits under heat stress was investigated. Results showed that wheat genotypes differed significantly in their response to high temperature. Single-strand conformation polymorphism (SSCP) analysis of targeted coding sequence of different HSP genes produced seven different haplotypes from HSP 16.9, while only two haplotypes were produced from HSP 23.5, HSP 90a and HSP 90b. Among 25 SSCP variants detected in HSP 16.9 targeted coding sequence, 12 were polymorphic and three of them were found significantly associated with canopy temperature (CT), relative water content (RWC), thousand grain weight (TGW) and normalized difference vegetation index (NDVI). These associated alleles explained 11.4 to 32.9% of the variation for individual trait. The association between HSP variants and these traits may provide new insight for HSPs potential contribution to thermo-tolerance which can be used for improvement of thermo-tolerance in wheat through marker assisted selection.

Cyp11A1 Canola plants under short time heat stress conditions

In order to investigate the high temperature tolerance of spring canola plants (Brassica napus L.) constitutively expressing cyp11A1 gene which encodes bovine cytochrome P450(scc) the growth features were analyzed under short time heat stress (42 degrees C) in growth chamber. Earlier it was documented that results of the heat tolerance test positively correlated with improvement of high temperature resistance in field trial. Higher relative water content (by 13%) and superoxide dismutase (SOD) activity, lower electrolyte leakage (up 1.4-fold) and smaller increase in chlorophyll a and carotenoid contents in cyp11A1 canola leaves in comparison with wild-type plants under stress allowed to conclude cyp11A1 plants are more tolerant to high temperature than the control ones. We suppose that SOD activity increase which revealed in our transgenic canola in normal condition plays the defining role in the biochemical alterations in plant metabolism for the thermotolerance improvement. SOD activity increment could be caused by heterologous cytochrome P450(scc) activity which resulted in the superoxide radical formation. Cyp11A1 canola plants might be resistant to the other stress conditions of different origin.