Search

Freezing tolerance of triticale is a major trait contributing to its winter hardiness. The identification of genomic regions – quantitative trait loci (QTLs) and molecular markers associated with freezing tolerance in winter hexaploid triticale was the aim of this study. For that purpose a new genetic linkage map was developed for the population of 92 doubled haploid lines derived from ‘Hewo’ × ‘Magnat’ F1 hybrid. Those lines, together with parents were subjected to freezing tolerance test three times during two winter seasons. Plants were grown and cold-hardened under natural fall/winter conditions and then subjected to freezing in controlled conditions. Freezing tolerance was assessed as the plants recovery (REC), the electrolyte leakage (EL) and chlorophyll fluorescence parameters (JIP) after freezing. Three consistent QTLs for several fluorescence parameters, electrolyte leakage and the percentage of the survived plants were identified with composite interval mapping (CIM) and single marker analysis (SMA). The first locus Qfr.hm-7A.1 explained 9 % of variation of both electrolyte leakage and plants recovery after freezing. Two QTLs explaining up to 12 % of variation in plants recovery and shared by selected chlorophyll fluorescence parameters were found on 4R and 5R chromosomes. Finally, main locus Qchl.hm-5A.1 was detected for chlorophyll fluorescence parameters that explained up to 19.6 % of phenotypic variation. The common QTLs located on chromosomes 7A.1, 4R and 5R, clearly indicated physiological and genetic relationship of the plant survival after freezing with the ability to maintain optimal photochemical activity of the photosystem II and preservation of the cell membranes integrity. The genes located in silico in the identified QTLs include those encoding transmembrane helix proteins like potassium channel and phosphoric ester hydrolase involved in response to osmotic stress as well as proteins involved in the regulation of the gene expression, chloroplast RNA processing and pyrimidine salvage pathway. Additionally, our results confirm that the JIP test is a valuable tool to evaluate freezing tolerance of triticale under unstable winter environments.

Combining to tolerance to biotic and abiotic stresses is important target for modern triticale breeding. Cultivation of varieties resistant to fungal pathogens is economically and environmentally important and may lead to reducing of the use of fungicides. Molecular markers are necessary for accumulation of advantageous alleles in the best genotypes by means of marker-assisted and genomic selection approaches. In present research, QTL regions associated with the powdery mildew resistance at adult plant stage were evaluated in order to provide the effective selection tools. Testing of DH population in multiple environments under natural infestation revealed 20 QTL on wheat (4A, 3B, 4B) and rye (2R, 4R, 5R, 6R) chromosomes. Regions explained 8.1% - 29.3% of phenotypic variation depending of the trait, localization and year of the experiment. Main QTL with effect exceeding 15% were found on chromosomes 3B, 4B, 2R, 5R and 6R. QTL and candidate genes located on chromosomes 4B, 2R, 5R and 6R are so far reported for the first time as regions associated with PM resistance in the adult triticale plants. Additionally, within all QTL, 21 candidate genes associated with the PM resistance were revealed. Predicted function of protein encoded by these genes include triggering a defense system which restricts the pathogen growth, enzyme activity, regulation of hormone-activated pathways, transcriptional corepressor complex and cell wall construction. Availability of QTL, molecular markers together with candidate genes linked with the powdery mildew resistance can be validated on triticale lines and varieties and then, used in MAS to improve modern breeding.

Plant overwintering may be affected in the future by climate change. Low-temperature waterlogging, associated with a predicted increase in rainfall during autumn and winter, can affect freezing tolerance, which is the main component of winter hardiness. The aim of this study was to elucidate the mechanism of change in freezing tolerance caused by low-temperature waterlogging in Lolium perenne, a cool-season grass that is well adapted to a cold climate. The work included: (i) a freezing tolerance test (plant regrowth after freezing), (ii) analysis of plant phytohormones production (abscisic acid [ABA] content and ethylene emission), (iii) measurement of leaf water content and stomatal conductance, (iv) carbohydrate analysis, and (v) analysis of Aco1, ABF2, and FT1 transcript accumulation. Freezing tolerance may be improved as a result of cold waterlogging. The mechanism of this change is reliant on multifaceted actions of phytohormones and carbohydrates, whereas ethylene may counteract ABA signaling. The regulation of senescence processes triggered by concerted action of phytohormones and glucose signaling may be an essential component of this mechanism.

Ten doubled haploid (DH) lines of winter barley with an increased range of freezing/drought tolerance were used to identify phytohormones involved in plant stress acclimation. Cold hardening and drought stress were applied at the most critical stages of plant development on young seedlings and heading plants, respectively. The level of the phytohormones was significantly higher at heading, more than 5-fold in respect of salicylic acid (SA) and total brassinosteroids (BRs) and 1.7-fold in the case of abscisic acid (ABA). Moreover, the spectrum of detectable BRs increased from one—homocastasterone (HCS)—found in seedlings to four BRs identified in heading plants [HCS, castasterone (CS), teasterone and dolicholide], with the last one detected for the first time in cereal species. To some extent freezing tolerance seems to be determined by native hormonal status as control seedlings of tolerant DH lines contained 1.4- and 2.3-fold lower amount of ABA and HCS and 2.3-fold higher amount of SA in comparison to freezing-sensitive ones. Such dependency was not observed in heading plants as significant variation in CS content was the only detected difference. Under stress treatments, tolerant DH lines accumulated significantly lower (75–81%) amount of ABA, which probably reflected lower stress intensity resulting from another defence strategy. In contrast, stress-induced significant almost 2-fold increase in HCS/CS and 2–3-fold decrease in SA content specific for tolerant DH lines of barley suggest the involvement of these molecules in freezing/drought defence. Detected correlations suggest their interaction with nonspecific peroxidase and low molecular weight antioxidants.

The increasing economic importance of triticale (×Triticosecale Wittm.) makes this synthetic hybrid cereal an interesting object of genetic studies. Genomic regions (QTL) of morphological winter triticale traits were determined using the mapping population of 89 doubled haploids lines (DHs) developed from F1 hybrid of cv. ‘Hewo’ and cv. ‘Magnat’ accompanied with the genetic map consisting of 20 linkage groups assigned to the A (7), B (7), and R (6) genomes (total of 3539 DArT, SNP-DArT and SSR markers, length of 4997.4 cM). Five independent experiments were performed in the field and greenhouse controlled conditions. A total of 12 major QTLs located on 2B, 5A, 5R, and 6B chromosomes connected to the stem length, the plant height, the spike length, the number of the productive spikelets per spike, the number of grains per spike, and the thousand kernel weight were identified by a composite interval mapping (CIM).

Mechanisms involved in the de-acclimation of herbaceous plants caused by warm periods during winter are poorly understood. This study identifies the genes associated with this mechanism in winter barley. Seedlings of eight accessions (four tolerant and four susceptible to de-acclimation cultivars and advanced breeding lines) were cold acclimated for three weeks and de-acclimated at 12 °C/5 °C (day/night) for one week. We performed differential expression analysis using RNA sequencing. In addition, reverse-transcription quantitative real-time PCR and enzyme activity analyses were used to investigate changes in the expression of selected genes. The number of transcripts with accumulation level changed in opposite directions during acclimation and de-acclimation was much lower than the number of transcripts with level changed exclusively during one of these processes. The de-acclimation-susceptible accessions showed changes in the expression of a higher number of functionally diverse genes during de-acclimation. Transcripts associated with stress response, especially oxidoreductases, were the most abundant in this group. The results provide novel evidence for the distinct molecular regulation of cold acclimation and de-acclimation. Upregulation of genes controlling developmental changes, typical for spring de-acclimation, was not observed during mid-winter de-acclimation. Mid-winter de-acclimation seems to be perceived as an opportunity to regenerate after stress. Unfortunately, it is competitive to remain in the cold-acclimated state. This study shows that the response to mid-winter de-acclimation is far more expansive in de-acclimation-susceptible cultivars, suggesting that a reduced response to the rising temperature is crucial for de-acclimation tolerance.

Lolium multiflorum/Festuca arundinacea introgression forms have been proved several times to be good models to identify key components of grass metabolism involved in the mechanisms of tolerance to water deficit. Here, for the first time, a relationship between photosynthetic and antioxidant capacities with respect to drought tolerance of these forms was analyzed in detail. Two closely related L. multiflorum/F. arundinacea introgression forms distinct in their ability to re-grow after cessation of prolonged water deficit in the field were selected and subjected to short-term drought in pots to dissect precisely mechanisms of drought tolerance in this group of plants. The studies revealed that the form with higher drought tolerance was characterized by earlier and higher accumulation of abscisic acid, more stable cellular membranes, and more balanced reactive oxygen species metabolism associated with a higher capacity of the antioxidant system under drought conditions. On the other hand, both introgression forms revealed the same levels of stomatal conductance, CO2 assimilation, and consequently, intrinsic water use efficiency under drought and recovery conditions. However, simultaneous higher adjustment of the Calvin cycle to water deficit and reduced CO2 availability, with respect to the accumulation and activity of plastid fructose-1,6-bisphosphate aldolase, were clearly visible in the form with higher drought tolerance.

Impact of photosynthetic and antioxidant capacities on drought tolerance of two closely related forage grasses, Festuca arundinacea and Festuca glaucescens, was deciphered. Within each species, two genotypes distinct in drought tolerance were subjected to a short-term drought, followed by a subsequent re-watering. The studies were focused on: (i) analysis of plant physiological performance, including: water uptake, abscisic acid (ABA) content, membrane integrity, gas exchange, and relative water content in leaf tissue, (ii) analysis of plant photosynthetic capacity (chlorophyll fluorescence, gene expression, protein accumulation, and activity of selected enzymes of the Calvin cycle), and (iii) analysis of plant antioxidant capacity (reactive oxygen species (ROS) generation, gene expression, protein accumulation and activity of selected enzymes). Though, F. arundinacea and F. glaucescens revealed different strategies in water uptake, and partially also in ABA signaling, their physiological reactions to drought and further re-watering, were similar. On the other hand, performance of the Calvin cycle and antioxidant system differed between the analyzed species under drought and re-watering periods. A stable efficiency of the Calvin cycle in F. arundinacea was crucial to maintain a balanced network of ROS/redox signaling, and consequently drought tolerance. The antioxidant capacity influenced mostly tolerance to stress in F. glaucescens.

Impact of photosynthetic and antioxidant capacities on drought tolerance of two closely related forage grasses, Festuca arundinacea and Festuca glaucescens, was deciphered. Within each species, two genotypes distinct in drought tolerance were subjected to a short-term drought, followed by a subsequent re-watering. The studies were focused on: (i) analysis of plant physiological performance, including: water uptake, abscisic acid (ABA) content, membrane integrity, gas exchange, and relative water content in leaf tissue; (ii) analysis of plant photosynthetic capacity (chlorophyll fluorescence; gene expression, protein accumulation, and activity of selected enzymes of the Calvin cycle); and (iii) analysis of plant antioxidant capacity (reactive oxygen species (ROS) generation; gene expression, protein accumulation and activity of selected enzymes). Though, F. arundinacea and F. glaucescens revealed different strategies in water uptake, and partially also in ABA signaling, their physiological reactions to drought and further re-watering, were similar. On the other hand, performance of the Calvin cycle and antioxidant system differed between the analyzed species under drought and re-watering periods. A stable efficiency of the Calvin cycle in F. arundinacea was crucial to maintain a balanced network of ROS/redox signaling, and consequently drought tolerance. The antioxidant capacity influenced mostly tolerance to stress in F. glaucescens.

Plant tolerance to environmental stress is determined by a very complicated network composed of many intra- and extracellular factors. The aim of this study was to select candidate genes involved in responses to freezing and drought in barley on the basis of previous proteomic studies and to analyze changes in their expression caused by application of both stress factors. Six candidate genes for freezing tolerance (namely the genes encoding elongation factor 1 alpha (EF1A), ferredoxin-NADP reductase, a 14-3-3a protein, &beta, fructofuranosidase, CBF2A and CBF4B) and six for drought tolerance (encoding transketolase, periplasmic serine protease, triosephosphate isomerase, a protein with a co-chaperon region (GroEs), pfam14200 and actin) were chosen arbitrarily on the basis of in silico bioinformatic analyses. The expression levels of these genes were measured under control and stress conditions in six DH (doubled haploid) lines with differing freezing and drought tolerance. The results of gene expression analysis confirmed the roles of the candidate genes preselected in this study on the basis of previous proteome analysis in contributing to the differences in freezing and drought tolerance observed in the studied population of DH lines of winter barley.

Under natural conditions, the defense responses of plants exposed to combined abiotic and biotic stress factors, which can randomly interact with each other, are in many aspects different from the response induced by an individual stress. Predicted climatic changes through affecting these simultaneously occurring interactions might change the microclimate surrounding plants, plants’ susceptibility, the range of host microorganisms (i.e., symbionts or pathogens), and their simultaneous interaction with plants. The influence of climate change on interactions between environmental stresses and plants can lead to positive or negative impacts of one stress on the others and cause changes in strategies adopted by plants—either negative (i.e., susceptibility) or positive (i.e., tolerance)—thus causing modifications of primary and secondary metabolism of plants. Primary metabolism plays a key role in plants’ adaptive/defense response through the influence on the modulation of secondary metabolism and the activation of the host’s various defense mechanisms. Alterations in primary and secondary metabolism might include changes in the availability of nutrients, metabolically active compounds, or in carbon (C) and nitrogen (N) metabolism and C/N balance.

Frost tolerance is the main component of winter-hardiness. To express this trait, plants sense low temperature, and respond by activating the process of cold acclimation. The molecular mechanisms of this acclimation have not been fully understood in the agronomically important group of forage grasses, including Lolium-Festuca species. Herein, the introgression forms of L. multiflorum/F. arundinacea distinct with respect to their frost tolerance, were used as models for the comprehensive, proteomic and physiological, research to recognize the crucial components of cold acclimation in forage grasses. The obtained results stressed the importance of photosynthetic performance under acclimation to low temperature. The stable level of photochemical processes after three weeks of cold acclimation in the introgression form with a higher level of frost tolerance, combined simultaneously with only slightly (but not significantly) decreased level of CO2 assimilation after that period, despite significantly lower stomatal conductance, indicated the capacity for that form to acclimate its photosynthesis to low temperature. This phenomenon was driven by the Calvin cycle efficiency, associated with revealed here accumulation profiles and activities of chloroplastic aldolase. The capacity to acclimate the photosynthetic machinery to cold could be one of the most crucial components of forage grass metabolism to improve frost tolerance.

Giant miscanthus (Miscanthus × giganteus) is a valuable plant for bioenergy production and carbon sequestration in low-input agriculture. The newly establishing plantations of miscanthus are, however, at risk during the winter because of the small size of the clones and limited ability of reserves storage in the rhizomes. Increased freezing tolerance of both shoots and rhizomes of giant miscant has been observed after cold acclimation, when plants are exposed to low, non-freezing temperatures. In plants cold acclimation is under complex genetic control that is orchestrated by C-repeat binding factors (CBFs). The present paper aimed the identification and characterisation of CBF-family transcription factors in miscanthus and determined their expression profiles at a low temperature. Three miscanthus genes, homologous to sorghum and maize CBF-family genes were identified. However, only one – MgCBF6 was expressed in the cold-acclimated leaves of miscanthus with an expression profile typical of a CBF-family gene. The ability of miscanthus to cold acclimate was also confirmed using an electrolyte leakage test and survival count. An increase in freezing tolerance was observed both in leaves and rhizomes but only when an increase in the freezing tolerance of plasma membranes was considered. It may be concluded that CBF-induced gene expression pathway may be induced in M. × giganteus while this species use the dormancy strategy for overwintering.

Key message Association mapping of drought-related traits in barley was used to increase the density of existing QTL maps without recreating mapping populations. Abstract We used 109 spring barley genotypes exhibiting high or low drought tolerance to elucidate the associations between diversity array technology sequencing (DArTseq) and single nucleotide polymorphism (SNP) markers and various physiological parameters related to plant responses to drought conditions. The study was performed in controlled conditions (growth chambers), drought tolerance was phenotyped in the four-leaf seedlings. We identified 58 associations including 34 new markers (i.e., 16 DArTseq and 18 SNP markers). The results for three markers were consistent with the data obtained in an earlier traditional biparental QTL mapping study. The regions neighboring markers on linkage group 2H contained the highest number of significant marker–trait associations. Five markers related to the photosynthetic activity of photosystem II were detected on chromosome 4H. The lowest number of associations were observed for the sequences neighboring DArT markers on linkage group 6H. A chromosome 3H region related to water use efficiency and net photosynthesis rate in both biparental QTL, and association study, may be particularly valuable, as these parameters correspond to the ability of plants to remain highly productive under water deficit stress. Our findings confirm that association mapping can increase the density of existing QTL maps without recreating mapping populations. Electronic supplementary material The online version of this article (doi:10.1007/s00122-017-2994-1) contains supplementary material, which is available to authorized users.

Festuca arundinacea and F. pratensis are the models in forage grasses to recognize the molecular basis of drought, salt and frost tolerance, respectively. Transcription profiles of plasma membrane intrinsic proteins (PIPs) and tonoplast intrinsic proteins (TIPs) aquaporin genes were obtained for leaves of Festuca species treated with different abiotic stimuli. F. arundinacea plants were exposed to drought and salt stress, whereas F. pratensis plants were cold-hardened. Changes in genes expression measured with use of real time qRT-PCR method were compared between two genotypes characterized with a significantly different level of each stress tolerance. Under drought the transcript level of PIP1;2 and TIP1;1 aquaporin decreased in both analyzed F. arundinacea genotypes, whereas for PIP2;1 only in a high drought tolerant plant. A salt treatment caused a reduction of PIP1;2 transcript level in a high salt tolerant genotype and an increase of TIP1;1 transcript abundance in both F. arundinacea genotypes, but it did not influence the expression of PIP2;1 aquaporin. During cold-hardening a decrease of PIP1;2, PIP2;1, and TIP1;1 aquaporin transcripts was observed, both in high and low frost tolerant genotypes. The obtained results revealed that the selected genotypes responded in a different way to abiotic stresses application. A reduced level of PIP1;2 transcript in F. arundinacea low drought tolerant genotype corresponded with a faster water loss and a lowering of photosynthesis efficiency and gas exchange during drought conditions. In F. pratensis, cold acclimation was associated with a lower level of aquaporin transcripts in both high and low frost tolerant genotypes. This is the first report on aquaporin transcriptional profiling under abiotic stress condition in forage grasses. Electronic supplementary material The online version of this article (doi:10.1007/s13353-017-0403-8) contains supplementary material, which is available to authorized users.

Climate warming can change the winter weather patterns. Warmer temperatures during winter result in a lower risk of extreme freezing events. On the other hand the predicted warm gaps during winter will decrease their freezing tolerance. Both contradict effects will affect winter survival but their resultant effect is unclear. In this paper, we demonstrate that climate warming may result in a decrease in winter survival of plants. A field study of winterhardiness of common wheat and triticale was established at 11 locations and repeated during three subsequent winters. The freezing tolerance of the plants was studied after controlled cold acclimation and de-acclimation using both plant survival analysis and chlorophyll fluorescence measurements. Cold deacclimation resistance was shown to be independent from cold acclimation ability. Further, cold deacclimation resistance appeared to be crucial for overwintering when deacclimation conditions occurred in the field. The shortening of uninterrupted cold acclimation may increase cold deacclimation efficiency, which could threaten plant survival during warmer winters. Measurements of chlorophyll fluorescence transient showed some differences triggered by freezing before and after deacclimation. We conclude that cold deacclimation resistance should be considered in the breeding of winter cereals and in future models of winter damage risk.

Perennial ryegrass (Lolium perenne L.) belongs to the common cultivated grass species in Central and Western Europe. Despite being considered to be susceptible to drought, it is frequently used for forming the turf in urban green areas. In such areas, the water deficit in soil is recognized as one of the most important environmental factors, which can limit plant growth. The basic aim of this work was to explore the mechanisms standing behind the changes in the photosynthetic apparatus performance of two perennial ryegrass turf varieties grown under drought stress using comprehensive in vivo chlorophyll fluorescence signal analyses and plant gas exchange measurements. Drought was applied after eight weeks of sowing by controlling the humidity of the roots ground medium at the levels of 30, 50, and 70% of the field water capacity. Measurements were carried out at four times: 0, 120, and 240 h after drought application and after recovery (refilling water to 70%). We found that the difference between the two tested varieties’ response resulted from a particular re-reduction of P700+ (reaction certer of PSI) that was caused by slower electron donation from P680. The difference in the rate of electron flow from Photosystem II (PSII) to PSI was also detected. The application of the combined tools (plants’ photosynthetic efficiency analysis and plant gas exchange measurements) allowed exploring and explaining the specific variety response to drought stress.

Photosynthetic acclimation to cold conditions is an important factor influencing freezing tolerance of plants. Photosynthetic enzyme activities increase as part of a photochemical mechanism underlying photosynthetic acclimation to low temperatures. Additionally, a non-photochemical mechanism may be activated to minimize photooxidative damage. The aim of this study was to test the hypothesis that differences in stomatal conductance in Hordeum vulgare plants with contrasting freezing tolerances induce various strategies for photosynthetic acclimation to cold stress. Different stomatal behaviors during the prehardening step resulted in diverse plant reactions to low-temperature stress. Plants with a relatively low freezing tolerance exhibited decreased stomatal conductance, resulting in decreased photochemical activity, faster induction of the non-photochemical mechanism, and downregulated expression of two Rubisco activase (RcaA) splicing variants. In contrast, plants with a relatively high freezing tolerance that underwent a prehardening step maintained the stomatal conductance at control level and exhibited delayed photochemical activity and RcaA expression decrease, and increased Rubisco activity, which increased net photosynthetic rate. Thus, in barley, the induction of photoinhibition avoidance (i.e., non-photochemical photoacclimation mechanism) is insufficient for an effective cold acclimation. An increase in cold-induced net photosynthetic rate due to open stomata is also necessary.

Common buckwheat (Fagopyrum esculentum Moench), a pseudocereal crop, produces a large number of flowers, but this does not guarantee high seed yields. This species demonstrates strong abortion of flowers and embryos. High temperatures during the generative growth phase result in an increase in the degeneration of embryo sacs. The aim of this study was to investigate proteomic changes in flowers and leaves of two common buckwheat accessions with different degrees of heat tolerance, Panda and PA15. Two-dimensional gel electrophoresis and mass spectrometry techniques were used to analyze the proteome profiles. Analyses were conducted for flower buds, open flowers capable of fertilization, and wilted flowers, as well as donor leaves, i.e., those growing closest to the inflorescences. High temperature up-regulated the expression of 182 proteins. The proteomic response to heat stress differed between the accessions and among their organs. In the Panda accession, we observed a change in abundance of 17, 13, 28, and 11 proteins, in buds, open and wilted flowers, and leaves, respectively. However, in the PA15 accession there were 34, 21, 63, and 21 such proteins, respectively. Fifteen heat-affected proteins were common to both accessions. The indole-3-glycerol phosphate synthase chloroplastic-like isoform X2 accumulated in the open flowers of the heat-sensitive cultivar Panda in response to high temperature, and may be a candidate protein as a marker of heat sensitivity in buckwheat plants.

Different pigments often occur together and affect photosynthetic characteristics of the respective leaf portions. In this study, photosynthetic activity in variegated leaves of five cultivars of the ornamental and medicinal plant, Coleus × hybridus hort., was estimated by image analysis and point data measurements of major chlorophyll (Chl) fluorescence parameters and related to the amount of photosynthetic pigments measured with a Chl meter or spectrophotometrically in leaf extracts. Significant differences in Chl and carotenoid (Car) contents were noticed among differentially pigmented sectors of a leaf and among the cultivars. Although the higher Chl concentration was noticed in purple parts compared to green parts of the leaves, the values of minimal and maximal fluorescence yield at the dark- and light-adapted state (F0, Fm, F0', Fm', respectively) were a little lower than those in the green sectors, indicating photoprotective effects provided by anthocyanins and Car, more abundant in the red parts. The lowest Chl and Car content was detected in creamy-yellow and pink sectors and this contributed to low F0, Fm, and Fm', maximal quantum yield of PSII photochemistry, and nonphotochemical and photochemical quenching but high PSII maximum efficiency and effective quantum yield of PSII photochemistry. Both methods of Chl fluorescence analysis revealed heterogeneity in capture, transfer, and dissipation of excitation energy but Chl fluorescence imaging was more suitable in examining very narrow pigmented leaf areas.

Alternative splicing of the Rubisco activase gene was shown to be a point for optimization of photosynthetic carbon assimilation. It can be expected to be a stress-regulated event that depends on plant freezing tolerance. The aim of the study was to examine the relationships among Rubisco activity, the expression of two Rubisco activase splicing variants and photoacclimation to low temperature. The experiment was performed on two Lolium perenne genotypes with contrasting levels of freezing tolerance. The study investigated the effect of pre-hardening (15°C) and cold acclimation (4°C) on net photosynthesis, photosystem II photochemical activity, Rubisco activity and the expression of two splicing variants of the Rubisco activase gene. The results showed an induction of Rubisco activity at both 15°C and 4°C only in a highly freezing-tolerant genotype. The enhanced Rubisco activity after pre-hardening corresponded to increased expression of the splicing variant representing the large isoform, while the increase in Rubisco activity during cold acclimation was due to the activation of both transcript variants. These boosts in Rubisco activity also corresponded to an activation of non-photochemical mechanism of photoacclimation induced at low temperature exclusively in the highly freezing-tolerant genotype. In conclusion, enhanced expression of Rubisco activase splicing variants caused an increase in Rubisco activity during pre-hardening and cold acclimation in the more freezing-tolerant Lolium perenne genotype. The induction of the transcript variant representing the large isoform may be an important element of increasing the carbon assimilation rate supporting the photochemical mechanism of photosynthetic acclimation to cold.

Increased precipitation expected during autumn and winter at higher latitudes in the northern hemisphere may lead to low temperature short-term waterlogging. The temperature of waterlogging was shown to be an important factor controlling plant reaction to this stress. Photosynthetic apparatus response to water excess in the soil at low temperature was examined in Festuca pratensis genotypes with contrasting freezing tolerance. The study was aimed to test the hypothesis whether changes in leaf water-soluble carbohydrate concentration brought about alterations in Rubisco activity may affect the photoacclimation to cold under water excess in the soil. The study investigated the effects of waterlogging during cold acclimation process on a set of chlorophyll a fluorescence parameters, water-soluble carbohydrates, expression of Rubisco activase gene, and Rubisco activity. It was shown that carbohydrate status affected by Rubisco activity might be crucial for the activation of non-photochemical mechanism of photoacclimation to cold under waterlogging. Altered Rubisco activity was only partially attributed to the expression of Rubisco activase gene. Additionally, low carbohydrate concentration in the leaves of waterlogged plants was the condition preventing sugar repression of photosynthesis, including RcaA expression. This indicates that sugar de-repression of photosynthetic genes may be considered a component of photosynthetic acclimation to cold under waterlogging.

Diversity arrays technology (DArT) marker sequences for barley were used for identifying new potential candidate genes for freezing tolerance (FT). We used quantitative trait loci (QTL) genetic linkage maps for FT and photosynthetic acclimation to cold for six- and two-row barley populations, and a set of 20 DArT markers obtained using the association mapping of parameters for photosynthetic acclimation to low temperatures in barley for the bioinformatics analyses. Several nucleotide and amino acid sequence, annotation databases and associated algorithms were used to identify the similarities of six of the marker sequences to potential genes involved in plant low temperature response. Gene ontology (GO) annotations based on similarities to database sequences were assigned to these marker sequences, and indicated potential involvement in signal transduction pathways in response to stress factors and epigenetic processes, as well as auxin transport mechanisms. Furthermore, relative gene expressions for three of six of new identified genes (Hv.ATPase, Hv.DDM1, and Hv.BIG) were assessed within four barley genotypes of different FT. A physiological assessment of FT was conducted based on plant survival rates in two field-laboratory and one laboratory experiments. The results suggested that plant survival rate after freezing but not the degree of freezing-induced leaf damage between the tested accessions can be correlated with the degree of low-temperature downregulation of the studied candidate genes, which encoded proteins involved in the control of plant growth and development. Additionally, candidate genes for qRT-PCR suitable for the analysis of cold acclimation response in barley were suggested after validation.

One hundred and nine accessions of spring barley seedlings were phenotyped under soil drought conditions. Chlorophyll fluorescence induction (OJIP) parameters, leaf water content, relative turgidity, net assimilation rate (P N), and water use efficiency (WUE) of plants were measured. All the tested lines were genotyped by means of DArT sequencing (DArTseq) technology. For association mapping a 11,780 polymorphic DArTseq and 4,725 DArTseq SNP markers were used. Our results revealed dissimilar patterns of the relationships between OJIP-parameters under control and drought conditions. A high level of correlation between parameters characterizing Photosystem's II (PSII) energy trapping efficiency (Fv/Fm) and photochemical events downstream of PSII reaction center (e.g., Performance Index-PICSo) was observed only in the case of drought-treated plants. Generally, OJIP parameters were correlated with leaf water content (less in control). This correlation was weaker with WUE, and absent with P N. Under drought stress, 6,252 genotype × phenotype associations, which passed false discovery rate (FDR) verification, were found between all the studied phenotypic characteristics (23, including 19 OJIP parameters) and 2,721 markers. On the other hand, only 282 associations passed FDR test in the control. They comprised 22 phenotypic parameters and 205 markers. Probing for gene annotations of sequences was performed for markers associated with Fv/Fm for both drought and control, markers were associated with studied traits in both control and drought, as well as for markers associated with both OJIP and other physiological parameters in drought. Our work allowed us to conclude that drought treatment differentiates the studied lines through the revealing of relationships between water content and the damages to PSII reaction centers or different components of PSII energy transfer chain. Moreover, the former was not connected with net photosynthesis rate.

Photosynthetic acclimation to cold conditions is an important factor influencing freezing tolerance of plants. Photosynthetic enzyme activities increase as part of a photochemical mechanism underlying photosynthetic acclimation to low temperatures. Additionally, a non-photochemical mechanism may be activated to minimize photooxidative damage. The aim of this study was to test the hypothesis that differences in stomatal conductance in Hordeum vulgare plants with contrasting freezing tolerances induce various strategies for photosynthetic acclimation to cold stress. Different stomatal behaviors during the prehardening step resulted in diverse plant reactions to low-temperature stress. Plants with a relatively low freezing tolerance exhibited decreased stomatal conductance, resulting in decreased photochemical activity, faster induction of the non-photochemical mechanism, and downregulated expression of two Rubisco activase (RcaA) splicing variants. In contrast, plants with a relatively high freezing tolerance that underwent a prehardening step maintained the stomatal conductance at control level and exhibited delayed photochemical activity and RcaA expression decrease, and increased Rubisco activity, which increased net photosynthetic rate. Thus, in barley, the induction of photoinhibition avoidance (i.e., non-photochemical photoacclimation mechanism) is insufficient for an effective cold acclimation. An increase in cold-induced net photosynthetic rate due to open stomata is also necessary.

In a ×Festulolium population (FuRs0357) of parental origin Lolium perenne × Festuca pratensis, selection of freezing tolerance by freezing tests on whole plants (FT) and chlorophyll a (Chl-a) fluorimetry on frozen detached leaves (CF) was assessed in high and low directions during two cycles of selection. The original population went through two cycles of random mating. All selections and non-selected intercrossed generations of the original population were established in field trials at a coastal site and a continental site in Norway. At the coastal site, analyses of Chl-a fluorimetry parameters and leaf growth on individual plants in autumn and winter hardiness observed in field plots in spring showed that the first-generation selections for high freezing tolerance were associated with winter hardiness and early growth cessation. The second-generation FT-selections for high freezing tolerance were also associated with winter hardiness, whereas the CF-high selections diverged toward high photosynthetic activity. Both low selections were correlated with high photosynthetic activity. There were smaller variations between generations in unselected generations of the original population. Low accumulated leaf growth and early growth cessation were observed in the second-generation FT-selection for high freezing tolerance, whereas high normalized difference vegetation index (NDVI) were seen in Chl-a selections. Both selection methods distinguished diverging selections with significantly different high and low freezing tolerance, but selection efficiency was comparable only for the first selection cycle. Moreover, due to mixed ploidy level in the original population, selection by FT and CF generated diploid and tetraploid plants, respectively, which intensified the response of selection, particularly in the diploid selections. Total dry matter yield (DMY) (mean of three annual cuts for 3 years) of the FT-high selections was lower than for the CF-selections. At coastal sites, selection intensity using freezing tests on whole plants should be adapted to actual climate conditions, to obtain genotypes that balance photosynthetic activity during autumn and good winter hardiness, making them persistent and high yielding.

The aim of the study was to assess the ef fects of low-temperature hardening (2 ◦ C) on the biochemical compounds and processes that can increase resistance of winter rape to inoculation with Leptosphaeria maculans spores. The study involved an evaluation of the entire pool of phenolic compounds, L-phenylalanine ammonia lyase (PAL) activity, excitation intensity for blue and green fluorescence, catalase (CAT) activity, respiration intensity and heat emission from leaf tissues. All the measurements were performed 24 and 72 hours after the inoculation. Low-temperature hardening, which preceded the inoculation of rape seedlings with spores of L. maculans, caused a significant increase in CAT activity and the level of phenolic compounds. The observed changes in PAL activity reflected the changes in phenolics content. The hardened plants showed a significantly higher intensity of blue fluorescence excitation at 24 and 72 hours after the inoculation, as compared to the non-hardened seedlings. Increased content of phenolic compounds and PAL and catalase activity triggered by the temperature of 2 ◦ C and maintained for 24 hours after the inoculation, may confirm the stimulating effect of the hardening temperature. Intensified emission of blue fluorescence indicating saturation of a cell wall with phenolic compounds makes the cell wall structure less stretchy, more tight and leakproof, and thereby hinders fungal growth through plant tissue.

Photoacclimation (PA) and freezing tolerance (FT) have been identified as closely related traits, due to common mechanisms of environmental control. In this study, diversity array technology (DArT) was used for identification of the quantitative trait loci (QTL) of FT and PA in winter barley. Simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers were subsequently used to saturate QTL regions. Two F2 mapping populations were created, for two-rowed (P44) and six-rowed barley (CaP). Different regions of the genome were responsible for differences in traits between parents in these two populations. Eleven QTLs were identified in the P44 population, including five typical for FT and PA, on chromosomes 2H, 3H and 7H. In the CaP population, only one QTL connected with PA and 10 connected with FT were found on all chromosomes except 2H. Our results demonstrate that different sets of markers should be applied in marker-assisted selection for FT in two- and six-rowed barley, as several loci determine FT at the level of biparental crosses.