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Drought and insufficient nutrient supply are the main limiting factors for field crop production, therefore, the present study aimed to investigate the responses of four registered soybean varieties to limited nitrogen (N) supply in combination with drought stress. Plants were grown in tubes filled with silica sand, under open-air conditions. Water shortage was initiated at the three-leaf stage by providing 50% of water, and N deficiency was obtained by reducing the N content of the half-strength Hoagland solution by 50%. N deficiency did not influence the grain yield, while it significantly increased the root dry biomass in Boglár and Pannónia Kincse. Compared with nitrogen supply, the effects of the water shortage were dominant in root development. The expression levels of the investigated genes also showed genotypic variations. The expression of the Inducible nitrate reductase1 gene increased under N-deficient conditions in Boglár and decreased in Pannónia Kincse under drought conditions. The expression level of the aldehyde dehydrogenase gene and abscisic acid 8’-hydroxylase 3 increased under combined stress conditions. Summarising the agronomic and physiological characteristics, Boglár and Sinara were sensitive to drought, Bagera was sensitive to N deficiency but produced the highest yield under limited watering in each nitrogen treatment, while Pannónia Kincse was tolerant to nitrogen deficiency under well-watered conditions.

Aegilops biuncialis is a promising gene source to improve salt tolerance of wheat via interspecific hybridization. In the present work, the salt stress responses of wheat-Ae. biuncialis addition lines were investigated during germination and in young plants to identify which Aegilops chromosomes can improve the salt tolerance of wheat. After salt treatments, the Aegilops parent and the addition lines 2M, 3M and 3M.4BS showed higher germination potential, shoot and root growth, better CO2 assimilation capacity and less chlorophyll degradation than the wheat parent. The Aegilops parent accumulated less Na in the roots due to an up-regulation of SOS1, SOS2 and HVP1 genes, while it contained higher amount of proline, fructose, glucose, galactose, maltose and raffinose. In the leaves, lower Na level was accompanied by high amount of proline and increased expression of NHX2 gene. The enhanced accumulation of sugars and proline was also observed in the roots of 3M and 3M.4BS addition lines. Typical mechanism of 2M addition line was the sequestration of Na into the vacuole due to the increased expression of HVP1 in the roots and NHX2 in the leaves. These results suggest the Aegilops chromosomes 2M and 3M can improve salt tolerance of wheat in different way.

Heat acclimation at non-lethal elevated temperatures may provide protection against a subsequent high temperature stress. The aim of the present work was to evaluate of the effects of heat acclimation in young wheat plants with different origins. Various photosynthetic parameters were used to characterise the physiological responses of plants to high temperature stress with or without heat acclimation at 30 °C. The elevated acclimating temperature did not induce either stomatal closure or photoinhibition. Certain genotypes were able to induce transpiration at acclimating temperature and did not reduce net assimilation. Heat tolerant genotypes could also close stomata faster when they were exposed to severe high temperatures. Heat acclimation could also be detected in various chlorophyll-a fluorescence induction parameters; however, these were less genotype-dependent, and less reflected the differences between varieties. Heat sensitivity and heat acclimation indexes provided useful tool to differentiate the heat responses of the different wheat genotypes. Further studies are needed to establish whether the heat acclimation processes and the differences between the genotypes can also be manifested in adult plants.

The herbicide tolerance levels of 49 Martonvásár inbred parents were examined in Martonvásár in a herbicide susceptibility trial in 2016. The normal dosage recommended in the permit documentations and double dosage were used for the 12 small-plot herbicide treatments performed in two repetitions. Spraying of early post-emergent herbicides was carried out in the 1–2-leaf stage, while post-emergent treatments were applied in the 7–8-leaf stage of maize. The extent of phytotoxicity was scored for the early post-emergent herbicides two and four weeks after treatments and for the post-emergent herbicides two weeks after treatments, respectively. Some of the herbicides examined are not approved in seed production; however it is important to know the reaction of maize parent genotypes for every type of herbicides. The active agent topramezone was withdrawn from the market in 2015, but it was included in the trials as its usage was allowed until stocks run out in 2016. The herbicide agents were examined as follows: mesotrione + S-metolachlor + terbutylazine; isoxaflutol + tiencarbazon methyl + cyprosulfamide; isoxaflutol + cyprosulfamide; mesotrione + terbuthylazine; tembotrione + isoxidifen-ethyl; mesotrione + nicosulfuron; prosulfu ron; nicosulfuron +prosulfuron + dicamba; bentazone + dicamba; nicosulfuron; topramezone; foramsulfuron + isoxadifen-ethyl.Among early post-emergent herbicides, isoxaflutol + cyprosulfamide caused the less phytotoxic damage in the genotypes. The large amount of precipitation during the spring facilitated the infiltration of the active ingredient S-metolachlor, used regularly and successfully also in seed production, into the root zone, resulting in phytotoxic symptoms on susceptible inbred lines at the time of the first inspection. These genotypes recovered by the end of the vegetation period. The spring weather was cooler than usual, retarding the development of maize and thus led to the slower fermentation of herbicide active ingredients, accordingly, all of the post-emergent herbicides caused visible phytotoxic symptoms on some of genotypes. The most severe damages were generally caused by the double dosage of nicosulfuron + prosulfuron + dicamba, nicosulfuron, and foramsulfuron + isoxadifen-ethyl.

In the present work changes in polyamine contents were investigated after various hydroponic polyamine treatments (putrescine, spermidine and spermine at 0.1, 0.3 and 0.5 mM concentrations) in two different crop species, wheat and maize. In contrast to putrescine, higher polyamines (spermidine and spermine) induced concentration-dependent oxidative damage in both crops, resulting in decreased biomass. The unfavourable effects of polyamines were more pronounced in the roots, and maize was more sensitive than wheat. The adverse effects of polyamine treatment were proportional to the accumulation of polyamine and the plant hormone salicylic acid in the leaves and roots of both plant species. Changes in polyamine content and catabolism during osmotic stress conditions were also studied after beneficial pre-treatment with putrescine. The greater positive effect of putrescine in wheat than in maize can be explained by differences in the polyamine metabolism under normal and osmotic stress conditions, and by relationship between polyamines and salicylic acid. The results demonstrated that changes in the polyamine pool are important for fine tuning of polyamine signalling, which influences the hormonal balance required if putrescine is to exert a protective effect under stress conditions.

Understanding the genetic diversity of Aegilops biuncialis, a valuable source of agronomical useful genes, may significantly facilitate the introgression breeding of wheat. The genetic diversity and population structure of 86 Ae. biuncialis genotypes were investigated by 32700 DArT markers with the simultaneous application of three statistical methods— neighbor-joining clustering, Principal Coordinate Analysis, and the Bayesian approach to classification. The collection of Ae. biuncialis accessions was divided into five groups that correlated well with their eco-geographic habitat: A (North Africa), B (mainly from Balkans), C (Kosovo and Near East), D (Turkey, Crimea, and Peloponnese), and E (Azerbaijan and the Levant region). The diversity between the Ae. biuncialis accessions for a phenological trait (heading time), which is of decisive importance in the adaptation of plants to different eco-geographical environments, was studied over 3 years. A comparison of the intraspecific variation in the heading time trait by means of analysis of variance and principal component analysis revealed four phenotypic categories showing association with the genetic structure and geographic distribution, except for minor differences. The detailed exploration of genetic and phenologic divergence provides an insight into the adaptation capacity of Ae. biuncialis, identifying promising genotypes that could be utilized for wheat improvement.

Plant development is continually fine-tuned based on environmental factors. How environmental perturbations are integrated into the developmental programs and how poststress adaptation is regulated remains an important topic to dissect. Vegetative to reproductive phase change is a very important developmental transition that is complexly regulated based on endogenous and exogenous cues. Proper timing of flowering is vital for reproductive success. It has been shown previously that AGAMOUS LIKE 16 (AGL16), a MADS-box transcription factor negatively regulates flowering time transition through FLOWERING LOCUS T (FT), a central downstream floral integrator. AGL16 itself is negatively regulated by the microRNA miR824. Here we present a comprehensive molecular analysis of miR824/AGL16 module changes in response to mild and recurring heat stress. We show that miR824 accumulates gradually in response to heat due to the combination of transient transcriptional induction and posttranscriptional stability. miR824 induction requires heat shock cis-elements and activity of the HSFA1 family and HSFA2 transcription factors. Parallel to miR824 induction, its target AGL16 is decreased, implying direct causality. AGL16 posttranscriptional repression during heat stress, however, is more complex, comprising of a miRNA-independent, and a miR824-dependent pathway. We also show that AGL16 expression is leaf vein-specific and overlaps with miR824 (and FT) expression. AGL16 downregulation in response to heat leads to a mild derepression of FT. Finally, we present evidence showing that heat stress regulation of miR824/AGL16 is conserved within Brassicaceae. In conclusion, due to the enhanced post-transcriptional stability of miR824, stable repression of AGL16 is achieved following heat stress. This may serve to fine-tune FT levels and alter flowering time transition. Stress-induced miR824, therefore, can act as a “posttranscriptional memory factor” to extend the acute impact of environmental fluctuations in the poststress period.

Moderate heat stress may provide protection against a subsequent severe high temperature stress in plants. However, the exact mechanisms of heat acclimation of wheat are still poorly understood. In the present work, two wheat varieties Ellvis and Soissons were exposed to a moderate elevated temperature at 30 °C, and the changes of certain protective mechanisms were investigated. Although the differences in the proline level between the genotypes were not substantial, it was approx. 2–3 times higher in the heat-treated plants than in the controls. After exposure to moderate elevated temperature, the activities of ascorbate peroxidase and catalase were also induced. Similarly, the amount of the free salicylic acid also increased after moderate heat stress, independently on the genotypes. The amount of the main polyamines, namely, putrescine, spermidine, and spermine either did not change or decreased after the same period. However, heat acclimation increased the level of 1,3-diaminopropane, in parallel with a polyamine oxidase gene, TaPAO. While the expression level of the peroxisomal polyamine oxidase gene TaperPAO hardly changed, TaPAO showed a substantial increase after 1 day, especially in Soissons, and at the end of the heat treatment was still significantly higher than in the controls. These suggest that signalling processes related to polyamine metabolisms or salicylic acid-related processes might also contribute to the higher heat tolerance induced by moderate heat stress. The variations in recorded measurements were mainly temperature dependent, and the effect of genotype was less pronounced than the effect of moderate heat treatment itself.

Wild relatives of wheat, such as Aegilops spp. are potential sources of genes conferring tolerance to drought stress. As drought stress affects seed composition, the main goal of the present study was to determine the effects of drought stress on the content and composition of the grain storage protein (gliadin (Gli), glutenin (Glu), unextractable polymeric proteins (UPP%) and dietary fiber (arabinoxylan, β-glucan) components of hexaploid bread wheat (T. aestivum) lines containing added chromosomes from Ae. biuncialis or Ae. geniculata. Both Aegilops parents have higher contents of protein and β-glucan and higher proportions of water-soluble arabinoxylans (determined as pentosans) than wheat when grown under both well-watered and drought stress conditions. In general, drought stress resulted in increased contents of protein and total pentosans in the addition lines, while the β-glucan content decreased in many of the addition lines. The differences found between the wheat/Aegilops addition lines and wheat parents under well-watered conditions were also manifested under drought stress conditions: Namely, elevated β-glucan content was found in addition lines containing chromosomes 5Ug, 7Ug and 7Mb, while chromosomes 1Ub and 1Mg affected the proportion of polymeric proteins (determined as Glu/Gli and UPP%, respectively) under both well-watered and drought stress conditions. Furthermore, the addition of chromosome 6Mg decreased the WE-pentosan content under both conditions. The grain composition of the Aegilops accessions was more stable under drought stress than that of wheat, and wheat lines with the added Aegilops chromosomes 2Mg and 5Mg also had more stable grain protein and pentosan contents. The negative effects of drought stress on both the physical and compositional properties of wheat were also reduced by the addition of these. These results suggest that the stability of the grain composition could be improved under drought stress conditions by the intraspecific hybridization of wheat with its wild relatives.

Young wheat plants (Triticum aestivum L. cv. Mv Béres) were exposed to 0 or 25 mM NaCl for 11 days (salt acclimation). Thereafter the plants were irrigated with 500 mM NaCl for 5 days (salt stress). Irrigating the plants with a low concentration of NaCl successfully led to a reduction in chlorotic symptoms and in the impairment of the photosynthetic processes when the plants were exposed to subsequent high-dose salt treatment. After exposure to a high concentration of NaCl there was no difference in leaf Na content between the salt-acclimated and non-acclimated plants, indicating that salt acclimation did not significantly modify Na transport to the shoots. While the polyamine level was lower in salt-treated plants than in the control, salt acclimation led to increased osmotic potential in the leaves. Similarly, the activities of certain antioxidant enzymes, namely glutathione reductase, catalase and ascorbate peroxidase, were significantly higher in salt-acclimated plants. The results also suggest that while SOS1, SOS2 or NHX2 do not play a decisive role in the salt acclimation processes in young wheat plants; another stress-related gene, WALI6, may contribute to the success of the salt acclimation processes. The present study suggested that the responses of wheat plants to acclimation with low level of salt and to treatment with high doses of salt may be fundamentally different.

Interspecific hybridization between bread wheat (Triticum aestivum, 2n = 42) and related species allows the transfer of agronomic and quality traits, whereby subsequent generations comprise an improved genetic background and can be directly applied in wheat breeding programmes. While wild relatives are frequently used as sources of agronomically favourable traits, cultivated species can also improve wheat quality and stress resistance. A salt-tolerant ‘Asakaze’/‘Manas’ 7H disomic addition line (2n = 44) with elevated β-glucan content, but with low fertility and an unstable genetic background was developed in an earlier wheat-barley prebreeding programme. The aim of the present study was to take this hybridization programme further and transfer the favourable barley traits into a more stable genetic background. Taking advantage of the breakage-fusion mechanism of univalent chromosomes, the ‘Rannaya’ winter wheat 7B monosomic line was used as female partner to the 7H addition line male, leading to the development of a compensating wheat/barley Robertsonian translocation line (7BS.7HL centric fusion, 2n = 42) exhibiting higher salt tolerance and elevated grain β-glucan content. Throughout the crossing programme, comprising the F1-F4 generations, genomic in situ hybridization, fluorescence in situ hybridization and chromosome-specific molecular markers were used to trace and identify the wheat and barley chromatin. Investigations on salt tolerance during germination and on the (1,3;1,4)-β-D-glucan (mixed-linkage glucan [MLG]) content of the seeds confirmed the salt tolerance and elevated grain MLG content of the translocation line, which can be directly applied in current wheat breeding programmes.

The phytotoxic effect of herbicides applied post-emergence was investigated in a herbicide sensitivity experiment set up on parental maize genotypes in Martonvásár. A total of 48 Martonvásár inbred lines and 12 single line crosses were included in small-plot experiments set up in two replications. Ten herbicides were applied at the normal authorised rate and at twice this quantity. Compounds intended for pre-emergence application were applied when maize was in the 3–4-leaf stage and post-emergence herbicides in the 7–8-leaf stage of development. The extent of phytotoxicity was scored two weeks after treatment. Some of the herbicides tested are not authorised for use in seed production fields, but it is important to know how the parental genotypes respond to all types of herbicides. Phytotoxic symptoms of varying intensity were only observed on a third of the 60 parental genotypes examined; the majority of the lines exhibited no reaction to any of the herbicides. Averaged over the 60 genotypes the level of phytotoxic damage was less than 10% for the single dose. When the double dose was applied somewhat more severe damage was induced by products containing Mesotrione + Nicosulfuron or Foramsulfuron + Isoxadifen-ethyl, but this was still below 15%. The herbicide dose had a three times stronger influence on the intensity of the symptoms than the type of herbicide. With the exception of Topramezone, there was a significant difference between the effects of the normal and double doses. The greatest dose effect differences, in decreasing order, were observed for Mesotrione + Nicosulfuron, Foramsulfuron + Isoxadifen-ethyl. Nicosulfuron and Mesotrione + Terbutylazine. The Mesotrione + Terbutylazine active ingredient combination only caused mild (

Cereal grain fiber is an important health-promoting component in the human diet. One option to improve dietary fiber content and composition in wheat is to introduce genes from its wild relatives Aegilops biuncialis and Aegilops geniculata. This study showed that the addition of chromosomes 2Ug, 4Ug, 5Ug, 7Ug, 2Mg, 5Mg, and 7Mg of Ae. geniculata and 3Ub, 2Mb, 3Mb, and 7Mb of Ae. biuncialis into bread wheat increased the seed protein content. Chromosomes 1Ug and 1Mg increased the proportion of polymeric glutenin proteins, while the addition of chromosomes 1Ub and 6Ub led to its decrease. Both Aegilops species had higher proportions of β-glucan compared to arabinoxylan (AX) than wheat lines, and elevated β-glucan content was also observed in wheat chromosome addition lines 5U, 7U, and 7M. The AX content in wheat was increased by the addition of chromosomes 5Ug, 7Ug, and 1Ub while water-soluble AX was increased by the addition of chromosomes 5U, 5M, and 7M, and to a lesser extent by chromosomes 3, 4, 6Ug, and 2Mb. Chromosomes 5Ug and 7Mb also affected the structure of wheat AX, as shown by the pattern of oligosaccharides released by digestion with endoxylanase. These results will help to map genomic regions responsible for edible fiber content in Aegilops and will contribute to the efficient transfer of wild alleles in introgression breeding programs to obtain wheat varieties with improved health benefits. Key Message: Addition of Aegilops U- and M-genome chromosomes 5 and 7 improves seed protein and fiber content and composition in wheat.

The salt stress responses of wheat–barley addition lines (2H, 3H, 3HS, 4H, 6H, 7H and 7HL) were compared to those of the parental genotypes wheat cv. Asakaze and barley cv. Manas and two other wheat genotypes [Chinese Spring (CS) and Mv9kr1] during germination and in young plants grown in hydroponic culture with or without salt treatment. Among the wheat genotypes frequently used for interspecific hybridization, Asakaze possesses relatively high salt tolerance, as indicated by the less pronounced reduction in germination % and in root and shoot growth and the retention of high leaf water content and photosynthetic activity, as compared to CS and Mv9kr1. The barley cv. Manas showed better salt tolerance than wheat cv. Asakaze, although Manas accumulated more Na in the root, but its transport to the shoots is restricted. Among the addition lines tested, the disomic addition line 7H and ditelosomic line 7HL exhibited higher salt tolerance both during germination and in the early developmental stages than the wheat parent, which may be related to the elevated osmotic adjustment capacity of these addition lines, similar to that found for barley cv. Manas. The paper also discusses the effects of other chromosomes on the salt stress response.

Osmotic stress responses of water content, photosynthetic parameters and biomass production were investigated in wheat-Aegilops biuncialis amphiploids and in wheat genotypes to clarify whether they can use to improve the drought tolerance of bread wheat. A decrease in the osmotic pressure of the medium resulted in considerable water loss, stomatal closure and a decreased CO2 assimilation rate for the wheat genotypes, while the changes in these parameters were moderate for the amphiploids. Maximal assimilation rate was maintained at high level even under severe osmotic stress in the amphiploids, while it decreased substantially in the wheat genotypes. Nevertheless, the effective quantum yield of PS II was higher and the quantum yield of non-photochemical quenching of PS II and PS I was lower for the amphiploids than for the wheat cultivars. Parallel with this, higher cyclic electron flow was detected in wheat than in the amphiploids. The elevated photosynthetic activity of amphiploids under osmotic stress conditions was manifested in higher biomass production by roots and shoots as compared to wheat genotypes. These results indicate that the drought-tolerant traits of Ae. biuncialis can be manifested in the wheat genetic background and these amphiploids are suitable genetic materials for improving drought tolerance of wheat.

Goat grasses (Aegilops spp.) contributed to the evolution of bread wheat and are important sources of genes and alleles for modern wheat improvement. However, their use in alien introgression breeding is hindered by poor knowledge of their genome structure and a lack of molecular tools. The analysis of large and complex genomes may be simplified by dissecting them into single chromosomes via flow cytometric sorting. In some species this is not possible due to similarities in relative DNA content among chromosomes within a karyotype. This work describes the distribution of GAA and ACG microsatellite repeats on chromosomes of the U, M, S and C genomes of Aegilops, and the use of microsatellite probes to label the chromosomes in suspension by fluorescence in situ hybridization (FISHIS). Bivariate flow cytometric analysis of chromosome DAPI fluorescence and fluorescence of FITC-labelled microsatellites made it possible to discriminate all chromosomes and sort them with negligible contamination by other chromosomes. DNA of purified chromosomes was used as a template for polymerase chain reation (PCR) using Conserved Orthologous Set (COS) markers with known positions on wheat A, B and D genomes. Wheat-Aegilops macrosyntenic comparisons using COS markers revealed significant rearrangements in the U and C genomes, while the M and S genomes exhibited structure similar to wheat. Purified chromosome fractions provided an attractive resource to investigate the structure and evolution of the Aegilops genomes, and the COS markers assigned to Aegilops chromosomes will facilitate alien gene introgression into wheat.

The Al tolerance of newly developed wheat/barley disomic addition, substitution and translocation lines carry chromosomes of three different barley cultivars was evaluated by comparing the root growth in solution containing 75 μM AlCl3 at pH 4.0 to that of known Al-tolerant and sensitive wheat genotypes. The wheat Asakaze komugi, barley Manas cultivars and their hybrid derivatives were found to have high levels of Al tolerance. The wheat line Mv9kr1, barley cultivar Igri and progenies of the hybrids were sensitive to Al. In most cases, the Al tolerance of the wheat/barley introgression lines derived from Al-sensitive wheat Mv9kr1 and barley Betzes with moderate Al tolerance was similar to that of the wheat parents, but the 2DS.2DL-1HS translocation line of Mv9kr1/Betzes exhibited more intensive root growth, while accumulating less Al than the parental lines. This indicates that either the lack of the distal part of chromosome 2DL or the presence of the distal part of 1HS improved the Al tolerance level.

The chilling tolerance of doubled haploid (DH) maize plants selected and regenerated from microspores exposed to prooxidants, paraquat or tert-butyl hydroperoxide was determined by monitoring cold-induced changes in the photosynthetic electron transport, CO2 assimilation processes and chlorophyll breakdown in young leaves after cold treatment (8°C for 5 days). The results were compared to those of the non-selected DH line and the original hybrid plants. Chilling stress caused a great reduction in the Fv/Fm, qP and ΔF/Fm’ fluorescence parameters, related to the photosynthetic electron transport processes, and in carbon assimilation, and resulted in chlorophyll breakdown. These changes were less extensive in the selected DH plants, which showed elevated antioxidant capacity both at ambient and at low temperature. Among the antioxidant enzymes tested, the activity of GR and GST was induced by chilling stress to the greatest extent. Correlations between cold-induced changes in the photosynthetic apparatus and the antioxidant capacity of the plants suggested that the better protection against oxidative stress induced by the elevated antioxidant capacity of the plants contributed to protecting the photosynthetic apparatus from cold.

To improve the abiotic stress tolerance of maize (Zea mays L.), doubled haploid (DH) plants were produced by in vitro selection of microspores exposed to tert-butyl hydroperoxide (t-BuOOH) as a powerful prooxidant This study investigated the tolerance of the progenies of t-BuOOH-selected DH lines to oxidative stress, cold and drought in controlled environment pot experiments by analyses of photosynthetic electron transport and CO2 assimilation processes, chlorophyll bleaching and lipid peroxidation of leaves. Our results demonstrated that the t-BuOOH-selected DH plants exhibited enhanced tolerance not only to oxidative stress-induced by t-BuOOH but also to cold and drought stresses. In addition, they showed elevated activities of antioxidant enzymes such as superoxide dismutase, ascorbate peroxidase, catalase, glutathione reductase and glutathione S-transferase when compared with the DH lines derived from microspores that were not exposed to t-BuOOH and to the original hybrid plants. The results suggest that the simultaneous up-regulation of several antioxidant enzymes may contribute to the oxidative and cold stress tolerance of the t-BuOOH-selected DH lines, and that the in vitro microspore selection represents a potential way to improve abiotic stress tolerance in maize.

To improve the oxidative stress tolerance of maize (Zea mays L.), doubled-haploid (DH) plants were regenerated from in vitro selected microspores exposed to paraquat. The efficiency of in vitro selection was tested by physiological and biochemical investigations, in which the tolerance to paraquat-mediated oxidative damage and antioxidant capacity of the second generation of fertile microspore-selected lines (DH2) were determined. In the leaves of paraquat-selected DH2 lines, reduced accumulation of reactive oxygen species, lower rates of chlorophyll bleaching, protein breakdown and membrane damage, but higher photosynthetic activity were detected on exposure to paraquat stress than either in the leaves of nonselected DH2 lines or in the original hybrid used as controls. The tolerant DH2 plants showed elevated activities of antioxidant enzymes such as super-oxide dismutase, ascorbate peroxidase, glutathione reductase, glutathione S-transferase, and catalase as compared to both controls. These results suggest that elevated antioxidant capacity may contribute to the paraquat tolerance of the paraquat-selected DH2 lines and that in vitro microspore selection represents a potential way to improve oxidative stress tolerance in maize.

A procedure has been developed for the cryopreservation of wheat female gametes. The procedure involves loading the cells with 25% concentrated vitrification solution consisting of 30% glycerol, 10% sucrose, 120 mM ascorbic acid (AA) and 5% propylene glycol (PG), dehydration in 80% concentrated vitrification solution, droplet vitrification and storage in liquid nitrogen, unloading and rehydration of the cells by gradual addition of isolation solution. Supplementation with AA significantly increased the proportion of viable egg cells after de- and rehydration. During the early phase of rehydration AA reduced the probability of membrane damage caused by rapid water uptake. Maintaining the temperature of the cells at 0°C during the de- and rehydration processes increased cell survival. Microscopic examination of the semi-thin sections of untreated and viable cryopreserved cells revealed that the vitrification process might cause changes in cell structure.

The in vitro selection of microspores and microspore-derived structures under Al stress is one way to improve the Al tolerance of crops. In our study, cytological alterations caused by Al were examined in anther cultures of a commercial wheat (Triticum aestivum L.) variety ‘Mv Palma’, and the efficiency of in vitro selection was demonstrated. Although the anther walls retarded the appearance of toxicity symptoms, cytological changes similar to those observed in root cells (inhibition of cell division, intense vacuolisation, occurrence of micronuclei and cell wall thickening) were detected in the microspores. The severity of Al toxicity and the efficiency of selection depended on the Al concentration and the mode of treatment. Single Al treatments (0.6 and especially 1.6 mM) allowed DH lines with increased Al tolerance to be selected. Repeated Al treatment severely inhibited the cell division of the microspores and it was lethal even at a concentration as low as 0.6 mM. The results show that microspore embryogenesis can be exploited for studying the cytological effect of Al and for increasing the Al tolerance of wheat.

In order to produce doubled-haploid maize plants tolerant of oxidative stress, in vitro microspore selection was carried out in anther culture with reactive oxygen species (ROS) progenitors such as paraquat, menadione, tert-butylhydroperoxide (t-BHP), and methionine combined with riboflavin. All the ROS progenitors reduced the anther induction, the formation of microspore-derived structures, and their regeneration potential. Abnormal cell divisions and progeny cell degradation could be observed during the development of microspores treated with ROS progenitors. Menadione and t-BHP influenced the microspore developmental pathway, as menadione induced the formation of embryoids, while t-BHP increased the proportion of calli in the microspore-derived structures. As the result of in vitro selection, 15, 10, 10, and 3 fertile doubled-haploid plants were obtained in cultures treated with paraquat, t-BHP, methionine combined with riboflavin, and menadione, respectively.

The formation of reactive oxygen species induced by aluminium stress was investigated in an Al-sensitive wheat (Triticum aestivum L.) genotype and in Al-tolerant lines developed by in vitro microspore selection. The roots of Al-tolerant plants were found to exhibit more intensive root growth, while accumulating less Al3+ and reactive oxygen species than Al-sensitive plants under Al stress condition. Aluminium accumulation was found chiefly in the root apex, while the formation of superoxides and peroxides was detected mainly in the elongation zone. Among the superoxide dismutase, ascorbate peroxidase (APX), catalase (Cat) and glutathione-S-transferase (GST) enzymes induced by Al stress, catalase and GST may play an important role in the detoxification of reactive oxygen species in Al-tolerant plants, since they were found to have higher activity than in the Al-sensitive plant.

Atrazine-resistant (AR) weeds have a modified Dl protein structure, with a Ser 264 →Gly mutation on the Dl protein, near the plastoquinone binding niche. The photosynthetic performance, the light response of the xanthophyll cycle and chlorophyll fluorescence quenching-related parameters were compared in attached leaves of susceptible (S) and AR biotypes of the C 3 dicot Chenopodium album L., Epilobium addnocaulon Hausskn., Erigeron canadensis L., Senecio vulgaris L. and Solanum nigrum L. and the C 4 dicot Amaranthus retroflexus L. grown under natural high-light conditions. No significant difference in CO 2 assimilation rate per leaf area unit was found between the S and AR biotypes of the investigated C 3 plants, whereas the AR biotype of A. retroflexus exhibited a relatively poor photosynthetic performance. The Dl protein mutant plants expressed a reduced activity of light-stimulated zeaxanthin formation. Neither the lower violaxanthin de-epoxidase activity nor the depletion of ascorbate seems to be the cause of the lower in vivo zeaxanthin formation in the AR plants. All the Dl mutant weeds had limited light-induced non-photochemical (NPQ) and photochemical (qp) quenching capacities, and displayed a higher photosensitivity, as characterized by the ratio (1-qp)/NPQ and a higher susceptibility to photoinhibition. Analysis of the chlorophyll fluorescence parameters showed that a lower proportion of excitation energy was allocated to PSII photochemistry, while a higher excess of excitation remained in the AR weeds relative to the S plants.

A new method for growing isolated wheat zygotes by co-cultivation with microspores isolated from the same species has been developed. Although the mortality of isolated zygotes within 6 h after the transfer into the nurse culture was relatively high (70%) in this method, the majority of the structures which survived developed into fertile plants (61%). Zygotic structures grown in these cultures were morphologically not as regular as embryos grown in planta, but they resembled them closely in cytological properties and developmental pattern.

The assembly of the photosynthetic apparatus was studied during the first six days of development of Fucus serratus L. embryos. HPLC analysis revealed that oospheres and zygotes contain the same photosynthetic pigments (i.e., chlorophyll a, chlorophyll c, fucoxanthin, violaxanthin, and β-carotene) as fully developed thalli. Total pigment amount increased after fertilization, mainly due to an active synthesis of Chl a and fucoxanthin. Spectral modifications revealing the progressive integration of Chl a and Chl c in the photosynthetic units are described. In particular, a distinct emission at 705 nm, reflecting the accumulation of LHC I, was clearly detected. The emission bands at 705 nm and 725 nm were characterized by 77 K excitation fluorescence measurements. Their spectra differed by the presence of a large band at approximately 550 nm due to fucoxanthin in the excitation spectrum of F705 nm. Room temperature variable fluorescence was first observed 30 h after fertilization indicating a functional Photosystem II electron transfer at this developmental stage.

Providing an adequate quantity and quality of food for the escalating human population under changing climatic conditions is currently a great challenge. In outdoor cultures, sunlight provides energy (through photosynthesis) for photosynthetic organisms. They also use light quality to sense and respond to their environment. To increase the production capacity, controlled growing systems using artificial lighting have been taken into consideration. Recent development of light-emitting diode (LED) technologies presents an enormous potential for improving plant growth and making systems more sustainable. This review uses selected examples to show how LED can mimic natural light to ensure the growth and development of photosynthetic organisms, and how changes in intensity and wavelength can manipulate the plant metabolism with the aim to produce functionalized foods.

The correlation between polyamine content and paraquat resistance in horseweed ( Erigeron canadensis L.) was investigated. Untreated paraquat-resistant and paraquat/atrazine-coresistant plants contained about two to three times higher amounts of polyamines, especially putrescine, compared to sensitive ones. A marked increase of polyamine content was detected after paraquat treatment in each biotype which decreased to the initial level in 6 h. Functional activity (characterized by variable fluorescence) of sensitive plants continuously decreased after paraquat treatment, while resistant biotypes recovered their activities after a transitory inhibition. Changes in polyamine content did not correlate with the changes in functional activity. Exogenously added 10 −4 mol L −1 putrescine combined with 10 −5 mol L −1 paraquat in detached sensitive leaves showed a slight protective effect, while paraquat-resistant and paraquat/atrazine-coresistant leaves remained unaffected. The polyamine level of the cycloheximide-treated or long-term cold-treated plants also changed independently from their functional activity. It can be concluded that the higher polyamine level of the treated resistant leaves and the increase of the polyamine content after paraquat treatment seem to be a general stress response rather than the specific reason for paraquat resistance.

The activities of ascorbate peroxidase (APX), glutathione reductase (GR), and catalase (CAT) were investigated in the rosette stage of paraquat-susceptible, paraquat-resistant, and paraquat/atrazine-coresistant biotypes of Conyza canadensis (L.) Cronq. plants of Hungarian origin, the latter with resistance factors (RF) of 160 and 650 to paraquat. The APX activity in the total leaf extracts of the untreated paraquat-resistant biotype was higher by 25% relative to the paraquat-susceptible and paraquat/atrazine-coresistant biotypes. The levels of GR in both paraquat-resistant biotypes were lower by about 20–25% compared with the susceptible biotype. The APX activity of these biotypes was not induced by paraquat treatment in leaves of sprayed intact plants, whereas the GR activity was slightly increased (by 15–25%). In a series of experiments, the paraquat RF and superoxide dismutase (SOD), APX, and GR activities were compared in the rosette and flowering stages of C. canadensis . An increased (RF ∼1000) level of paraquat resistance was observed in the flowering stage, but only the SOD activity level was enhanced (by 50%) in the paraquat-resistant biotype. Foliar application of diethyldithiocarbamate (DDC, an in vivo inhibitor of Cu/Zn-SOD) or amitrole (AM, an in vivo inhibitor of CAT) caused a marked (50–70%) reversible loss of SOD activity in the chloroplast extract and an 80–90% irreversible decrease in CAT activity in the total cell extracts in all biotypes. However, the paraquat-resistant plants survived the combined treatment with DDC + paraquat or AM + paraquat, and the transient character of paraquat inhibition remained unaffected. The results collectively suggest that the oxygen radical detoxifying pathway with SOD and the enzymes of the ascorbate-glutathione cycle or CAT alone does not explain the very high level of paraquat resistance of the investigated C. canadensis plants.

The role of salicylic acid in Cd tolerance has attracted more attention recently but no information is available on the efficiency of different forms of salicylic acid. The aim was thus to investigate whether both the acid and salt forms of salicylic acid provide protection against Cd stress and to compare their mode of action. Young maize plants were grown under controlled environmental conditions. One group of 10-day-old seedlings were treated with 0.5 mM SA or NaSA for 1 day then half of the pants were treated with 0.5 mM Cd for 1 day. Another group of seedlings was treated with 0.5 mM CdSO4 for 1 day without pre-treatment with SA or NaSA, while a third group was treated simultaneously with Cd and either SA or NaSA. Both salicylic acid forms reduced the Cd accumulation in the roots. Treatment with the acidic form meliorated the Cd accumulation in the leaves, while Na-salicylate increased the phytochelatin level in the roots and the amount of salicylic acid in the leaves. Furthermore, increased antioxidant enzyme activity was mainly induced by the acid form, while glutathione-related redox changes were influenced mostly by the salt form. The acidic and salt forms of salicylic acid affected the two antioxidant systems in different ways, and the influence of these two forms on the distribution and detoxification of Cd also differed. The present results also draw attention to the fact that generalisations about the stress protective mechanisms induced by salicylic acid are misleading since different forms of SA may exert different effects on the plants via separate mechanisms.

Summary The xanthophyll cycle and in vivo photoinhibition were investigated in the herbicide-susceptible (S), paraquat-resistant (PQ-R), atrazine-resistant (Atr-R) and paraquat-atrazine co-resistant (PQAtr-R) biotypes of Conyza canadensis using both low-(LLG) and high-light-grown (HLG) plants. HPLC revealed markedly reduced levels of zeaxanthin under photoinhibitory conditions in atrazine-resistant (Atr-R and PQAtr-R) biotypes as compared with the S and PQ-R biotypes. The variable chlorophyll fluorescence (F v ) decrease and constant fluorescence (F o ) increase revealed an enhanced susceptibility to in vivo photoinhibition in Atr-R and PQAtr-R biotypes. LLG plants were more susceptible than HLG plants to in vivo photoinhibitory treatment. It is suggested that the increased susceptibility to photoinhibition in Atr-R and PQAtr-R biotypes of C. canadensis is a consequence not only of D1 protein mutation, but also possibly of a lower rate of violaxanthin to zeaxanthin de-epoxidation.

Summary Photosynthetic responses of paraquat/atrazine coresistant (PqAR) and only paraquat resistant (PqR) biotypes of horseweed [ Conyza canadensis (L.) Cronq.] on diquat (Reglone) treatment were investigated. Both resistant biotypes showed a transitory inhibition of photosynthetic activity characterized by in vivo CO 2 fixation and F v in the first 2 hours after spraying of the intact plants with 0.5 mM diquat. Similar to the kinetics observed after paraquat spraying of the resistant plants this transitory inhibition was also followed by a gradual recovery. Comparison of paraquat and diquat treatment of paraquat/atrazine coresistant plants showed that recovery after diquat spraying was slower. Ultrastructure of chloroplasts from resistant plants treated by floating the leaves on 10 μM diquat solution showed only moderate changes, while chloroplasts from sensitive Conyza plants seemed to be severely damaged. Paraquat/atrazine coresistant Conyza biotypes of different origin exhibited different, but relatively high resistance against diquat (resistance factor of PqAR 1 = 50, PqAR 2 = 102). Resistance factors for paraquat were higher (PqAR 1 = 210, PqAR 2 = 650).

The excess of the absorbed energy must be sufficiently dissipated to maintain the integrity of the photosynthetic apparatus. One of these protective energy dissipating mechanisms the xanthophyll epoxidation cycle working on the lumenal side of grana thylakoids recently has been thoroughly investigated in many species and under a wide range of environmental and physiological conditions [1,2,3,4].

Atrazine-resistant (AR) weed biotypes have a Ser264-Gly mutation on the D1 protein, causing a decreased binding of atrazine and QB, and some structural/functional changes in PSII complexes (1). The photosynthetic function of AR plants has been reported to be more sensitive to a high light intensity (2–5). Adverse effects of photoinhibition (PI) on the photosynthetic apparatus occur when the capacity of photoprotection (damage and repair) of the plant is exceeded. It has been suggested that a higher susceptibility to the PI of AR biotypes may be caused by an altered DI protein turnover (2) and/or by the limited conversion of xanthophyll cycle components (4,5). A well-studied photoprotective mechanism in plants is associated with the xanthophyll cycle activity and non-photochemical quenching. The increase in the amount of zeaxanthin seems to correlate with an increase in non-photochemical fluorescence quenching of chlorophyll, and is suggested to be a mechanism for the protection of PSII against PI via non-radiative energy dissipation (6).

The Ser264ÆGly substitution on the D1 protein is accompanied by a higher photosensitivity of the mutant plant. This may be due to an increased D1 protein turnover and/or to a lower xanthophyll cycle activity in vivo. The relative importance of these two photoprotective mechanisms in wild and D1 protein mutant biotypes of Erigeron canadensis L. was established by using dithiothreitol and streptomycin. Moreover, the interconversion of violaxan-thin to zeaxanthin via antheraxanthin was studied in isolated thylakoids and in intact leaves treated with paraquat. Streptomycin caused a more severe decrease in the optimal quantum yield (Fv/Fm) of PS II and a large increase in the initial fluorescence yield (Fo) in the mutant compared to the wild biotype. In the fluorescence-quenching parameters of the wild-type leaves, dithiothreitol caused alterations similar to those observed in the mutant plant without dithiothreitol. A lowered activity of the xanthophyll cycle was detected in the mutant biotype compared to the wild-type in vivo. However, under in vitro, conditions which were optimal for violaxanthin de-epoxidation, or when paraquat was used on intact leaves to accelerate the electron transport, violaxanthin could readily be converted to zeaxanthin even in the mutant plants. This demonstrates that neither the decrease in the enzymatic activity of violaxanthin de-epoxidase nor the low availability of violaxanthin is responsible for the low zeaxanthin formation under in vivo conditions. It is presumed that, in vivo, the D1 protein mutation results in slower electron transport, a smaller DpH and lower zeaxanthin formation, and thereby in alterations in the defensive strategies against high light illumination.

RbcS antisense DNA mutants of tobacco have reduced amounts of ribulose bisphosphate carboxylase oxygenase (Rubisco). We found that carotenoid and chlorophyll contents decrease in parallel as Rubisco is decreased, however, pigment levels are not significantly altered until Rubisco levels are reduced sharply. The mutants have normal Chi a /Chi b ratios and normal plastid ultrastructures, suggesting that reductions in Rubisco do not dramatically alter the composition of the thylakoid membranes. Nevertheless, chlorophyll fluorescence measurements, in which developmentally homogenous leaves were sampled, showed that there is reduced photosynthetic capacity of PSII and an enhanced photosensitivity in the mutants, especially in transgenics with severe reductions in Rubisco content. Support for this conclusion comes from several observations; 1) light saturation occurs at a lower light inten­ sity in the mutants, resulting in an earlier closure of PS II (lower photochemical quenching); 2) the mutants have reduced photosynthetic efficiency (lower ΔF/Fm'); and 3) the mutants have a slower recovery of Fv/Fm. We found that acclimation to increasing light intensies in the mutants appears to involve an enhanced inactivation of PSII reaction centers as well as an increased activation of photoprotective mechanisms, notably an engagement of the xanthophyll cycle at lower than normal light intensities. We conclude that the photosensitivity of the antisense mutants is due, in part, to a limitation in Rubisco activation state.