Lipids are vital cellular constituents in plant, and lipid peroxidation metabolites are critical defence substances in plants. In this study, mass spectrometry along with projections to latent structures discriminant analysis (PLS-DA) was used to detect lipid metabolism changes in Arabidopsis thaliana in response to oligochitosan (an effective resistance elicitor for the control of plant diseases). The PLS-DA showed that lipid metabolites of Arabidopsis thaliana were influenced by oligochitosan treatment. The total content of oxylipin containing monogalactosyldiacylglycerols, oxylipin-containing digalactosyldiacylglycerols, and oxylipin-containing phosphatidylglycerols increased firstly (after 1 h), and then decreased with the increase of oligochitosan treatment duration. In contrast, the total content of monogalactosyldiacylglycerols, phosphatidylcholines, phosphatidyl-ethanolamine, and phosphatidylglycerols decreased firstly, and then increased with the increase of oligochitosan treatment duration. The amounts of free fatty acids (C16:2, C16:3, C18:2, and C18:3) were lower after treatment with oligochitosan for 1, 3, and 6 h than in the control, while the production of volatile organic compounds such as 2-hexenal (except for 3 h) and nonanal was higher than in the control. In conclusion, lipid metabolites of Arabidopsis thaliana were influenced by oligochitosan treatment, and the synthesized lipids and oxylipin-containing lipids were remodelled and free fatty acids was metabolized to volatile organic compounds.

Silicon has been widely reported to have a beneficial effect on improving plant tolerance to biotic and abiotic stresses. However, the mechanisms of Si in mediating responses to simultaneous salt and drought stresses are still poorly understood. Glycyrrhiza uralensis Fisch. is classified as a non-Si accumulator and suffered from salt and drought stresses. In this study, we investigated the long-term application of Si on Si content in G. uralensis roots, stems and leaves, leaf anatomy, ultrastructure, chlorophyll (Chl) content, gas exchange characteristics, relative water content, and growth of two-year-old plants under different salt and drought stresses. Silicon application resulted in a higher Si uptake in G. uralensis roots and more Si accumulation in leaves (especially deposition of Si on cell walls), and Si counteracted the adverse effects induced by salt and drought stresses on the leaf anatomy and ultrastructure. In plants treated with Si, a higher chlorophyll content, net photosynthetic rate and relative water content led to a higher growth rate and dry mass under salt and drought stresses compared with corresponding non-Si treated plants.

Festulolium are grasses formed through interspecific hybridisation of ryegrass (Lolium) and fescue (Festuca) species. The Lolium-Festuca genome complex represents a vast array of heterogeneous and largely outbreeding grass species that have evolved, diverged, and adapted, allowing their world-wide colonisation of temperate grasslands. While strategies for grass improvement have focused primarily on intraspecific breeding and, in particular, on the agronomically desirable species Lolium perenne and Lolium multiflorum, a growing interest has emerged in interspecific hybrids as alternatives. The principal driver has been the increased appreciation of the capability of wide hybridisation to extend phenotypic variation beyond the ranges available within a single species. Lolium and Festuca species share complementary and desirable traits, and the prime aim in Festulolium (Festuca × Lolium) cultivar development has been to combine the agronomic performance of Lolium and the stress resistance of Festuca species. Advances in Festulolium development are timely, and support strategies aimed at delivering a more sustainable future for livestock agriculture, with grass cultivars that are persistent and productive. Festulolium hybrids occur naturally, including examples that demonstrate extreme heterosis with adaptations sufficient to sustain growth in harsh conditions. However, they are largely sterile and their perpetuity depends mainly on vegetative propagation. Synthetic Festulolium hybrids suitable for plant breeding require genome stability and fertility, sufficient for a cost-effective seed production. To this end, suitable amphiploid and introgression-breeding approaches have been developed. Herein, we provide detailed selected highlights in the research and breeding of Festulolium. In addition, recognising the multifunctional properties of grasslands and the development of enabling technologies that permit their study, we review additional benefits likely to accrue from Festulolium that may mitigate climate change effects and provide valuable ecosystem services.

The utility of plant tissue culture for the mass propagation of trees is well known, but continuous in vitro multiplication of plant material may increase the possibility of somaclonal variation; therefore, it is essential to evaluate the genetic integrity of regenerants from species-specific in vitro protocols prior to mass production and implementation. The objectives of this study were: 1) to determine the effect of 2,4-dichlorophenoxyacetic acid (2,4-D) concentration over two cycles of secondary somatic embryogenesis in Magnolia dealbata; and 2) to verify the genetic stability of the regenerants obtained. The embryogenic response was not significantly affected by the concentration of 2,4-D but did vary across cycles of induction. The addition of 4.52 μM 2,4-D induced the highest total number of embryos (100.5), the mean number of somatic embryos (25.1) and somatic embryos per explant (80.6). In both 2,4-D concentration (2.26 or 4.52 μM), genetic integrity between the donor and the propagated clones was 0.90, and the low genetic instability (≤ 0.10 in both PGR treatments) might be due to effect of cyclic somatic embryogenesis or the different response of the explants at stress in in vitro culture conditions. However, it is necessary to examine more cell lines and somatic embryogenesis cycles.

Salicylic acid (SA) is an important plant hormone involved in the activation of defense responses against environmental stresses. However, there are still large of unsolved mysteries about how SA pretreatment affects the establishment of plant stress tolerance. In this study, application of SA at different concentrations and different times were conducted to investigate their effects on the response of Arabidopsis seedlings to salt stress. The pretreatment with 10 or 20 μM SA for more than 6 h promoted Arabidopsis seedlings resistance to salt stress. On the other hand, pretreatment with 200 μM SA reduced Arabidopsis resistance to salt stress and aggravated oxidative damage to the seedlings. At all concentrations used, SA pretreatment inhibited the total respiration and promoted reactive oxygen species (ROS) generation. However, the ROS content in 10 or 20 μM SA pretreated seedlings decreased to the basal level within 6 h and high activities of antioxidant enzymes and alternative oxidase were maintained. Notably, the SA-enhanced salt stress resistance was significantly impaired by blocking alternative oxidase (AOX) pathway. Our findings indicate that SA-mediated salt stress response is in a dose- and time-dependent manner and that the effects were related to the induction of AOX capacity and antioxidant system.

London plane (Platanus acerifolia Wild.) is a famous landscape plant because of its numerous desirable traits except the abundant pollens and seed hairs, which not only pollute the environment but also affect human health. To resolve these problems, we herein isolated and functionally analyzed the promoter of PlacSEP1.1, an orthologous gene of Arabidopsis SEPALLATA1, and investigated the potential usability for cell ablation strategies to engineer reproductive sterility in plants. A 2130 bp 5' upstream region of PlacSEP1.1 was isolated and termed pPlacSEP1.1. Putative motif detections show that there were several types of motifs in pPlacSEP1.1 including core promoter elements, tissue-specific expression regulatory elements, and some negative regulatory elements. β-Glucuronidase histochemical and quantitative assay showed that pPlacSEP1.1 of all deletions was active in all detected tissues except the shortest deletion D5 in roots. In order to test whether pPlacSEP1.1 could be used for London plane sterility breeding with a cytotoxic gene Barnase, the pPlacSEP1.1::Barnase and pPlacSEP1.1::Barnase-mic35S-Barstar vectors were constructed and transformed into tobacco. The pPlacSEP1.1::Barnase transgenic tobacco showed serious defects with respect to vegetative development and died within a couple of weeks after transplantation. On the other hand, most pPlacSEP1.1::Barnase-mic35S-Barstar transgenic tobacco showed normal vegetative growth and inflorescence, and flower development prevented phenotype.

Water deficit is one of the main environmental factors, and the recognition of plant response to drought stress seems to be crucial for the yield improvement of the important crops. The aim of the study was to investigate the effect of prolonged drought stress on dry matter yield, gas exchange parameters, and relative chlorophyll content in the leaves of Festulolium braunii and Trifolium repens grown in pure stands and in mixture. In the pot experiment, different levels of soil moisture (well-watered conditions and drought stress) and of cultivation method (pure stand and mixture) have been applied. The study has shown that all measured parameters were affected by drought stress. Dry mass yield, net photosynthetic rate (P_N), transpiration rate (E), and stomatal conductance were significantly lower under drought stress than under well-watered conditions in all treatment types. T. repens showed the strongest response to stress, whereas F. braunii the weakest when both those species were grown in the pure stand. Under drought conditions, the yield of the mixture was similar to that of F. braunii cultivated in the pure stand, and significantly higher than that of T. repens cultivated in the pure stand. It was also found that under the stress, P_N and E of both species in the the mixture was higher than in the pure stand. The highest water use efficiency was observed in F. braunii grown in the mixture. Our research shows, that in regions with limited rainfall, T. repens is more useful for growing in a mixture with F. braunii, than in the pure stand.

Columnar apple is a valuable resource for genetic improvement of cultivated apples due to its special tree architecture. Strigolactones (SLs) are a novel class of plant hormones controlling shoot branching. The content of SLs is higher in columnar apple than in standard apples. In this study, the members of major gene families involved in SLs biosynthesis and signaling were identified from apple genomic sequences and their expression profiles were characterized in columnar and standard apples using reverse transcription quantitative polymerase chain reactions. In comparison with standard apple, the higher expressions of MORE AXILLARY GROWTH genes MdMAX3-1 and MdMAX4-4 were detected in both buds and shoots of columnar apple but the expression of DWARF gene MdD53-4 showed a lower expression in columnar apple. Overexpression of Columnar gene MdCo31 in tobacco increased SLs content and weakened the inhibition of SLs signal transduction by increasing expression of MAX3 and down-regulating the transcription of D53. Thus MdCo31 could be a strong candidate gene for the control of columnar habit.

Imidazolinone herbicides combined with imidazolinone resistant (IMI-R) crops provide a tool for solving the important problem of the occurrence of weeds during the early growth stages of sunflower. These herbicides inhibit the synthesis of branched chain amino acids by interrupting the key enzyme acetohydroxyacid synthase (AHAS). We studied the imazamox detoxification in an IMI-R sunflower hybrid together with plant growth and photosynthetic performance. Inhibition of photosynthesis and growth were observed as initial effects of imazamox application. A slight decrease in AHAS activity was also noticed. These effects disappeared within two weeks after application. A fast and well-functioning detoxification mechanism for the herbicide, of which the content decreased for about 90 % at 14 d after application, seems to be responsible for this. The activity of the xenobiotic detoxifying enzyme glutathione S-transferases (GSTs) significantly increased after imazamox application. Our results suggest that the metabolite glutathione serves as an auxiliary tool for imazamox detoxification through conjugation reactions realized by the GSTs, thereby taking part in the non-target mechanisms of resistance in IMI-R sunflower hybrids.

Climate change calls for new methods and plant materials to breed crops adapted to new environmental conditions. Sustainable forage and amenity grass production during periods of severe drought and heat waves during summer, and unequal distribution of precipitation over the year will require drought-tolerant genotypes. However, high-yielding ryegrasses (Lolium spp.), which are the most commonly used grass species, suffer during abiotic stresses. Introgression of drought and heat tolerance from closely related fescues (Festuca spp.) offers an opportunity to develop superior hybrid cultivars to mitigate the negative impact of climate change. Intergeneric cross-hybridization and the development of Festulolium (Festuca × Lolium) hybrids was initiated 100 years ago and resulted in registration of almost one hundred cultivars. For a long time, their genome composition was not known and was debated by breeders and geneticists. In the last three decades, molecular cytogenetic and genomic approaches have enabled their detailed characterization. These studies revealed a gradual replacement of Festuca chromosomes by those of Lolium in consecutive generations leading to an almost complete elimination of Festuca chromatin in the introgression forms. On the other hand, amphiploid cultivars seem to be more stable with the optimal proportions of the Lolium to Festuca genomes at about 2:1. In this mini review, we discuss recent advances in the analysis of the genome composition of Festulolium hybrids with a specific focus on genome (in)stability.

Drought resistance in plants can be associated with four different strategies to cope with water stress. These strategies are classified as drought escape, avoidance, tolerance, and recovery. The expression of each strategy depends on plant species and its genetic potential, but also on the environmental conditions, including the stress intensity and duration. Often, prolonged drought conditions are associated with drought escape or avoidance, whereas short but severe drought periods induce drought tolerance. To analyze the components of drought resistance in forage grasses, we applied two Lolium multiflorum/Festuca arundinacea introgression forms into a comprehensive research. Obtained results clearly show that the response of plants to severe short term drought conditions with limited rhizosphere did not reflect their response to long progressive drought conditions, which did not limit root growth. The BC4-INT-40 introgression form with extensive and deep roots was characterized by a more efficient drought avoidance and regeneration mechanisms under long-term drought, whereas the BC4-INT-66 form with shorter roots revealed a lower productivity and re-growth capacity under the prolonged drought. On the other hand, this form had also a better photosynthetic performance under short and intensive drought conditions with a limited space for root development.

Light and temperature are two essential environmental cues for plants, helping to optimize plant body architecture and physiology. To sense a broad spectrum of sun radiation spanning from UV-B to far-red wavelength, plants are equipped with a sophisticated array of photoreceptors, including phytochromes, cryptochromes, phototropins, Zeitlupes, and UV-B photoreceptor UVR8. On the contrary, since the thermodynamic effects extensively affect the molecular and supramolecular structures, it is difficult to identify the entry point or initial receptor of temperature. Even so, several putative temperature sensors have been proposed, such as calcium ion channels, H2A.Z, and the thermodynamic change of plasma membrane fluidity. Considering that many processes in plant respond to irradiance and temperature, scientists devote to finding out the converge point of these environmental cues. As a typical example, circadian rhythm is such an integration point, which receives the signal input of both irradiance and temperature. The updating evidence shows, as an important photoreceptor, phytochrome B acts as temperature sensors via a thermodynamic active state revision. These findings suggest that the studies on light and temperature receptors in plants should not be separated. Their extensive convergence during signalling provides a new direction for understanding the stimuli perception mechanisms.

To clarify the effects of the introduction of Festuca pratensis-derived genomic regions in a tetraploid Lolium perenne genomic background, we analyzed quantitative trait loci (QTLs) for winter hardiness and eight other agronomic traits using two mapping populations generated by sequential backcrosses of two different amphiploid F. pratensis × L. perenne hybrids to tetraploid L. perenne. We constructed two interspecific linkage maps: FLBC2A (143 DNA markers, 301.2 cM total) and FLBC2C (126 markers, 256.2 cM), based on F. pratensis-derived alleles, with 72 markers in common. Interval mapping detected QTLs for seven traits in FLBC2A and for six traits in FLBC2C. In both populations, QTLs for winter hardiness with a positive effect of the F. pratensis allele were detected around common markers on linkage group (LG) 1. In FLBC2A, other additional QTLs for winter hardiness with a positive effect of the F. pratensis allele were detected on LG4 and LG5, and one with a negative effect was detected on LG2. In both populations, a QTL for heading date was detected on LG7. Quantitative trait loci for vigor after planting and stubble width were detected on different LGs between populations. Quantitative trait loci for dry matter yield before winter, plant growth habit, inflorescence length, number of spikelets per inflorescence, and leaf width were detected in only one population. Population-dependent QTLs thought to be involved in intraspecific variation in addition to interspecific one. On the other hand, QTLs found in the two different populations could lead to the development of versatile markers for Festulolium breeding.

Strelitzia reginae is a popular cut flower that has blue petals and orange sepals. Flower color is an important plant trait; however, little is known about its molecular mechanisms in S. reginae. In this study, cDNA libraries were constructed for blue petals and orange sepals of S. reginae. A total of 75 487 unigenes were obtained from transcriptome sequencing and de novo assembly, of which 41.86 % were annotated by public databases. Ultra-high performance liquid chromatography analysis revealed that anthocyanins were the main pigment in blue petals, and that carotenoids controled pigment formation in the orange sepals. Using a system analysis-based approach, 73 and 29 candidate genes related to anthocyanin and carotenoid biosyntheses were identified, respectively. Among these, chalcone synthase 2, chalcone isomerase 1, flavanone 3-hydroxylase 1, flavonoid 3',5'-hydroxylase 1, dihydroflavonol 4-reductase 1, anthocyanidin synthase 1, and anthocyanidin 3-O-glucosyltransferase 1 were considered to be important in regulating the formation of blue petals, and phytoene synthase 1, phytoene desaturaser 1, ζ-carotene desaturase 1, lycopene β-cyclase 3, and β-carotene hydroxylase 2 might play important roles in orange sepal formation. This study improves our understanding of flower color and provides evidence for future molecular breeding of ornamental plants based on flower color modifications.

Pyramiding transgenes of interest is one of the strategies to engineer multiple stress resistance in crop plants. Transgenic plants which stably express different genes can be hybridized to bring these genes together in one plant. Transgenic rice (Oryza sativa L. cv. ASD 16) plants harbouring genes Xa21 (conferring bacterial blight resistance), tlp (conferring resistance to sheath blight), or gna (conferring resistance to brown planthopper) were used in hybridization experiments. Sexual hybridization was carried out in two different gene combinations: Xa21 × gna and tlp × gna. Molecular analyses were carried out to confirm the presence of transgenes. In F₁ generation, lines harbouring either gene in each of the cross-combination were selected and forwarded to F₂ generation. The presence of genes in F₂ generation was confirmed by PCR, Southern blot hybridization, and Western blotting. The F₂ progeneis harbouring Xa21 and gna exhibited resistance against bacterial blight and moderate resistance against brown planthopper. Similarly, the F₂ lines of tlp and gna combination provided resistance against sheath blight and moderate resistance against brown planthopper. The level of resistance observed in pyramided lines for insect or pathogens was comparable to the resistance observed in their parental lines. Our study shows that pyramiding genes by hybridization between transgenic plants could be one of the strategies to develop cultivars with multiple biotic stress resistances.

This study investigated the mitigating effects of spermidine (Spd) application on salinity-induced ion inbalance, physiological properties, and the expression of some genes in foxtail millet (Setaria italica L.). We observed 30-d-old seedlings maintained at a half-strength Hoagland solution (control), 1.0 % NaCl, 10, 20, and 40 μM Spd, and 10, 20, and 40 μM Spd + 1.0 % (m/v) NaCl for 14 d. The results show that salt stress significantly inhibited plant growth, and this was significantly ameliorated by Spd. The mass of the shoots and roots, content of chlorophyll a and chlorophyll b, root activity, and K⁺ content were higher whereas Na⁺ content, Na⁺/K⁺ ratio, relative electrolyte leakage, glutathione content, H₂O₂ content, activity of glutathione reductase (GR), and catalase (CAT) were lower after application of Spd in comparison with NaCl alone. The expression of GR, ascorbate peroxidase, CAT, and superoxide dismutase genes also significantly decreased in salt stressed plants with Spd. This study has proved the role of Spd in alleviating salt stress in foxtail millet and identified that 20 μM Spd was most effective.

Melatonin (N-acetyl-5-methoxytryptamine) is an essential molecule which regulates plant growth and development and alleviates the damaging effects of abiotic stresses. To evaluate the important functions of melatonin in response to salinity stress, the effects of exogenous melatonin on the antioxidant system and growth of tomato (Solanum lycopersicum L.) under 150 mM NaCl stress were investigated. The application of 100 μM melatonin compensated the growth inhibition caused by salt-stress. Melatonin treated seedlings had an increased fresh and dry masses of shoots and roots. The application of 1 - 200 µM melatonin notably enhanced the relative chlorophyll content (SPAD index), root characteristics, and gas exchange in tomato seedlings subjected to salt stress compared to seedlings treated with salt stress alone. Moreover, melatonin pretreatment minimized accumulation of reactive oxygen species and improved activities of antioxidative enzymes including catalase, superoxide dismutase, glutathione reductase, and ascorbate peroxidase.

Dramatic accumulation of proline is a common physiological response in plants exposed to various abiotic stresses. Accumulation of proline could be due to de novo synthesis, decreased degradation, lower utilization, or hydrolysis of proteins. Extensive intercellular proline transport occurs between the cytosol, chloroplasts, and mitochondria due to its compartmentalized metabolism. Although all functions of proline in stress tolerance are still a matter of debate, it is suggested that proline contributes to stabilization of sub-cellular structures, scavenging free radicals, and buffering cellular redox potential. It also chelates heavy metals, modulates cellular functions, and even triggers gene expression. Apparently, proline acts as stress-related signal exhibiting cross tolerance to a range of different stresses. Besides these significant roles, its metabolism is found to be coupled to several key pathways such as pentose phosphate, tricarboxylic acid, or urea cycles and contributes to, i.e., purine synthesis and the phenylpropanoid pathway. Although the molecular basis of regulation of proline metabolism is still largely obscure, the genetic engineering of proline content could lead to new opportunities to achieve plant stress tolerance.

Reverse transcription quantitative PCR is a widely used method to detect gene expressions. To obtain accurate expression results, the selection of proper reference genes is important and necessary. However, related works concerning reference gene selection have not been carried on many plant species, especially endangered ones. The aim of the present study was to select dependable reference genes for expression normalization of an endangered plant species with medicinal and ornamental value: Sinocalycanthus chinensis (Calycanthaceae). Nine reference genes with stable expressions were chosen for further analysis according to transcriptomic sequencing data from S. chinensis. The expression stability of these candidate genes in inner and outer petals at different floral developmental stages and in many different tissues was then analyzed with the geNorm and NormFinder software. The reference genes were evaluated by normalizing the expression of the anthocyanidin synthase gene in the outer and inner petals at different floral developmental stages to further verify the expression stability of these genes. Elongation factor 1-alpha (ScEF) and 50S ribosomal protein L27 were found to be the two most stable genes in the overall ranking of all the samples and different tissues. Furthermore, ScEF and protein phosphatase 2A were stably expressed in all petal samples. Moreover, among the nine candidate reference genes, phosphoglycerate kinase performed poorly in all sample sets. Our results will help to obtain reliable expression data in molecular studies of S. chinensis.

Worldwide, a relevant surface of arable lands is facing salt stress, and this surface is increasing continuously due to both natural and anthropogenic activities. Nitric oxide (NO) is a small, gaseous molecule with a plethora of physiological roles in plants. In addition to its normal physiological functions, NO protects plants subjected to different environmental cues including salinity. For example, NO mediates photosynthesis and stomatal conductance, stimulates the activity of Na⁺/H⁺ antiport in tonoplast, promotes the biosynthesis of osmolytes, and counteracts overaccumulation of reactive oxygen species in plant cells under salt stress. Exogenous NO is also beneficial for plants subjected to salinity, in which it increases salinity tolerance via growth promotion, reversing oxidative damage, and maintaining ion homeostasis. This review provides a comprehensive picture of the NO-mediated mechanisms in plants, resulting in salinity tolerance with a particular focus on the photosynthetic processes, the antioxidant patterns as well as the cross-talk with other regulatory compounds in plant cells.

Transient heterologous gene expression in two model plant species, Nicotiana benthamiana and N. excelsior, has been used to study the localization of the heterologous Δ9 acyl-lipid desaturase (Δ9 desaturase) of Synechococcus vulcanus in different cell compartments and its functional activity in the cases of the cytosol, chloroplast, and endoplasmic reticulum (ER) localization. The functional activity and substrate specificity of the heterologous desaturase under the conditions of transient expression have been confirmed by comparison of fatty acid (FA) profiles. The Δ9 desaturase, responsible for the synthesis of oleic and palmitoleic acids, has also been shown to strongly promote the accumulation of polyunsaturated FAs. The results convincingly demonstrate that the Δ9 desaturase of the thermophilic cyanobacterium transiently expressed in two Nicotiana species considerably alters lipid metabolism in their leaves towards a higher FA unsaturation. The functional activity of Δ9 desaturase depends on both the model plant species, N. benthamiana or N. excelsior, and the cellular localization of the enzyme. The method of transient expression of heterologous genes in plants is highly effective, inexpensive, and not time-consuming, which makes it attractive for estimating the functional activity and/or substrate specificity of heterologous desaturases.

Cadmium (Cd) is a highly toxic and widespread soil pollutant, which negatively affects various aspects of plant growth and physiology. Here, the role of photosynthesis in response to Cd was investigated in the Cd-tolerant pea (Pisum sativum L.) mutant SGECd^t. The wild type SGE and the mutant SGECd^t were grown in a hydroponic solution supplemented with 1, 3, or 4 µM CdCl₂ for 12 d. Root and shoot biomasses of the Cd-treated SGECd^t were significantly higher than of SGE. Cadmium had little effect on the quantum yield of photosystem II (φPSII) and chlorophyll content of intact leaves of both pea genotypes. However, when leaf slices were taken from Cd-exposed plants and incubated with high Cd concentrations, the SGECd^t mutant showed 1.5 - 2 times higher φPSII values than SGE, with genotypic differences maximal at 0.1 and 1 mM CdCl₂. In contrast, when leaf slices were taken from plants previously unexposed to Cd, both pea genotypes exhibited similar φPSII values. Cadmium content in leaves and mesophyll protoplasts of Cd-treated SGECd^t were about 2 - 3 times higher than in SGE ones. The mutant leaves and mesophyll protoplasts had also higher Ca, Mg, Mn, and Zn content. Thus, SGECd^t acclimated to Cd during growth in the Cd-supplemented nutrient solution by developing a molecular mechanism related to photosynthetic integrity. A higher foliar nutrient content likely allows enhanced photosynthesis by counteracting the damage of leaves caused by Cd.

Temperature is an important factor for growth, development, productivity and geographical distribution of many plants (Nahar et al. 2015). Chilling stress is a major abiotic stress of crop production in Northeast China. Chilling stress exposure has been shown to enhance production of reactive oxygen species (ROS) and oxidative stress occurs (Nahar et al. 2015). The ROS, which include superoxide radical (O₂^*-), hydrogen peroxide (H₂O₂), hydroxyl radical (.OH), and singlet oxygen (¹O₂), cause damage to structural proteins, nucleic acids, enzymes, cell membranes, and other essential molecules involved in plant metabolism (Sharma et al. 2012, Nahar et al. 2015). Plants have developed mechanisms to tolerate environmental stress conditions through various physiological adaptations, including non-enzymatic and enzymatic ROS scavenging pathways (Hossain et al. 2010, Sharma et al. 2012, Nahar et al. 2015). Non-enzymic components of the antioxidative defense system include reduced ascorbate (AsA) and reduced glutathione (GSH) as well as osmotic adjustment substances as proline, soluble sugars, and soluble proteins which protect membrane integrity and cellular components from dehydration (Özlem and Ekmekçi 2011). The enzymatic antioxidants comprise superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX), glutathione reductase (GR), etc. (Gill and Tuteja 2010, Hossain et al. 2010). These enzymes, through step-by-step reaction, scavenge ROS with AsA and GSH as electron acceptors (Gill and Tuteja 2010). Nahar et al. (2015) has established that low temperature stress increased H₂O₂ and MDA content. Exogenous spermidine (Spd) in low temperature treatment increases the content of AsA and GSH, decreases the content of oxidized ascorbate (DHA) and oxidized glutathione GSSG, and improves activity of APX and GR. Various strategies are being employed in order to minimize the adverse effects of environmental stresses in plants. Exogenously applied plant growth regulators (PGRs) is an effective, facile, and practical technique to enhance tolerance of crops, and this approach has been used widely in recent years. One of the PGRs is 5-aminolevulinic acid, or 5-amino-4-oxo-pentanoic acid (ALA), which has a relative molecular mass of 131, and it is an essential precursor of tetrapyrrole compounds including chlorophyll, heme, and phytochrome (Balestrasse et al. 2010) and its formation may be the rate limiting step. Hotta et al. (1997a,b) observed that low dosage of ALA has plant growth regulator properties, such as promoting chlorophyll biosynthesis and enhancing photosynthesis (Memon et al. 2009), responding to environmental stresses (Korkmaz and Korkmaz 2009, Korkmaz et al. 2010, Zhang et al. 2012, Dan et al. 2013, Fu et al. 2016), and promoting recovery of growth after herbicide applications (Zhang et al. 2008). High dosages of ALA can cause the accumulation of several chlorophyll synthesis intermediates, but also production of ROS leading to oxidative stress (Balestrasse et al. 2010). Materials and methods Plants and chilling stress: Mung be

Allopolyploidy is long recognized as an essential driving force in plant evolution. Recent studies have demonstrated that small RNAs, including microRNAs (miRNAs), play important roles in the process of allopolyploidy. However, the question that how the distinct parent-of-origin miRNAs are maintained in allopolyploids and how these small RNAs affect gene expression and phenotype remain to be answered. Therefore, we investigated the miRNA expression profiles of a synthesized allotetraploid, Cucumis ×hytivus and its parents. The different developmental stages of leaves of C.×hytivus showing contrasting leaf colour were compared as well. Following high-throughput sequencing, 546 conserved d 287 novel miRNAs were identified. The expressions of nine miRNAs obtained by real-time quantitative PCR were consistent with the sequencing results. We detected that 15 miRNAs were divergently expressed between the parent species, and 23 miRNAs were differentially expressed in C. ×hytivus compared to either of its parents or both, which suggests the significant effect of allopolyploidization on miRNAs accumulation. Additionally, 26 conserved and 13 novel miRNAs differed in expression between the young and mature leaves of C. ×hytivus, indicating an essential role of miRNA-mediated regulation of leaf development following allopolyploidization. This study enriches the context of allopolyploidy effect on miRNAs and lays a foundation for the elucidation of the miRNA-mediated regulatory mechanism in phenotypic variation in allopolyploids.

Cellulose is the most abundant polysaccharide produced by plants. In the form of rigid microfibrils surrounding the cells, cellulose constitutes the load-bearing cell wall element that controls cell growth and shape. Cellulose microfibrils are laid down outside the cell by the multimeric plasma membrane-inserted cellulose synthase complexes (CSCs), which move along underlying cortical microtubules (CMTs). In plants, CSCs are shaped as rosettes with six lobes symmetrically arranged in a hexagonal structure. In Arabidopsis, the CSC is composed of at least three functionally non-redundant cellulose synthase (CESA) glycosyltransferases in both primary and secondary cell walls. The number, organization, and interactions of CESA proteins within the CSC have been debated for many years on the basis of numerous lines of evidence provided by electron microscopy, biochemical and genetic approaches, spectroscopic techniques, as well as computational modeling. The Arabidopsis thaliana model was extremely useful in elucidating the molecular composition of CSC and enabled to elucidate the specialized functions of distinct AtCESA isoforms. Several additional, non-CESA proteins involved in cellulose synthesis and its regulation were also identified in Arabidopsis. This review outlines the latest findings on CSC organization, trafficking, and plant-specific proteins directly associated with the complex and interconnecting CESAs with CMTs.

The response of functional traits of plants to external environment can influence their competitive ability because these functional traits are required for the acquisition of resources. The overuse of silver nanoparticles (AgNPs) has gained attention due to their environmental toxicity. This study aimed to examine the effects of AgNPs with different concentrations and particle sizes on functional traits of wheat. It was observed that AgNPs significantly reduced the plant height and so decrease its competitive ability. Ag ions decreased leaf chlorophyll and nitrogen content and specific leaf area more than AgNPs, but the opposite was true for leaf length, single leaf fresh mass, and shoot fresh mass. Hence, the toxicity of AgNPs may be higher than that of Ag ions in some cases. In this study, leaf chlorophyll and nitrogen content decreased with increasing concentration of AgNPs (with size 30 nm). The AgNPs with smaller particle size exerted higher toxicity on leaf chlorophyll and N content than those with larger particle size at the same concentration. However, AgNPs with larger particle size reduced more aboveground fresh mass than those with smaller particle size at the same concentration.

Plants are severely affected by many biotic stresses, which cause a reduction in crop quality and quantity. One of the strategies to manage biotic stresses is the generation of transgenic plant lines that can be used as biosensors. These biosensor plants can trigger an early warning upon any pathogen infection. Two promoters with β-glucuronidase reporter gene fusions were constructed. The first contained the flagellin sensing 2 gene promoter, whereas the second contained synthetic promoter containing four repeats of cis-acting elements from the pathogen-related protein 1 gene and two transcription enhancers from the 35S promoter. Transformed leaves were treated with a phytohormone salicylic acid to mimic the occurrence of biotic stress. Validation of reporter gene expression induced from both constructs in transformed potato leaves displayed an increase upon salicylic acid treatment. The results reflect that both constructs could serve in the production of potato biotic stress biosensors.

The miR399 is a conserved microRNA (miRNA) family, and it has been characterized as an essential regulator of phosphorus transport in plants. However, the biological function of miR399 in Cunninghamia lanceolata is still largely unclear. In this study, the comparison of mature miR399 sequence revealed a high similarity between Arabidopsis thaliana and C. lanceolate, and the pre-miR399 was capable of forming a typical stem-loop hairpin structure. A gene PHOSPHATE 2 (PHO2) was identified as a target of cln-miR399 using 5' rapid amplification of cDNA ends. Furthermore, the relationship between cln-miR399 and PHO2 was further confirmed through a transient co-expression of both genes in Nicotiana benthamiana. To examine the function of miR399 in Arabidopsis, miR399-overexpressing transgenic Arabidopsis thaliana was acquired using Agrobacterium-mediated approach. Real-time PCR showed that the amount of cln-MIR399 transcripts was higher in miR399-overexpressing plants than in wild-type plants, which was accompanied with down-regulation of expression of its target gene AtPHO2. The P content was 1.40 to 1.56-fold higher in the leaves of three transgenic lines than in wild type plants. However, the P content in the roots of the three transgenic lines was 24.5 - 37.2 % less than that in wild type plants. Moreover, the transcriptions of three phosphate transporter genes (PHT1, PHT2, and PHT3) were up-regulated in roots of miR399-overexpressing Arabidopsis plants. Interestingly, the transgenic lines exhibited retarded growth under normal P conditions compared with the wild type. Our findings demonstrate that cln-miR399 may play crucial roles in P transport and plant growth via regulation of its target gene PHO2.

In plants, auxin/indoleacetic acid (Aux/IAA) proteins are transcriptional regulators, which regulate developmental process and responses to phytohormones and stress treatments. A previous study has shown that the rice Aux/IAA transcription factor gene OsIAA18 is induced by salt and osmotic stresses. However, little is known about the regulatory functions of this gene. In this study, the OsIAA18 gene was successfully cloned from rice. Subcellular localization analysis in onion epidermal cells indicated that OsIAA18 was localized to the nucleus. Expression analysis in yeast showed that the full length OsIAA18 exhibited transcriptional activation. Heterologous expression of OsIAA18 significantly enhanced salt and osmotic tolerance in transgenic Arabidopsis plants. Real-time quantitative PCR analysis showed that constitutive expression of OsIAA18 up-regulated genes involved in abscisic acid (ABA) biosynthesis, proline biosynthesis, stress responses, and reactive oxygen species scavenging under salt and osmotic stresses. Enzymatic analyses found that the transgenic plants had higher 9-cis-epoxycarotenoid dioxygenase, pyrroline-5-carboxylate synthase, superoxide dismutase, and peroxidase activities than wild-type plants under salt and osmotic stresses. In the transgenic plants, ABA and proline content significantly increased, whereas H₂O₂ and malondialdehyde content significantly decreased. In addition, the transgenic plants had also a lower electrolyte leakage and water loss rate. These overall results indicate that the OsIAA18 gene is involved in enhancing salt and osmotic tolerance in transgenic Arabidopsis plants. The OsIAA18 gene has a potential to be used to enhance the tolerance to abiotic stresses in other plant species.

γ-Aminobutyric acid (GABA) regulates plant tolerance to abiotic stresses; however, a transcriptomic change and key stress-related genes induced by GABA have not been investigated in plants during a prolonged period of salt stress. Roots of creeping bentgrass (Agrostis stolonifera) cv. Penncross were pretreated with or without 0.5 mM GABA solution for 2 days and then subjected to salt stress for 20 days (150 mM NaCl solution for 3 d, 200 mM NaCl for another 3 d, and 250 mM NaCl for 14 d) in controlled growth chambers. The application of GABA significantly increased GABA content in roots and alleviated a salt-stress induced decrease in GABA content in leaves. This was associated with a significant increase in salt tolerance as demonstrated by a significantly higher leaf relative water content, photochemical efficiency, performance index on absorption basis, and lower electrolyte leakage in GABA-pretreated plants as compared to untreated plants under salt stress. Transcriptomic analysis found that GABA-induced salt tolerance was closely associated with saccharide, amino acid, and lipid metabolism. The GABA upregulated key differentially expressed genes including cytochrome P450 (CYP450), zinc transporter 29 (ZTP29), alpha-amylase 3 (AMY3), 3-ketoacyl-CoA synthase 6 (KCS6), aldehyde oxidase (AO), acetyl-CoA carboxylase 1 (ACC1), and magnesium-chelatase (Mg-CHT) involved in zinc homeostasis, starch degradation, and the biosynthesis of wax, fatty acid, chlorophyll, and abscisic acid, which could contribute to GABA-regulated salt tolerance. Current findings prove that GABA application is an efficient approach to enhance salt tolerance of creeping bentgrass during a prolonged period of salt stress and also provide valuable information to better understand key candidate genes and regulatory pathways of GABA-induced salt tolerance in plants.