γ-Aminobutyric acid (GABA) priming alleviates acid-aluminum toxicity to roots of creeping bentgrass via enhancements in antioxidant defense and organic metabolites remodeling.

MAIN CONCLUSION: γ-Aminobutyric acid alleviates acid-aluminum toxicity to roots associated with enhanced antioxidant metabolism as well as accumulation and transportation of citric and malic acids. Aluminum (Al) toxicity has become the main limiting factor for crop growth and development in acidic soils and is further being aggravated worldwide due to continuous industrial pollution. The current study was designed to examine effects of GABA priming on alleviating acid-Al toxicity in terms of root growth, antioxidant defense, citrate and malate metabolisms, and extensive metabolites remodeling in roots under acidic conditions. Thirty-seven-day-old creeping bentgrass (Agrostis stolonifera) plants were used as test materials. Roots priming with or without 0.5 mM GABA for 3 days were cultivated in standard nutrient solution for 15 days as control or subjected to nutrient solution containing 5 mM AlCl3·6H2O for 15 days as acid-Al stress treatment. Roots were sampled for determinations of root characteristics, physiological and biochemical parameters, and metabolomics. GABA priming significantly alleviated acid-Al-induced root growth inhibition and oxidative damage, despite it promoted the accumulation of Al in roots. Analysis of metabolomics showed that GABA priming significantly increased accumulations of organic acids, amino acids, carbohydrates, and other metabolites in roots under acid-Al stress. In addition, GABA priming also significantly up-regulated key genes related to accumulation and transportation of malic and citric acids in roots under acid-Al stress. GABA-regulated metabolites participated in tricarboxylic acid cycle, GABA shunt, antioxidant defense system, and lipid metabolism, which played positive roles in reactive oxygen species scavenging, energy conversion, osmotic adjustment, and Al ion chelation in roots.

Arsenic uptake by Agrostis capillaris, as related to its genotypic diversity in the area of historical ore mining and processing.

Common bentgrass Agrostis capillaris L. is known as tolerant to toxic elements. A hypothesis was examined that its ecotypes growing in historically polluted sites show a limited arsenic uptake and have genetic features that distinguish them from commercially available cultivars. The study was conducted in Zloty Stok, a historical area of arsenic mining. Additionally, two commercial cultivars were grown in pots with arsenic-rich soils. Based on arsenic concentrations in plant roots and shoots, bioconcentration and translocation factors BCF and TF were calculated. Commercial cultivars indicated many times higher BCF shoots and TF values compared to field plants. DNA analysis of leaf blades showed a clear distinction between the plants growing in some sites and patches in the field, and also a gene overlap between the plants in the field and commercial forms. The research did not allow for identification of ecotypes with exceptionally limited arsenic uptake. Moreover, there were no significant differences between the genotypic characteristics of plants growing in polluted sites and those poorly tolerant grown from commercially available seeds. Apparently, other factors, and not genetically determined features, are responsible for A. capillaris tolerance to arsenic in Zloty Stok.

Rhizobacteria-enhanced drought tolerance and post-drought recovery of creeping bentgrass involving differential modulation of leaf and root metabolism.

Rhizobacteria that produce 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase (ACCd) that inhibits ethylene production may mitigate stress damages. The objectives of this study were to examine whether a novel strain of ACCd-producing bacteria, Paraburkholderia aspalathi "WSF23," promotes plant tolerance to drought stress and post-stress recovery and determine changes in metabolic profiles in leaves and roots associated with the positive ACCd-bacteria effects in cool-season perennial grass species. Creeping bentgrass (Agrostis Stolonifera L. cv. "Penncross") plants were inoculated with P. aspalathi "WSF23" and exposed to drought by withholding irrigation for 35 days, followed by re-watering for 15 days in growth chambers. Inoculated plants demonstrated increased turf quality, canopy density, and root growth during drought stress and more rapid re-growth upon re-watering. Metabolomic analysis demonstrated that inoculation with P. aspalathi "WSF 23" increased the content of metabolites in the metabolic pathways related to stress defense, including osmoregulation, cell wall stability, and antioxidant protection in both leaves and roots, as well as nitrogen metabolism in roots of creeping bentgrass exposed to drought stress. The promotion of post-stress recovery by P. aspalathi "WSF 23" was mainly associated with enhanced carbohydrate and pyrimidine metabolism and zeatin biosynthesis pathways in leaves and increased carbohydrates, biosynthesis of DNA and proteins, cellular metabolism, and TCA cycle activity in roots. These results provide insights into the metabolic pathways regulated by "WSF23," with the PGPR conferring improvements in drought stress tolerance and post-drought recovery in a perennial grass species.

Chitosan (CTS) Alleviates Heat-Induced Leaf Senescence in Creeping Bentgrass by Regulating Chlorophyll Metabolism, Antioxidant Defense, and the Heat Shock Pathway.

Chitosan (CTS) is a deacetylated derivative of chitin that is involved in adaptive response to abiotic stresses. However, the regulatory role of CTS in heat tolerance is still not fully understood in plants, especially in grass species. The aim of this study was to investigate whether the CTS could reduce heat-induced senescence and damage to creeping bentgrass associated with alterations in antioxidant defense, chlorophyll (Chl) metabolism, and the heat shock pathway. Plants were pretreated exogenously with or without CTS (0.1 g L-1) before being exposed to normal (23/18 °C) or high-temperature (38/33 °C) conditions for 15 days. Heat stress induced detrimental effects, including declines in leaf relative water content and photochemical efficiency, but significantly increased reactive oxygen species (ROS) accumulation, membrane lipid peroxidation, and Chl loss in leaves. The exogenous application of CTS significantly alleviated heat-induced damage in creeping bentgrass leaves by ameliorating water balance, ROS scavenging, the maintenance of Chl metabolism, and photosynthesis. Compared to untreated plants under heat stress, CTS-treated creeping bentgrass exhibited a significantly higher transcription level of genes involved in Chl biosynthesis (AsPBGD and AsCHLH), as well as a lower expression level of Chl degradation-related gene (AsPPH) and senescence-associated genes (AsSAG12, AsSAG39, Asl20, and Ash36), thus reducing leaf senescence and enhancing photosynthetic performance under heat stress. In addition, the foliar application of CTS significantly improved antioxidant enzyme activities (SOD, CAT, POD, and APX), thereby effectively reducing heat-induced oxidative damage. Furthermore, heat tolerance regulated by the CTS in creeping bentgrass was also associated with the heat shock pathway, since AsHSFA-6a and AsHSP82 were significantly up-regulated by the CTS during heat stress. The potential mechanisms of CTS-regulated thermotolerance associated with other metabolic pathways still need to be further studied in grass species.

Effects of biochar, ochre and manure amendments associated with a metallicolous ecotype of Agrostis capillaris on As and Pb stabilization of a former mine technosol.

Metal(loid) soil pollution is a major environmental and health issue, requiring these areas to be remediated, for example through phytoremediation processes. In order to allow proper plant establishment and growth, amendments must be applied to highly contaminated and poorly fertile soils. Amendments are diverse, but many studies have shown the beneficial effects of biochar, manure and ochre, although studies on their combined use are scarce. Moreover, no studies have evaluated the effect of these combined amendments on endemic plant growth. Endemic plants growing on contaminated soils showed higher tolerance toward pollutants compared to plants coming from unpolluted areas. Therefore, the aim of the present study was to evaluate both the effect of amendments (single or combined) on the physicochemical properties of a former mining technosol, and the growth and metal(loid) accumulation ability of endemic Agrostis capillaris plants. This study revealed an improvement in the soil physicochemical properties following the application of amendments, with combined amendments showing better results than the application of just one. On top of this, Agrostis plants performed better on the amended technosols, especially the ones receiving manure, due to its high nutrient content. Finally, based on soil properties, plant growth and the metal(loid) accumulation profile, the use of biochar combined with manure seems to be the most appropriate treatment. Indeed, this treatment showed an improvement in both soil fertility and plant growth. Moreover, Agrostis plants grown in these conditions were among those showing higher root metal(loid) concentration associated with a lower translocation toward aerial parts.

Chitosan regulates metabolic balance, polyamine accumulation, and Na+ transport contributing to salt tolerance in creeping bentgrass.

BACKGROUND: Chitosan (CTS), a natural polysaccharide, exhibits multiple functions of stress adaptation regulation in plants. However, effects and mechanism of CTS on alleviating salt stress damage are still not fully understood. Objectives of this study were to investigate the function of CTS on improving salt tolerance associated with metabolic balance, polyamine (PAs) accumulation, and Na+ transport in creeping bentgrass (Agrostis stolonifera). RESULTS: CTS pretreatment significantly alleviated declines in relative water content, photosynthesis, photochemical efficiency, and water use efficiency in leaves under salt stress. Exogenous CTS increased endogenous PAs accumulation, antioxidant enzyme (SOD, POD, and CAT) activities, and sucrose accumulation and metabolism through the activation of sucrose synthase and pyruvate kinase activities, and inhibition of invertase activity. The CTS also improved total amino acids, glutamic acid, and γ-aminobutyric acid (GABA) accumulation. In addition, CTS-pretreated plants exhibited significantly higher Na+ content in roots and lower Na+ accumulation in leaves then untreated plants in response to salt stress. However, CTS had no significant effects on K+/Na+ ratio. Importantly, CTS enhanced salt overly sensitive (SOS) pathways and also up-regulated the expression of AsHKT1 and genes (AsNHX4, AsNHX5, and AsNHX6) encoding Na+/H+ exchangers under salt stress. CONCLUSIONS: The application of CTS increased antioxidant enzyme activities, thereby reducing oxidative damage to roots and leaves. CTS-induced increases in sucrose and GABA accumulation and metabolism played important roles in osmotic adjustment and energy metabolism during salt stress. The CTS also enhanced SOS pathway associated with Na+ excretion from cytosol into rhizosphere, increased AsHKT1 expression inhibiting Na+ transport to the photosynthetic tissues, and also up-regulated the expression of AsNHX4, AsNHX5, and AsNHX6 promoting the capacity of Na+ compartmentalization in roots and leaves under salt stress. In addition, CTS-induced PAs accumulation could be an important regulatory mechanism contributing to enhanced salt tolerance. These findings reveal new functions of CTS on regulating Na+ transport, enhancing sugars and amino acids metabolism for osmotic adjustment and energy supply, and increasing PAs accumulation when creeping bentgrass responds to salt stress.

A physiological approach to study the competition ability of the grassland species Trifolium pratense and Agrostis capillaris.

The response of plant species to external factors depends partly on the interaction with the environment and with the other species that coexist in the same ecosystem. Several studies have investigated the main traits that determine the competitive capacity of plant species, and although the relevance of the traits is not clear, traits both from belowground and aboveground have been observed. In this paper, we grew Trifolium pratense and Agrostis capillaris in intra- and interspecific competition, analyzing the photosynthetic metabolism and nitrogen uptake, among other variables. The results indicated that T. pratense possesses better competition ability due to the higher competitive performance for soil resources compared to A. capillaris, explained by a higher root biomass and a higher nitrogen uptake rate in the former than in the latter. These traits permitted T. pratense to show higher photosynthetic rate than A. capillaris when both species were grown in mixture. Furthermore, the interspecific competition provoked A. capillaris to activate its antioxidant metabolism, through SOD activity, to detoxify the reactive oxygen species generated due to its lower capacity for using the photochemical energy absorbed. In this experiment, we conclude that the competitiveness seems to be more related with soil resources competition than with light competition, and that the photosynthetic rate decline in A. capillaris is more a secondary effect as a consequence of nitrogen limitation.

γ-Aminobutyric Acid Enhances Heat Tolerance Associated with the Change of Proteomic Profiling in Creeping Bentgrass.

γ-Aminobutyric acid (GABA) participates in the regulation of adaptability to abiotic stress in plants. The objectives of this study were to investigate the effects of GABA priming on improving thermotolerance in creeping bentgrass (Agrostis stolonifera) based on analyses of physiology and proteome using iTRAQ technology. GABA-treated plants maintained significantly higher endogenous GABA content, photochemical efficiency, performance index on absorption basis, membrane stability, and osmotic adjustment (OA) than untreated plants during a prolonged period of heat stress (18 days), which indicated beneficial effects of GABA on alleviating heat damage. Protein profiles showed that plants were able to regulate some common metabolic processes including porphyrin and chlorophyll metabolism, glutathione metabolism, pyruvate metabolism, carbon fixation, and amino acid metabolism for heat acclimation. It is noteworthy that the GABA application particularly regulated arachidonic acid metabolism and phenylpropanoid biosynthesis related to better thermotolerance. In response to heat stress, the GABA priming significantly increased the abundances of Cu/ZnSOD and APX4 that were consistent with superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities. The GABA-upregulated proteins in relation to antioxidant defense (Cu/ZnSOD and APX4) for the reactive oxygen species scavenging, heat shock response (HSP90, HSP70, and HSP16.9) for preventing denatured proteins aggregation, stabilizing abnormal proteins, promoting protein maturation and assembly, sugars, and amino acids metabolism (PFK5, ATP-dependent 6-phosphofructokinase 5; FK2, fructokinase 2; BFRUCT, ß-fructofuranosidase; RFS2, galactinol-sucrose galactosyltransferase 2; ASN2, asparagine synthetase 2) for OA and energy metabolism, and transcription factor (C2H2 ZNF, C2H2 zinc-finger protein) for the activation of stress-defensive genes could play vital roles in establishing thermotolerance. Current findings provide an illuminating insight into the new function of GABA on enhancing adaptability to heat stress in plants.

AsHSP26.8a, a creeping bentgrass small heat shock protein integrates different signaling pathways to modulate plant abiotic stress response.

BACKGROUND: Small heat shock proteins (sHSPs) are critical for plant response to biotic and abiotic stresses, especially heat stress. They have also been implicated in various aspects of plant development. However, the acting mechanisms of the sHSPs in plants, especially in perennial grass species, remain largely elusive. RESULTS: In this study, AsHSP26.8a, a novel chloroplast-localized sHSP gene from creeping bentgrass (Agrostis stolonifera L.) was cloned and its role in plant response to environmental stress was studied. AsHSP26.8a encodes a protein of 26.8 kDa. Its expression was strongly induced in both leaf and root tissues by heat stress. Transgenic Arabidopsis plants overexpressing AsHSP26.8a displayed reduced tolerance to heat stress. Furthermore, overexpression of AsHSP26.8a resulted in hypersensitivity to hormone ABA and salinity stress. Global gene expression analysis revealed AsHSP26.8a-modulated expression of heat-shock transcription factor gene, and the involvement of AsHSP26.8a in ABA-dependent and -independent as well as other stress signaling pathways. CONCLUSIONS: Our results suggest that AsHSP26.8a may negatively regulate plant response to various abiotic stresses through modulating ABA and other stress signaling pathways.

iTRAQ-based proteomics reveals key role of γ-aminobutyric acid (GABA) in regulating drought tolerance in perennial creeping bentgrass (Agrostis stolonifera).

γ-Aminobutyric acid (GABA), a non-proteinaceous amino acid, modulates plant growth and stress tolerance. However, the potential role of GABA in regulating key metabolic pathways and stress-defensive proteins against drought in plants has never been explored. Creeping bentgrass (Agrostis stolonifera) plants were pretreated with or without GABA and then subjected to water stress for 8 days in controlled growth chambers (23/19 °C, day/night). Physiological analysis showed that elevated endogenous GABA level via exogenous GABA application significantly mitigated water stress damage to creeping bentgrass, as manifested by increased leaf relative water content, water use efficiency, osmotic adjustment (OA), photochemical efficiency (Fv/Fm), net photosynthetic rate, and reduced oxidative damage. iTRAQ-based proteomics found that enhanced chaperones accumulation, carbohydrates, amino acids, and energy metabolism played important roles in protein protection, OA, energy maintenance, and metabolic balance, which is important adaptive response to drought stress in creeping bentgrass. The GABA further promoted energy production and conversion, antioxidant defense, and DHN3 accumulation that were essential for energy requirement, ROS-scavenging, and the prevention of cell dehydration in leaf during drought stress. In addition, GABA-treated plants maintained significantly higher abundance of dicarboxylate transporter 2.1, ATP-dependent zinc metalloprotease, receptor-like protein kinase HERK1, o-acyltransferase WSD1, omega-6 fatty acid desaturase, and two-component response regulator ORR21 than untreated plants under drought stress. The result provides new evidences that GABA-induced drought tolerance is possibly involved in the improvement of nitrogen recycling, protection of photosystem II, mitigation of drought-depressed cell elongation, wax biosynthesis, fatty acid desaturase, and delaying leaf senescence in creeping bentgrass.

Regulation of Heat Shock Factor Pathways by γ-aminobutyric Acid (GABA) Associated with Thermotolerance of Creeping Bentgrass.

Activation and enhancement of heat shock factor (HSF) pathways are important adaptive responses to heat stress in plants. The γ-aminobutyric acid (GABA) plays an important role in regulating heat tolerance, but it is unclear whether GABA-induced thermotolerance is associated with activation of HSF pathways in plants. In this study, the changes of endogenous GABA level affecting physiological responses and genes involved in HSF pathways were investigated in creeping bentgrass during heat stress. The increase in endogenous GABA content induced by exogenous application of GABA effectively alleviated heat damage, as reflected by higher leaf relative water content, cell membrane stability, photosynthesis, and lower oxidative damage. Contrarily, the inhibition of GABA accumulation by the application of GABA biosynthesis inhibitor further aggravated heat damage. Transcriptional analyses showed that exogenous GABA could significantly upregulate transcript levels of genes encoding heat shock factor HSFs (HSFA-6a, HSFA-2c, and HSFB-2b), heat shock proteins (HSP17.8, HSP26.7, HSP70, and HSP90.1-b1), and ascorbate peroxidase 3 (APX3), whereas the inhibition of GABA biosynthesis depressed these genes expression under heat stress. Our results indicate GABA regulates thermotolerance associated with activation and enhancement of HSF pathways in creeping bentgrass.

Use of wastes from the pulp and paper industry for the remediation of soils degraded by mining activities: Chemical, biochemical and ecotoxicological effects.

Fly ash (FA) from biomass combustion and biological sludge (S), both wastes from the pulp and paper industry, were granulated in different proportions (90% FA+10% S, and 70% FA+30% S w/w, dry weight basis, dw) and used to recover the functionality of soils affected by mining activities (Aljustrel, Iberian Pyrite Belt), with and without the application of municipal solid waste compost (MSWC). Application doses of both mixtures were 2.5, 5.0 and 10% (w/w, dw). These materials corrected soil acidity to circumneutral values and increased extractable P and K concentrations. A significant increase in soil organic matter (from 0.6 to 0.8-1.5% w/w, dw) and N content (from 0.04 to 0.09-0.12% w/w, dw) was also observed, but only when MSWC was applied. The soil was already heavily contaminated with Cu, Pb and Zn and the application of amendments did not increase their pseudo-total concentrations. The CaCl2 extractable fractions of both Cu and Zn decreased to very low values. The improvement in soil quality, compared to fertilizer only treatment, was further evidenced by the increase in some soil enzymatic activities (dehydrogenase, ß-glucosidase and cellulase), with a better response for the granules with the higher proportion of biological sludge, as well as by the decrease in the soil-water extract toxicity towards different organisms (Daphnia magna, Thamnocephalus platyurus, and Pseudokirchneriella subcapitata). Agrostis tenuis germinated and grew during the first month only in the amended pots, but, after that, a considerable phytotoxic effect was evident. This was mainly attributed to salt stress or to some specific ionic toxicity. In conclusion, to establish a long-term plant cover in mining soils amended with biomass ash-based materials, the selection of plants with higher resistance to salinity and/or the stabilization of the amendments, to reduce their soluble salt content, is recommended.

Recent developments in metabolomics-based research in understanding transgenic grass metabolism.

BACKGROUND: Transgenic herbicide-resistant (HR) turfgrass together with its associated, broad spectrum herbicides promise cheap, selective and efficient weed control by excluding infested weeds resulting in turf lawn with high uniformity and aesthetic value. The concept of this "weeding program" initiated from modern biotechnology has been widely implemented in several principal crops including maize, soybean, canola and cotton as early as the 1990s. Transgenic HR turfgrass classified as a genetically modified organism (GMO) has undoubtedly caused public concern with respect to its biosafety and legalities similar to well-established HR crops. Nevertheless, applying metabolomics-based approaches which focuses on the identification of the global metabolic state of a biological system in response to either internal or external stimuli can also provide a comprehensive characterization of transgenic grass metabolism and its involvement in biosecurity and public perception. AIM OF REVIEW: This review summaries the recent applications of metabolomics applied to HR crops to predict the molecular and physiological phenotypes of HR turfgrass species, glyphosate-resistant Kentucky bluegrass (Poa pratensis L.) and glufosinate-resistant creeping bentgrass (Agrotis stonifera L.). Additionally, this review also presents background knowledge with respect to the application of metabolomics, transformation of HR crops and its biosafety concerns, turfgrass botanical knowledge and its economic and aesthetic value. KEY SCIENTIFIC CONCEPTS OF REVIEW: The purpose of this review is to demonstrate the molecular and physiological phenotypes of HR turfgrass based on several lines of evidence primarily derived from metabolomics data applied to HR crops to identify alterations on HR turfgrass metabolism as a result of genetic modification that confers resistant traits.

Transgenic creeping bentgrass overexpressing Osa-miR393a exhibits altered plant development and improved multiple stress tolerance.

MicroRNA393 (miR393) has been implicated in plant growth, development and multiple stress responses in annual species such as Arabidopsis and rice. However, the role of miR393 in perennial grasses remains unexplored. Creeping bentgrass (Agrostis stolonifera L.) is an environmentally and economically important C3 cool-season perennial turfgrass. Understanding how miR393 functions in this representative turf species would allow the development of novel strategies in genetically engineering grass species for improved abiotic stress tolerance. We have generated and characterized transgenic creeping bentgrass plants overexpressing rice pri-miR393a (Osa-miR393a). We found that Osa-miR393a transgenics had fewer, but longer tillers, enhanced drought stress tolerance associated with reduced stomata density and denser cuticles, improved salt stress tolerance associated with increased uptake of potassium and enhanced heat stress tolerance associated with induced expression of small heat-shock protein in comparison with wild-type controls. We also identified two targets of miR393, AsAFB2 and AsTIR1, whose expression is repressed in transgenics. Taken together, our results revealed the distinctive roles of miR393/target module in plant development and stress responses between creeping bentgrass and other annual species, suggesting that miR393 would be a promising candidate for generating superior crop cultivars with enhanced multiple stress tolerance, thus contributing to agricultural productivity.

Strategy of optical path of daylight signal into tissues in cold-season turfgrasses using small, concave silica bodies.

Plants incorporate inorganic materials (biominerals), such as silica, into their various components. Plants belonging to the order Poales, like rice plants and turfgrasses, show comparatively high rates of silicon accumulation, mainly in the form of silica bodies. This work aims to determine the shapes and roles of these silica bodies by microscopic observation and optical simulation. We have previously found convex silica bodies on the leaves of rice plants and hot-season turfgrasses (adapted to hot-seasons). These silica bodies enabled light reflection and ensured reduction of the photonic density of states, which presumably prevented the leaves from overheating, as suggested by theoretical optical analyses. The silica bodies have been considered to have the functions of reinforcement of the plant body. The present work deals with cold-season turfgrasses, which were found to have markedly different silica bodies, cuboids with a concave top surface. They presumably acted as small windows for introducing light into the tissues, including the vascular bundles in the leaves. The area of the silica bodies was calculated to be about 5% of the total surface area of epidermis, which limits the thermal radiation of the silica bodies. We found that the light signal introduced through the silica bodies diffused in the organs even reaching the vascular bundles, the physiological functions of this phenomena remain as future problems. Light signal in this case is not related with energy which heat the plant but sensing outer circumstances to respond to them.

2, 3-Butanediol activated disease-resistance of creeping bentgrass by inducing phytohormone and antioxidant responses.

Brown patch, caused by Rhizoctonia solani, is a serious disease in Agrostis stolonifera. 2, 3-butanediol (2, 3-BD) is the major component of volatile organic compounds and was found to initiate induced systemic resistance (ISR). To investigate the induced resistance mechanism of 2, 3-BD, we examined the effects of resistance by area affected, along with changes in the content of phytohormones (Zeatin (ZT), Abscisic Acid (ABA) and Indole-3-Acetic Acid (IAA)), the activities of three phenylpropanoid metabolic enzymes (Phenylalaninammo-Nialyase (PAL), Chalcone Isomerase (CHI) and 4-Coumarate:Coenzyme A Ligase (4CL)) and the level of secondary metabolites (total phenols, flavonoid and lignin). The result showed that 2, 3-BD treatment at 250 µmoL/L had the best induction effect with the area affected decreased from 95% of the control to 55%. Compared to the controls, treatment with 250 µmoL/L 2, 3-BD induced higher levels of PAL, CHI and 4CL activity and increased total phenols, flavonoid and lignin levels. While 2, 3-BD treatment decreased the content of ZT and ABA but increased the content of IAA compared to controls. This study provides a basis for elucidating the mechanism of 2, 3-BD as a new plant disease control agent.

Role of Plant Volatiles in Host Plant Recognition by Listronotus maculicollis (Coleoptera: Curculionidae).

The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby, is an economically important pest of short cut turfgrass. Annual bluegrass, Poa annua L., is the most preferred and suitable host for ABW oviposition, larval survival and development. We investigated the involvement of grass volatiles in ABW host plant preference under laboratory and field conditions. First, ovipositional and feeding preferences of ABW adults were studied in a sensory deprivation experiment. Clear evidence of involvement of olfaction in host recognition by ABW was demonstrated. Poa annua was preferred for oviposition over three bentgrasses, Agrostis spp., but weevils with blocked antennae did not exhibit significant preferences. ABW behavioral responses to volatiles emitted by Agrostis spp. and P. annua were examined in Y-tube olfactometer assays. Poa annua was attractive to ABW females and preferred to Agrostis spp. cultivars in Y-tube assays. Headspace volatiles emitted by P. annua and four cultivars of Agrostis stolonifera L. and two each of A. capillaris L. and A. canina L. were extracted, identified and compared. No P. annua specific volatiles were found, but Agrostis spp. tended to have larger quantities of terpenoids than P. annua. (Z)-3-hexenyl acetate, phenyl ethyl alcohol and their combination were the most attractive compounds to ABW females in laboratory Y-tube assays. The combination of these compounds as a trap bait in field experiments attracted adults during the spring migration, but was ineffective once the adults were on the short-mown turfgrass. Hence, their usefulness for monitoring weevil populations needs further investigation.

The direction of carbon and nitrogen fluxes between ramets in Agrostis stolonifera changes during ontogeny under simulated competition for light.

Resource sharing is universal among connected ramets of clonal plants and is driven both by the developmental status of the ramets and the resource gradients. Above-ground competition forms spatial light gradients, but the role of resource sharing in such competition is unclear. We examined translocation of resources between mother and daughter ramets of Agrostis stolonifera under light heterogeneity throughout ramet ontogeny. We labelled ramets with 13C and 15N to estimate the bidirectional translocation of resources at three developmental stages of the daughters. In addition, we compared the final biomass of integrated and severed ramets in order to estimate the effect of integration on growth. Young developing daughters were supported by carbon, whereas nitrogen was only translocated towards daughters at the beginning of rooting, regardless of the light conditions. Shading of mothers was a major determinant of resource translocation between developed ramets, with carbon being preferentially moved to daughters from shaded mothers while nitrogen translocation was limited from daughters to shaded mothers. Surprisingly, the absolute amounts of translocated resources did not decline during development. Growth of daughters was enhanced by integration regardless of the shading. Overall, A. stolonifera maximizes the resource translocation pattern in order to enable it to spread from unfavourable habitats, rather than compensating for light heterogeneity among ramets.

Unravelling the bioherbicide potential of Eucalyptus globulus Labill: Biochemistry and effects of its aqueous extract.

In the worldwide search for new strategies in sustainable weed management, the use of plant species able to produce and release phytotoxic compounds into the environment could be an effective alternative to synthetic herbicides. Eucalyptus globulus Labill. is known to be a source of biologically active compounds responsible for its phytotoxic and allelopathic properties. Our previous results demonstrated the bioherbicide potential of eucalyptus leaves incorporated into the soil as a green manure, probably through the release of phytotoxins into the soil solution. Thus, the aims of this study were to understand the phytotoxicity of the eucalyptus leaves aqueous extract applied in pre- and post-emergence, and to identify and quantify its potentially phytotoxic water-soluble compounds. The effects were tested on the germination and early growth of the model target species Lactuca sativa and Agrostis stolonifera, and on physiological parameters of L. sativa adult plants after watering or spraying application. Dose-response curves and ED50 and ED80 values for eucalyptus aqueous extracts revealed pre-emergence inhibitory effects on both target species, effects being comparable to the herbicide metolachlor. While spraying treatment reduced the aerial and root biomass and increased the dry weight/fresh weight ratio of lettuce adult plants, watering application reduced protein contents and chlorophyll concentrations with respect to control, reflecting different modes of action depending on the site of phytotoxin entry. Via HPLC analyses, a total of 8 phenolic compounds (chlorogenic, two ρ-coumaric derivatives, ellagic, hyperoside, rutin, quercitrin, and kaempferol 3-O-glucoside) and other 5 low weight organic acids (citric, malic, shikimic, succinic and fumaric acids) were obtained from aqueous extract, the latter being identified for the first time in E. globulus. Despite some phytotoxic effects were found on lettuce adult plants, the use of eucalyptus aqueous extract would be discarded in post-emergence, whereas it was promising as a pre-emergence bioherbicide.

Transcriptional regulation of hormone-synthesis and signaling pathways by overexpressing cytokinin-synthesis contributes to improved drought tolerance in creeping bentgrass.

The objective of this study was to investigate transcriptomic changes and molecular factors regulated by cytokinins that may contribute to improved drought tolerance in creeping bentgrass (Agrostis stolonifera) overexpressing adenine isopentenyltransferase (ipt). Wild-type (WT) and ipt-transgenic plants were maintained well irrigated or exposed to 21 days of drought stress in growth chambers. Transcriptomic analysis conducted by RNA-seq revealed 661 and 648 upregulated and 764 and 862 downregulated drought-responsive genes (DRGs) in the WT and ipt-transgenic plants, respectively, under drought stress using adjusted P-value of 0.001 and log2 fold change. Gene ontology (GO) term classification showed that a greater number of DRGs were found in ipt-transgenic plants than in WT plants pertaining to biological functions including metabolic process, cellular process, cell structure and growth, macromolecular complex, and binding and catalytic activity, whereas fewer DRGs were found in ipt-transgenic plants than in WT plants pertaining to response to stimulus and antioxidant activity. Furthermore, plant hormone signal transduction pathway analysis revealed three downregulated transcripts [type B - Arabidopsis response regulators (B-ARR), ABA-responsive element binding factor (ABF) and pyrabactin resistance/like (PYR/PYL)] and two upregulated transcripts (BIN2 and JAZ) that were significantly differentiated between ipt-transgenic and WT plants under drought stress, which are particularly interesting for further investigation of molecular mechanisms of hormone-regulation of drought tolerance.