Discovery of a Hybrid Molecule with Phytotoxic Activity by Genome Mining, Heterologous Expression, and OSMAC Strategy.

Genome mining in association with the OSMAC (one strain, many compounds) approach provides a feasible strategy to extend the chemical diversity and novelty of natural products. In this study, we identified the biosynthetic gene cluster (BGC) of restricticin, a promising antifungal agent featuring a reactive primary amine, from the fungus Aspergillus sclerotiorum LZDX-33-4 by genome mining. Combining heterologous expression and the OSMAC strategy resulted in the production of a new hybrid product (1), along with N-acetyl-restricticin (2) and restricticinol (3). The structure of 1 was determined by spectroscopic data, including optical rotation and electronic circular dichroism (ECD) calculations, for configurational assignment. Compound 1 represents a fusion of restricticin and phytotoxic cichorin. The biosynthetic pathway of 1 was proposed, in which the condensation of a primary amine of restricticin with a precursor of cichorine was postulated. Compound 1 at 5 mM concentration inhibited the growth of the shoots and roots of Lolium perenne, Festuca arundinacea, and Lactuca sativa with inhibitory rates of 71.3 and 88.7% for L. perenne, 79.4 and 73.0% for F. arundinacea, and 58.2 and 52.9% for L. sativa. In addition, compound 1 at 25 µg/mL showed moderate antifungal activity against Fusarium fujikuroi and Trichoderma harzianum with inhibition rates of 22.6 and 31.6%, respectively. These results suggest that heterologous expression in conjunction with the OSMAC approach provides a promising strategy to extend the metabolite novelty due to the incorporation of endogenous metabolites from the host strain with exogenous compounds, leading to the production of more complex compounds and the acquisition of new physiological functions.

Adequate water supply enhances seedling growth and metabolism in Festuca kryloviana: insights from physiological and transcriptomic analys.

BACKGROUND: Festuca kryloviana is a significant native grass species in the Qinghai Lake region, and its low emergence rate is a primary factor limiting the successful establishment of cultivated grasslands. The region's arid and low-rainfall climate characteristics result in reduced soil moisture content at the surface. Despite the recognized impact of water availability on plant growth, the specific role of moisture in seedling development remains not fully elucidated. This study aims to investigate the germination rate and seedling growth velocity of F. kryloviana seeds under varying moisture conditions, and to integrate physiological and transcriptomic analyses of seedlings under these conditions to reveal the mechanisms by which water influences seedling development. RESULTS: The emergence rate of F. kryloviana seedlings exhibited an initial increase followed by a decrease with increasing moisture content. The highest emergence rate, reaching 75%, was observed under 20% soil moisture conditions. By the eighth day of the experiment, the lengths of the plumules and radicles under the optimal emergence rate (full water, FW) were 21.82% and 10.87% longer, respectively, than those under closely matching the soil moisture content during the background survey (stress water, SW). The differential development of seedlings under varying moisture regimes is attributed to sugar metabolism within the seeds and the accumulation of abscisic acid (ABA). At FW conditions, enhanced sugar metabolism, which generates more energy for seedling development, is facilitated by higher activities of α-amylase, sucrose synthase, and trehalose-6-phosphate synthase compared to SW conditions. This is reflected at the transcriptomic level with upregulated expression of the α-amylase (AMY2) gene and trehalose-6-phosphate synthase (TPS6), while genes associated with ABA signaling and transduction are downregulated. Additionally, under FW conditions, the expression of genes related to the chloroplast thylakoid photosystems, such as photosystem II (PSII) and photosystem I (PSI), is upregulated, enhancing the seedlings' light-capturing ability and photosynthetic efficiency, thereby improving their autotrophic capacity. Furthermore, FW treatment enhances the expression of the non-enzymatic antioxidant system, promoting metabolism within the seeds. In contrast, SW treatment increases the activity of the enzymatic antioxidant system, including peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT), to cope with water stress. CONCLUSIONS: Our experiment systematically evaluated the impact of moisture conditions on the growth and development of F. kryloviana seedlings. Physiological and transcriptomic data collectively indicate that adequate water (20%) supply enhances seedling growth and development by reducing ABA levels and increasing α-amylase activity within seeds, thereby boosting sugar metabolism and promoting the growth of seedling, which in turn leads to an improved emergence rate. Considering water management in future cultivation practices may be a crucial strategy for enhancing the successful establishment of F. kryloviana in grassland ecosystems.

Phenotypic screening of seed retention and histological analysis of the abscission zone in Festuca pratensis and Lolium perenne.

BACKGROUND: Seed retention is the basic prerequisite for seed harvest. However, only little breeding progress has been achieved for this trait in the major forage grasses. The aim of this study was to evaluate the potential of plant genetic resources of the important fodder grasses Festuca pratensis Huds. and Lolium perenne L. as source for seed retention in the breeding process. Furthermore, the morphology of the abscission zone, where shattering occurs, was studied on the cell tissue level in different developmental stages of contrasting accessions. RESULTS: 150 and 286 accessions of Festuca pratensis and Lolium perenne were screened for seed retention, respectively. Contrasting accessions were selected to be tested in a second year. We found a great variation in seed retention in Festuca pratensis and Lolium perenne, ranging from 13 to 71% (average: 35%) and 12 to 94% (average: 49%), respectively, in the first year. Seed retention was generally lower in the second year. Cultivars were within the accessions with highest seed retention in Festuca pratensis, but had lower seed retention than ecotypes in Lolium perenne. Field-shattered seeds had a lower thousand grain weight than retained seeds. Cell layers of the abscission zone appeared already in early seed stages and were nested within each other in accessions with high seed retention, while there were two to three superimposed layers in accessions with low seed retention. CONCLUSIONS: Plant genetic resources of Lolium perenne might be a valuable source for breeding varieties with high seed retention. However, simultaneous selection for high seed weight is necessary for developing successful commercial cultivars.

Chemical Basis for Determining the Allelopathic Potential of Invasive Plant Wall Barley (Hordeum murinum L. subsp. murinum).

The study investigated compounds present in the invasive grass Hordeum murinum L. subsp. murinum and tested the allelopathic potential of this plant against common meadow species Festuca rubra L. and Trifolium repens L. Gas chromatography-mass spectrometry (GC-MS) performed separately on the ears and stalks with leaves of wall barley revealed 32 compounds, including secondary metabolites, that may play an important role in allelopathy. Two compounds, N-butylbenzenesulfonamide (NBBS) and diphenylsulfone (DDS), were described for the first time for wall barley and the Poaceae family. The presence of 6,10,14-trimethylpentadecan-2-one (TMP) has also been documented. Aqueous extracts of H. murinum organs (ears and stalks with leaves) at concentrations of 2.5%, 5%, and 7.5% were used to evaluate its allelopathic potential. Compared to the control, all extracts inhibited germination and early growth stages of meadow species. The inhibitory effect was strongest at the highest concentration for both the underground and aboveground parts of the seedlings of the meadow species tested. Comparing the allelopathic effect, Trifolium repens proved to be more sensitive. In light of the results of the study, the removal of wall barley biomass appears to be important for the restoration of habitats where this species occurs due to its allelopathic potential.

Reciprocal allopolyploid grasses (Festuca × Lolium) display stable patterns of genome dominance.

Allopolyploidization entailing the merger of two distinct genomes in a single hybrid organism, is an important process in plant evolution and a valuable tool in breeding programs. Newly established hybrids often experience massive genomic perturbations, including karyotype reshuffling and gene expression modifications. These phenomena may be asymmetric with respect to the two progenitors, with one of the parental genomes being "dominant." Such "genome dominance" can manifest in several ways, including biased homoeolog gene expression and expression level dominance. Here we employed a k-mer-based approach to study gene expression in reciprocal Festuca pratensis Huds. × Lolium multiflorum Lam. allopolyploid grasses. Our study revealed significantly more genes where expression mimicked that of the Lolium parent compared with the Festuca parent. This genome dominance was heritable to successive generation and its direction was only slightly modified by environmental conditions and plant age. Our results suggest that Lolium genome dominance was at least partially caused by its more efficient trans-acting gene expression regulatory factors. Unraveling the mechanisms responsible for propagation of parent-specific traits in hybrid crops contributes to our understanding of allopolyploid genome evolution and opens a way to targeted breeding strategies.

A comparison of shared patterns of differential gene expression and gene ontologies in response to water-stress in roots and leaves of four diverse genotypes of Lolium and Festuca spp. temperate pasture grasses.

Ryegrasses (Lolium spp.) and fescues (Festuca spp.) are closely related and widely cultivated perennial forage grasses. As such, resilience in the face of abiotic stresses is an important component of their traits. We have compared patterns of differentially expressed genes (DEGs) in roots and leaves of two perennial ryegrass genotypes and a single genotype of each of a festulolium (predominantly Italian ryegrass) and meadow fescue with the onset of water stress, focussing on overall patterns of DEGs and gene ontology terms (GOs) shared by all four genotypes. Plants were established in a growing medium of vermiculite watered with nutrient solution. Leaf and root material were sampled at 35% (saturation) and, as the medium dried, at 15%, 5% and 1% estimated water contents (EWCs) and RNA extracted. Differential gene expression was evaluated comparing the EWC sampling points from RNAseq data using a combination of analysis methods. For all genotypes, the greatest numbers of DEGs were identified in the 35/1 and 5/1 comparisons in both leaves and roots. In total, 566 leaf and 643 root DEGs were common to all 4 genotypes, though a third of these leaf DEGs were not regulated in the same up/down direction in all 4 genotypes. For roots, the equivalent figure was 1% of the DEGs. GO terms shared by all four genotypes were often enriched by both up- and down-regulated DEGs in the leaf, whereas generally, only by either up- or down-regulated DEGs in the root. Overall, up-regulated leaf DEGs tended to be more genotype-specific than down-regulated leaf DEGs or root DEGs and were also associated with fewer GOs. On average, only 5-15% of the DEGs enriching common GO terms were shared by all 4 genotypes, suggesting considerable variation in DEGs between related genotypes in enacting similar biological processes.

Soil chemical legacies trigger species-specific and context-dependent root responses in later arriving plants.

Soil legacies play an important role for the creation of priority effects. However, we still poorly understand to what extent the metabolome found in the soil solution of a plant community is conditioned by its species composition and whether soil chemical legacies affect subsequent species during assembly. To test these hypotheses, we collected soil solutions from forb or grass communities and evaluated how the metabolome of these soil solutions affected the growth, biomass allocation and functional traits of a forb (Dianthus deltoides) and a grass species (Festuca rubra). Results showed that the metabolomes found in the soil solutions of forb and grass communities differed in composition and chemical diversity. While soil chemical legacies did not have any effect on F. rubra, root foraging by D. deltoides decreased when plants received the soil solution from a grass or a forb community. Structural equation modelling showed that reduced soil exploration by D. deltoides arose via either a root growth-dependent pathway (forb metabolome) or a root trait-dependent pathway (grass metabolome). Reduced root foraging was not connected to a decrease in total N uptake. Our findings reveal that soil chemical legacies can create belowground priority effects by affecting root foraging in later arriving plants.

Genome-wide small RNA profiling reveals tiller development in tall fescue (Festuca arundinacea Schreb).

BACKGROUND: Tall fescue (Festuca arundinacea Schreb.) is a major cool-season forage and turfgrass species. The low tiller density and size dramatically limits its turf performance and forage yield. MicroRNAs (miRNA)-genes modules play critical roles in tiller development in plants. In this study, a genome-wide small RNA profiling was carried out in two tall fescue genotypes contrasting for tillering production ('Ch-3', high tiller production rate and 'Ch-5', low tiller production rate) and two types of tissue samples at different tillering development stage (Pre-tillering, grass before tillering; Tillering, grass after tillering). 'Ch-3', 'Ch-5', Pre-tillering, and Tillering samples were analyzed using high-throughput RNA sequencing. RESULTS: A total of 222 million high-quality clean reads were generated and 208 miRNAs were discovered, including 148 known miRNAs belonging to 70 families and 60 novel ones. Furthermore, 18 miRNAs were involved in tall fescue tiller development process. Among them, 14 miRNAs displayed increased abundance in both Ch-3 and Tillering plants compared with that in Ch-5 and Pre-tillering plants and were positive with tillering, while another four miRNAs were negative with tiller development. Out of the three miRNAs osa-miR156a, zma-miR528a-3p and osa-miR444b.2, the rest of 15 miRNAs were newfound and associated with tiller development in plants. Based on our previous full-length transcriptome analysis in tall fescue, 28,927 potential target genes were discovered for all identified miRNAs. Most of the 212 target genes of the 18 miRNAs were dominantly enriched into "ubiquitin-mediated proteolysis", "phagosome", "fatty acid biosynthesis", "oxidative phosphorylation", and "biosynthesis of unsaturated fatty acids" KEGG pathways. In addition, bdi-miR167e-3p targets two kinase proteins EIF2AK4 and IRAK4, and osa-miR397a targets auxin response factor 5, which may be the significant miRNA-genes controllers in tillering development. CONCLUSIONS: This is the first genome-wide miRNA profiles analysis to identify regulators involved in tiller development in cool-season turfgrass. Tillering related 18 miRNAs and their 212 target genes provide novel information for the understanding of the molecular mechanisms of miRNA-genes mediated tiller development in cool-season turfgrass.

High temperature damage to fatty acids and carbohydrate metabolism in tall fescue by coupling deep transcriptome and metabolome analysis.

High temperature damage impairs the growth of tall fescue by inhibiting secondary metabolites. Little is known about the regulation pattern of the fatty acids and carbohydrate metabolism at the whole-transcriptome level in tall fescue under high temperature stress. Here, two tall fescue genotypes, heat tolerant PI578718 and heat sensitive PI234881 were subjected to high temperature stress for 36 h. PI 578718 showed higher SPAD chloroplast value, lower EL and leaf injury than PI 234881 during the first 36 h high-temperature stress. Furthermore, by transcriptomic analysis, 121 genes were found to be induced during the second energy production phase in tall fescue exposed to high-temperature conditions, indicating that there may be one energy-sensing system in cool-season turfgrass to adapt high-temperature conditions. PI 578718 showed higher differentially expressed unigenes involved in fatty acids and carbohydrate metabolism compared with PI 234881 for 36 h heat stress. Interestingly, a metabolomic analysis using GC-MS uncovered that the sugars and sugar alcohol accounted for more than 65.06% of the total 41 metabolites content and high-temperature elevated the rate to 82.89-91.16% in PI 578718. High-temperature damage decreased the rate of fatty acid in the total 41 metabolites content and PI 578718 showed lower content than in PI 234881, which might be attributed to the down-regulated genes in fatty acid biosynthesis pathway in tall fescue. The integration of deep transcriptome and metabolome analyses provides systems-wide datasets to facilitate the identification of crucial regulation factors in cool-season turfgrass in response to high-temperature damage.

Impact of endophyte inoculation on the morphological identity of cultivars of Lolium perenne (L) and Festuca arundinacea (Schreb.).

Grass endophytes have been shown to confer enhanced environmental resilience to symbiont cultivars with reports of modified growth. If inoculating with an endophyte (E+) made an accession morphologically distinct from its registered endophyte free (E-) accession, there could be protection and ownership issues for testing authorities and breeders. This study investigated if, in official Plant Breeders Rights (PBR) field trials, the morphological characteristics of E+and E- accessions of perennial ryegrass and tall fescue cultivars were sufficiently modified to designate them as mutually distinct and also distinct from their definitive accessions (Def), held by the testing authorities. Testing perennial ryegrass on 17 characters at 2 sites generated 48,960 observations and for tall fescue on 9 characters at 1 site, 12,960 observations (each for 3 accessions of 4 cultivars × 60 plants × 2 growing cycles). Distinctness required a p < 0.01 difference in a single character from the combined over years analysis (COYD). A few significant differences were recorded between E- and E+accessions. Cultivar Carn E+ was smaller than Carn E- for Infloresence Length (p < 0.01) in both years but COYD analysis (p < 0.05) was insufficient to declare distinctiveness. Overall, the number of observed differences between E-/E+ accessions was less or similar to the number expected purely by chance. In contrast, comparisons between Def and E- or E+ accessions showed a number of significant differences that were substantially more numerous than expected by chance. These results showed no conclusive evidence of endophyte inclusion creating false PBR distinctions but unexpectedly, several E- and E+ accessions were distinguished from their official definitive stock.

Species Differences in Phenology Shape Coexistence.

Ecological theory produces opposing predictions about whether differences in the timing of life-history transitions, or "phenology," promote or limit coexistence. Phenological separation is predicted to create temporal niche differences, increasing coexistence, yet phenological separation could also competitively favor one species, increasing fitness differences and hindering coexistence. We experimentally manipulated relative germination timing, a critical phenological event, of two annual grass species, Vulpia microstachys and V. octoflora, to test these contrasting predictions. We parameterized a competition model to estimate within-season niche differences, fitness differences, and coexistence and to estimate coexistence when year-to-year fluctuations of germination timing occur. Increasing germination separation caused parallel changes in niche and fitness differences, with the net effect of weakening within-year coexistence. Both species experienced a competitive advantage by germinating earlier, and a 4-day head start allowed the generally inferior competitor to exclude the otherwise superior competitor. The overall consequence of germination separation was to limit coexistence within a given year, although year-to-year variation in the relative timing of germination was sufficient to support long-term coexistence. Our results clarify how phenological differences structure competitive interactions and highlight the need to quantify year-to-year variation in these differences to better understand species coexistence.

Comparative study of alleviation effects of DMTU and PCIB on root growth inhibition in two tall fescue varieties under cadmium stress.

In plants, tolerance to cadmium (Cd) stress is closely related to indole-3-acetic acid (IAA) and hydrogen peroxide (H2O2). However, it is unclear whether Cd-resistant and -sensitive varieties respond differently to Cd stress. In this study, the effects of dimethylthiourea (DMTU, a H2O2 scavenger) and p-chlorophenoxy isobutyric acid (PCIB, an IAA signaling inhibitor) on root growth, endogenous hormones and antioxidant system were investigated to decipher how DMTU and PCIB treatments alleviate the inhibition of root elongation in Cd-resistant (Commander) and -sensitive (Crossfire III) tall fescue varieties under Cd stress. Both varieties subjected to 10 µM Cd treatments for 12 h presented a substantial decrease in root elongation coupled with a reduction in brassinosteroid (BR) and zeatin riboside (ZR) contents, but the changes in IAA and abscisic acid (ABA) contents under Cd stress were opposite in the two varieties. In addition, the H2O2 content and antioxidant enzyme activities significantly increased in both varieties. However, pretreatment with PCIB or DMTU mitigated the inhibition of root elongation caused by Cd, accompanied by the significant changes of aforementioned physiological parameters. PCIB significantly reduced the IAA content in 'Commander', while DMTU significantly increased the IAA content in 'Crossfire III' and effectively relieved the inhibition of root elongation. But both treatments decreased the Cd-induced H2O2 accumulation. These results indicated that DMTU or PCIB can alleviate the Cd-inhibited root elongation in two varieties whose resistance differed under Cd stress, but they presented differences in the response of hormones, especially IAA, which may be due to the different adaptation mechanisms of two varieties in response to Cd stress.

Kikuyu pastures associated with tall fescue grazed in autumn in small-scale dairy systems in the highlands of Mexico.

Intensive grazing increases the profitability and sustainability of small-scale dairy systems by reducing feeding costs. Kikuyu grass is a subtropical species from East Africa that has similar performance compared with temperate grasses when grazed by dairy cows in these systems during the summer rainy season but reduces growth and quality at low temperatures, when temperate species may have an advantage. The objective was to evaluate intensive grazing of kikuyu pastures (KYKY) alone or in association with two varieties of endophyte-free tall fescue, TF-33 (TF33) and Cajun II (CAJN), during the summer-autumn transition period when low temperatures set in, by lactating cows in small-scale dairy farms. Pasture variables were analysed with a split-plot design for sward height, net herbage accumulation and chemical composition, in vitro digestibility of organic matter, and estimated metabolisable energy content of herbage and concentrate. Experimental design for animal variables was a 3 × 3 Latin Square repeated three times with nine Holstein cows and 14 days experimental periods. Cows received 4.65 kg DM/day of a 16% CP commercial concentrate. Milk yield and composition, live weight, and body condition score were recorded. There were no significant differences (P > 0.05) for sward height and net herbage accumulation, nor for important components of chemical composition of herbages. There were no significant differences (P > 0.05) for milk yield (19 kg/cow/day) and composition, although differences (P < 0.05) were detected for live weight and body condition score. The conclusion is that there is no advantage of associating tall fescue with kikuyu in summer-autumn transition period for small-scale dairy systems.

Effect of 24-epibrassinolide on reactive oxygen species and antioxidative defense systems in tall fescue plants under lead stress.

Lead is one of the most hazardous pollutants to both the environment as well as human beings. As one of the approaches to enhance phytoremediation, brassinosteroids are predicted as a potential candidate phytohormone for assisted phytoremediation. Few studies have focused on the physiological regulations of tall fescue plants (Festuca arundinacea Schreb.), a potential phytoremediation species, for its responses to applications of brassinosteroids under lead stress. Therefore, the objectives of this study were to investigate the effects of foliar application of 24-epibrassinolide, a brassinosteroids analogue, on reactive oxygen species accumulation and antioxidative defense systems of tall fescue when exposed to lead, and ultimately its potential to be used in phytoremediation. When exposed to lead (1000 mg/kg) for 80 d, decreases in shoot and root biomass of tall fescue biomass as well as chlorophyll and carotenoid productions were found. Foliar application of 24-epibrassinolide at three rates and five applications every 7 d improved the biomass of both shoots and roots, and increased the photosynthetic pigments. The improved lead tolerance in tall fescue plants after 24-epibrassinolide applications was associated with reduced H2O2 and O2.- accumulations and increased antioxidative enzyme activities including superoxide dismutase, catalase, and guaiacol peroxidase. Additionally, osmoprotectants increased and lipid peroxidation decreased. Ultimately, foliar applications of 24-epibrassinolide enhanced the lead recovery rate of tall fescue plants, proving its potential role in phytoremediation for soil contaminated with heavy metals such as lead.

Assisted phytostabilization of soil from a former military area with mineral amendments.

Due to the presence of toxic pollutants, soils in former military areas need remedial actions with environmentally friendly methods. Greenhouse experiments were conducted to investigate the aided phytostabilization of multi-heavy metals (HMs), i.e. Cd, Cr, Cu, Ni, Pb, Zn, in post-military soil by Festuca rubra and three mineral amendments (diatomite, dolomite and halloysite). The amendments were applied at 0 and 3.0% to each pot filled with 5 kg of polluted soil. After seven weeks of the phytostabilization, selected soil properties, biomass yield of F. rubra and immobilization of HMs by their accumulation in plant and redistribution among individual fractions in soil were determined. In addition, ecotoxicology parameters of non-amended and amended soil were established using Phytotoxkit (Sinapsis alba) and Ostracodtoxkit (Heterocypris incongruens) tests. The addition of halloysite significantly increased F. rubra biomass. Diatomite significantly increased both the Cd, Cu, Pb and Cr concentrations in the roots and the pH of the soil. The application of halloysite significantly decreased the Cd and Zn contents of the soil after the completion of the experiment. Dolomite and halloysite were more effective in HM immobilization in soil by decreasing their content in an exchangeable fraction than diatomite. These soil amendments significantly differentiated the length of S. alba roots and had a positive effect on the development of H. incongruens.

Gibberellic acid inhibition of tillering in tall fescue involving crosstalks with cytokinins and transcriptional regulation of genes controlling axillary bud outgrowth.

Tiller production in grass species is controlled by both axillary bud initiation and bud outgrowth, which may be regulated by plant hormones. However, how gibberellic acid (GA) affects tillering in perennial grass species is still unclear. This study aims to elucidate the roles and the underlying mechanisms of GA in regulating tiller development. Tall fescue seedlings were treated with different concentrations of GA3 by foliar application, dose-dependent inhibitory effects of GA on tiller production were observed. GA3 (25 µM) slowed down the transition from axillary buds to tillers by specifically inhibiting the outgrowth of axillary buds. GA-inhibition of tillering were not related to endogenous content for auxin or strigolactone, but was mainly due to the antagonistic interaction with cytokinins (CK), as shown by the decreased CK content and up-regulation expression of CK degradation genes in GA3-treated plants. Furthermore, GA could act through regulating the expression of FaTB1 specifically expressed in axillary buds to repress bud outgrowth. These results provide insights for the regulatory mechanisms of GA for tiller bud outgrowth through crosstalks with CK and signaling of FaTB1 expression.

Interactions between decabromodiphenyl ether and lead in soil-plant system.

Pot experiments were conducted under abiotic conditions to investigate the interactive influence of decabromodiphenyl ether (BDE-209) and lead (Pb) on the seed germination, germ length, root exudation and physiological characteristics of tall fescue (Festuca arundinaceae), and the uptake, accumulation of Pb and BDE-209 in the plant tissues. Results show that seed germination and germ length were impacted by Pb but less influenced by BDE-209. BDE-209 spiking (10 and 50 mg/L) could alleviate the toxicity of high Pb concentration on seed germination and growth. The chlorophyll content was significantly increased at 500 mg/kg Pb but declined at 2000 mg/kg Pb. Low-level Pb contamination (500 mg/kg) activated antioxidase activity; however, 2000 mg/kg Pb significantly reduced the antioxidase activity. Plant biomass slightly decreased at 500 mg/kg Pb but significantly declined at 2000 mg/kg Pb. The addition of a moderate dosage of BDE-209 (10-50 mg/kg) lessened Pb phytotoxicity, leading to improved plant growth relative to the case of Pb spiking alone. The exudate secretion was significantly enhanced by Pb addition, but BDE-209 spiking only caused slightly increased secretion. Pb could interfere with BDE-209 adsorption and translocation of tall fescue by affecting physiological behavior of the plant, but BDE-209 exhibited little influence on the Pb fate in the plant. Overall, BDE-209 had slight interference on the impact of Pb towards tall fescue. The results demonstrate the complex interactive effects of organic pollutants and heavy metals in the soil-plant system.

Interaction between Cd and Zn on Metal Accumulation, Translocation and Mineral Nutrition in Tall Fescue (Festuca arundinacea).

Tall fescue (Festuca arundinacea), an accumulator that is able to accumulate and excrete cadmium (Cd), has attracted much attention for its possible use in phytoremediation of heavy metal contaminated soils. In the present study, the interaction between Cd and Zn, and their uptake, translocation and accumulation under external Cd and Zn treatment in tall fescue were investigated. The concentrations of K, Ca, Mg in xylem sap under Cd and Zn treatment were measured to determine the level of mineral nutrients and their relationship with Cd alleviation. The result showed that Cd and Zn antagonized each other in the roots, while Cd antagonized Zn and Zn synergized Cd in the shoots of tall fescue. Compared with Cd only treatment, the concentrations of Ca, Mg and K in xylem sap increased after the addition of Zn, and they increased the most in the guttation. This result indicated that the addition of Zn facilitates the level of mineral elements to alleviate Cd toxicity, which might be used to improve the phytoremediation efficiency of Cd contaminated soils by tall fescue.

The Impact of Endophyte Infection, Seed Aging, and Imbibition on Selected Sugar Metabolite Concentrations in Seed.

This study investigated effects of seed aging and imbibition on sugar metabolite concentrations in Epichloë endophyte-infected and endophyte-free seed of tall fescue ( Festuca arundinacea Schreb.). Two treatments, namely, accelerated aging and imbibition, were applied to the seeds, with embryo and endosperm tissues analyzed separately. Gas chromatography with flame ionization detection was employed for analysis of sugar metabolites within the seed tissues. Mannitol, ribitol, and trehalose were more abundant in embryo than endosperm tissues and were identified at consistently higher concentrations within endophyte-infected compared to endophyte-free seeds. The ratio of raffinose to sucrose significantly increased with seed aging in both endophyte-free and endophyte-infected embryo tissues, while significantly lower concentrations of trehalose were detected in tissues dissected from aged-seed regardless of endophyte status. This research provides fundamental insight into the metabolic details of endophyte survival in seed and provides a first evaluation of key carbohydrates present in the fungal-plant symbiosis.

Transcriptome sequencing of Festulolium accessions under salt stress.

OBJECTIVES: The objective of this study was to establish transcriptome assemblies of Festulolium hybrids under salt stress, and identify genes regulated across the hybrids in response to salt stress. The development of transcriptome assemblies for Festulolium hybrids and cataloguing of genes regulated under salt stress will facilitate further downstream studies. RESULTS: Plants were grown at three salt concentrations (0.5%, 1% and 1.5%) and phenotypic and transcriptomic data was collected. Salt stress was confirmed by progressive loss of green leaves as salt concentration increased from 0 to 1.5%. We generated de-novo transcriptome assemblies for two Festulolium pabulare festucoid genotypes, for a single Festulolium braunii genotype, and a single F. pabulare loloid genotype. We also identified 1555 transcripts that were up regulated and 1264 transcripts that were down regulated in response to salt stress in the Festulolium hybrids. Some of the identified transcripts showed significant sequence similarity with genes known to be regulated during salt and other abiotic stresses.