The genome of Vicia sativa ssp. amphicarpa provides insights into the role of terpenoids in antimicrobial resistance within subterranean fruits.

Vicia sativa ssp. amphicarpa is a unique forage crop capable of simultaneously producing fruits above and below ground, representing a typical amphicarpic plant. In this study, we sequenced and assembled seven pseudo-chromosomes of the genome of V. sativa ssp. amphicarpa (n = 7) yielding a genome size of 1.59 Gb, with a total annotation of 48 932 protein-coding genes. Long terminal repeat (LTR) elements constituted 62.28% of the genome, significantly contributing to the expansion of genome size. Phylogenetic analysis revealed that the divergence between V. sativa ssp. amphicarpa and V. sativa was around 0.88 million years ago (MYA). Comparative transcriptomic and metabolomic analysis of aerial and subterranean pod shells showed biosynthesis of terpenoids in the subterranean pod shells indicating a correlation between the antimicrobial activity of subterranean pod shells and the biosynthesis of terpenoids. Furthermore, functional validation indicates that overexpression of VsTPS5 and VsTPS16 enhances terpenoid biosynthesis for antibacterial activity. Metabolomic analysis suggests the involvement of terpenoids in the antimicrobial properties of subterranean pod shells. Deciphering the genome of V. sativa ssp. amphicarpa elucidated the molecular mechanisms behind the antimicrobial properties of subterranean fruits in amphicarpic plants, providing valuable insights for the study of amphicarpic plant biology.

Endophytes Alleviate Drought-Derived Oxidative Damage in Achnatherum inebrians Plants Through Increasing Antioxidants and Regulating Host Stress Responses.

Endophytes generally increase antioxidant contents of plants subjected to environmental stresses. However, the mechanisms by which endophytes alter the accumulation of antioxidants in plant tissues are not entirely clear. We hypothesized that, in stress situations, endophytes would simultaneously reduce oxidative damage and increase antioxidant contents of plants and that the accumulation of antioxidants would be a consequence of the endophyte ability to regulate the expression of plant antioxidant genes. We investigated the effects of the fungal endophyte Epichloë gansuensis (C.J. Li & Nan) on oxidative damage, antioxidant contents, and expression of representative genes associated with antioxidant pathways in Achnatherum inebrians (Hance) Keng plants subjected to low (15%) and high (60%) soil moisture conditions. Gene expression levels were measured using RNA-seq. As expected, the endophyte reduced the oxidative damage by 17.55% and increased the antioxidant contents by 53.14% (on average) in plants subjected to low soil moisture. In line with the accumulation of antioxidants in plant tissues, the endophyte increased the expression of most plant genes associated with the biosynthesis of antioxidants (e.g., MIOX, crtB, gpx) while it reduced the expression of plant genes related to the metabolization of antioxidants (e.g., GST, PRODH, ALDH). Our findings suggest that endophyte ability of increasing antioxidant contents in plants may reduce the oxidative damage caused by stresses and that the fungal regulation of plant antioxidants would partly explain the accumulation of these compounds in plant tissues.

Leaf morphology, functional trait and altitude response in perennial vetch (Vicia unijuga A. Braun), alfalfa (Medicago sativa L.) and sainfoin (Onobrychis viciifolia Scop.).

MAIN CONCLUSION: Species have plasticity across altitude gradients in leaf morphology and function, and their response to high altitude conditions was mainly reflected in leaf cell metabolism and gas exchange. Leaf morphological and functional adaptation to altitude has received research attention in recent years, but there are no studies for forage legumes. Here we report differences in 39 leaf morphology and functional traits of three leguminous forages (alfalfa, sainfoin and perennial vetch) at three sites in Gansu Province, China, ranging from 1768 to 3074 m altitude to provide information for potential use in breeding programmes. With increasing altitude, plant water status increased, reflecting increase in soil water content and decreased average temperature, which lead to leaf intercellular CO2 concentration. Stomatal conductance and evapotranspiration increased significantly but water-use efficiency decreased. At high altitude, ΦPSII decreased but non-photochemical quenching and chlorophyll a:b ratio increased while spongy mesophyll tissue and leaf thickness increased. These changes may be due to UV or low-temperature damage of leaf protein and metabolic cost of plant protection or defence responses. Contrary to many other studies, leaf mass per area decreased significantly at higher altitude. This was consistent with predictions under the worldwide leaf economic spectrum on the basis that soil nutrients increased with increasing altitude. The key species differences were more irregularly shaped epidermal cells and larger stomatal size in perennial vetch compared to alfalfa or sainfoin that enhanced gas exchange and photosynthesis by generating mechanical force, increasing guard cell turgor, and promoting stomatal operation. The lower adaxial stomatal density also enhanced water-use efficiency. These adaptations might confer perennial vetch an advantage in environments with extreme diurnal temperature fluctuation or in frigid conditions.

Arbuscular mycorrhizal fungus changes alfalfa (Medicago sativa) metabolites in response to leaf spot (Phoma medicaginis) infection, with subsequent effects on pea aphid (Acyrthosiphon pisum) behavior.

Plant disease occurs simultaneously with insect attack. Arbuscular mycorrhizal fungi (AMF) modify plant biotic stress response. Arbuscular mycorrhizal fungi and pathogens may modify plant volatile organic compound (VOC) production and insect behavior. Nevertheless, such effects are rarely studied, particularly for mesocosms where component organisms interact with each other. Plant-mediated effects of leaf pathogen (Phoma medicaginis) infection on aphid (Acyrthosiphon pisum) infestation, and role of AMF (Rhizophagus intraradices) in modifying these interactions were elucidated in a glasshouse experiment. We evaluated alfalfa disease occurrence, photosynthesis, phytohormones, trypsin inhibitor (TI) and total phenol response to pathogen and aphid attack, with or without AMF, and aphid behavior towards VOCs from AMF inoculated and non-mycorrhizal alfalfa, with or without pathogen infection. AM fungus enhanced alfalfa resistance to pathogen and aphid infestation. Plant biomass, root : shoot ratio, net photosynthetic rate, transpiration rate, stomatal conductance, salicylic acid, and TI were significantly increased in AM-inoculated alfalfa. Arbuscular mycorrhizal fungi and pathogen significantly changed alfalfa VOCs. Aphids preferred VOCs of AM-inoculated and nonpathogen-infected to nonmycorrhizal and pathogen-infected alfalfa. We propose that AMF alter plant response to multiple biotic stresses in ways both beneficial and harmful to the plant host, providing a basis for strategies to manage pathogens and herbivore pests.

Microbial carbon use efficiency in different ecosystems: A meta-analysis based on a biogeochemical equilibrium model.

Soil microbial carbon use efficiency (CUE) is a crucial parameter that can be used to evaluate the partitioning of soil carbon (C) between microbial growth and respiration. However, general patterns of microbial CUE among terrestrial ecosystems (e.g., farmland, grassland, and forest) remain controversial. To address this knowledge gap, data from 41 study sites (n = 197 soil samples) including 58 farmlands, 95 forests, and 44 grasslands were collected and analyzed to estimate microbial CUEs using a biogeochemical equilibrium model. We also evaluated the metabolic limitations of microbial growth using an enzyme vector model and the drivers of CUE across different ecosystems. The CUEs obtained from soils of farmland, forest, and grassland ecosystems were significantly different with means of 0.39, 0.33, and 0.42, respectively, illustrating that grassland soils exhibited higher microbial C sequestration potentials (p < .05). Microbial metabolic limitations were also distinct in these ecosystems, and carbon limitation was dominant exhibiting strong negative effects on CUE. Exoenzyme stoichiometry played a greater role in impacting CUE values than soil elemental stoichiometry within each ecosystem. Specifically, soil exoenzymatic ratios of C:phosphorus (P) acquisition activities (EEAC:P ) and the exoenzymatic ratio of C:nitrogen (N) acquisition activities (EEAC:N ) imparted strong negative effects on soil microbial CUE in grassland and forest ecosystems, respectively. But in farmland soils, EEAC:P exhibited greater positive effects, showing that resource constraints could regulate microbial resource allocation with discriminating patterns across terrestrial ecosystems. Furthermore, mean annual temperature (MAT) rather than mean annual precipitation (MAP) was a critical climate factor affecting CUE, and soil pH as a major factor remained positive to drive the changes in microbial CUE within ecosystems. This research illustrates a conceptual framework of microbial CUEs in terrestrial ecosystems and provides the theoretical evidence to improve soil microbial C sequestration capacity in response to global change.

Drought Eliminates the Difference in Root Trait Plasticity and Mycorrhizal Responsiveness of Two Semiarid Grassland Species with Contrasting Root System.

Root traits and arbuscular mycorrhizal (AM) fungi are important in determining the access of plants to soil resources. However, whether plants with different root systems (i.e., taproot vs. fibrous-root) exhibit different root trait plasticity and mycorrhizal responsiveness under drought remains largely unexplored. Tap-rooted Lespedeza davurica and fibrous-rooted Stipa bungeana were grown in monocultures in sterilized and live soils, followed by a drought treatment. Biomass, root traits, root colonization by AM fungi, and nutrient availability were evaluated. Drought decreased biomass and root diameter but increased the root:shoot ratio (RSR), specific root length (SRL), soil NO3--N, and available P for the two species. Under control and drought conditions, soil sterilization significantly increased the RSR, SRL, and soil NO3--N for L. davurica, but this only occurs under drought condition for S. bungeana. Soil sterilization significantly reduced AM fungal root colonization of both species, but drought significantly increased it in live soil. In water-abundant conditions, tap-rooted L. davurica may depend more on AM fungi than fibrous-rooted S. bungeana; however, under drought conditions, AM fungi are of equal importance in favoring both plant species to forage soil resources. These findings provide new insights for understanding the resource utilization strategies under climate change.

Pathogen Identification and Factors Influencing Infection Frequency and Severity of Fungal Rust in Four Native Grasses in Hulunber Grassland, China.

A serious rust infection present in 2014 and 2015 on the dominant grass species (Leymus chinensis) in the Hulunber grassland of Inner Mongolia, China, and also present on three other grass species (Agropyron cristatum [wheat grass], Bromus inermis, and Festuca ovina) was investigated. Field surveys, laboratory determination of morphological characteristics, pathogenicity tests, and molecular identification methods were integrated to identify two rust-causing pathogens on L. chinensis. It was found that Puccinia elymi was the major pathogen of L. chinensis, and also infected A. cristatum and F. ovina. This is the first report of P. elymi on A. cristatum in China. P. striiformis caused stripe rust on L. chinensis and B. inermis. The incidence and severity of rust infection increased through the growing season, presumably from asexual spread by urediniospores, and was higher on grass species phylogenetically more closely related to common crop hosts of the pathogens. High host grass density and presence of a potential alternate host for P. elymi, Thalictrum squarrosum, were two further factors promoting rust incidence. These results provide insight into ecological factors linked to the rust epidemic and provide a theoretical basis for the formulation of control strategies.

Integrative Analyses of Transcriptomes and Metabolomes Reveal Associated Genes and Metabolites with Flowering Regulation in Common Vetch (Vicia sativa L.).

As an important source of protein for livestock and human consumption, Vicia sativa is cultivated worldwide, but its seed production is hampered at high altitudes because of the short frost-free period. Flowering represents the transition from a vegetative to a reproductive period, and early flowering benefits plant seed production at high altitudes. However, the molecular mechanisms of flowering regulation in V. sativa remain elusive. In the present study, two V. sativa accessions with different flowering characteristics were used: Lan3 (early-flowering) was cultivated by our laboratory, and 503 (late-flowering) was selected from 222 V. sativa accessions after three years of field experiments. The shoot samples (shoot tip length = 10 cm) of these two accessions were collected 63, 70, and 77 days after sowing, and the molecular regulatory mechanism of the flowering process was identified by integrative analyses of the transcriptomes and metabolomes. Kyoto Encyclopedia of Genes and Genomes enrichment showed that the synthesis and signal transduction of plant hormone pathways were the most enriched pathways in 4274 differentially expressed genes (DEGs) and in 259 differential metabolites between Lan3 and 503. Moreover, the contents of three metabolites related to salicylic acid biosynthesis and the transcription levels of two DEGs related to salicylic acid signal transduction in Lan3 were higher than those in 503. Further verification in various accessions indicated that salicylic acid metabolism may be involved in the flowering regulation process of V. sativa. These findings provide valuable information for understanding the flowering mechanism and for promoting breeding research in V. sativa.

The genome of Cleistogenes songorica provides a blueprint for functional dissection of dimorphic flower differentiation and drought adaptability.

Cleistogenes songorica (2n = 4x = 40) is a desert grass with a unique dimorphic flowering mechanism and an ability to survive extreme drought. Little is known about the genetics underlying drought tolerance and its reproductive adaptability. Here, we sequenced and assembled a high-quality chromosome-level C. songorica genome (contig N50 = 21.28 Mb). Complete assemblies of all telomeres, and of ten chromosomes were derived. C. songorica underwent a recent tetraploidization (~19 million years ago) and four major chromosomal rearrangements. Expanded genes were significantly enriched in fatty acid elongation, phenylpropanoid biosynthesis, starch and sucrose metabolism, and circadian rhythm pathways. By comparative transcriptomic analysis we found that conserved drought tolerance related genes were expanded. Transcription of CsMYB genes was associated with differential development of chasmogamous and cleistogamous flowers, as well as drought tolerance. Furthermore, we found that regulation modules encompassing miRNA, transcription factors and target genes are involved in dimorphic flower development, validated by overexpression of CsAP2_9 and its targeted miR172 in rice. Our findings enable further understanding of the mechanisms of drought tolerance and flowering in C. songorica, and provide new insights into the adaptability of native grass species in evolution, along with potential resources for trait improvement in agronomically important species.

Elucidating the Molecular Mechanisms by which Seed-Borne Endophytic Fungi, Epichloë gansuensis, Increases the Tolerance of Achnatherum inebrians to NaCl Stress.

Seed-borne endophyte Epichloë gansuensis enhance NaCl tolerance in Achnatherum inebrians and increase its biomass. However, the molecular mechanism by which E. gansuensis increases the tolerance of host grasses to NaCl stress is unclear. Hence, we firstly explored the full-length transcriptome information of A. inebrians by PacBio RS II. In this work, we obtained 738,588 full-length non-chimeric reads, 36,105 transcript sequences and 27,202 complete CDSs from A. inebrians. We identified 3558 transcription factors (TFs), 15,945 simple sequence repeats and 963 long non-coding RNAs of A. inebrians. The present results show that 2464 and 1817 genes were differentially expressed by E. gansuensis in the leaves of E+ and E- plants at 0 mM and 200 mM NaCl concentrations, respectively. In addition, NaCl stress significantly regulated 4919 DEGs and 502 DEGs in the leaves of E+ and E- plants, respectively. Transcripts associated with photosynthesis, plant hormone signal transduction, amino acids metabolism, flavonoid biosynthetic process and WRKY TFs were differentially expressed by E. gansuensis; importantly, E. gansuensis up-regulated biology processes (brassinosteroid biosynthesis, oxidation-reduction, cellular calcium ion homeostasis, carotene biosynthesis, positive regulation of proteasomal ubiquitin-dependent protein catabolism and proanthocyanidin biosynthesis) of host grass under NaCl stress, which indicated an increase in the ability of host grasses' adaptation to NaCl stress. In conclusion, our study demonstrates the molecular mechanism for E. gansuensis to increase the tolerance to salt stress in the host, which provides a theoretical basis for the molecular breed to create salt-tolerant forage with endophytes.

Effects of cultivar and growing degree day accumulations on forage partitioning and nutritive value of common vetch (Vicia sativa L.) on the Tibetan plateau.

BACKGROUND: The use of common vetch in grassland-livestock systems has expanded greatly within recent years, partly because of its value as a high-quality forage crop but also to improve the soil nitrogen availability. In-field estimation of forage yield potential and nutritional characteristics is required for providing management decision to farmers on how to optimize the management and use of common vetch forages. The aim of this work was to study changes in forage partitioning and nutritive value responses of a late-maturing and an early maturing cultivar of common vetch in a two-year study on the Tibetan Plateau. RESULTS: This study provided evidence for differential patterns of forage accumulation for common vetch with contrasting maturity over 2 years. The late-maturing cultivar exhibited greater forage yield and a lower proportion of pods, compared to the early maturing cultivar. There was a tendency towards lower forage nutritive value with the late-maturing cultivar. Regressions of nutritive value parameters of common vetch forages on growing degree days were explained by the cubic (P < 0.001) models, all with high coefficients of determination (R2 ≥ 0.792). CONCLUSION: This study shows that the late-maturing cultivar harvested at end of the pod-filling stage produces high forage yield, increasing the availability of high-quality forage for ruminants, thereby improving the self-sufficiency of farmers, in terms of forage yield and high-concentration protein. For early maturing cultivars, it may be better to harvest at the early flowering stage for better nutritive value and in part to enable a subsequent double crop of oat. © 2020 Society of Chemical Industry.

Response of Plant Fungal Diseases to Beef Cattle Grazing Intensity in Hulunber Grassland.

The effects of grazing by large herbivores on biodiversity and ecosystem functioning have been extensively studied, whereas how grazing influence plant diseases, especially in natural grasslands, remains poorly understood. Therefore, we undertook a field study regarding a grazing trial in a temperate meadow steppe grassland to investigate mechanisms underlying grazing-host-pathogen interactions. The effects of cattle grazing at different grazing intensities of 0, 0.23, 0.34, 0.46, 0.69, and 0.92 AU/ha (where 1 AU= 500 kg of adult cattle) on the microenvironment, vegetation characteristics, and occurrence of diseases were evaluated. At the population level, the effects of grazing on grassland vegetation characteristics and disease varied with grassland plant species. Compared with nongrazing, grazing directly decreased the average density, coverage, and disease incidence of palatable and edible forages by 51.4, 62.4, and 82.4% in the 0.92 AU/ha treatment but increased the occurrence and prevalence of disease in remaining small herbs by 752.1%. At the community level, with the increase of grazing intensity, the pathogen load of the whole community in grassland was positively related to host coverage. In addition, there was a trend toward increased microtemperature and decreased microhumidity with increased grazing. Although occurrence of plant diseases in natural grasslands is influenced by a range of factors, comprehensive analysis highlighted the major role that cattle grazing intensity plays in the occurrence of plant diseases in natural grasslands. In addition to its direct effect, grazing also indirectly affects disease occurrence by shifting plant community structure and the microenvironment. However, direct effects of grazing intensity affected disease occurrence more than indirect effects.

Phytophthora cactorum as a Pathogen Associated with Root Rot on Alfalfa (Medicago sativa) in China.

Alfalfa (Medicago sativa) is the largest grown pasture crop in China due to its economic and ecological importance. During the summer season from June to August in 2018, stunted plants was frequently observed in alfalfa fields that have been established for two years in Jinchang, Gansu Province. The disease incidence of root rot ranged from 40% to 50%. Taproots of stunted plants showed red-brown to dark brown discolorations, and lateral roots were poorly developed. Shoots wilted with rotted taproots and lateral roots in severely affected plants. Twenty symptomatic plants were collected and transported to the laboratory for pathogen isolations. Roots were washed under running tap water, cut into 2 to 3 mm pieces (40 pieces each plant), and then sterilized in 75% ethanol for 2 mins followed by three times washing with autoclaved distilled water. Surface dried pieces on autoclaved filter papers were put onto water agar and also a Phytophthora selective medium P5ARP(H) (Jeffers and Martin 1986). The plates were incubated at 22°C for 3 to 5 days and then the growing hypha were subcultured onto potato dextrose agar (PDA). Thirty-two Phytophthora-like isolates were obtained and showed similar morphologies on PDA. Five isolates picked randomly were purified by single-hyphal-tip and plugs (4 to 5 mm) from PDA cultures were incubated in petri dishes with autoclaved distilled water at 22°C for 5 days. Sporangia, chlamydospores and oospores were examined. Sporangia were usually ovoid and sometimes appeared ellipsoid, with the length of 30.5-39.1 µm and width of 23.4-27.8 µm. The diameter of chlamydospores was 29.6 to 42.5µm. Oospores had a diameter of 23.6 to 30.2 µm. The isolates were tentatively identified as P. cactorum based on these morphology characteristics (Montealegre et al. 2016). DNA of these isolates were extracted and PCR amplifications of the rDNA ITS region and cytochrome oxidase subunit I (Cox I) (Kroon et al. 2004) were conducted. Sequences of these isolates were then compared with reference sequences in GenBank using BLAST search. The 866-bp ITS sequences had a sequence identity of 99% to 100% with P. cactorum (e.g. accession nos. EU662221, KJ128036). In addition, the 663-bp CoxI sequences showed 100% sequence identity with three P. cactorum isolates (accession nos. AB688156, HQ708234, EU660851). The ITS and CoxI sequences of one representative isolate Phy.c2 have been deposited in GenBank with the accession no. MT280033 and MT344138, respectively. Pathogenicity of the five isolates (Phy.c1-Phy.c5) were determined on two-week-old alfalfa seedlings (cv. Longdong) grown from seeds. Inoculums were prepared by subculturing agar plugs from edges of PDA cultures into the flask with autoclaved millet seeds, and incubated at 22°C in darkness for two weeks and shaken by hand every two days to ensure uniform colonization. Seedlings were transplanted into pots (12 cm x 12 cm) filled with autoclaved potting mix infested with millet-seed inoculum of each isolate at a rate of 0.5% (w/w). Control seedlings for comparison were transplanted into pots with uninfested potting mix. There were five seedlings per pot and twelve replicate pots for both inoculated and noninoculated treatments, and pots were kept under controlled environment room (22°C, 12 h photoperiod and 65% relative humidity) that were watered every two days to free draining. 87%~92% of the inoculated plants showed stunted symptoms with poorly developed and brown-discoloured roots three weeks after inoculation while the control plants were healthy with no root disease symptoms. To fulfil Koch's postulates, re-isolated cultures from discoloured root tissues were confirmed as the inoculated isolates by morphological examination and ITS sequencing. The five-purified isolates were submitted to the Grassland Culture Collection Center, Lanzhou University, with the accession nos. LZU-MsR-Phy.c1-Phy.c5. To our knowledge, this is the first report of P. cactorum as a pathogen of root rot on alfalfa in China. Phytophthora spp. has been reported causing root rot on alfalfa in America, Australia and Canada, and other legumes such as chickpea, and many other crops worldwide (Musial et al. 2005; Tan and Tan 1986; Vandemark and Barker 2003), and P. cactorum was reported as a root rot pathogen on lavender in China (Chen et al. 2017). P. cactorum may be a significant pathogen associated with root rot in major commercial alfalfa-producing areas in China where are based on flood-irrigation during the growth season.

Rhizophagus intraradices promotes alfalfa (Medicago sativa) defense against pea aphids (Acyrthosiphon pisum) revealed by RNA-Seq analysis.

Pea aphids (Acyrthosiphon pisum) are one of the most important insect pests of alfalfa (Medicago sativa). Arbuscular mycorrhizal (AM) fungi are important microorganisms of the agroecosystem that promote plant growth and improve plant resistance to abiotic and biotic stress. Little information is available on AM fungi-regulated defense responses of alfalfa to pea aphids. To better understand how alfalfa responds and to evaluate the impact of an AM fungus on aphid infestation, transcriptome sequencing was done and physiological parameters were analyzed. Our experiments showed that Rhizophagus intraradices can regulate plant response to aphids by promoting growth and increasing plant peroxidase (POD) and catalase (CAT) activities and salicylic acid (SA) concentration after aphid infestation. Transcriptome analysis showed that R. intraradices increased the expression of resistance-related genes, such as "WRKY transcription factor" and "Kunitz trypsin inhibitor." Additionally, GO terms "chitinase activity," "peroxidase activity," "defense response," and "response to biotic stimulus," and KEGG pathways "phenylpropanoid biosynthesis" and "phenylalanine metabolism" were significantly enriched in mycorrhizal fungus-inoculated plants and aphid-infested plants. These findings will improve our understanding about the impact of this AM fungus on alfalfa response to aphid feeding and will provide the basis for further research on plant defense against aphids.

Morphological fractions, chemical composition and in vitro digestibility of stover of four common vetch varieties grown on the Tibetan plateau.

This study evaluated the varietal variability and interrelationship of the grain and stover yields, and stover quality characteristics of four common vetch varieties (three improved varieties and one local variety) used on the Tibetan Plateau. Stover quality attributes determined included chemical composition, in vitro true digestibility of dry matter (IVTD) and neutral detergent fibre (dNDF). The improved varieties were superior to the local variety in grain yield, harvest index and potential utility index (PUI). The wide ranges in the proportions of leaf (24.3%-34.6%), pod (22.0%-45.6%) and stem (29.5%-44.7%) fractions were observed. Significant varietal differences (p < .001) were observed for the whole-stover content of crude protein [CP; 78.1-101 g/kg dry matter (DM)], neutral detergent fibre (aNDF; 520-564 g/kg DM), acid detergent fibre (ADF; 520-564 g/kg DM), IVTD (597-658 g/kg DM) and dNDF (305-384 g/kg aNDF). The leaf fraction had the highest levels of CP, IVTD and dNDF, while the highest contents of aNDF and ADF were observed in the stem fraction. Canonical correlation analysis showed that significant (p < .001) correlations between the quality attributes of whole stover and proportions and quality attributes of stover morphological fractions. Pearson correlation between the grain and stover yield was negative and significant (r = -.295; p = .001), while the correlations between the grain/stover yield and stover quality traits were positive or non-significant. Ranking of the varieties differed when grain yield, PUI and stover quality scores were investigated. Of the varieties tested, Lanjian No.2 has the best potential for use as a ruminant feed, as indicated by PUI and stover quality.

Role of Epichloë Endophytes in Defense Responses of Cool-Season Grasses to Pathogens: A Review.

Various cool-season grasses are infected by Epichloë endophyte, and this symbiotic relationship is always of benefit to the host grass due to an increased resistance to abiotic and biotic stresses. Fungal diseases adversely affect the yield, quality, and economic benefits of rangelands, which affects the production of animal husbandry. Therefore, it is imperative to breed resistant cultivars and to better understand the role of fungal endophytes in order to protect grasses against pathogens. The present review introduces research regarding how these endophytes affect the growth of pathogens in vitro and how they change the resistance of host plants to plant diseases. From the perspective of physical defense, changes in physiological indexes, and secretion of chemical compounds, we summarize the potential mechanisms by which endophytes are able to enhance the disease resistance of a host grass. Through these, we aim to establish a solid theoretical foundation for plant disease control and disease resistance breeding by application of fungal endophytes. A broader understanding of fungal endophyte effects on hosts could create a new opportunity for managing or introducing fungal symbioses in both agronomic or non-agronomic ecosystems.

Multiple Regulatory Networks Are Activated during Cold Stress in Medicago sativa L.

Cultivated alfalfa (Medicago sativa L.) is one of the most important perennial legume forages in the world, and it has considerable potential as a valuable forage crop for livestock. However, the molecular mechanisms underlying alfalfa responses to cold stress are largely unknown. In this study, the transcriptome changes in alfalfa under cold stress at 4 °C for 2, 6, 24, and 48 h (three replicates for each time point) were analyzed using the high-throughput sequencing platform, BGISEQ-500, resulting in the identification of 50,809 annotated unigenes and 5283 differentially expressed genes (DEGs). Metabolic pathway enrichment analysis demonstrated that the DEGs were involved in carbohydrate metabolism, photosynthesis, plant hormone signal transduction, and the biosynthesis of amino acids. Moreover, the physiological changes of glutathione and proline content, catalase, and peroxidase activity were in accordance with dynamic transcript profiles of the relevant genes. Additionally, some transcription factors might play important roles in the alfalfa response to cold stress, as determined by the expression pattern of the related genes during 48 h of cold stress treatment. These findings provide valuable information for identifying and characterizing important components in the cold signaling network in alfalfa and enhancing the understanding of the molecular mechanisms underlying alfalfa responses to cold stress.

Effects of feeding drunken horse grass infected with Epichloë gansuensis endophyte on animal performance, clinical symptoms and physiological parameters in sheep.

BACKGROUND: Many reports showed that grass-endophyte symbiosis induced livestock poisoned. Yet, there is no study evaluating clinical symptoms and physiological parameters in sheep fed Epichloë gansuensis endophyte-infected grass. The objective of the present study was to investigate these indexes by feeding sheep with endophyte-infected A. inebrians (E+ Group) or endophyte-free A. inebrians (E- Group) drunken horse grass or alfalfa hay (Control Group). RESULTS: The Epichloë endophyte caused obvious toxicity symptoms in the sheep fed E+ A. inebrians, with 1 of the 5 sheep having died by the 35th day. The feed intake and body weight gain of the E+ Group were significantly less than the E- and control groups (P < 0.05). Serum concentrations of alanine aminotransferase (ALT, 45.5 mmol/L) and aspartate aminotransferase for the E+ group (AST, 139.3 mmol/L) were significantly (P < 0.05) greater than for the E- (ALT, 31.2 mmol/L; AST, 78.6 mmol/L) and control (ALT, 32.6 mmol/L; AST, 56.6 mmol/L) groups at the fifth week; serum concentration of creatinine for the E+ group (63.8 mmol/L) was also significantly (P < 0.05) greater than for E- (56.6 mmol/L) and control groups (58.5 mmol/L). Meanwhile, urine biochemical indices for the E+ group indicated that ketone and occult blood were significantly (P < 0.05) elevated compared to the other groups while urine pH values were significantly (P < 0.05) acidic. The relative weight of heart, brain, liver, lung and kidney for Group E+ were almost two fold more than the other groups, but uterus weight was about half that found for Group E- or Control. CONCLUSIONS: We conclude that the Epichloë endophyte infection is the cause of A. inebrians toxicity to sheep. Interestingly, none of the measured parameters differed significantly between E- and the control groups, which implied that drunken horse grass could be utilized efficiently by sheep when not infected by the Epichloë endophyte.

Highly oxygenated triterpenoids from the roots of Schisandra chinensis and their anti-inflammatory activities.

A new highly oxygenated triterpenoid, schinchinenlactone D (1), and three known compounds (2-4) were isolated from the roots of Schisandra chinensis. Their structures were determined by combining the spectroscopic analysis with the theoretical computations. The anti-inflammatory activities of compounds 1-4 were evaluated, and compound 3 exhibits the most significant activity in the inhibition of NO production with an IC50 value of 10.6 µM.

Genome-Wide Analysis of Codon Usage Bias in Epichloë festucae.

Analysis of codon usage data has both practical and theoretical applications in understanding the basics of molecular biology. Differences in codon usage patterns among genes reflect variations in local base compositional biases and the intensity of natural selection. Recently, there have been several reports related to codon usage in fungi, but little is known about codon usage bias in Epichloë endophytes. The present study aimed to assess codon usage patterns and biases in 4870 sequences from Epichloë festucae, which may be helpful in revealing the constraint factors such as mutation or selection pressure and improving the bioreactor on the cloning, expression, and characterization of some special genes. The GC content with 56.41% is higher than the AT content (43.59%) in E. festucae. The results of neutrality and effective number of codons plot analyses showed that both mutational bias and natural selection play roles in shaping codon usage in this species. We found that gene length is strongly correlated with codon usage and may contribute to the codon usage patterns observed in genes. Nucleotide composition and gene expression levels also shape codon usage bias in E. festucae. E. festucae exhibits codon usage bias based on the relative synonymous codon usage (RSCU) values of 61 sense codons, with 25 codons showing an RSCU larger than 1. In addition, we identified 27 optimal codons that end in a G or C.