Cool-season Grasses Research Articles

Ontogenetic and trans-generational dynamics of a vertically transmitted fungal symbiont in an annual host plant in ozone-polluted settings.

Tropospheric ozone is an abiotic stress of increasing importance in the context of global climate change. This greenhouse gas is a potent phytotoxic molecule with demonstrated negative effects on crop yield and natural ecosystems. Recently, oxidative stress has been proposed as a mechanism that could regulate the interaction between cool-season grasses and Epichloë endophytes. We hypothesized that exposure of Lolium multiflorum plants, hosting endophytes to an ozone-polluted environment at different ontogenetic phases, would impact the trans-generational dynamics of the vertically transmitted fungal symbiont. Here, we found that the ozone-induced stress on the mother plants did not affect the endophyte vertical transmission but it impaired the persistence of the fungus in the seed exposed to artificial ageing. Endophyte longevity in seed was reduced by exposure of the mother plant to ozone. Although ozone exposure did not influence either the endophyte mycelial concentration or their compound defences (loline alkaloids), a positive correlation was observed between host fitness and the concentration of endophyte-derived defence compounds. This suggests that fungal defences in grass seeds were not all produced in situ but remobilized from the vegetative tissues. Our study reveals ozone trans-generational effects on the persistence of a beneficial symbiont in a host grass.

Yield, persistence, forage nutritive value, and preference of perennial ryegrass under grazing

AbstractPerennial ryegrass (PR) is a quality and high yielding forage that tends to lack winter hardiness to persist in the upper Midwest. The objective was to evaluate yield, stand persistence, forage nutritive value, and preference of experimental populations of winter hardy PR grown in monoculture and in mixture with white clover under livestock grazing. Research was conducted in St. Paul and Grand Rapids, MN. Meadow fescue (MF), tall fescue (TF), orchardgrass (OG), a reference PR, and two experimental populations of PR were grazed every 32 days by either horses (St. Paul) or cow‐calf pairs (Grand Rapids). Prior to grazing, forage yield and quality samples were collected, and post‐grazing, fields were visually assessed for the percentage of forage removal to determine preference. Yields of all PR (6.9 to 7.6 Mg ha−1) were similar, but lower compared to other cool‐season grasses (CSG). Persistence was similar among all CSG in St. Paul; however, it was lower for all PR in Grand Rapids compared to other CSG. All PR were highly preferred by both horses (≥74% removal) and cattle (≥70% removal) and were consistently among the highest for crude protein (CP; 200 g kg−1), equine digestible energy (DE; 2.35 Mcal kg−1) and bovine metabolizable energy (ME; 2.6 Mcal kg−1), moderate for nonstructural carbohydrates (NSC; 136 g kg−1), and among the lowest for neutral detergent fiber (NDF; 518 g kg−1). Experimental populations of PR produced forage of high nutritive value, but yield and stand persistence may be less compared to other CSG at some locations.

The intensity of water stress dictates whether the N status of temperate-type perennial grass swards is affected by drought

Abstract Temperate-type, summer-active cool-season perennial grasses are frequently exposed to dry spells during the warm season due to the high atmospheric demand and fast soil desiccation. Since water availability is essential for sward nitrogen (N) uptake and most mineral N is located in the topsoil, water stress may hinder sward N nutrition if it reduces N supply. In hydro-halomorphic soils, imbalances between soil N supply and sward N demand during summer could be further aggravated by, first, the limited water holding capacity of the topsoil, and second, potential availability of water from deep, N-poor soil horizons that are kept moist by a shallow water table. The aim of this work was to quantify the effect of N and water availability on N status and productivity of temperate-type perennial swards growing in a hydro-halomorphic soil of the Flooding Pampa. During two consecutive years, shoot and root dynamics, as well as radiation capture, N nutrition index (NNI), soil water availability and leaf water potential were followed over six weeks long mid-summer regrowths of old stands (>9 years) of temperate-type tall fescue [Lolium arundinaceum (Schreb.) Darbysh.] sown on a typical Natraqualf soil in Balcarce, Argentina. Swards were either rainfed or irrigated and fertilized with either 40 or 200 kg de N ha−1. Root systems were large (5.8–9.3 Mg DM roots ha−1), but concentrated mostly in the 0.2 m topsoil (66%–75% of root biomass or length). Despite invariably high available water below 0.4 m throughout both experimental periods, rainfed swards faced transient episodes of water stress whenever the upper soil layer dried out. Tall fescue swards were strongly co-limited by water and N during summer, showing comparable, very high responses to both N addition and irrigation. Further, forage yield showed synergistic increases when swards received both supplemental water and N simultaneously. This was mainly due to consistent synergistic responses in radiation use efficiency and leaf lifespan. Sward NNI was depressed only by an historically severe drought but was not altered when water stress intensity was lower, close to average for the season in the area. Therefore, the major causes of typically low summer productivity of temperate-type tall fescue growing in hydro-halomorphic soils of the Flooding Pampa are the intrinsic effects of low N and low water availability, with only uncommonly extreme water stress episodes reducing yield indirectly through an aggravation of crop N status.

Evaluating winter annual grass control and native species establishment following applications of indaziflam on rangeland

AbstractIndaziflam, a PRE herbicide option for weed management on rangeland and natural areas, provides long-term control of invasive winter annual grasses (IWAGs). Because indaziflam only provides PRE control of IWAGs, POST herbicides such as glyphosate can be mixed with indaziflam to control germinated IWAG seedlings. Field trials were conducted at three sites on the Colorado Front Range to evaluate glyphosate dose required to provide adequate POST IWAG control and compare long-term downy brome (Bromus tectorum L.), Japanese brome (Bromus arvensis L.), and feral rye (Secale cereale L.) control with indaziflam and imazapic. Two of the three sites were void of desirable species, so species establishment through drill seeding was assessed, while the remnant native plant response was assessed at the third site. Herbicide applications were made March 2014 through April 2015, and two sites were drill seeded with native species 9 mo after herbicide application. Yearly visual control evaluations, biomass of all plant species, and drilled species stand counts were collected. Glyphosate at 474 g ae ha−1 reduced B. tectorum biomass to zero, while glyphosate at 631 g ae ha−1 was needed to reduce biomass to near zero at the S. cereale site. At all three sites, only indaziflam treatments had significant reductions in IWAG biomass compared with the nontreated check at 3 yr after treatment (YAT). By 3 YAT in the drill-seeded sites, cool-season grass frequency ranged from 37% to 69% within indaziflam treatments (73 and 102 g ai ha−1), while imazapic treatments ranged from 0% to 26% cool-season grass frequency. In the site with a remnant native plant community, indaziflam treatments resulted in a 3- to 4-fold increase in native grass biomass. These results indicate that the multiyear IWAG control provided by indaziflam can aid in desirable species reestablishment through drill seeding or response of the remnant plant community.

SMRT and Illumina RNA sequencing reveal novel insights into the heat stress response and crosstalk with leaf senescence in tall fescue

BackgroundAs a cool-season grass species, tall fescue (Festuca arundinacea) is challenged by increasing temperatures. Heat acclimation or activation of leaf senescence, are two main strategies when tall fescue is exposed to heat stress (HS). However, lacking a genome sequence, the complexity of hexaploidy nature, and the short read of second-generation sequencing hinder a comprehensive understanding of the mechanism. This study aims to characterize the molecular mechanism of heat adaptation and heat-induced senescence at transcriptional and post-transcriptional levels.ResultsTranscriptome of heat-treated (1 h and 72 h) and senescent leaves of tall fescue were generated by combining single-molecular real-time and Illumina sequencing. In total, 4076; 6917, and 11,918 differentially expressed genes (DEGs) were induced by short- and long-term heat stress (HS), and senescence, respectively. Venn and bioinformatics analyses of DEGs showed that short-term HS strongly activated heat shock proteins (Hsps) and heat shock factors (Hsfs), as well as specifically activated FK506-binding proteins (FKBPs), calcium signaling genes, glutathione S-transferase genes, photosynthesis-related genes, and phytohormone signaling genes. By contrast, long-term HS shared most of DEGs with senescence, including the up-regulated chlorophyll catabolic genes, phytohormone synthesis/degradation genes, stress-related genes, and NACs, and the down-regulated photosynthesis-related genes, FKBPs, and catalases. Subsequently, transient overexpression in tobacco showed that FaHsfA2a (up-regulated specifically by short-term HS) reduced cell membrane damages caused by HS, but FaNAC029 and FaNAM-B1 (up-regulated by long-term HS and senescence) increased the damages. Besides, alternative splicing was widely observed in HS and senescence responsive genes, including Hsps, Hsfs, and phytohormone signaling/synthesis genes.ConclusionsThe short-term HS can stimulate gene responses and improve thermotolerance, but long-term HS is a damage and may accelerate leaf senescence. These results contribute to our understanding of the molecular mechanism underlying heat adaptation and heat-induced senescence.

A nuclear protein NsiA from Epichloë festucae interacts with a MAP kinase MpkB and regulates the expression of genes required for symbiotic infection and hyphal cell fusion.

The endophytic fungus Epichloë festucae systemically colonizes the intercellular spaces of cool-season grasses to establish a mutualistic symbiosis. Hyphal growth of the endophyte within the host plant is tightly regulated and synchronized with the growth of the host plant. A genetic screen to identify symbiotic genes identified mutant FR405 that had an antagonistic interaction with the host plant. Perennial ryegrass infected with the FR405 mutant were stunted and underwent premature senescence and death. The disrupted gene in FR405 encodes a nuclear-localized protein, designated as NsiA for nuclear protein for symbiotic infection. Like previously isolated symbiotic mutants the nsiA mutant is defective in hyphal cell fusion. NsiA interacts with Ste12, a C2H2 zinc-finger transcription factor, and a MAP kinase MpkB. Both are known as essential components for cell fusion in other fungal species. In E. festucae, MpkB, but not Ste12, is essential for cell fusion. Expression of several genes required for cell fusion and symbiosis, including proA/adv-1, pro41/ham-6, ham7, ham8, and ham9 were downregulated in the nsiA mutant. However, the NsiA ortholog in Neurospora crassa was not essential for hyphal cell fusion. These results demonstrate that the roles of NsiA and Ste12 orthologs in hyphal cell fusion are distinctive between fungal species.

Long-Term Yield and Quality Performance of Perennial Energy Grasses (Agropyron spp.) on Marginal Land

The cultivation of perennial grasses is one of the most desirable alternatives as energy feedstock, but it is difficult to achieve competitive yields under Mediterranean marginal conditions. The aim of this study was to assess the performance of three cool-season grasses (Agropyron spp.) from an agronomic and energetic point of view by comparing the dry matter (DM) yields, rain use efficiency, chemical composition, and biomass quality over an eight-year period in Spain under marginal rainfed conditions. The tall wheatgrass (Agropyron elongatum (Host) Beauv.) cultivars, Alkar (4.8 Mg DM·ha−1) and Jose (4.7 Mg DM·ha−1), achieved the highest yields. Productions below 0.5 Mg DM·ha−1 were obtained when rainfall was lower than 150 mm between March and June. The biomass obtained from the tested grasses showed relatively high contents of ash, silicon, and alkali elements. Net calorific values ranged between 16.7 and 18.5 MJ·kg−1 db. Differences in the composition among species and cultivars are not likely to affect their combustion behavior from a practical point of view. The ash content, as well as the concentrations of K, S, Na, and Cl, tended to decrease over the years. The results offered would be very useful for the implementation of this type of crop in marginal land.

Soil nitrate leaching under grazed cool-season grass pastures of the North Central US.

Nitrate leaching from agricultural cropping systems contributes to widespread and devastating eutrophication of water bodies globally. In the North Central USA, this problem is acute, with millions of dollars spent annually in efforts to clean up recreational and drinking water. The frequent soil disturbance and exogenous nitrogen (N) amendments applied in annual cropping systems make them major sources of ground- and surface-water nitrate pollution. Perennial grasslands under managed livestock grazing have been touted for their ability to retain soils and nutrients while simultaneously providing milk and meat to society. The present study provides an evaluation of the peer-reviewed literature addressing nitrate leaching loads beneath corn, pasture and prairie in temperate humid and sub-humid regions of the US, with a focus on cool-season grass pastures. Inputs of exogenous N to these agroecosystems comes from wet and dry deposition, livestock manure from imported feed, biological fixation and inorganic N fertilizer. Nitrate loads were highest beneath corn and lowest beneath restored prairie and switchgrass managed for bioenergy. Cool-season grass pastures had relatively low levels of nitrate leaching loads where little or no N was applied. However, where grazed perennial grasslands had inorganic N applied, nitrate leaching loads rivaled those of corn in some cases. When producing milk and meat from livestock, grazed perennial cool-season grass pastures should reduce nitrate leaching loads compared to growing corn that is used to feed livestock in confinement. However, cool-season grass pastures can lose significant nitrate to leaching with moderate- to high-levels of exogenous N inputs. © 2020 Society of Chemical Industry.

Cover Crop Performance between Plastic-mulched Beds: Impacts on Weeds and Soil Resources

Growing a cover crop living mulch between plastic-mulched beds may reduce soil erosion while providing other agroecosystem services. However, information regarding the relative differences among living mulch species to maximize services and minimize competition for nutrients and water in adjacent plastic-mulched beds is limited. A 2-year experiment in Michigan evaluated nine living mulch species for biomass production, in-season weed suppression, and potential for cash crop competition. Species included three warm season grasses {Italian ryegrass [Lolium perenne L. ssp. multiflorum (Lam.) Husnot], teff [Eragrostis tef (Zuccagni) Trotter, and sudangrass [Sorghum bicolor (L.) Moench ssp. drummondii (Nees ex Steud.) de Wet & Harlan]}; three cool season grasses [barley (Hordeum vulgare L.), rye (Secale cereale L.), and wheat (Triticum aestivum L.)]; and three clover species grown in combination with rye {Dutch white clover (Trifolium repens L.), New Zealand white clover (T. repens L.) and yellow blossom sweet clover [Melilotus officinalis (L.) Lam.]}. Although all living mulch treatments significantly reduced in-season weed biomass relative to the weedy control in 2018, weeds were generally a dominant component of total biomass in all living mulch treatments other than teff. Weed biomass was negatively correlated with living mulch biomass, and teff exhibited both the greatest biomass and weed suppression in both years. However, despite spatial and physical separation, all living mulches demonstrated the potential to compete with a cash crop by reducing soil inorganic nitrogen and moisture levels in adjacent plastic mulch–covered beds. Growers interested in integrating living mulches into plasticulture systems must consider desired benefits such as enhanced weed suppression, soil quality, and harvesting conditions alongside potential risks to cash crop yields.

Development of SCAR markers related to heat tolerance in Kentucky bluegrass

As a high-quality cool-season grass, Kentucky bluegrass (Poa pratensis) is facing increasing threat of high temperature, so improving its heat tolerance (HT) has become an important breeding target. In this study, the HT of 84 materials was identified in the artificial climate chamber, and 15 most heat-tolerant and 15 most heat-sensitive materials were selected respectively to construct two DNA pools. There was a significant difference in high-temperature tolerance time between the plants in the two pools, which was more than 22 days. A total of 304 sequence-related amplified polymorphism (SRAP) and 88 simple sequence repeat (SSR) markers were used to screen the polymorphic bands between the two pools. Then, these bands were transformed into sequence characterized amplified region (SCAR) markers, and finally 12 SCAR dominant markers related to HT were obtained, which could detect the heat-sensitive materials efficiently. Among them, S-me8×em2 and S-me52×em5 had the best identification effect, and the consistency between the absence of these two markers and the heat-sensitive phenotype was 87%. According to the comparison with NCBI database, the sequences of 12 SCAR markers had no homology with known HT related genes. Next, we would further verify the HT identification efficiency of these SCAR markers in single plants within materials, and try to use them in molecular marker-assisted breeding.

Winter Annual Rye Seeding Date Influence on Nitrogen Recovery and Ammonia Volatilization from Late Fall Surface-Applied Manure

Dairy farmers in the northeast face challenges in the application of manure in fall and on-time planting of cool-season grasses to maximize recovery of residual N and nutrients released from fall applied manure. Ammonia emission from animal manure is a serious environmental concern and can be reduced if cover crop is integrated in the farming system. On-time planting of cover crops can reduce ammonia volatilization from fall, surface-applied manure, and prevents N loss to leaching. A two-year study was conducted in 2015 and 2016 to investigate if time of planting of winter annual rye (Secale cereale L.) along with late fall application of manure when air temperature is low can influence ammonia emission and preserve nitrogen (N) to meet the N requirement of forage rape. Three planting dates (16 September, 30 September, and 14 October) of rye cover crop with two manure application treatments including late-fall application and no manure were assessed for mitigating ammonia volatilization, and also yield and recovery of N by forage rape (Brassica napus L.). The highest rates of ammonia volatilization were detected in the first 24 hours after manure spreading regardless of the treatment. The result indicated that cover crop use significantly limited volatilization compared with no cover crop. The earliest planting date produced 3823 kg ha−1 dry matter of winter rye cover crop that was 16 and 35 percent higher than second and third dates of planting, respectively. The manured cover crop accumulated 132 kg N ha−1 when planted early. However, biomass yield of forage rape was more when planted after all cover crop treatments with manure application. Prior to forage planting, the nitrate-N content in all three soil depths (0–20, 20–40, and 40–60 cm) in the plots with manure was higher than plots with no manure. No significant differences in forage rape yield was detected among winter rye planting dates; however, forage rape planted after winter rye was higher than after no-cover crop. The results of this study suggest that when immediate incorporation of manure into soil is not feasible, establishing cover crop early and then applying manure in the late fall, is a practical management to limit nonpoint source pollution from ammonia loss.

Protocol for Agrobacterium-mediated transformation of tall fescue and future perspective on the application of genome editing.

Tall fescue (Festuca arundinacea Schreb.) is a major cool-season perennial grass grown for forage and turf. We have obtained transgenic tall fescue by Agrobacterium-mediated transformation to improve agronomically important traits. In our protocol, we use embryogenic calli derived from not only mature seeds but also shoot tips. Although tall fescue cultivars consist of various genotypes with different genetic variation, we can produce transgenic plants at any time with calli induced from shoot tips of in vitro-maintained responsive genotypes. When the hygromycin phosphotransferase gene is used as a selectable marker, transformants are selected by incubation with 100 mg l-1 hygromycin in both selection and regeneration media. Since tall fescue is an anemophilous species, the cultivation of transgenic plants poses the risk of transgenic pollen flow. Recently, it has been reported that genome-edited plants without the integration of foreign DNA fragments can be produced by an Agrobacterium-mediated transient gene expression system. We hope that our protocol will contribute to production of transgene-free genome-edited tall fescue.

Role of nitrogen and herbicides in integrated management of mugwort (Artemisia vulgaris) in cool-season forage grasses

AbstractMugwort (Artemisia vulgaris L.) is becoming increasingly problematic in cool-season pastures and grasslands. A 3-yr field experiment evaluated different rates of nitrogen and herbicides for A. vulgaris management in a permanent grassland. The main plot had three nitrogen rates, 0, 62, and 124 kg N ha−1; the subplot had three herbicides, aminopyralid, clopyralid, and glyphosate; and the sub-subplot had three herbicide rates, aminopyralid (61, 122, and 244 g ae ha−1), clopyralid (140, 280, and 560 g ae ha−1), and glyphosate (552, 1,104, and 2,208 g ae ha−1). Results revealed that nitrogen had no effect on A. vulgaris control, rhizome biomass, and stem density. However, cool-season grass biomass was the highest (7,126 kg ha−1) in the plots that received 124 kg N ha−1 and 244 g ae ha−1 of aminopyralid. Only glyphosate caused grass injury, which varied from 65% to 100% depending upon application rate. By 9 mo after initial herbicide treatment (MAIT), A. vulgaris was controlled 60% to 98% with aminopyralid at ≥61 g ae ha−1 or glyphosate at ≥552 g ae ha−1. By 21 MAIT, aminopyralid at ≥122 g ae ha−1 or glyphosate at ≥1,104 g ae ha−1 resulted in >95% reduction in A. vulgaris stem density and rhizome biomass and provided ≥98% visual control. By 33 MAIT, complete control of A. vulgaris was confirmed in plots treated with aminopyralid at ≥122 g ae ha−1 or glyphosate at ≥1,104 g ae ha−1. Clopyralid was not effective; A. vulgaris control was <40% even after three annual applications at 560 g ae ha−1. Results indicate that integration of nitrogen fertilization with aminopyralid did not improve A. vulgaris control, but was advantageous in enhancing cool-season grass productivity.

Comparison of cool and warm season perennial grasses for biomass yield, quality, and energy balance in two contrasting semiarid environments

Perennial grasses are valuable biomass feedstocks due to their potentially high biomass yield and environmental benefits. This study aimed to compare 7 cool season: tall wheatgrass, bulbous canary grass, reed canary grass, tall fescue, perennial ryegrass, orchard grass, and smooth brome grass and 3 warm season perennial grasses: miscanthus, switchgrass, and giant reed for biomass yield, quality, and energy balance at 2 harvesting periods (autumn and winter to early spring) over 3 growing seasons in cold (Cankiri, Turkey) and warm (Adana, Turkey) semiarid environments. Harvests in winter and spring both led to considerable yield losses with all of the grass species in both locations, while it promoted biomass quality by reducing the moisture and ash contents of the warm season grasses in both locations, and the cool season grasses only in Cankiri. In contrast, autumn harvest provided significantly lower moisture and ash contents in the cool season grasses grown in Adana over the 3 years, where it was observed that, on average, giant reed exhibited the highest biomass productivity in both locations. Furthermore, bulbous canary grass and tall wheatgrass were found to be the most productive species in the study among the cool season grasses. Additionally, bulbous canary grass in Adana and tall wheatgrass in Cankiri achieved considerably higher energy ratios than the warm season grasses, mainly as a result of a lack of energy input for irrigation. In this sense, these 2 cool season grasses may be considered as alternative bioenergy crops in semiarid environments for sustainable biomass production.

Molecular mechanisms in grass-Epichloë interactions: towards endophyte driven farming to improve plant fitness and immunity.

All plants harbor many microbial species including bacteria and fungi in their tissues. The interactions between the plant and these microbes could be symbiotic, mutualistic, parasitic or commensalistic. Mutualistic microorganisms are endophytic in nature and are known to play a role in plant growth, development and fitness. Endophytes display complex diversity depending upon the agro-climatic conditions and this diversity could be exploited for crop improvement and sustainable agriculture. Plant-endophyte partnerships are highly specific, several genetic and molecular cascades play a key role in colonization of endophytes in host plants leading to rapid changes in host and endophyte metabolism. This results in the accumulation of secondary metabolites, which play an important role in plant defense against biotic and abiotic stress conditions. Alkaloids are one of the important class of metabolites produced by Epichloë genus and other related classes of endophytes and confer protection against insect and mammalian herbivory. In this context, this review discusses the evolutionary aspects of the Epichloë genus along with key molecular mechanisms determining the lifestyle of Epichloë endophytes in host system. Novel hypothesis is proposed to outline the initial cellular signaling events during colonization of Epichloë in cool season grasses. Complex clustering of alkaloid biosynthetic genes and molecular mechanisms involved in the production of alkaloids have been elaborated in detail. The natural defense and advantages of the endophyte derived metabolites have also been extensively discussed. Finally, this review highlights the importance of endophyte-arbitrated plant immunity to develop novel approaches for eco-friendly agriculture.

Role of Epichloë Endophytes in Improving Host Grass Resistance Ability and Soil Properties.

The past decade has witnessed significant advances in understanding the interaction between grasses and systemic fungal endophytes of the genus Epichloë, with evidence that plants have evolved multiple strategies to cope with abiotic stresses by reprogramming physiological responses. Soil nutrients directly affect plant growth, while soil microbes are also closely connected to plant growth and health. Epichloë endophytes could affect soil fertility by modifying soil nutrient contents and soil microbial diversity. Therefore, we analyze recent advances in our understanding of the role of Epichloë endophytes under the various abiotic stresses and the role of grass-Epichloë symbiosis on soil fertility. Various cool-season grasses are infected by Epichloë species, which contribute to health, growth, persistence, and seed survival of host grasses by regulating key systems, including photosynthesis, osmotic regulation, and antioxidants and activity of key enzymes of host physiology processes under abiotic stresses. The Epichloë endophyte offers significant prospects to magnify the crop yield, plant resistance, and food safety in ecological systems by modulating soil physiochemical properties and soil microbes. The enhancing resistance of host grasses to abiotic stresses by an Epichloë endophyte is a complex manifestation of different physiological and biochemical events through regulating soil properties and soil microbes by the fungal endophyte. The Epichloë-mediated mechanisms underlying regulation of abiotic stress responses are involved in osmotic adjustment, antioxidant machinery, photosynthetic system, and activity of key enzymes critical in developing plant adaptation strategies to abiotic stress. Therefore, the Epichloë endophytes are an attractive choice in increasing resistance of plants to abiotic stresses and are also a good candidate for improving soil fertility and regulating microbial diversity to improve plant growth.

Long-term nitrate removal in three riparian buffers: 21 years of data from the Bear Creek watershed in central Iowa, USA

Riparian buffers are a conservation practice that increases vegetation diversity on the agricultural landscape while providing environmental benefits. This study specifically focused on the ability of riparian buffers to remove nitrate from shallow groundwater. There are many studies that assessed nitrate removal within buffers, but not many have a long-term, continuous data set that can analyze for variation in nitrate removal rates over time. Here we report on 21 years of nitrate well data, from 1996 through 2017, for three buffers in the Bear Creek watershed in central Iowa. These buffers are named using abbreviations to help keep landowners anonymous (e.g. RN, RS, and ST). Studied buffers RS and ST showed greater nitrate reduction (or removal) after 10 and 6 years of its establishment, respectively. Buffer RN did not experience a significant nitrate removal increase with time, but instead had higher nitrate removal rates when compared to buffers RS and ST of 10.3 g NO3−-N m−1 day−1 from the start of this study. From this data, we suggest that past land management played a major role in the responses observed. RN had previously been established in cool-season grasses for grazing before being converted to a buffer, while RS and ST had been managed in a corn and soybean rotation. RN was thought to have higher denitrification immediately with increased labile soil carbon input and enhanced soil aggregation due to the grassland perennials, while buffer vegetation establishment increased soil carbon inputs and soil aggregation over time for RS and ST. These nitrate removal trends would not have been observed without access to long-term, continuous data. This study highlighted the importance of long-term data sets and the need to assess conservation practices over time to determine their longevity and efficiency with time.

Comparative analysis of the skim milk and milk fat globule membrane proteomes produced by Jersey cows grazing pastures with different plant species diversity

The objective of this experiment was to identify and characterize the bovine milk proteome within the skim milk fraction and milk fat globule membrane (MFGM)-associated fraction from 16 organically certified lactating Jersey cows after a short term of grazing pastures with or without annual forage crops (AFC). Cows were offered a partial mixed ration (~60% of dry matter intake) and approximately 40% of their total dry matter intake as herbage. Eight cows were offered a cool-season grass–legume herbage (GLH), which included orchardgrass (Dactylis glomerata), timothy (Phleum pratense), Kentucky bluegrass (Poa pratensis), and white clover (Trifolium repens). The other 8 cows were offered the same GLH strip-tilled with the AFC, including oat (Avena sativa), millet (Pennisetum glaucum), teff (Eragrostis tef), buckwheat (Fagopyrum esculentum), and chickling vetch (Lathyrus sativus). Milk samples were collected from each cow during a.m. and p.m. milkings on d 19 to 21 of grazing, and composite milk samples per cow were analyzed for (1) the high-abundance milk protein profile, (2) the skim milk low-abundance protein-enriched proteome, and (3) the MFGM proteome. Of the 443 proteins identified in the skim and MFGM proteomes, 433 were included in statistical analysis, including 68 proteins identified in the skim milk fraction and 365 in the MFGM-associated fraction. Analysis of the skim and MFGM proteomes encompassed unique gene ontology profiles and proportions of functional classifications. In response to diet, αS1-casein as well as 8 low-abundance proteins were present in higher concentration or abundance in milk from cows grazing the GLH strip-tilled with the AFC compared with milk from cows grazing GLH, suggesting that even short-term grazing of pastures including some AFC may affect the milk proteome.

Transcriptome analysis on responses of orchardgrass (Dactylis glomerata L.) leaves to a short term flooding

BackgroundOrchardgrass (Dactylis glomerata L.) is a popular cool-season perennial grass with a high production value, and orchardgrass seed is the fourth top-selling forage grass seed in the world. However, its yield and quality are often affected by flooding. To date, the molecular responses of orchardgrass to flooding were poorly understood.ResultsHere, we performed mRNA-seq to explore the transcriptomic responses of orchardgrass to a short term flooding (8 h and 24 h). There were 1454 and 565 differentially expressed genes identified in the 8 h and 24 h of flooding, respectively, compared to well control. GO functional enrichment analysis showed that oxidoreductase activity and oxidation-reduction process were highly present, suggesting that flooding induced the response to oxygen stress. Pathways enrichment analysis highlights the importance of glutathione metabolism, peroxidase, glycolysis and plant hormone signal transduction in response to flooding acclimation. Besides, the ROS clearance system is activated by significantly expressed glutathione S-transferase and genes encoding SOD and CAT (CAT1 and CDS2). The significant positive correlation between RNA sequencing data and a qPCR analysis indicated that the identified genes were credible.ConclusionIn the process of orchardgrass response to flooding stress, multiple differential genes and biological processes have participated in its acclimation to flooding, especially the biological processes involved in the removal of ROS. These results provide a basis for further research on the adaptation mechanism of orchardgrass to flood tolerance.

A Tale of Two Grass Species: Temperature Affects the Symbiosis of a Mutualistic Epichloë Endophyte in Both Tall Fescue and Perennial Ryegrass.

Many cool-season grasses form permanent, mutualistic symbioses with asexual Epichloë endophytes. These fungal symbionts often perform a protective role within the association as many strains produce secondary metabolites that deter certain mammalian and invertebrate herbivores. Although initially a serious issue for agriculture, due to mammalian toxins that manifested in major animal health issues, selected strains that provide abiotic stress protection to plants with minimal ill effects to livestock are now commercialized and routinely used to enhance pasture performance in many farming systems. These fungal endophytes and their grass hosts have coevolved over millions of years, and it is now generally accepted that most taxonomic groupings of Epichloë are confined to forming compatible associations (i.e., symptomless associations) with related grass genera within a tribe. The most desired compounds associated with Epichloë festucae var. lolii, an endophyte species associated with perennial ryegrass, are peramine and epoxy-janthitrems. No other major secondary metabolites with invertebrate bioactivity have been identified within this association. However, other agriculturally beneficial compounds, such as lolines, have been discovered in related endophyte species that form associations with fescue grasses. A rationale therefore existed to develop novel grass-endophyte associations between loline-producing endophytes originally isolated from tall fescue with elite cultivars of perennial ryegrass to achieve a wider spectrum of insect bioactivity. A suitable loline-producing endophyte strain of Epichloë sp. FaTG-3 was selected and inoculated into perennial ryegrass. We hypothesed that endophyte transmission frequency, endophyte mycelial biomass and endophyte-derived alkaloid production would differ between the original tall fescue host and the artificial association. Consistent with our hypothesis, our data strongly suggest that plant species significantly affected the plant-endophyte association. This effect became more apparent for transmission frequency and endophyte biomass as the plants matured. Overall, the viable endophyte infection frequency was greater in the tall fescue host than in perennial ryegrass, at all sampling dates. Additionally, temperature was found to be a significant factor affecting endophyte transmission frequency, endophyte mycelial biomass and alkaloid production. Implications for the development of novel grass-endophyte associations are discussed.