Genome evolution and initial breeding of the Triticeae grass Leymus chinensis dominating the Eurasian Steppe.

Leymus chinensis, a dominant perennial grass in the Eurasian Steppe, is well known for its remarkable adaptability and forage quality. Hardly any breeding has been done on the grass, limiting its potential in ecological restoration and forage productivity. To enable genetic improvement of the untapped, important species, we obtained a 7.85-Gb high-quality genome of L. chinensis with a particularly long contig N50 (318.49 Mb). Its allotetraploid genome is estimated to originate 5.29 million years ago (MYA) from a cross between the Ns-subgenome relating to Psathyrostachys and the unknown Xm-subgenome. Multiple bursts of transposons during 0.433-1.842 MYA after genome allopolyploidization, which involved predominantly the Tekay and Angela of LTR retrotransposons, contributed to its genome expansion and complexity. With the genome resource available, we successfully developed a genetic transformation system as well as the gene-editing pipeline in L. chinensis. We knocked out the monocot-specific miR528 using CRISPR/Cas9, resulting in the improvement of yield-related traits with increases in the tiller number and growth rate. Our research provides valuable genomic resources for Triticeae evolutionary studies and presents a conceptual framework illustrating the utilization of genomic information and genome editing to accelerate the improvement of wild L. chinensis with features such as polyploidization and self-incompatibility.

MsWRKY33 increases alfalfa (Medicago sativa L.) salt stress tolerance through altering the ROS scavenger via activating MsERF5 transcription.

Alfalfa (Medicago sativa L.) is considered to be the most important forage crop on a global scale. Nevertheless, soil salinity significantly decreases productivity, seriously threatening food security worldwide. One viable strategy is to explore salt stress-responsive factors and elucidate their underlying molecular mechanism, and utilize them in further alfalfa breeding. In the present study, we designated MsWRKY33 as a representative salt stress-responsive factor preferentially expressed in alfalfa roots and leaves. Subsequently, it was demonstrated that MsWRKY33 was localized in the cell nucleus, and functioned as a transcriptional activator of the W-box element. Transgenic alfalfa overexpressing MsWRKY33 displayed enhanced salt stress tolerance and antioxidant activities with no significant difference in other agronomic traits. Transcriptome profiling of MsWRKY33 transgenic alfalfa under control and salt treatment unveiled significantly altered expression of reactive oxygen species (ROS) scavenger genes in transgenic alfalfa. Subsequent examination revealed that MsWRKY33 binded to the promoter of MsERF5, activating its expression and consequently fine-tuning the ROS-scavenging enzyme activity. Furthermore, MsWRKY33 interacted with the functional fragment of MsCaMBP25, which participates in Ca2+ signaling transduction. Collectively, this research offers new insight into the molecular mechanism of alfalfa salt stress tolerance and highlights the potential utility of MsWRKY33 in alfalfa breeding.

Contamination of hay and haylage with enteric bacteria and selected antibiotic resistance genes following fertilization with dairy manure or biosolids.

The present study evaluated if enteric bacteria or antibiotic resistance genes carried in fecal amendments contaminate the hay at harvest, representing a potential route of exposure to ruminants that consume the hay. In the field experiments, dairy manure was applied to a hay field for three successive growing seasons, and biosolids were applied to a hay field for one growing season. Various enteric bacteria in the amendments were enumerated by viable plate count, and selected gene targets were quantified by qPCR. Key findings include the following: at harvest, hay receiving dairy manure or biosolids did not carry more viable enteric bacteria than hay from unamended control plots. The fermentation of hay did not result in a detectable increase in viable enteric bacteria. The application of dairy manure or biosolids resulted in a few gene targets being more abundant in hay during the first harvest. Fermentation of hay resulted in an increase in the abundance of gene targets, but this occurred with hay from both the amended and control plots. Overall, the application of fecal amendments resulted in an increase in the abundance of some gene targets associated with antibiotic resistance in the first cut hay.

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.

Forage Crop Lolium multiflorum Assisted Synthesis of AgNPs and Their Bioactivities Against Poultry Pathogenic Bacteria in In Vitro.

Italian ryegrass is one of main feed for livestock animals/birds. It has potential antioxidant metabolites that can improve their health and protect them against various infectious diseases. In this work, we studied synthesis of silver nanoparticles assisted by forage crop Lolium multiflorum as a green synthesis way. Potential antibacterial efficacy of these synthesized nanosized silver nanoparticles against poultry pathogenic bacteria was then studied. Aqueous extract of IRG was used as reducing agent for bio-reduction of silver salt to convert Ag+ to Ag0 metallic nano-silver. Size, shape, metallic composition, functional group, and crystalline nature of these synthesized silver nanoparticles were then characterized using UV-Vis spectrophotometer, FESEM, EDX, FT-IT, and XRD, respectively. In addition, antibacterial effects of these synthesized AgNPs against poultry pathogenic bacteria were evaluated by agar well diffusion method. UV-Vis spectra showed strong absorption peak of 440-450 nm with differ reaction time ranging from 30 min to 24 h. FESEM measurements revealed particles sizes of around 20-100 nm, majority of which were spherical in shape while a few were irregular. These biosynthesized silver nanoparticles using IRG extract exhibited strong antibacterial activities against poultry pathogenic microorganisms, including Pseudomonas aeruginosa, Salmonella typhi, Escherichia coli, and Bacillus subtilis. Overall results confirmed that IRG plant extract possessed potential bioactive compounds for converting silver ions into nanosized silver at room temperature without needing any external chemical for redox reaction. In addition, such synthesized AgNPs showed strong antibacterial activities against pathogenic bacteria responsible for infectious diseases in poultry.

Persistence of endophytic fungi in cultivars of Lolium perenne grown from seeds stored for 22 years.

PREMISE OF THE STUDY: Genetic resources for forage crops often consist of seeds of specific species and cultivars in cold storage for future use in breeding and selection programs. Temperate grasses such as Lolium perenne, used worldwide for forage and turf, produce seeds commonly infected by hyphae of an endophytic fungus (Epichloë festucae var. lolii). This research determined whether endophytes could persist and infect seedlings of L. perenne emerging from seeds stored for over two decades. METHODS: Endophyte-infected seeds (>90% infected) of four cultivars were obtained in 1994 and stored dry in plastic bags at 4°C. Seed germination was tested after 12 yr (for two cultivars) and after 18 and 22 yr (for all cultivars). Seedling leaf sheaths were excised, stained, and examined at 400× for endophytic hyphae to quantify infection frequency (% plants infected) and intensity (mean number of endophytic hyphae per field of view). KEY RESULTS: Seed germination after 22 yr depended on cultivar, ranging from 53 to 78%. Between 58 and 73% of plants grown from seeds stored for 22 yr still contained viable endophytic hyphae. Infection intensity remained at original levels for 18 yr in one cultivar; however, in all cultivars, infection intensity declined significantly between 18 and 22 yr. CONCLUSIONS: Persistence of the grass seed-endophyte symbiosis for over 20 yr surpasses all prior records of endophyte longevity within stored seeds. Storage of germplasm of cool-season grass cultivars that contain potentially beneficial fungal endophytes should be possible for several decades under dry, cold conditions.

Salinity-mediated cyanogenesis in white clover (Trifolium repens) affects trophic interactions.

BACKGROUND AND AIMS: Increasing soil salinity poses a major plant stress in agro-ecosystems worldwide. Surprisingly little is known about the quantitative effect of elevated salinity on secondary metabolism in many agricultural crops. Such salt-mediated changes in defence-associated compounds may significantly alter the quality of food and forage plants as well as their resistance against pests. In the present study, the effects of soil salinity on cyanogenesis in white clover (Trifolium repens), a forage crop of international importance, are analysed. METHODS: Experimental clonal plants were exposed to five levels of soil salinity, and cyanogenic potential (HCNp, total amount of accumulated cyanide in a given plant tissue), ß-glucosidase activity, soluble protein concentration and biomass production were quantified. The attractiveness of plant material grown under the different salt treatments was tested using cafeteria-style feeding trials with a generalist (grey garden slug, Deroceras reticulatum) and a specialist (clover leaf weevil, Hypera punctata) herbivore. KEY RESULTS: Salt treatment resulted in an upregulation of HCNp, whereas ß-glucosidase activity and soluble protein concentration showed no significant variation among treatments. Leaf area consumption of both herbivore species was negatively correlated with HCNp, indicating bottom-up effects of salinity-mediated changes in HCNp on plant consumers. CONCLUSIONS: The results suggest that soil salinity leads to an upregulation of cyanogenesis in white clover, which results in enhanced resistance against two different natural herbivores. The potential implications for such salinity-mediated changes in plant defence for livestock grazing remain to be tested.

Alfalfa weevils (Coleoptera: Curculionidae) in the western United States are resistant to multiple type II pyrethroid insecticides.

The alfalfa weevil (Hypera postica Gyllenhal (Coleoptera: Curculionidae)), a key pest of alfalfa (Medicago sativa L. (Fabales: Fabacae)) across the US, has developed resistance to pyrethroids lambda-cyhalothrin and zeta-cypermethrin in at least 6 western US states. Unfortunately, 6 pyrethroid active ingredients represent most commercial insecticides registered for alfalfa weevil control in forage alfalfa systems. Thus, the loss of efficacy of this mode of action group due to multiple resistance represents a significant agricultural challenge because of a limited registered alternative mode of actions. To evaluate the extent and severity of resistance among pyrethroids around the United States, laboratory bioassays using larvae from Arizona, California, Montana, Oregon, Washington, and Wyoming, including both the Egyptian and western strains, were conducted. Results indicated that similar degrees of resistance among type II pyrethroids as determined by both laboratory bioassays and field trials exist. The LC50 values of alpha-cypermethrin, beta-cyfluthrin and zeta-cypermethrin produced significant correlations with the LC50 values of lambda-cyhalothrin. In contrast, resistance did not include type I pyrethroid, bifenthrin (registered for seed alfalfa production), whose LC50 values yielded a slope not significantly different from zero when correlated with lambda-cyhalothrin. Field trials conducted in Arizona, Montana, and Washington corroborated laboratory results, as commercial formulations with type II pyrethroid active ingredients failed to adequately control alfalfa weevils resistant to lambda-cyhalothrin. Integrated resistance management recommendations are discussed.

Eliminating explanations for Maladera formosae (Coleoptera: Scarabaeidae) preponderance in sandy soil.

Most field corn in the United States receives a neonicotinoid seed treatment for the management of early-season, soil-dwelling insect pests. Grubs of Maladera formosae (Brenske) (Coleoptera: Scarabaeidae) have been reported feeding on young field corn with both low and high rates of clothianidin seed treatments in Indiana, Michigan, and Ohio. Anecdotally, these infestations are restricted to sandy soils in the region. The purpose of this study was to (1) evaluate whether grub populations in corn are restricted to sandy soils, (2) assess whether soil type influences M. formosae survival, and (3) determine whether soil type affects clothianidin uptake by the plant, possibly explaining the observed differences in M. formosae abundance by soil type. We observed nearly 10-times more grubs in sand (>80% sand content) than loam (<80% sand content) soil within a single corn field. Grub survival to adult was not influenced by soil type. We then compared the concentrations of clothianidin seed treatment in the roots and shoots of corn seedlings grown in either sand or loam soil over time. Similar amounts of the active ingredient were found in the roots and shoots of corn grown in both soil types. Within 2 week, the clothianidin concentrations in both soil types had significantly declined in roots and shoots and were no different from the no-insecticide control. These findings suggest that factors other than insecticide exposure contribute to the higher abundance of M. formosae larvae in sand relative to loam soils, even within the same field.

On-farm harvest timing effects on alfalfa weevil across the Intermountain West region of the United States.

Alfalfa (Medicago sativa L.) is an economically important commodity in the Intermountain Western United States. A major concern for alfalfa producers in this region is the alfalfa weevil (Hypera postica Gyllenhal). Insecticide resistance development coupled with regulatory changes in pesticide use has resulted in renewed interest by producers in non-chemical control methods such as cultural control. One such cultural control method is early harvest, which consists of producers timing their harvests early in the season to decrease alfalfa weevil damage. This method is thought to be effective by exposing weevil larvae to adverse conditions before significant damage occurs. Still, early harvest can be difficult to employ because recommendations are often vague. To better understand how early harvest impacts both alfalfa weevils and their natural enemies and how producers are using this method across the Intermountain Western United States, we conducted a study in alfalfa production fields in Colorado, Montana, and Wyoming over three growing seasons. We determined that the timing of the initial alfalfa harvest spanned more than 1 month across fields, and alfalfa plant stage at harvest ranged from late vegetative to early bloom. Harvest was more impactful on reducing alfalfa weevil densities the earlier it was implemented. Removing windrows in a timely manner is likely useful to further decrease alfalfa weevil densities. Harvest timing was not associated with parasitism rates of alfalfa weevil, but higher parasitism rates were associated with lower post-harvest alfalfa weevil densities. This work has increased our understanding of early harvest in an on-farm setting and to improve recommendations for producers across the Intermountain Western United States.

A novel insecticide, isocycloseram, shows promise as an alternative to chlorpyrifos against a direct pest of peanut, Diabrotica undecimpunctata howardi (Coleoptera: Chrysomelidae).

Larvae of the southern corn rootworm (SCR) Diabrotica undecimpunctata howardi Barber (Coleoptera: Chrysomelidae) are primary pests of peanut in the Virginia-Carolina region of the United States, and are relatively sporadic pests in southern states such as Georgia, Alabama, and Florida. Peanuts have strict quality standards which, when they are not met, can diminish crop value by more than 65%. Management of direct pests like SCR is therefore crucial to maintaining the economic viability of the crop. The soil-dwelling nature of SCR larvae complicates management due to difficulties associated with monitoring and predicting infestations. Nonchemical management options are limited in this system; preventative insecticide applications are the most reliable management strategy for at-risk fields. Chlorpyrifos was the standard product for larval SCR management in peanut until its registration was revoked in 2022, leaving no effective chemical management option for larvae. We tested a novel insecticide, isocycloseram, for its ability to reduce pod scarring, pod penetration, and non-SCR pod damage in field studies conducted in Suffolk, Virginia in 2020-2022. Overall injury was low in 2020 and 2022, and in 2022 there was not a significant effect of treatment. In 2021, 2 simulated chemigation applications of isocycloseram in July significantly reduced pod scarring and overall pod injury relative to chlorpyrifos and the untreated control. Our results suggest that isocycloseram may become an effective option for managing SCR in peanut, although more work is needed to understand the mechanisms by which it is effective as a soil-applied insecticide.

Effects of leaf properties on the counts of microbes on the leaf surfaces of wheat, rye and triticale.

Currently, our understanding of the effects of glaucousness on the population sizes of microbial communities on leaf surfaces is limited. The objective of this study was to investigate the influence of glaucousness and chemical composition on the colony-forming unit (CFU) counts of microbes on leaf surfaces. Various leaf morphological and physicochemical features, microbial CFU counts and glaucousness on the leaf surfaces of wheat (Triticum aestivum var. Shimai No.1), rye (Secale cereale var. Gannong No.1) and triticale (Triticale wittmack var. Gannong No.2) were assessed. The results showed that larger CFU counts of lactic acid bacteria (LAB) and yeasts were found on the non-glaucous leaf surfaces in wheat than on the glaucous leaf surfaces in rye and triticale. The CFU counts of LAB and yeasts were negatively correlated with the amount of soluble wax (P < 0.05), and positively correlated with the contents of starch and fructose in leaf tissue (P < 0.05), soluble and reduced sugars, soluble protein and free amino acids on leaf surfaces (P < 0.05). The CFU counts of microbes on leaf surfaces were mainly influenced by the level of available carbon sources and glaucousness of leaf surfaces.

Phenotypic Diversity in Domesticated and Wild Timothy Grass, and Closely Related Species for Forage Breeding.

Timothy grass (Phleum pratense L.) is one of the most important forage crops in temperate regions. Forage production, however, faces many challenges, and new cultivars adapted to a changing climate are needed. Wild populations and relatives of timothy may serve as valuable genetic resources in the breeding of improved cultivars. The aim of our study is to provide knowledge about the phenotypic diversity in domesticated (cultivars, breeding lines and landraces) and wild timothy and two closely related species, P. nodosum (lowland species) and P. alpinum, (high altitude species) to identify potential genetic resources. A total of 244 accessions of timothy and the two related species were studied for growth (plant height, fresh and dry weight) and plant development (days to stem elongation, days to booting and days to heading) in the field and in a greenhouse. We found a large diversity in development and growth between the three Phleum species, as well as between the accessions within each species. Timothy showed the highest growth, but no significant difference was found between wild accessions and cultivars of timothy in fresh and dry weight. However, these two groups of accessions showed significant differences in plant development, where timothy cultivars as a group reached flowering earlier than the wild accessions. This suggests that there has not been a strong directional selection towards increased yield during the domestication and breeding of timothy; rather, timothy has been changed for other traits such as earlier heading. Principal component analysis and cluster analysis based on all traits revealed distinct clusters. Accessions falling within the same cluster showed similarities in the development and growth rather than the type of accession. The large diversity found in this study shows the potential of using timothy accessions as genetic resources in crosses with existing cultivars. Also, accessions of P. nodosum with favorable traits can be candidates for the domestication of a novel forage crop, and the high-altitude relative P. alpinum may be a source of genes for the development of more cold and stresstolerant cultivars.

Effects of autotoxicity and allelopathy on seed germination and seedling growth in Medicago truncatula.

Autotoxicity is a form of intraspecific allelopathy, in which a plant species inhibits the establishment or growth of the same species through the release of toxic chemical compounds into the environment. The phenomenon of autotoxicity in crops is best traced in alfalfa (Medicago sativa). A close relative of alfalfa, M. truncatula, has been developed into an excellent model species for leguminous plants. However, it is not known whether M. truncatula has autotoxicity. In this study, M. truncatula root exudates showed a negative impact on the growth of M. truncatula seedlings, indicating autotoxicity. Detailed analyses with plant extracts from M. truncatula and alfalfa revealed varying degrees of suppression effects in the two species. The extracts negatively affected seed germination potential, germination rate, radicle length, hypocotyl length, synthetic allelopathic effect index, plant height, root growth, fresh weight, dry weight, net photosynthetic rate, transpiration rate, and stomatal conductance in both M. truncatula and alfalfa. The results demonstrated that autotoxicity and allelopathic effects exist in M. truncatula. This opens up a new way to use M. truncatula as a model species to carry out in-depth studies of autotoxicity and allelopathy to elucidate biochemical pathways of allelochemicals and molecular networks controlling biosynthesis of the chemicals.

Alfalfa Weevil (Coleoptera: Curculionidae) Resistance to Lambda-cyhalothrin in the Western United States.

Forage alfalfa (Medicago sativa L. [Fabales: Fabaceae]) is a key agricultural commodity of the western region of the United States. The key insect pest of alfalfa, Hypera postica Gyllenhal (Coleoptera: Curculionidae), has developed resistance to the most common class of insecticide used to manage its damage. Alfalfa weevil samples from 71 commercial alfalfa fields located in Arizona, California, Montana, Oregon, Washington, and Wyoming were assayed for susceptibility to lambda-cyhalothrin during 2020-2022 using a laboratory concentration-response assay. Seventeen field sites representing all six states were highly resistant to lambda-cyhalothrin (resistance ratios > 79.6) and bioassay mortality often did not exceed 50% even at the highest concentration tested (3.30 µg/cm2 in 2020 and 10.00 µg/cm2 in 2021-2022). Field sites assayed with more than one pyrethroid active ingredient indicated likely cross-resistance between lambda-cyhalothrin and zeta-cypermethrin (type II pyrethroids) and variable and/or limited potential cross-resistance to permethrin (type I pyrethroid). Thirty-two field sites representing five states were susceptible to lambda-cyhalothrin (resistance ratios ranging from 1 to 20). While resistance is widespread, integrated resistance management strategies including rotating mode of action groups, applying chemical control tactics only when economic thresholds have been met, and utilizing cultural control tactics can be employed to slow the further development of resistance.

Assessment of the Genetic Distinctiveness and Uniformity of Pre-Basic Seed Stocks of Italian Ryegrass Varieties.

Lolium multiflorum Lam., commonly known as Italian ryegrass, is a forage grass mostly valued for its high palatability and digestibility, along with its high productivity. However, Italian ryegrass has an outbreeding nature and therefore has high genetic heterogeneity within each variety. Consequently, the exclusive use of morphological descriptors in the existing varietal identification and registration process based on the Distinctness, Uniformity, and Stability (DUS) test results in an inadequately precise assessment. The primary objective of this work was to effectively test whether the uniformity observed at the phenological level within each population of Italian ryegrass was confirmed at the genetic level through an SSR marker analysis. In this research, using 12 polymorphic SSR loci, we analyzed 672 samples belonging to 14 different Italian ryegrass commercial varieties to determine the pairwise genetic similarity (GS), verified the distribution of genetic diversity within and among varieties, and investigated the population structure. Although the fourteen commercial varieties did not show elevated genetic differentiation, with only 13% of the total variation attributable to among-cultivar genetic variation, when analyzed as a core, each variety constitutes a genetic cluster on its own, resulting in distinct characteristics from the others, except for two varieties. In this way, by combining a genetic tool with the traditional morphological approach, we were able to limit biases linked to the environmental effect of field trials, assessing the real source of diversity among varieties and concretely answering the key requisites of the Plant Variety Protection (PVP) system.

Application of Magnesium and Calcium Sulfate on Growth and Physiology of Forage Crops under Long-Term Salinity Stress.

Soil salinity is major threat to crop growth and reducing cultivated land areas and salt-resistant crops have been required to sustain agriculture in salinized areas. This original research was performed to determine the effectiveness of MgSO4 (MS) and CaSO4 (CS) for each species and assess changes in the physiology and growth of fodder crops after short and long-term salt stress. Six treatments (CON (control); NaCl (NaCl 100 mM); 1 MS (1 mM MgSO4 + 100 mM NaCl); 2 MS (2 mM MgSO4 + 100 mM NaCl); 7.5 CS (7.5 mM CaSO4 + 100 mM NaCl); and 10 CS (10 mM CaSO4 + 100 mM NaCl)) were applied to Red clover (Trifolium pratense) and Tall fescue (Festuca arundinacea) under greenhouse conditions. Cultivars were evaluated based on their dry weights, physiological parameters, forage quality, and ion concentrations. The biomass of both species decreased significantly under NaCl treatments and increased under the MS and CS treatments compared to solely salinity treatments. Salinity caused a decrease in the photosynthetic rate, but compared to CON, the MS and CS treatments yielded superior results. Moreover, the Na+/K+ ratio increased as Na+ concentration increased but crop quality (CP, NDF, ADF) did not show significant differences under salinity. Overall, we concluded that these T. pratense and F. arundinacea species demonstrated various responses to salinity, MS, and CS by different physiological and morphological parameters and it turned out to be efficient under salinity stress.

Appropriate N fertilizer addition mitigates N2O emissions from forage crop fields.

Forage crops are widely cultivated as livestock feed to relieve grazing pressure in agro-pastoral regions with arid climates. However, gaseous losses of soil nitrogen (N) following N fertilizer application have been considerable in response to the pursuit of increased crop yield. A two-year experiment was carried out in a typical saline field under a temperate continental arid climate to investigate the effect of N application rate on N2O emissions from barley (Hordeum vulgare L.), corngrass (Zea mays × Zea Mexicana), rye (Secale cereale L.), and sorghum-sudangrass hybrid (Sorghum bicolor × Sorghum sudanense). The dynamics of N2O emissions, hay yield, and crude protein (CP) yield were measured under four N application rates (0, 150, 200, and 250 kg ha-1) in 2016 and 2017. An N2O emission peak was observed for all crop species five days after each N application. Cumulative N2O fluxes in the growing season ranged from 0.66 to 2.40 kg ha-1 and responded exponentially to N application rate. Emission factors of N2O showed a linear increase with N application rate for all crop species, but the linear slopes significantly differed between barley or rye and corngrass and sorghum-sudangrass hybrid. The hay and CP yields of all forage grasses significantly increased with the increase of N application rate from 0 to 200 kg ha-1. Barley and rye with lower hay and CP yields showed higher N2O emission intensities. The increased level of N2O emission intensity was higher from 200 to 250 kg ha-1 than from 150 to 200 kg ha-1. At N application rates of 200 and 250 kg ha-1, CP yield had a significantly negative correlation with cumulative N2O emission and explained 50.5% and 62.9% of the variation, respectively. In conclusion, ~200 kg ha-1 is the optimal N rate for forage crops to minimize N2O emission while maintaining yield in continental arid regions.

Transcriptomic and Metabolomic Analyses Reveal the Differential Regulatory Mechanisms of Compound Material on the Responses of Brassica campestris to Saline and Alkaline Stresses.

Oilseed rape not only has the function of improve saline and alkaline soils, but also alleviate the local feed shortage. However, medium- and high-degree soil salinization and alkalinization always inhibit the growth of oilseed rape. Studies have shown that compound material can improve the tolerance to saline and alkaline stress of crops, but the difference in the regulation mechanism of compound material on oilseed rape in saline and alkaline soils is not clear. This study explored the difference through determining the leaf ion contents, physiological indexes, transcriptomics, and metabolomics of oilseed rape in salinized soil (NaCl 8 g kg-1) and alkalinized soil (Na2CO3 8 g kg-1) at full flowering stage, respectively after the application of compound material. The results showed that in salinized and alkalinized soil, the compound material upregulated the genes related to the regulation of potassium ion transport, and changed the amino acid metabolic pathway, which reduced the contents of Na+, malondialdehyde (MDA), and relative conductivity (REC) in leaves, and increased the contents of K+ and Mg2+ and the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). However, there were differences in the regulation mechanism of compound material in salinized and alkalinized soil. In salinized soil, the compound material improved the tolerance of oilseed rape to saline stress by upregulating transcription factors mannose-1-phosphate guanylyltransferase (GPMM) and Glutamine--fructose-6-phosphate transaminase (GFPT) and downregulating phosphomannomutase (PMM) to change nucleotide metabolism pathway and lipid metabolism pathway. In alkalized soil, the compound material improved the tolerance of oilseed rape to alkaline stress by upregulating transcription factors Phenylalanine ammonia lyase (PAL) to change the biosynthesis pathway of other secondary metabolites. Therefore, the compound material can improve the tolerance of oilseed rape to saline and alkaline stress by regulating the genetic adaptability and apparent plasticity, but the mechanisms were different. This study provides a practical method for the ecological environment restoration and the development of animal husbandry.

Effect of planting and mowing cover crops as livestock feed on soil quality and pear production.

Introduction: The increasing demand for animal-products has led to an increasing demand for livestock feed. Using cover crop as green manure in orchards is an effective measure to improve fruit yield and quality. However, the effect of mowing cover forage crops as livestock feed on soil quality and crop production is unclear. Method: Therefore, a 4-year field experiment, which included two treatments, was conducted in pear orchards in Luniao County, China: natural grass (NG) and planting and mowing forage crop ryegrass as livestock feed (MF). Results: Under MF treatment, most soil nutrient content, especially alkalihydrolysable N (AN), total phosphate (TP), available phosphate (AP), and microbial biomass phosphate (MBP), had decreased significantly (P<0.05), while ß-D-glucosidase (BG, C-cycle enzyme) and soil C limitation at 10-20 cm depth and P limitation at subsoil (20-40 cm) was increased. In addition, the soil bacterial community component in topsoil (0-10 cm and 10-20 cm) and fungal community component in topsoil and subsoil were changed in the MF treatment. Network analysis showed that MF treatment had a lower edge number in topsoil but the community edge numbers increased from 12794 in NG to 13676 in MF in subsoil. The average weight degree of the three soil layers in MF treatment were reduced, but the modularity had increased than that in NG. For crop production, MF treatment was 1.39 times higher in pear yield and titratable acids (AC) reduced from 0.19% to 0.13% compared with NG. These changes were more associated with the indicators at the subsoil, especially for TP, AN, pH, and F-NMDS1 (non-metric multidimensional scaling (NMDS) axis 1 of fungi). Discussion: These results provide data support for the feasibility of planting and mowing forage crops as livestock feed on orchards as well as a new idea for the integration of crop and livestock.