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O-coumaric acid (OCA), as a significant phenolic allelochemical found in hairy vetch (Vicia villosa Roth.), that can hinder the growth of alfalfa (Medicago sativa L.), particularly the growth of alfalfa roots. Nonetheless, the mechanism by which OCA inhibits alfalfa root growth remains unclear. In this study, a liquid chromatography tandem mass spectrometry (LC-MS/MS)-based quantitative proteomics analysis was carried out to identify differentially accumulated proteins (DAPs) under OCA treatment. The findings indicated that 680 proteins were DAPs in comparison to the control group. Of those, 333 proteins were up-regulated while 347 proteins were down-regulated. The enrichment analysis unveiled the significance of these DAPs in multiple biological and molecular processes, particularly in ribosome, phenylpropanoid biosynthesis, glutathione metabolism, glycolysis/gluconeogenesis and flavonoid biosynthesis. The majority of DAPs reside in the cytoplasm (36.62%), nucleus (20.59%) and extracellular space (14.12%). In addition, phenylalanine deaminase was identified as a potential chemical-induced regulation target associated with plant lignin formation. DAPs were mainly enriched in flavonoid biosynthesis pathways, which were related to plant root size. Using the UPLC-ESI-MS/MS technique and database, a total of 87 flavonoid metabolites were discovered. The metabolites were predominantly enriched for biosynthesizing naringenin chalcone, which was linked to plant lignin formation, aligning with the enrichment outcomes of DAPs. Consequently, it was deduced that OCA impacted the structure of cell walls by mediating the synthesis of lignin in alfalfa roots, subsequently inducing wilt. Furthermore, a range of proteins have been identified as potential candidates for the breeding of alfalfa strains with enhanced stress tolerance. [ABSTRACT FROM AUTHOR]

Foraging is one of the most fundamental activities contributing to the maximization of an animal's fitness, and thus herbivores must optimize their diet selection and intake to meet their nutrient demands for survival, growth, and reproduction. Using plant DNA barcoding, we determined diet composition of five subpopulations of adult female pronghorn antelope (Antilocapra americana) grazing rangelands in southern and southeastern Idaho, USA. Fecal samples were collected for two years (2018–2019), and across metabolically-important adult female life history stages (late gestation, early lactation, breeding season). Plant DNA barcoding yielded 137 detected species within pronghorn diets across subpopulations and sampling periods with forbs being the most abundant. Pronghorn dietary functional group composition ranged from 52.2–60.3% from forbs followed by shrubs (22.6–28.2%), graminoids (8.7–15.7%), and legumes (5.5–9.6%). Dietary protein intake was also highest from forbs and ranged from 32.4–62.4% followed by graminoids (1.2–43.1%), shrubs (18.7–21.3%), and legumes (2.6–7.4%). We found significant intra- and interannual differences in the mean number of genera-based plant detections in pronghorn diets. Dietary protein intake of cultivated legumes (e.g., alfalfa [Medicago sativa] and sainfoin [Onobrychis viciifolia]) was lower than expected, ranging from <1.0–30.8%, suggesting that even within an agricultural-dominated landscape, factors other than plant nutritional composition contributed to pronghorn diets. Although the plant DNA barcoding technique exhibits limitations, it demonstrated potential for elucidating pronghorn dietary species richness, particularly for plants consumed in small proportions, as well as for observing temporal fluctuations in functional group composition and dietary protein intake explained through the interplay between environmental factors, plant chemical composition, and the animals' physiological needs. [ABSTRACT FROM AUTHOR]

Animals raised in environments that prevent natural foraging opportunities may have difficulty adapting to novelty, such as feeding and management changes. Our objective was to evaluate how early provision and presentation of forage in dairy calves affected response to novel TMR (total mixed ration; grain and alfalfa) at weaning. Holstein heifer calves were housed individually in a covered outdoor hutch with an attached uncovered wire-fenced pen on sand bedding. Calves were fed a diet of starter grain and milk replacer (5.7–8.4L/d step-up) via a bottle (Control, n = 9) or given additional access to mountaingrass hay presented either in a bucket (Bucket, n = 9), or PVC pipe feeder (Pipe, n = 9). Treatments were applied from birth through 50 d of age, when step-down weaning began. All calves had 3 buckets and a pipe feeder provided in their uncovered pen area. On d 50, each calf was briefly blocked inside their hutch. TMR was put in the 3rd bucket that previously contained hay (Bucket) or was empty (Control, Pipe). The calf was released from the hutch and video-recorded for 30 min. Neophobia towards TMR was affected by prior experience with presentation: Bucket calves began eating TMR faster than Pipe and Control (P≤0.012) and showed the fewest number of startle responses (P = 0.004). Intake was similar across groups (P = 0.978), suggesting this apparent neophobia was transient, but Control calves took longer to eat than Bucket (P<0.001) and Pipe (P = 0.070) calves and were less likely to give up on eating to lie down instead. These results suggest that previous experience with hay improves processing ability when presented with novel TMR. Overall, response to a novel feed is affected by both early life experience, such as opportunities to process forage, and the presentation of the feed itself. Calves also appear motivated to access forage, evidenced by transient neophobia, high intake, and persistence in feeding by naïve calves. [ABSTRACT FROM AUTHOR]

Since the release of genetically engineered (GE) crops, there has been increased concern about the introduction of GE genes into non-GE fields of a crop and their spread to feral or wild cross-compatible relatives. More recently, attention has been given to the differential impact of distinct pollinators on gene flow, with the goal of developing isolation distances associated with specific managed pollinators. To examine the differential impact of bee species on gene movement, we quantified the relationship between the probability of getting a GE seed in a pod, and the order in which a flower was visited, or the cumulative distance traveled by a bee in a foraging bout. We refer to these relationships as 'seed curves' and compare these seeds curves among three bee species. The experiments used Medicago sativa L. plants carrying three copies of the glyphosate resistance (GR) allele as pollen donors (M. sativa is a tetraploid), such that each pollen grain carried the GR allele, and conventional plants as pollen recipients. Different foraging metrics, including the number of GR seeds produced over a foraging bout, were also quantified and contrasted among bee species. The lowest number of GR seeds set per foraging bout, and the GR seeds set at the shortest distances, were produced following leafcutting bee visits. In contrast, GR seeds were found at the longest distances following bumble bee visits. Values for honey bees were intermediate. The ranking of bee species based on seed curves correlated well with field-based gene flow estimates. Thus, differential seed curves of bee species, which describe patterns of seed production within foraging bouts, translated into distinct abilities of bee species to move genes at a landscape level. Bee behavior at a local scale (foraging bout) helps predict gene flow and the spread of GE genes at the landscape scale. [ABSTRACT FROM AUTHOR]

This document is a correction notice for an article titled "Influence of different irrigation methods on the alfalfa rhizosphere soil fungal communities in an arid region." The correction addresses errors in the author affiliations. The correct affiliations are provided, which include the Sichuan Provincial Key Laboratory for Development and Utilization of Characteristic Horticultural Biological Resources at Chengdu Normal University in China, as well as the College of Life Sciences at Shihezi University in Xinjiang Province, China. The correction is reported by the authors themselves. [Extracted from the article]

Xinjiang is the largest arid and saline agricultural region in China. The common irrigation methods in this area are traditional flood irrigation and drip irrigation. In this study, we investigated the effects of these two irrigation methods on the fungal diversity, community structures, and functions in alfalfa rhizosphere soil as well as the associated environmental factors in northern Tianshan Mountain (Xinjiang, China). Soil enzyme activities (urease and neutral phosphatase) were significantly higher in the drip-irrigated alfalfa rhizosphere soil than in the flood-irrigated alfalfa rhizosphere soil, whereas the fungal alpha diversity in the drip-irrigated alfalfa rhizosphere soil was significantly lower than that in the flood-irrigated alfalfa rhizosphere soil. Six dominant fungal phyla were identified (>0.1%), with Ascomycota being the most abundant in all soils, followed by Basidiomycota (5.47%), Mortierellomycota (1.07%), Glomeromycota (0.55%), Rozellomycota (0.27%), and Chytridiomycota (0.14%). Ascomycota and Glomeromycota species were significantly less abundant in drip-irrigated alfalfa rhizosphere soil than in flood-irrigated alfalfa rhizosphere soil. A LEFSe analysis identified Cladosporiaceae (20.8%) species as the most abundant marker fungi in drip-irrigated alfalfa rhizosphere soil. Of the 13 fungal functional groups identified on the basis of the functional annotation using the FUNGuild database, Ectomycorrhizal (22.29%) was the primary functional group. Compared with flood irrigation, drip irrigation significantly decreased the relative abundance of Ectomycorrhizal and Arbuscular_Mycorrhizal, while increasing the relative abundance of Plant_Pathogen, although not significantly (P = 0.19). Available potassium was revealed to be the main environmental factor influencing soil enzyme activities, fungal alpha diversity, fungal community structures, and fungal functions in response to the different irrigation methods. In conclusion, drip irrigation may be more appropriate than flood irrigation in the Tianshan dryland agricultural area for enhancing soil enzyme activities, but it may also increase the abundance of plant pathogenic fungi in the soil. [ABSTRACT FROM AUTHOR]

Excess copper (Cu) in soil due to industrial and agricultural practices can result in reduced plant growth. Excess Cu resulted in severely retarded root growth with severe discoloration of Alfalfa (Medicago sativa) and Medicago truncatula. Growth in the presence of hydrogen peroxide resulted in similar symptoms that could be partially recovered by the addition of the reductant ascorbic acid revealing damage was likely due to oxidative stress. The addition of proanthocyanidins (PAs) in the presence of Cu prevented much of the damage, including plant growth and restoration of lignin synthesis which was inhibited in the presence of excess Cu. Transcriptome analyses of the impact of excess Cu and the amelioration after PAs treatment revealed that changes were enriched in functions associated with the cell wall and extracellular processes, indicating that inhibition of cell wall synthesis was likely the reason for retarded growth. Excess Cu appeared to induce a strong defense response, along with alterations in the expression of a number of genes encoding transcription factors, notably related to ethylene signaling. The addition of PAs greatly reduced this response, and also induced novel genes that likely help ameliorate the effects of excess Cu. These included induction of genes involved in the last step of ascorbic acid biosynthesis and of enzymes involved in cell wall synthesis. Combined, these results show that excess Cu causes severe oxidative stress damage and inhibition of cell wall synthesis, which can be relieved by the addition of PAs. [ABSTRACT FROM AUTHOR]

The LIM (Lin-11, Isl-1 and Mec-3 domains) family is a key transcription factor widely distributed in animals and plants. The LIM proteins in plants are involved in the regulation of a variety of biological processes, including cytoskeletal organization, the development of secondary cell walls, and cell differentiation. It has been identified and analyzed in many species. However, the systematic identification and analysis of the LIM genes family have not yet been reported in alfalfa (Medicago sativa L.). Based on the genome-wide data of alfalfa, a total of 21 LIM genes were identified and named MsLIM01-MsLIM21. Comprehensive analysis of the chromosome location, physicochemical properties of the protein, evolutionary relationship, conserved motifs, and responses to abiotic stresses of the LIM gene family in alfalfa using bioinformatics methods. The results showed that these MsLIM genes were distributed unequally on 21 of the 32 chromosomes in alfalfa. Gene duplication analysis showed that segmental duplications were the major contributors to the expansion of the alfalfa LIM family. Based on phylogenetic analyses, the LIM gene family of alfalfa can be divided into four subfamilies: αLIM subfamily, βLIM subfamily, γLIM subfamily, and δLIM subfamily, and approximately all the LIM genes within the same subfamily shared similar gene structure. The 21 MsLIM genes of alfalfa contain 10 Motifs, of which Motif1 and Motif3 are the conserved motifs shared by these genes. Furthermore, the analysis of cis-regulatory elements indicated that regulatory elements related to transcription, cell cycle, development, hormone, and stress response are abundant in the promoter sequence of MsLIM genes. Real-time quantitative PCR demonstrated that MsLIM gene expression is induced by low temperature and salt. The present study serves as a basic foundation for future functional studies on the alfalfa LIM family. [ABSTRACT FROM AUTHOR]

Alfalfa (Medicago sativa L.) is an important legume crop for forage, agriculture, and environment in the world. Ascorbic acid (AsA) plays positive roles in plants. However, its effects on germination and salt-tolerance of alfalfa are unknown. The effects of AsA applications on seed germination and seedling salt-tolerance of alfalfa were investigated. The results revealed that 0.1 and 1 mmol L-1 of exogenous AsA increased germination, amylase, and protease, as well as seedling length, fresh weight (FW), dry weight (DW), and endogenous AsA both in the shoots and roots, except that 1 mmol L-1 AsA reduced the activities of α-amylase, β-amylase and protease on day 3. However, 10 and 100 mmol L-1 AsA inhibited these parameters and even caused serious rot. It indicates that 0.1 mmol L-1 AsA has the optimal effects, whereas 100 mmol L-1 AsA has the worst impacts. Another part of the results showed that 0.1 mmol L-1 AsA not only enhanced stem elongation, FW and DW, but also increased chlorophyll and carotenoids both under non-stress and 150 mmol L-1 NaCl stress. Furthermore, 0.1 mmol L-1 AsA mitigated the damages of membrane permeability, malondialdehyde, and excessive reactive oxygen species (ROS) and ions both in the shoots and roots under 150 mmol L-1 NaCl stress. Hence, 0.1 mmol L-1 AsA improves growth and induces salt-tolerance by inhibiting excessive ROS, down-regulating the ion toxicity and up-regulating the antioxidant system. The principal component analysis included two main components both in the shoots and roots, and it explained the results well. In summary, the optimum concentration of 0.1 mmol L-1 AsA can be implemented to improve the seed germination and seedling growth of alfalfa under salt stress. [ABSTRACT FROM AUTHOR]

In insect-pollinated crops, gene flow is affected by numerous factors including crop characteristics, mating system, life history, pollinators, and planting management practices. Previous studies have concentrated on the impact of distance between genetically engineered (GE) and conventional fields on adventitious presence (AP) which represents the unwanted presence of a GE gene. Variables other than distance, however, may affect AP. In addition, some AP is often present in the parent seed lots used to establish conventional fields. To identify variables that influence the proportion of AP in conventional alfalfa fields, we performed variable selection regression analyses. Analyses based on a sample-level and a field-level analysis gave similar, though not identical results. For the sample-level model, distance from the GE field explained 66% of the variance in AP, confirming its importance in affecting AP. The area of GE fields within the pollinator foraging range explained an additional 30% of the variation in AP in the model. The density of alfalfa leafcutting bee domiciles influenced AP in both models. To minimize AP in conventional alfalfa seed fields, management practices should focus on optimizing isolation distances while also considering the size of the GE pollen pool within the pollinator foraging range, and the foraging behavior of pollinators. [ABSTRACT FROM AUTHOR]

We previously reported that feeding Se-biofortified alfalfa hay to weaned beef calves in a preconditioning program decreases morbidity and mortality during the feedlot period. To understand the mode of action by which supranutritional Se supplementation supports calf health, we examined the effect of agronomic Se-biofortification on nasal microbiome and fecal parasites. Recently weaned Angus-cross beef calves (n = 30) were randomly assigned to two groups and fed an alfalfa hay-based diet for 9 weeks in a preconditioning program. Alfalfa hay was harvested from fields fertilized with sodium selenate at a rate of 0 or 90 g Se/ha. Calculated Se intake from dietary sources was 1.09 and 27.45 mg Se/calf per day for calves consuming alfalfa hay with Se concentrations of 0.06 and 3.47 mg Se/kg dry matter, respectively. Feeding Se-biofortified alfalfa hay for 9 weeks was effective at increasing whole-blood Se concentrations (556 ± 11 vs 140 ± 11 ng/mL; P < 0.001) and increasing body weight (PTreatment, = 0.03) in weaned beef calves. Slaughter yield grades were higher for calves that had been fed Se-enriched alfalfa hay during the preconditioning period (PTreatment = 0.008). No significant differences were observed in fecal parasite load, which remained low. The nasal microbiome and microbiota diversity within calves and across calves expanded from weaning (week 0) to the feedlot period (week 12), which was promoted by feeding Se-biofortified alfalfa hay. Especially concerning was the expansion of nasal Mycoplasmataceae in the feedlot, which reached over 50% of the total microbiota in some calves. In conclusion, we identified dietary Se-biofortified alfalfa hay as a potential promoter of nasal microbiome genome and microbiota diversity, which may explain in part high-Se benefits for prevention of bovine respiratory disease complex in beef calves. [ABSTRACT FROM AUTHOR]

Alfalfa is a forage legume commonly associated with ruminant livestock production that may be a potential source of health-promoting phytochemicals. Anecdotal evidence from producers suggests that later cuttings of alfalfa may be more beneficial to non-ruminants; however, published literature varies greatly in measured outcomes, supplement form, and cutting. The objective of this study was to measure body weight, average daily feed intake, host immunity, and the colon microbiota composition in mice fed hay, aqueous, and chloroform extracts of early (1st) and late (5th) cutting alfalfa before and after challenge with Citrobacter rodentium. Prior to inoculation, alfalfa supplementation did not have a significant impact on body weight or feed intake, but 5th cutting alfalfa was shown to improve body weight at 5- and 6-days post-infection compared to 1st cutting alfalfa (P = 0.02 and 0.01). Combined with the observation that both chloroform extracts improved mouse body weight compared to control diets in later stages of C. rodentium infection led to detailed analyses of the immune system and colon microbiota in mice fed 1st and 5th cutting chloroform extracts. Immediately following inoculation, 5th cutting chloroform extracts significantly reduced the relative abundance of C. rodentium (P = 0.02) and did not display the early lymphocyte recruitment observed in 1st cutting extract. In later timepoints, both chloroform extracts maintained lower splenic B-cell and macrophage populations while increasing the relative abundance of potentially beneficially genera such as Turicibacter (P = 0.02). At 21dpi, only 5th cutting chloroform extracts increased the relative abundance of beneficial Akkermansia compared to the control diet (P = 0.02). These results suggest that lipid soluble compounds enriched in late-cutting alfalfa modulate pathogen colonization and early immune responses to Citrobacter rodentium, contributing to protective effects on body weight. [ABSTRACT FROM AUTHOR]

Perennial crops in agricultural systems can increase sustainability and the magnitude of ecosystem services, but yield may depend upon biotic context, including soil mutualists, pathogens and cropping diversity. These biotic factors themselves may interact with abiotic factors such as drought. We tested whether perennial crop yield depended on soil microbes, water availability and crop diversity by testing monocultures and mixtures of three perennial crop species: a novel perennial grain (intermediate wheatgrass—Thinopyrum intermedium-- that produces the perennial grain Kernza®), a potential perennial oilseed crop (Silphium intregrifolium), and alfalfa (Medicago sativa). Perennial crop performance depended upon both water regime and the presence of living soil, most likely the arbuscular mycorrhizal (AM) fungi in the whole soil inoculum from a long term perennial monoculture and from an undisturbed native remnant prairie. Specifically, both Silphium and alfalfa strongly benefited from AM fungi. The presence of native prairie AM fungi had a greater benefit to Silphium in dry pots and alfalfa in wet pots than AM fungi present in the perennial monoculture soil. Kernza did not benefit from AM fungi. Crop mixtures that included Kernza overyielded, but overyielding depended upon inoculation. Specifically, mixtures with Kernza overyielded most strongly in sterile soil as Kernza compensated for poor growth of Silphium and alfalfa. This study identifies the importance of soil biota and the context dependence of benefits of native microbes and the overyielding of mixtures in perennial crops. [ABSTRACT FROM AUTHOR]

Late embryogenesis abundant (LEA) proteins are widely involved in many adverse conditions among plants. In this study, we isolated a LEA4 gene from alfalfa (Medicago sativa L.) termed MsLEA4-4 via a homology cloning strategy. MsLEA4-4 encodes 166 amino acids, and the structural analysis showed that the gene contained five repeating TAQAAKEKTQQ amino acid motifs. There were a large number of α-helix in MsLEA4-4, and belongs to hydrophilic amino acid. Subcellular localization analysis showed that MsLEA4-4 was localized in the nucleus. The MsLEA4-4 promoter consisted of G-box and A-box elements, abscisic acid-responsive elements (ABREs), photo regulation and photoperiodic-controlling cis-acting elements, and endosperm expression motifs. The MsLEA4-4 overexpressing in Arabidopsis conferred late-germination phenotypes. Resistance of the overexpressed plants to abiotic stress significantly outperformed the wild-type (WT) plants. Under salt stress and abscisic acid treatment, with more lateral roots and higher chlorophyll content, the overexpressed plants has a higher survival rate measured against WT. Compared to those in the WT plants, the levels of soluble sugar and the activity of various antioxidant enzymes were elevated in the overexpressed plants, whereas the levels of proline and malondialdehyde were significantly reduced. The expression levels of several genes such as ABF3, ABI5, NCED5, and NCED9 increased markedly in the overexpressed plants compared to the WT under osmotic stress. [ABSTRACT FROM AUTHOR]

Rumen protected amino acids inclusion in ewes' diets has been proposed to enhance their innate immunity. The objective of this work was to determine the impact of dietary supplementation with rumen-protected methionine or lysine, as well as with a combination of these amino acids in two different ratios, on the expression of selected key-genes (NLRs, MyD88, TRIF, MAPK-1, IRF-3, JunD, TRAF-3, IRF-5, IL-1α, IL-10, IKK-α, STAT-3 and HO-1). Thus, sixty Chios dairy ewes (Ovis aries) were assigned to one of the following five dietary treatments (12 animals/ treatment): A: basal diet consist of concentrates, wheat straw and alfalfa hay (control group); B: basal diet +6.0 g/head rumen-protected methionine; C: basal diet + 5.0 g/head rumen-protected lysine; D: basal diet +6.0 g/head rumen-protected methionine + 5.0 g/head rumen-protected lysine and E: basal diet +12.0 g/head rumen-protected methionine + 5.0 g/head rumen-protected lysine. The results revealed a significant downregulation of relative transcript level of the IL-1α gene in the neutrophils of C and in monocytes of D ewes compared with the control. Significantly lower mRNA transcript accumulation was also observed for the MyD88 gene in the neutrophils of ewes fed with lysine only (C). The mRNA relative expression levels of JunD gene were highly induced in the monocytes, while those of IL-10 and HO-1 genes were declined in the neutrophils of ewes fed with the C and D diets compared with the control. Lower transcript levels of STAT-3 gene were observed in the neutrophils of ewes fed with either C or with E diets in comparison with the control. In conclusion, our results suggest that the dietary supplementation of ewes with rumen-protected amino acids, down regulate the expression of some genes involved in the pro-inflammatory signalling. [ABSTRACT FROM AUTHOR]

This study investigated the associations between abundance of rumen ciliate protozoa and the proportion of the main bioactive fatty acids related to rumen biohydrogenation, as 18:0, t10-18:1, t11-18:1, c9,t11-18:2, 18:3n-3 and 18:2 n-6, in rumen and meat of growing lambs, using data derived from 3 production experiments. A global correlation analysis and a linear regression analysis considering the effect of the experiment were performed. Ten of the 86 lambs involved in the experiments did not present ciliate cells in rumen liquor and the remaining lambs presented an average of 1.35 × 106ciliates / ml rumen liquor. From the nine genera of ciliates identified, Entodinium was the most abundant, averaging 1.17 × 106 cells / ml of rumen liquor. A large variation among lambs was observed for both rumen concentration and community structure of ciliates. Rumen t11-18:1 (P < 0.001) and meat deposition of t11-18:1 (P < 0.001) and of c9,t11-18:2 (P < 0.001) increased linearly with total ciliates, whereas the t10/t11 ratio in rumen (P = 0.002) and in meat (P = 0.036) decreased linearly. Entodiniomorphids seems to be strongly related with meat deposition oft11-18:1 and c9,t11-18:2 and with the reduction of the trans-10 shifted pathway. Completeness of RBH decreased linearly with Holotrichs (P = 0.029), Entodiniomorphids (P = 0.029), Isotricha (P = 0.011) and Epidinium (P = 0.027) abundances. Rumen 18:0 also decreased linearly with increasing counts of total ciliates (P = 0.015), Holotrichs (P = 0.020), Entodiniomorphids (P = 0.010) and Isotricha (P = 0.014). Rumen protozoa were positively linked with the deposition of healthy bioactive FA and simultaneously negatively associated with the occurrence of trans-10 shift. [ABSTRACT FROM AUTHOR]

Flavonoids are one of the largest secondary metabolite groups, which are widely present in plants. Flavonoids include anthocyanins, proanthocyanidins, flavonols and isoflavones. In particular, proanthocyanidins possess beneficial effects for ruminant animals in preventing lethal pasture bloat. As a major legume forage, alfalfa (Medicago sativa) contains little proanthocyanidins in foliage to combat bloat. In an attempt to improve proanthocyanidin content in alfalfa foliage, we over-expressed two MYB transcription factors (CsMYB5-1 and CsMYB5-2) from tea plant that is rich in proanthocyanidins. We showed that, via targeted metabolite and transcript analyses, the transgenic alfalfa plants accumulated higher levels of flavonoids in stems/leaves than the control, in particular anthocyanins and proanthocyanidins. Over-expression of CsMYB5-1 and CsMYB5-2 induced the expression levels of genes involved in flavonoid pathway, especially anthocyanin/proanthocyanidin-specific pathway genes DFR, ANS and ANR in stems/leaves. Both anthocyanin/proanthocyanidin content and the expression levels of several genes were conversely decreased in flowers of the transgenic lines than in control. Our results indicated that CsMYB5-1 and CsMYB5-2 differently regulate anthocyanins/proanthocyanidins in stems/leaves and flowers. Our study provides a guide for increasing anthocyanin/proanthocyanidin accumulation in foliage of legume forage corps by genetic engineering. These results also suggest that it is feasible to cultivate new varieties for forage production to potentially solve pasture bloat, by introducing transcription factors from typical plants with high proanthocyanidin level. [ABSTRACT FROM AUTHOR]

Landscape complexity influences soybean aphid suppression by generalist predators in North America, but the role of adjacent habitats as sources of these predators has not been studied directly. We quantified movement of aphidophagous predators between soybean and five adjacent habitats common in Manitoba using bi-directional Malaise traps. To test the contribution of predators from neighboring habitats to soybean aphid suppression, we performed experimental manipulations in adjacent soybean and alfalfa fields and monitored the movement of sevenspotted lady beetles, Coccinella septempunctata, using mark-release-recapture experiments. The identity of adjacent habitats affected the net movement of predators into soybean. The most abundant predators were hover flies (Diptera: Syrphidae), moving from woodlands to soybean. Similar (but non-significant) trends were found for lady beetles, minute pirate bugs, and green and brown lacewings. There was also a net movement of hover flies and green lacewings from soybean to canola. Lady beetles showed higher bidirectional movement in alfalfa and wheat borders than in woodland and canola borders in a high lady beetle abundance year. Soybean aphid populations in predator exclusion cages were 21- to 122- fold higher than populations exposed to predators, both in alfalfa and soybean fields. Aerial predators provide similar levels of aphid suppression as aerial and epigeal predators combined. Mark-release-recapture experiments showed high dispersal of C. septempunctata between soybean and alfalfa, with a net movement towards alfalfa, probably due to the lack of aphids in soybean. These results demonstrate that predator assemblages from both soybeans and alfalfa can suppress soybean aphids. Our findings indicate that the type of adjacent habitat and predator identity affect the directionality of predator movement into soybean. This study suggests that information on predator movement can be used to design the distribution of crops and natural habitats in agricultural landscapes that maximize pest control services. [ABSTRACT FROM AUTHOR]

Diet composed of smaller particles can improve feed intake, digestibility, and animal growth or health, but in ruminant species can reduce rumination and buffering–the loss of which may inhibit fermentation and digestibility. However, the explicit effect of particle size on the rumen microbiota remains untested, despite their crucial role in digestion. We evaluated the effects of reduced particle size on rumen microbiota by feeding long-stem (loose) alfalfa hay compared to a ground and pelleted version of the same alfalfa in yearling sheep wethers during a two-week experimental period. In situ digestibility of the pelleted diet was greater at 48 h compared with loose hay; however, distribution of residual fecal particle sizes in sheep did not differ between the dietary treatments at any time point (day 7 or 14). Both average daily gain and feed efficiency were greater for the wethers consuming the pelleted diet. Observed bacterial richness was very low at the end of the adaptation period and increased over the course of the study, suggesting the rumen bacterial community was still in flux after two weeks of adaptation. The pelleted-hay diet group had a greater increase in bacterial richness, including common fibrolytic rumen inhabitants. The pelleted diet was positively associated with several Succiniclasticum, a Prevotella, and uncultured taxa in the Ruminococcaceae and Rickenellaceae families and Bacteroidales order. Pelleting an alfalfa hay diet for sheep does shift the rumen microbiome, though the interplay of diet particle size, retention and gastrointestinal transit time, microbial fermentative and hydrolytic activity, and host growth or health is still largely unexplored. [ABSTRACT FROM AUTHOR]

The correlations between growth and wool traits in response to canola and flaxseed oil supplementation were evaluated in Australian prime lambs. Sixty dual-purpose prime lambs including purebred Merino and crossbred lambs were allocated to one of five treatments of lucerne hay basal diet supplemented with isocaloric and isonitrogenous wheat-based pellets. Treatments were: no oil inclusion (Control); 2.5% canola oil; 5% canola oil; 2.5% flaxseed oil and 5% flaxseed oil, with lamb groups balanced by breed and gender. Each lamb was daily supplemented with 1kg of pellets and had free access to lucerne hay and water throughout the 7-week feeding trial, after a 3-week adaptation. Individual animal basal and supplementary pellet feed intakes were recorded daily, while body conformation traits, body condition scores and liveweights were measured on days 0, 21, 35 and 49. The lambs were dye-banded on the mid-side and shorn before commencing the feeding trial and mid-side wool samples were collected from the same dye-banded area of each lamb at the end of the experiment. Correlations between wool quality traits and lamb performance were non-significant (P>0.05). Oil supplementation had no detrimental effect on lamb growth and wool quality traits (P > 0.05). Gender significantly affected wither height gain and fibre diameter. There were significant interactions between oil supplementation and lamb breed on chest girth. The correlations between clean fleece yield (CFY) and other wool quality traits were moderate ranging from 0.29 to 0.55. Moderate to high correlations between fibre diameter (FD) and other wool quality traits were detected (0.46–0.99) with the strongest relationship between FD and wool spinning fineness (SF). The relationship between CFY and wool comfort factor (CF) were positive, while negative relationships between CFY and the others were observed. A combination of 5% oil supplementation and genetics is an effective and strategic management tool for enhancing feed efficiency and growth performance without negative effects on wool quality in dual-purpose lamb production. This is a good outcome for dual-purpose sheep farmers. It essentially means the absorbed nutrients in supplemented lambs yielded good growth performance without any detrimental impact on wool quality; a win-win case of nutrient partitioning into the synthesis of muscle and wool without compromising either traits. [ABSTRACT FROM AUTHOR]

Information on disease process and pathogenicity mechanisms is important for understanding plant disease. Spring black stem and leaf spot caused by the necrotrophic pathogen Phoma medicaginis var. medicaginis Malbr. & Roum causes large losses to alfalfa. However, till now, little is known about alfalfa-P. medicagnis interactions and the pathogenicity mechanisms of the pathogen. Here, alfalfa inoculated with P. medicaginis was subjected to GC-MS based metabolic profiling. The metabolic response in P. medicaginis-inoculated and mock-inoculated alfalfa leaves was assessed at 2, 4, 6, 8, 12, 16, 20, 24, 26 and 28 days post inoculation. In total, 101 peaks were detected in the control and inoculated groups, from which 70 metabolites were tentatively identified. Using multivariate analysis, 16 significantly regulated compounds, including amino acids, nitrogen-containing compounds and organic acids, polyols, fatty acids, and sugars were tentatively identified (Variable importance values, VIP>1.0 and p <0.05). Among these metabolites, the levels of malate, 5-oxoproline, palmitic acid and stearic acid were increased significantly in P. medicaginis-infected alfalfa leaves compared to the controls. In contrast, the levels ofγ-aminobutyric acid and 2-pyrrolidinone were significantly decreased in infected leaves compared to the controls. Further metabolic pathway analysis of the 16 significantly regulated compounds demonstrated that glycolysis, the tricarboxylic acid cycle, and β-oxidation of fatty acids were significantly induced in the alfalfa leaves at later stages of P. medicaginis infection. The strong induction of tricarboxylic acid cycle pathways at later infection stages caused by the pathogen may induce senescence in the alfalfa leaves, leading to plant death. However, intermediate metabolites of these metabolic pathways, and inositol phosphate, glutathione, the metabolic pathways of some amino acids accumulated rapidly and strongly at early stages of infection, which may enhance the ability of alfalfa to resist necrotrophic P. medicaginis disease. Understanding metabolic pathways is essential for understanding pathogenesis. [ABSTRACT FROM AUTHOR]

MYB transcription factors are important regulators of the plant response to abiotic stress. Their participation in the salinity stress of the key forage legume species alfalfa (Medicago sativa) was investigated here by comparing the transcriptomes of the two cultivars Dryland (DL) and Sundory (SD), which differed with respect to their ability to tolerate salinity stress. When challenged by the stress, DL plants were better able than SD ones to scavenge reactive oxygen species. A large number of genes encoding transcription regulators, signal transducers and proteins involved in both primary and secondary metabolism were differentially transcribed in the two cultivars, especially when plants were subjected to salinity stress. The set of induced genes included 17 MYB family of transcription factors, all of which were subsequently isolated. The effect of constitutively expressing these genes on the salinity tolerance expressed by Arabidopsis thaliana was investigated. The introduction of MsMYB4 significantly increased the plants’ salinity tolerance in an abscisic acid-dependent manner. A sub-cellular localization experiment and a transactivation assay indicated that MsMYB4 was deposited in the nucleus and was able to activate transcription in yeast. Based on this information, we propose that the MsMYB4 products is likely directly involved in alfalfa’s response to salinity stress. [ABSTRACT FROM AUTHOR]

The world's annual output of rabbits is over 1.2 billion, therefore this sector is also one of the sources of greenhouse gases in livestock production. One hundred-twenty New Zealand rabbits were allocated into four treatments, five replicates in each treatment and six rabbits in each replicate to examine the effect of grinding alfalfa hay to different sizes on growth performance, methane production and cecal archaeal populations. The particle sizes of the alfalfa meal in the four treatment diets were 2500, 1000, 100 and 10 μm, while the other ingredients were ground through a 2.5 mm sieve. The average daily gain (ADG) and average daily feed intake (ADFI) increased (P<0.001) as the particle size decreased, but the feed conversion ratio (FCR) was not affected (P = 0.305). The digestibility of neutral detergent fiber (NDF) (P = 0.006) and acid detergent fiber (ADF) (P<0.006) increased while the greatest digestibility of crude protein (CP) was obtained in 1000 um group (P = 0.015). The rabbits produced more methane (CH4, L/kgBM0.75/d) with decreasing alfalfa particle size (P<0.001). The molar proportion of acetic acid and propionic acid decreased (P<0.001) at the cost of butyric acid (P<0.001). The greatest villus height:crypt depth ratio were obtained in 1000 μm group, and the decrease in the alfalfa hay particle size decreased the jejunum and ilem villus height:crypt depth ratio (P<0.05). The gastric muscular and mucosal thickness decreased with decreasing alfalfa particle size (P<0.05). Archaea diversity decreased with decreasing alfalfa particle size, and the relative abundance of genus Methanobrevibacter increased (P<0.001) while the genus Methanosphaera decreased (P<0.001). It is concluded that a finer particle size favors the growth of genus Methanobrevibacter, which produces more methane but promotes the growth performance of rabbits. [ABSTRACT FROM AUTHOR]

Viral infections of alfalfa are widespread in major cultivation areas and their impact on alfalfa production may be underestimated. A new viral species, provisionally named alfalfa virus F (AVF), was identified using a virion-associated nucleic acid (VANA) metagenomics-based approach in alfalfa (Medicago sativa L.) samples collected in Southern France. The nucleotide sequence of the viral genome was determined by de-novo assembly of VANA reads and by 5’/3’ RACE with viral RNA extracted from enriched viral particles or with total RNA, respectively. The virus shares the greatest degree of overall sequence identity (~78%) with Medicago sativa marafivirus 1 (MsMV1) recently deduced from alfalfa transcriptomic data. The tentative nucleotide sequence of the AVF coat protein shares ~83% identity with the corresponding region of MsMV1. A sequence search of the predicted single large ORF encoding a polyprotein of 235kDa in the Pfam database resulted in identification of five domains, characteristic of the genus Marafivirus, family Tymoviridae. The AVF genome also contains a conserved “marafibox”, a 16-nt consensus sequence present in all known marafiviruses. Phylogenetic analysis of the complete nucleotide sequences of AVF and other viruses of the family Tymoviridae grouped AVF in the same cluster with MsMV1. In addition to 5’ and 3’ terminal extensions, the identity of the virus was confirmed by RT-PCRs with primers derived from VANA-contigs, transmission electron microscopy with virus-infected tissues and transient expression of the viral coat protein gene using a heterologous virus-based vector. Based on the criteria demarcating species in the genus Marafivirus that include overall sequence identity less than 80% and coat protein identity less than 90%, we propose that AVF represents a distinct viral species in the genus Marafivirus, family Tymoviridae. [ABSTRACT FROM AUTHOR]

The role of rhizosphere microbial communities in the degradation of hydrocarbons remains poorly understood and is a field of active study. We used high throughput sequencing to explore the rhizosphere microbial diversity in the alfalfa and barley planted oil contaminated soil samples. The analysis of 16s rRNA sequences showed Proteobacteria to be the most enriched (45.9%) followed by Bacteriodetes (21.4%) and Actinobacteria (10.4%) phyla. The results also indicated differences in the microbial diversity among the oil contaminated planted soil samples. The oil contaminated planted soil samples showed a higher richness in the microbial flora when compared to that of untreated samples, as indicated by the Chao1 indices. However, the trend was different for the diversity measure, where oil contaminated barley planted soil samples showed slightly lower diversity indices. While the clustering of soil samples grouped the oil contaminated samples within and across the plant types, the clean sandy soil samples formed a separate group. The oil contaminated rhizosphere soil showed an enrichment of known oil-degrading genera, such as Alcanivorax and Aequorivita, later being specifically enriched in the contaminated soil samples planted with barley. Overall, we found a few well known oil-degrading bacterial groups to be enriched in the oil contaminated planted soil samples compared to the untreated samples. Further, phyla such as Thermi and Gemmatimonadetes showed an enrichment in the oil contaminated soil samples, indicating their potential role in hydrocarbon degradation. The findings of the current study will be useful in understanding the rhizosphere microflora responsible for oil degradation and thus can help in designing appropriate phytoremediation strategies for oil contaminated lands. [ABSTRACT FROM AUTHOR]

A suitable irrigation method adopted to arid desert conditions, including a special soil structure and specialized plants, has been continuously studied and improved. A field study was conducted in the Awei irrigation area of Aletai in Xinjiang in 2015 and 2016 to investigate the applicability of shallow subsurface drip irrigation (SSDI) in an arid desert area. A completely randomized block design with three replications and three treatments for drip tape subsurface depths at 5, 10, 20 cm was established. The results indicated that the vertical distribution of the soil moisture of subsurface drip irrigation (SDI, buried depth at 20 cm) was mainly concentrated at 0–60 cm, while SSDI (buried depth at 5 and 10 cm) was concentrated at 0–30 cm. However, the roots distributions were concentrated at 0–30 cm for SDI and SSDI. The chlorophyll content and water consumption intensity for alfalfa first increased and then decreased in arid desert conditions. The dry yield and water use efficiency (WUE) of SSDI (buried depth at 10 cm) were higher than those of SDI. The SSDI was practical in arid desert conditions and the recommended buried depth was 10 cm. [ABSTRACT FROM AUTHOR]

The WRKY transcription factors play an important role in the regulation of transcriptional reprogramming associated with plant abiotic stress responses. In this study, the WRKY transcription factor MsWRKY11, containing the plant-specific WRKY zinc finger DNA–binding motif, was isolated from alfalfa. The MsWRKY11 gene was detected in all plant tissues (root, stem, leaf, flower, and fruit), with high expression in root and leaf tissues. MsWRKY11 was upregulated in response to a variety of abiotic stresses, including salinity, alkalinity, cold, abscisic acid, and drought. Overexpression of MsWRKY11 in soybean enhanced the salt tolerance at the seedling stage. Transgenic soybean had a better salt-tolerant phenotype, and the hypocotyls were significantly longer than those of wild-type seeds after salt treatment. Furthermore, MsWRKY11 overexpression increased the contents of chlorophyll, proline, soluble sugar, superoxide dismutase, and catalase, but reduced the relative electrical conductivity and the contents of malonaldehyde, H2O2, and O2-. Plant height, pods per plant, seeds per plant, and 100-seed weight of transgenic MsWRKY11 soybean were higher than those of wild-type soybean, especially OX2. Results of the salt experiment showed that MsWRKY11 is involved in salt stress responses, and its overexpression improves salt tolerance in soybean. [ABSTRACT FROM AUTHOR]

Bacterial stem blight caused by Pseudomonas syringae pv. syringae is a common disease of alfalfa (Medicago sativa L). Little is known about host-pathogen interactions and host defense mechanisms. Here, individual resistant and susceptible plants were selected from cultivars Maverick and ZG9830 and used for transcript profiling at 24 and 72 hours after inoculation (hai) with the isolate PssALF3. Bioinformatic analysis revealed a number of differentially expressed genes (DEGs) in resistant and susceptible genotypes. Although resistant plants from each cultivar produced a hypersensitive response, transcriptome analyses indicated that they respond differently at the molecular level. The number of DEGs was higher in resistant plants of ZG9830 at 24 hai than in Maverick, suggesting that ZG9830 plants had a more rapid effector triggered immune response. Unique up-regulated genes in resistant ZG9830 plants included genes encoding putative nematode resistance HSPRO2-like proteins, orthologs for the rice Xa21 and soybean Rpg1-b resistance genes, and TIR-containing R genes lacking both NBS and LRR domains. The suite of R genes up-regulated in resistant Maverick plants had an over-representation of R genes in the CC-NBS-LRR family including two genes for atypical CCR domains and a putative ortholog of the Arabidopsis RPM1 gene. Resistance in both cultivars appears to be mediated primarily by WRKY family transcription factors and expression of genes involved in protein phosphorylation, regulation of transcription, defense response including synthesis of isoflavonoids, and oxidation-reduction processes. These results will further the identification of mechanisms involved in resistance to facilitate selection of parent populations and development of commercial varieties. [ABSTRACT FROM AUTHOR]

Fall dormancy (FD) determines the adaptation of an alfalfa variety and affects alfalfa production and quality. However, the molecular mechanism underlying FD remains poorly understood. Here, 44 genes regulating FD were identified by comparison of the transcriptomes from leaves of Maverick (fall-dormant alfalfa) and CUF101(non-fall-dormant), during FD and non-FD and were classified them depending on their function. The transcription of IAA-amino acid hydrolase ILR1-like 1, abscisic acid receptor PYL8, and monogalactosyldiacylglycerol synthase-3 in Maverick leaves was regulated by daylength and temperature, and the transcription of the abscisic acid receptor PYL8 was mainly affected by daylength. The changes in the expression of these genes and the abundance of their messenger RNA (mRNA) in Maverick leaves differed from those in CUF101 leaves, as evidenced by the correlation analysis of their mRNA abundance profiles obtained from April to October. The present findings suggested that these genes are involved in regulating FD in alfalfa. [ABSTRACT FROM AUTHOR]

Conservation tillage is expected to have a positive effect on soil physical properties, soil Carbon (C) storage, while reducing fuel, labour and machinery costs. However, reduced tillage could increase soil nitrous oxide (N2O) emissions and offset the expected gains from increased C sequestration. To date, conservation tillage is barely practiced or studied in Bosnia and Herzegovina (BH). Here, we report a field study on the short-term effects of reduced (RT) and no tillage (NT) on N2O emission dynamics, yield-scaled N2O emissions, soil structure and the economics of cereal production, as compared with conventional tillage (CT). The field experiment was conducted in the Sarajevo region on a clayey loam under typical climatic conditions for humid, continental BH. N2O emissions were monitored in a Maize-Barley rotation over two cropping seasons. Soil structure was studied at the end of the second season. In the much wetter 2014, N2O emission were in the order of CT > RT > NT, while in the drier 2015, the order was RT > CT > NT. The emission factors were within or slightly above the uncertainty range of the IPCC Tier 1 factor, if taking account for the N input from the cover crop (alfalfa) preceding the first experimental year. Saturated soils in spring, formation of soil crusts and occasional droughts adversely affected yields, particularly in the second year (barley). In 2014, yield-scaled N2O emissions ranged from 83.2 to 161.7 g N Mg-1 grain (corn) but were much greater in the second year due to crop failure (barley). RT had the smallest yield-scaled N2O emission in both years. NT resulted in economically inacceptable returns, due to the increased costs of weed control and low yields in both years. The reduced number of operations in RT reduced production costs and generated positive net returns. Therefore, RT could potentially provide agronomic and environmental benefits in crop production in BH. [ABSTRACT FROM AUTHOR]

In nature, plants are often exposed to multiple stress factors at the same time. Yet, little is known about how plants modulate their physiology to counteract simultaneous abiotic and biotic stresses, such as soil salinity and insect herbivory. In this study, insect performance bioassays, phytohormone measurements, quantification of transcripts, and protein determination were employed to study the phenotypic variations of two alfalfa (Medicago sativa) cultivars in response to insect Spodoptera litura feeding under normal and salt stress condition. When being cultivated in normal soil, the salt-tolerant alfalfa cultivar Zhongmu-1 exhibited lower insect resistance than did the salt-sensitive cultivar Xinjiang Daye. Under salinity stress, the defense responses of Xinjiang Daye were repressed, whereas Zhongmu-1 did not show changes in resistance levels. It is likely that salinity influenced the resistance of Xinjiang Daye through suppressing the accumulation of jasmonic acid-isoleucine (JA-Ile), which is the bioactive hormone inducing herbivore defense responses, leading to attenuated trypsin proteinase inhibitor (TPI) activity. Furthermore, exogenous ABA supplementation suppressed the insect herbivory-induced JA/JA-Ile accumulation and levels of JAR1 (jasmonate resistant 1) and TPI, and further decreased the resistance of Xinjiang Daye, whereas Zhongmu-1 showed very little response to the increased ABA level. We propose a mechanism, in which high levels of abscisic acid induced by salt treatment may affect the expression levels of JAR1 and consequently decrease JA-Ile accumulation and thus partly suppress the defense of Xinjiang Daye against insects under salt stress. This study provides new insight into the mechanism by which alfalfa responds to concurrent abiotic and biotic stresses. [ABSTRACT FROM AUTHOR]

To investigate the effects of calcium propionate (CaP) supplementation on performance, the development of the internal organ, and gastrointestinal tract of calves at various stages of growth period, 54 male Jersey calves (age = 7 ± 1 d, body weight(BW) = 23.1 ± 1.2 kg) were randomly allocated to three treatment groups. While control calves were fed basis dietary with no additives (0CaP), other treatment calves were fed basis dietary supplementation with CaP at 50 (5CaP) or 100 (10CaP) g kg−1 dry matter. The experiment lasted 160 d and was divided into three feeding stages: Stage 1 (d 0 to 30), Stage 2 (d 31 to 90), and Stage 3 (d 91 to160). Six calves from each group were randomly selected and slaughtered on days 30, 90, and 160 when at the conclusion of each experimental feeding stage. The BW of calves increased with 10CaP after feeding 90 d, whereas it increased with 5CaP and 10CaP at feeding 120d and 160d compared to 0CaP. The 10CaP group improved average daily gain (ADG) of calves at stage 2, and d120-160 of stage 3 compared with the 0CaP group. The ADG of 5CaP was greater than the 0CaP group only at 120–160 d of stage 3 compared with the 0CaP group. The results of feed efficiency were in agreed with ADG as no dry matter intake difference at all stages of growth period. The 10CaP treatment exhibited the greatest spleen weight among the treatment at the end of the experiment; the liver weight of the 5CaP and 10CaP calves at feeding 90 d and of the 10CaP calves at feeding 160 d and were greater than those of the 0CaP animals. The CaP at the tested doses increased the rumen weight after feeding 90d of Jersey calves, and also improved the development of intestine. In conclusion, dietary supplementation with calcium propionate at the tested doses caused a beneficial effect in the growth performance and gastrointestinal tract traits of Jersey calves, thus to add 10% CaP before feeding 90 days was better and 5% CaP supplementation was expected at the period for feeding 90 to160 d. [ABSTRACT FROM AUTHOR]

In an effort to axenically culture the previously uncultivable populations of the rhizobacteria of Lucerne (Medicago sativa L.), we propose plant-only teabags culture media to mimic the nutritional matrix available in the rhizosphere. Here, we show that culture media prepared from Lucerne powder teabags substantially increased the cultivability of Lucerne rhizobacteria compared with a standard nutrient agar, where we found that the cultivable populations significantly increased by up to 60% of the total bacterial numbers as estimated by Quantitative Real-time Polymerase Chain Reaction (qRT-PCR). Cluster analysis of 16S rDNA Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) of cultivable Colony-Forming Units (CFUs) revealed a more distinct composition and separation of bacterial populations recovered on the plant-only teabags culture media than those developed on a standard nutrient agar. Further, the new plant medium gave preference to the micro-symbiont Sinorhizobium meliloti, and succeeded in isolating a number of not-yet-cultured bacteria, most closely matched to Novosphingobium sp., Lysobacter sp. and Pedobacter sp. The present study may encourage other researchers to consider moving from the well-established standard culture media to the challenging new plant-only culture media. Such a move may reveal previously hidden members of rhizobacteria, and help to further explore their potential environmental impacts. [ABSTRACT FROM AUTHOR]

Selenium (Se) is an essential trace mineral important for immune function and overall health of cattle. The nasopharyngeal microbiota in cattle plays an important role in overall respiratory health, especially when stresses associated with weaning, transport, and adaptation to a feedlot affect the normal respiratory defenses. Recent evidence suggests that cattle diagnosed with bovine respiratory disease complex have significantly less bacterial diversity. The objective of this study was to determine whether feeding weaned beef calves Se-enriched alfalfa (Medicago sativa) hay for 9 weeks in a preconditioning program prior to entering the feedlot alters nasal microbiota. Recently weaned beef calves (n = 45) were blocked by sex and body weight, randomly assigned to 3 treatment groups with 3 pens of 5 calves per treatment group, and fed an alfalfa hay based diet for 9 weeks. Alfalfa hay was harvested from fields fertilized with sodium selenate at a rate of 0, 45.0 or 89.9 g Se/ha. Blood samples were collected biweekly and analyzed for whole-blood Se concentrations. Nasal swabs were collected during week 9 from one or two calves from each pen (total n = 16). Calculated Se intake from dietary sources was 3.0, 15.6, and 32.2 mg Se/head/day for calves consuming alfalfa hay with Se concentrations of 0.34 to 2.42 and 5.17 mg Se/kg dry matter, respectively. Whole-blood Se concentrations after 8 weeks of feeding Se-fertilized alfalfa hay were dependent upon Se-application rates (0, 45.0, or 89.9 g Se/ha) and were 155, 345, and 504 ng/mL (PLinear < 0.0001). Microbial DNA was extracted from nasal swabs and amplified and sequenced. Alpha rarefaction curves comparing the species richness (observed OTUs) and overall diversity (Chao1, Observed OTU, and Shannon index) between calves fed selenium-biofortified alfalfa hay compared with control calves showed that Se-supplementation tended to be associated with an enriched nasal microbiota. ANOSIM of unweighted UniFrac distances showed that calves fed high Se-biofortified alfalfa hay clustered separately when compared with control calves in the PCoA plot (R = 0.216, P = 0.04). The bacterial orders Lactobacillales and Flavobacteriales were increased in healthy control calves compared with Clostridiales and Bacteroidales being increased in calves fed Se-biofortified alfalfa hay. Although there were strong trends, no significant differences were noted for any of the bacterial taxa. Based upon these findings, we suggest that weaned beef calves fed Se-biofortified hay tend to have an enriched nasal microbiota. Feeding Se-biofortified alfalfa hay to weaned beef calves prior to entering the feedlot is a strategy for increasing nasopharyngeal microbial diversity. [ABSTRACT FROM AUTHOR]

A new species of the family Alphaflexiviridae provisionally named alfalfa virus S (AVS) was discovered in alfalfa samples originating from Sudan. A complete nucleotide sequence of the viral genome consisting of 8,349 nucleotides excluding the 3’ poly(A) tail was determined by high throughput sequencing (HTS) on an Illumina platform. NCBI BLAST searches revealed that the virus shares the greatest degree of sequence identity with members of the family Alphaflexiviridae, genus Allexivirus. The AVS genome contains six computationally-predicted open reading frames (ORF) encoding viral replication protein, triple gene block protein 1 (TGB1), TGB2, TGB3-like protein, unknown 38.4 kDa protein resembling serine-rich 40 kDa protein characteristic for allexiviruses, and coat protein (CP). AVS lacks a clear 3’ proximal ORF that encodes a nucleic acid-binding protein typical for allexiviruses. The identity of the virus was confirmed by RT-PCR with primers derived from the HTS-generated sequence, dot blot hybridization with DIG-labeled virus-specific RNA probes, and Western blot analysis with antibodies produced against a peptide derived from the CP sequence. Transmission electron microscopic observations of the infected tissues showed the presence of filamentous particles similar to allexiviruses in their length and appearance. To the best of our knowledge, this is the first report on the identification of a putative allexivirus in alfalfa (Medicago sativa). The genome sequence of AVS has been deposited in NCBI GenBank on 03/02/2016 as accession № KY696659. [ABSTRACT FROM AUTHOR]

Many communities in the Midwestern United States obtain their drinking water from shallow alluvial wells that are vulnerable to contamination by NO3-N from the surrounding agricultural landscape. The objective of this research was to assess cropping systems with the potential to produce a reasonable return for farmers while simultaneously reducing the risk of NO3-N movement into these shallow aquifers. From 2009 to 2013 we conducted a field experiment in northwest Iowa in which we evaluated five cropping systems for residual (late fall) soil NO3-N content and profitability. Soil samples were taken annually from the top 30 cm of the soil profile in June and August, and from the top 180 cm in November (late fall). The November samples were divided into 30 cm increments for analysis. Average residual NO3-N content in the top 180 cm of the soil profile following the 2010 to 2013 cropping years was 134 kg ha-1 for continuous maize (Zea mays L.) with a cereal rye (Secale cereale L.) cover crop, 18 kg ha-1 for perennial grass, 60 kg ha-1 for a three year oat (Avena sativa L.)-alfalfa (Medicago sativa L.)-maize rotation, 85 kg ha-1 for a two year oat/red clover (Trifolium pratense L.)-maize rotation, and 90 kg ha-1 for a three year soybean (Glycine max (L.) Merr.)-winter wheat (Triticum aestivum L.)-maize rotation. However, residual NO3-N in the 90 to 180 cm increment of the soil profile was not significantly higher in the oat-alfalfa-maize cropping system than the perennial grass system. For 2010 to 2013, average profit ($ ha-1 yr-1) was 531 for continuous corn, 347 for soybean-winter wheat-maize, 264 for oat-alfalfa-maize, 140 for oat/red clover-maize, and -384 (loss) for perennial grass. Considering both residual soil NO3-N and profitability data, the oat-alfalfa-maize rotation performed the best in this setting. However, given current economic pressures widespread adoption is likely to require changes in public policy. [ABSTRACT FROM AUTHOR]

The structure and the activity of proteins are often regulated by transient or stable post- translational modifications (PTM). Different from well-known, abundant modifications such as phosphorylation and glycosylation some modifications are limited to one or a few proteins across a broad range of related species. Although few examples of the latter type are known, the evolutionary conservation of these modifications and the enzymes responsible for their synthesis suggest an important physiological role. Here, the first observation of a new, fold-directing PTM is described. During the analysis of alfalfa cell wall proteins a -2Da mass shift was observed on phenylalanine residues in the repeated tetrapeptide FxxY of the beta-subunit of polygalacturonase. This modular protein is known to be involved in developmental and stress-responsive processes. The presence of this modification was confirmed using in-house and external datasets acquired by different commonly used techniques in proteome studies. Based on these analyses it was found that all identified phenylalanine residues in the sequence FxxY of this protein were modified to α,β-didehydro-Phe (ΔPhe). Besides showing the reproducible identification of ΔPhe in different species arguments that substantiate the fold-determining role of ΔPhe are given. [ABSTRACT FROM AUTHOR]

Rhizobia-legume symbiosis is the most well researched biological nitrogen fixation system. Coating legume seeds with rhizobia is now a recognized practical measure for improving the production of legume corp. However, the efficacy of some commercial rhizobia inoculants cannot be guaranteed in China due to the low rate of live rhizobia in these products. A greenhouse experiment was conducted to assess the effects of different rhizobial inoculant formulations on alfalfa productivity and nitrogen fixation. Two rhizobia strains, (ACCC17631 and ACCC17676), that are effective partners with alfalfa variety Zhongmu No. 1 were assessed with different concentrations of ammonium molybdate in seed-coat formulations with two different coating adhesives. Our study showed that the growth, nodulation, and nitrogen fixation ability of the plants inoculated with the ACCC17631 rhizobial strain were greatest when the ammonium molybdate application was0.2% of the formulation. An ammonium molybdate concentration of 0.1% was most beneficial to the growth of the plants inoculated with the ACCC17676 rhizobial strain. The sodium carboxymethyl cellulose and sodium alginate, used as coating adhesives, did not have a significant effect on alfalfa biomass and nitrogen fixation. However, the addition of skimmed milk to the adhesive improved nitrogenase activity. These results demonstrate that a new rhizobial seed-coat formulation benefitted alfalfa nodulation and yield. [ABSTRACT FROM AUTHOR]

Genetic progress for forage quality has been poor in alfalfa (Medicago sativa L.), the most-grown forage legume worldwide. This study aimed at exploring opportunities for marker-assisted selection (MAS) and genomic selection of forage quality traits based on breeding values of parent plants. Some 154 genotypes from a broadly-based reference population were genotyped by genotyping-by-sequencing (GBS), and phenotyped for leaf-to-stem ratio, leaf and stem contents of protein, neutral detergent fiber (NDF) and acid detergent lignin (ADL), and leaf and stem NDF digestibility after 24 hours (NDFD), of their dense-planted half-sib progenies in three growing conditions (summer harvest, full irrigation; summer harvest, suspended irrigation; autumn harvest). Trait-marker analyses were performed on progeny values averaged over conditions, owing to modest germplasm × condition interaction. Genomic selection exploited 11,450 polymorphic SNP markers, whereas a subset of 8,494 M. truncatula-aligned markers were used for a genome-wide association study (GWAS). GWAS confirmed the polygenic control of quality traits and, in agreement with phenotypic correlations, indicated substantially different genetic control of a given trait in stems and leaves. It detected several SNPs in different annotated genes that were highly linked to stem protein content. Also, it identified a small genomic region on chromosome 8 with high concentration of annotated genes associated with leaf ADL, including one gene probably involved in the lignin pathway. Three genomic selection models, i.e., Ridge-regression BLUP, Bayes B and Bayesian Lasso, displayed similar prediction accuracy, whereas SVR-lin was less accurate. Accuracy values were moderate (0.3–0.4) for stem NDFD and leaf protein content, modest for leaf ADL and NDFD, and low to very low for the other traits. Along with previous results for the same germplasm set, this study indicates that GBS data can be exploited to improve both quality traits (by genomic selection or MAS) and forage yield. [ABSTRACT FROM AUTHOR]

Plant species diversity may benefit natural grassland productivity, but its effect in managed grassland systems is not well understood. A four-year multispecies grassland experiment was conducted to investigate the effect of species diversity–legumes and non-leguminous forbs–on productivity, persistence and sward quality under cutting or grazing regimes and with or without slurry application. Three mixtures were established– 3-mix: grass, red and white clover, 10-mix: 3-mix plus birdsfoot trefoil and six non-leguminous forbs, and 12-mix: 10-mix plus lucerne and festulolium. Species diversity increased sward production and yield persistence under cutting regime. The 12-mix had the highest yield from the second year onwards and no statistically significant yield reduction over four years, while annual yields in the 3-mix and 10-mix decreased significantly with increasing grassland age. The higher yield in the 12-mix was mainly due to the inclusion of high-yielding lucerne. The 10-mix and 12-mix had lower proportions of unsown species than the 3-mix, the difference being dependent on grassland age. Generally, the 3-mix had higher concentrations of in-vitro organic matter digestibility (IVOMD), neutral detergent fiber (NDF) and crude protein (CP), and a lower concentration of ash than the 10-mix and 12-mix. Slurry application increased annual yield production by 10% and changed the botanical composition, increasing the proportion of grass and decreasing the proportion of legumes. Compared to cutting, grazing increased forage production by 9% per cut on average and lowered legume and forb proportions in the mixtures, but yields did not differ among the three mixtures. Overall, our results suggest that species diversity increases sward productivity and persistence only under an ungrazed cutting regime. We conclude that increasing species diversity by selecting appropriate species with compatible management is key to achieving both high yields and high persistence in managed grasslands. [ABSTRACT FROM AUTHOR]

Common leaf spot (caused by Pseudopeziza medicaginis), rust (caused by Uromyces striatus), Leptosphaerulina leaf spot (caused by Leptosphaerulina briosiana) and Cercospora leaf spot (caused by Cercospora medicaginis) are the four common types of alfalfa leaf diseases. Timely and accurate diagnoses of these diseases are critical for disease management, alfalfa quality control and the healthy development of the alfalfa industry. In this study, the identification and diagnosis of the four types of alfalfa leaf diseases were investigated using pattern recognition algorithms based on image-processing technology. A sub-image with one or multiple typical lesions was obtained by artificial cutting from each acquired digital disease image. Then the sub-images were segmented using twelve lesion segmentation methods integrated with clustering algorithms (including K_means clustering, fuzzy C-means clustering and K_median clustering) and supervised classification algorithms (including logistic regression analysis, Naive Bayes algorithm, classification and regression tree, and linear discriminant analysis). After a comprehensive comparison, the segmentation method integrating the K_median clustering algorithm and linear discriminant analysis was chosen to obtain lesion images. After the lesion segmentation using this method, a total of 129 texture, color and shape features were extracted from the lesion images. Based on the features selected using three methods (ReliefF, 1R and correlation-based feature selection), disease recognition models were built using three supervised learning methods, including the random forest, support vector machine (SVM) and K-nearest neighbor methods. A comparison of the recognition results of the models was conducted. The results showed that when the ReliefF method was used for feature selection, the SVM model built with the most important 45 features (selected from a total of 129 features) was the optimal model. For this SVM model, the recognition accuracies of the training set and the testing set were 97.64% and 94.74%, respectively. Semi-supervised models for disease recognition were built based on the 45 effective features that were used for building the optimal SVM model. For the optimal semi-supervised models built with three ratios of labeled to unlabeled samples in the training set, the recognition accuracies of the training set and the testing set were both approximately 80%. The results indicated that image recognition of the four alfalfa leaf diseases can be implemented with high accuracy. This study provides a feasible solution for lesion image segmentation and image recognition of alfalfa leaf disease. [ABSTRACT FROM AUTHOR]

The effect of cropping system on the distribution of organic carbon (OC) and nitrogen (N) in soil aggregates has not been well addressed, which is important for understanding the sequestration of OC and N in agricultural soils. We analyzed the distribution of OC and N associated with soil aggregates in three unfertilized cropping systems in a 27-year field experiment: continuously cropped alfalfa, continuously cropped wheat and a legume-grain rotation. The objectives were to understand the effect of cropping system on the distribution of OC and N in aggregates and to examine the relationships between the changes in OC and N stocks in total soils and in aggregates. The cropping systems increased the stocks of OC and N in total soils (0–40 cm) at mean rates of 15.6 g OC m-2 yr-1 and 1.2 g N m-2 yr-1 relative to a fallow control. The continuous cropping of alfalfa produced the largest increases at the 0–20 cm depth. The OC and N stocks in total soils were significantly correlated with the changes in the >0.053 mm aggregates. 27-year of cropping increased OC stocks in the >0.053 mm size class of aggregates and N stocks in the >0.25 mm size class but decreased OC stocks in the <0.053 mm size class and N stocks in the <0.25 mm size class. The increases in OC and N stocks in these aggregates accounted for 99.5 and 98.7% of the total increases, respectively, in the continuous alfalfa system. The increases in the OC and N stocks associated with the >0.25 mm aggregate size class accounted for more than 97% of the total increases in the continuous wheat and the legume-grain rotation systems. These results suggested that long-term cropping has the potential to sequester OC and N in soils and that the increases in soil OC and N stocks were mainly due to increases associated with aggregates >0.053 mm. [ABSTRACT FROM AUTHOR]

Understanding the temporal and spatial variability in a crop yield is viewed as one of the key steps in the implementation of precision agriculture practices. Therefore, a study on a center pivot irrigated 23.5 ha field in Saudi Arabia was conducted to assess the variability in alfalfa yield using Landsat-8 imagery and a hay yield monitor data. In addition, the study was designed to also explore the potential of predicting the alfalfa yield using vegetation indices. A calibrated yield monitor mounted on a large rectangular hay baler was used to measure the actual alfalfa yield for four alfalfa harvests performed in the period from October 2013 to May 2014. A total of 18 Landsat-8 images, representing different crop growth stages, were used to derive different vegetation indices (VIs). Data from the yield monitor was used to generate yield maps, which illustrated a definite spatial variation in alfalfa yield across the experimental field for the four studied harvests as indicated by the high spatial correlation values (0.75 to 0.97) and the low P-values (4.7E-103 to 8.9E-27). The yield monitor-measured alfalfa actual yield was compared to the predicted yield form the Vis. Results of the study showed that there was a correlation between actual and predicted yield. The highest correlations were observed between actual yield and the predicted using NIR reflectance, SAVI and NDVI with maximum correlation coefficients of 0.69, 0.68 and 0.63, respectively. [ABSTRACT FROM AUTHOR]

From the perspective of an herbivorous insect, conspecific host plants are not identical, and intraspecific variation in host nutritional quality or defensive capacity might mediate spatially variable outcomes in plant-insect interactions. Here we explore this possibility in the context of an ongoing host breadth expansion of a native butterfly (the Melissa blue, Lycaeides melissa) onto an exotic host plant (alfalfa, Medicago sativa). We examine variation among seven alfalfa populations that differed in terms of colonization by L. melissa; specifically, we examined variation in phytochemistry, foliar protein, and plant population genetic structure, as well as responses of caterpillars and adult butterflies to foliage from the same populations. Regional patterns of alfalfa colonization by L. melissa were well predicted by phytochemical variation, and colonized patches of alfalfa showed a similar level of inter-individual phytochemical diversity. However, phytochemical variation was a poor predictor of larval performance, despite the fact that survival and weight gain differed dramatically among caterpillars reared on plants from different alfalfa populations. Moreover, we observed a mismatch between alfalfa supporting the best larval performance and alfalfa favored by ovipositing females. Thus, the axes of plant variation that mediate interactions with L. melissa depend upon herbivore life history stage, which raises important issues for our understanding of adaptation to novel resources by an organism with a complex life history. [ABSTRACT FROM AUTHOR]