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Frass, the residual material resulting from the bioconversion of organic matter by black soldier fly larvae (BSFL), has gained attention as a sustainable alternative to conventional fertilizers due to its high nutrient content. Additionally, frass has been found to possess antifungal properties, which can help control plant pathogens affecting horticultural crops. In this study, frass from BSFL reared on the Gainesville diet, a universally employed reference/control diet, was investigated in vitro for its effect on the growth of seven important fungal/oomycete pathogens. Dual culture overlay assays clearly showed that fresh Gainesville diet extract, as well as BSFL frass extract derived from this diet, contained microorganisms producing compound(s) that strongly inhibit(s) the mycelial growth of fungal/oomycete plant pathogens. Fungi and bacteria were then isolated from the fresh Gainesville diet and BSFL frass using the serial dilution technique. Among the different fungi/bacteria isolated, the isolate GV1-11 in Gainesville diet and FGV15-6 in frass demonstrated strong antifungal/anti-oomycete activity. Both isolates were genetically identified by whole-genome sequencing as Bacillus velezensis, a bacterium used as a biocontrol agent, strongly suggesting that B. velezensis, which is present in the Gainesville diet, can survive the process of BSFL rearing and is one of the key factors contributing to the observed antifungal and anti-oomycete activity in the resulting frass. This work underlines the importance of the inherent microbial characteristics of feedstocks on the antifungal/anti-oomycete activity of frass and points out the possibility of exploiting frass to control plant pathogens affecting horticultural crops.
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Alfalfa (Medicago sativa L.) cultivars developed for improved digestibility by conventional breeding or genetically modified (GM) have not been evaluated in eastern Canada. This study compared yield, nutritive value, and profitability of two reduced-lignin GM cultivars, three conventionally selected for low lignin or high-pectin, and one population with improved stem degradability, with two commercially adapted cultivars as controls. Alfalfa was harvested at the early bud (intensive management) or early flower (extensive management) stage of development during the first post-seeding year at three sites and the second post-seeding year at one site. All cultivars/population had similar annual dry matter (DM) yield, except for lower DM yield (−20%) of the improved stem degradability population. Conventionally selected cultivars for improved digestibility did not differ from control cultivars for in vitro DM digestibility (IVTD) and neutral detergent fiber digestibility (NDFd). Reduced-lignin GM cultivars, however, had a greater IVTD and NDFd (+10%), and less lignin (-10%) than control cultivars at the same stage of development. Reduced-lignin GM cultivars under extensive management had similar NDFd and greater annual DM yield (+1 to 2 Mg DM ha−1), but reduced total digestible nutrients (−41 to 44 g kg−1 DM), in comparison to control cultivars under intensive management. Using partial budget analysis the comparison of more digestible GM cultivars to the control cultivars resulted in an increase in annual farm net profits ranging from CAD$7.4 to $79.6 cow−1 yr−1 depending on stage of development at harvest. Further investigations are needed to quantify performance of dairy cows fed cultivars with improved digestibility.
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In the current study, a new hybrid machine learning (ML)-based model was developed by integrating a convolution neural network (CNN) with a random forest (RF) to forecast pesticide use on golf courses in Québec, Canada. Three main groups of independent variables were used to estimate pesticide use on golf courses, expressed as actual active ingredient rate (AAIR): (i) coordinates (i.e., longitude and latitude of the golf course), (ii) characteristics of the golf courses (i.e., pesticide type and the number of holes), and (iii) meteorological variables (i.e., total precipitation, P, and average temperature, T). The meteorological variables were collected from the Google Earth Engine by developing a JavaScript-based Code. On the basis of the different periods of total precipitation and average temperature, four different scenarios were defined. A data bank with more than 40,000 samples was used to calibrate and validate the developed model such that 70% of all samples were randomly selected to calibrate the model, while the remainder of the samples (i.e., 30%) that did not have any role in calibration were employed to validate the model’s generalizability. A comparison of different scenarios indicated that the model that considered the longitude and latitude of the golf course, pesticide type, and the number of holes in golf courses as well as total precipitation and average temperature from May to November as inputs (R = 0.997; NSE = 0.997; RMSE = 0.046; MAE = 0.026; NRMSE = 0.454; and PBIAS (%) = −0.443) outperformed the other models. Moreover, the sensitivity analysis result indicated that the total precipitation was the most critical variable in AAIR forecasting, while the average temperature, pesticide types, and the number of holes were ranked second to fourth, respectively.
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Background Pseudozyma flocculosa is a highly efficient biocontrol agent (BCA) of powdery mildews whose mode of action remains elusive. It is known to secrete unique effectors during its interaction with powdery mildews but effectors have never been shown to be part of the arsenal of a BCA. Here, we characterize the role of the effector Pf2826 released by Pseudozyma flocculosa during its tripartite interaction with barley and the pathogen fungus Blumeria graminis f. sp. hordei. Results We utilized CRISPR-Cas9-based genome editing and confirmed that secreted P. flocculosa effector Pf2826 is required for full biocontrol activity. We monitored the localization of the effector Pf2826 with C-terminal mCherry tag and found it localized around the haustoria and on powdery mildew spores. His-tagged Pf2826 recombinant protein was expressed, purified, and used as bait in a pull-down assay from total proteins extracted during the tripartite interaction. Potential interactors were identified by LC–MS/MS analysis after removing unspecific interactions found in the negative controls. A two-way yeast two-hybrid assay validated that Pf2826 interacted with barley pathogenesis-related (PR) proteins HvPR1a and chitinase and with an effector protein from powdery mildew. Conclusions In contrast to the usual modes of action of competition, parasitism, and antibiosis ascribed to BCAs, this study shows that effector pf2826 plays a vital role in the biocontrol activity of P. flocculosa by interacting with plant PR proteins and a powdery mildew effector, altering the host–pathogen interaction.
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Potato (Solanum tuberosum L.) crops are often cultivated in coarse-textured soils with low soil organic matter (SOM), and high nitrate leaching risk. Incorporating shrub willow chips into soil could enhance soil properties, while temporally immobilizing N and thus reducing N leaching. We performed a laboratory incubation study and a field experiment to evaluate the effects of shrub willow chips applied at increasing rates in the fall after the potato harvest on C, N and P cycling, soil pH and moisture, and on barley (Hordeum vulgare L.) yield in the following year. In comparison with the control, willow chip incorporation at the rates of 40 Mg ha-1 and 60 Mg ha-1 increased total C content, but it did not affect the activity of C cycling enzymes. Willow chip addition at these rates also induced nitrate immobilization and reduced barley grain yield and total N uptake, but increased the activity of N cycling enzymes (β-1,4-N-acetylglucosaminidase and leucine aminopeptidase). Mehlich-3 extractable P content and phosphomonoesterase activity were not affected by willow chip addition. Our results suggest that shrub willow chips increased total organic C and immobilized N following their incorporation and can thus mitigate nitrate leaching after the potato harvest. The N immobilization was short-lived and was not observed over second winter. We recommend to seed a forage legume in the spring following shrub willow chip incorporation. Willow chip incorporation is an effective means of increasing soil organic carbon.
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Broccoli is a vegetable that offers valuable components, such as glucosinolates (GLS), flavonoids, and hydroxycinnamic acids (HCA), for our daily food intake. These substances have been associated with reducing the risk of cancer and cardiovascular diseases (CVD). Broccoli florets are also highly perishable, given their elevated respiration rates and their sensitivity to ethylene. Experiments have been carried out on broccoli to investigate the consequences of abiotic stress post-harvest treatments in retarding the senescence process. Nevertheless, the influence of these treatments on the phytochemicals of broccoli has not been extensively examined. Florets of broccoli (Brassica oleracea) were exposed to an atmosphere consisting of 10,000 µL.L−1 ethanol at room temperature for 30 min and 120 min. The exposure to methyl jasmonate (MeJA) treatments was carried out at room temperature using 1 µL.L−1 for 45 min and 180 min. The yellowing of florets was delayed using 10,000 µL.L−1 of ethanol at both exposure times as compared to untreated florets, and the chlorophyll titers were also superior with both doses over the control. The total phenols of the florets increased by 15% and 18% with the application of the hormetic and high doses, respectively, throughout the storage period compared to unexposed broccoli. The GLS and HCA yields were also increased by both ethanol doses. The exposure of florets to 1 µL.L1 MeJA for 45 min resulted in delayed yellowing of florets; however, longer exposures resulted in yellowing after 21 d and significantly (p < 0.05) increased respiration rates relative to untreated florets. Overall antioxidant capacity of the florets was significantly reduced with both doses of methyl jasmonate; however, HCA titers were increased at both doses. The amount of total glucobrassicins within broccoli was increased following exposure of florets to both doses, but no significant differences in glucoraphanin content were observed. As a conclusion, the ethanol treatment could indeed delay senescence and lead to the induction of phytochemicals. In contrast, MeJA’s effect on quality is not quite substantial; it can, however, be used to improve the phytochemical content of florets, particularly indole-type GLS.
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Wildflowers in uncultivated field margin are important resources supporting both pest and non-pest thrips populations in agroecosystems. Environmental factors related to wildflowers have never been used as predictors to model pest thrips density in adjacent crops. Wild and cultivated flowers were sampled in strawberry field agroecosystem in Orléans Island, near Quebec City, Canada. Pest flower thrips Frankliniella intonsa and F. tritici had a wide wildflower host range, with preferences toward Leucanthemum vulgare, Trifolium pratense, Sonchus asper, and Cichorium intybus. Regression modeling revealed significant positive relationships between pest thrips density on specific wildflowers (Vicia cracca, Sinapis arvensis, S. asper, C. intybus, L. vulgare) and their density in strawberry flowers. Furthermore, thrips density within the crop decreased with distance from uncultivated field margin. Regarding crop management, knowing the associations between thrips pests and preferred wild flora as well as their spatial distribution in strawberry fields is an undeniable advantage.
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Plant biostimulants are substances or microorganisms aimed at promoting plant growth by increasing the mineral nutrition efficiency, tolerance to environmental stress, and crop quality traits. This new category of crop inputs has been capturing the interest of both researchers and agriculture takeholders in light of the promising effects they could have on crop productivity and sustainability. This study investigated a variety of biostimulants for their effect on germination rates, plant health, chlorophyll fluorescence parameters, SPAD index, and growth of baby leaf lettuce and Batavia lettuce submitted to biotic (absence/presence of Pythium ultimum in the growing medium) or abiotic (0, 40, 80, and 120 mM NaCl L−1 concentrations and −0.5, −2, −4, and −6 kPa water potentials) stresses when grown in a greenhouse under conventional and organic cultivation. The results obtained show that lettuce response to biostimulants was influenced by the type or level of stress applied and the growing system used. The effects of the tested biostimulants varied from strongly detrimental to strongly beneficial.
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Narrow-spectrum insecticides are currently used to control populations of spruce budworm, Choristoneura fumiferana Clemens (Lepidoptera: Tortricidae), in eastern Canada. However, these could have nontarget impacts on other caterpillars – some of which may serve as alternative or alternate hosts to key parasitoids – that are also susceptible to control tactics. This study was conducted to determine how the insecticides, Bacillus thuringiensis variety kurstaki (Btk) and tebufenozide, used to control spruce budworm populations, impact caterpillar communities and associated parasitism rates. Post-treatment field sampling of caterpillars was conducted in 2018 and 2019 in New Brunswick, Canada, at sites treated with either Btk or tebufenozide and at control sites. Caterpillar species richness and abundance, community structure, and parasitism rates were assessed using molecular analyses for 659 collected caterpillars. We found that insecticide applications had no significant impact on abundance, species richness, or parasitism rate relative to the measurements made in the control sites. Nonetheless, a significantly higher caterpillar abundance and lower parasitism rate occurred in Btk-treated sites than in tebufenozide-treated sites. Overall, however, Btk and tebufenozide treatments did not negatively affect the non-budworm caterpillar community under the present conditions of low caterpillar densities, suggesting that parasitoids have alternative and alternate hosts after treatments that target the spruce budworm.
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Phaeodactylum tricornutum has been highly studied for its potential as a platform for metabolic engineering. While the possible applications of extrachromosomal expression via an episome have been investigated, there is still a lack of information concerning its efficacy and limitations. Therefore, we studied the episome expression system in P. tricornutum, aiming to elucidate its limitations regarding heterologous protein production and episome rearrangement events. Our objectives were to screen positive transconjugants by fluorescent signal indicating as a proxy for the production of the proteins of interest that could be used for vanillin synthesis, and to characterize the transconjugants by flow cytometry and whole plasmid sequencing. We designed an episome harboring an expression cassette that consisted of the enhanced green-fluorescent-protein (eGFP) linked by Thosea asigna virus 2A self-cleaving peptide (T2A) to a fusion protein of enoyl-CoA hydratase/aldolase (ech) and feruloyl-CoA synthetase (fcs), both from Streptomyces sp. strain V-1. This construction resulted in a percentage of fluorescent transconjugants lower than 10 % and it presented rearranged episomes in the fluorescent and the non-fluorescent transconjugants. The replacement of the fusion protein ech-fcs in the expression cassette with the fluorescent protein mCherry increased the percentage of eGFP fluorescent transconjugants over 80 % suggesting a toxicity of the ech-fcs gene expression and in turn forcing selection for rearranged episomes. A comparison of flow cytometry results and sequencing analysis demonstrated that a successful transformation with an unaltered expression cassette could lead to diatoms that do not produce the protein. On the other hand, transconjugants with mutations or rearrangements in the genes encoding the fusion ech-fcs protein led to fluorescent signal detection. Here, we show that using fluorescent reporters can mislead the selection of positive transconjugants by not being able to identify rearrangements in the genes of interest, and intact cassettes can lack fluorescent signal due to lack of heterologous protein production.
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Understanding movements of Drosophila suzukii (Diptera: Drosophilidae) into berry fruit crops from wild-fruit hosts in borders of semi-natural agroecosystems, such as lowbush blueberry, is important to determining harvesting time and managing wild-fruit diversity near crops. This study aimed to inventory the wild-fruit hosts associated with lowbush blueberry (Ericaceae) production in Québec, Canada, near the limits of lowbush blueberry’s eastern North American range. We also tested the hypothesis that fruit traits of berry fruit species present in or near lowbush blueberry fields might explain field infestation levels. Flies of both sexes emerged from five wild-fruit species in 2016, to which six more species were added in 2017. The most productive wild-fruit hosts were Cornus canadensis (Cornaceae) and Aralia hispida (Araliaceae) (> 100 emerged flies/100 g of fruit) in 2016, to which Prunus pensylvanica (Rosaceae) and Rubus idaeus (Rosaceae) were added in 2017. Among nine variables considered in statistical modelling (fruit size, sugar content, and reflected colour bands) and taxonomic family (a nominal variable), none could explain field infestation levels over two years. In lab tests comparing fruits of Cornus canadensis, a common weed within fields, and lowbush blueberries, mated D. suzukii females laid twice as many eggs on blueberries, but both species were equally suitable for development.
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In this study, a crude ethanolic extract made of silver maple (Acer saccharinum L.) leaves (SML) was evaluated for antibacterial activity against the phytopathogenic bacteria Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato, and Xanthomonas fragariae. The extract was shown to cause important inhibition zones against the three bacteria in the disc diffusion assays, revealing its antibacterial activity. The minimal inhibitory concentration (MIC) of the extract was determined thereafter for each bacterium. The extract showed the same MIC value (1.56 mg·mL−1) for the three bacteria. Using a semipreparative high-performance liquid chromatography system, crude ethanolic SML extract was divided in 15 fractions and each fraction was tested for antibacterial activity against X. fragariae with the disc diffusion assay. Among the six fractions causing an inhibition zone, fraction 10 caused the largest inhibition. Fraction 10 was further analyzed by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. According to quadrupole time-of-flight mass spectrometry, the main peak of fraction 10 was identified as a galloyl-bis-HHDP-glucose (C41H28O26, 936.6454 g·mol−1) isomer, an ellagitannin known for antibacterial activity and for stimulating plant natural defenses. The study opens new avenues of research on the valorization of SML and on the control of plant diseases caused by bacteria in organic and conventional production of horticultural crops.
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Blackleg, caused by Pectobacterium spp. and Dickeya spp., is an important disease of potatoes. During the period from November 20 and March 2021, stems of potato plants showing necrosis and rot symptoms, and chlorotic leaves, were collected from commercial production areas of the Mayabeque province of Cuba.
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Tomato (Solanum lycopersicum L.) is one of the most important vegetables in the world. Its extensive cultivation has made this plant the target of many viral, fungal, and bacterial diseases. Among them, the bacterial canker of tomato caused by Clavibacter michiganensis (Cm) has been named one of the most devastating diseases affecting the tomato industry worldwide. It can significantly reduce the yields and profitability of this crop. One of the big challenges we found when working with Cm and trying to characterize the virulence of different isolates was the lack of a consensus methodology to inoculate tomato plants, how to fertilize them and characterize Cm virulence. This research aimed to identify an artificial inoculation method to induce bacterial canker on tomato plants in greenhouse conditions to homogenize the results of different studies with Cm. We compared two inoculation methods, including the scalpel and syringe method with two levels of fertilization, low and high fertilization. After evaluating several variables like the percentage of necrotic leaves and the height of the plants, the results showed that the syringe inoculation with low fertilization was the most effective inoculation method allowing the development of a multilevel scale that can be used to study the interaction between tomato plants and Cm isolates.
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Syrphid flies (Diptera, Syrphidae) play important ecological roles as pollinators, pest control agents, and decomposers. Communities are influenced by environmental variables such as land use. However, these interactions are poorly studied in urban landscapes. We used pan traps in community gardens, cemeteries, and parks in Montreal, Canada, to investigate syrphid fly communities in an urban landscape and describe how site surface area and heat island cover shape these communities. We estimated the species richness of communities and the occupancy patterns of three syrphid functional groups (insectivores, terrestrial saprophagous, and aquatic saprophagous). We captured 1,791 specimens from 48 species. Species richness increased with site surface area but did not vary with the level of urbanisation. The occupancy of the three groups did not change with site surface area. Syrphid flies of the terrestrial saprophagous group were less likely to occur on sites with high urbanisation, but the occupancy of insectivore and aquatic saprophagous groups did not vary with the variable.Implications for insect conservation Our results highlight that cities can harbour diversified syrphid fly communities and that certain functional groups are tolerant to urbanisation pressures. Our study suggests that conservation measures targeting syrphid flies in cities must be adapted to functional traits and larval requirements.
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Fusarium head blight (FHB), caused by Fusarium graminearum, is one of the most destructive wheat diseases worldwide. FHB infection can dramatically reduce grain yield and quality due to mycotoxins contamination. Wheat resistance to FHB is quantitatively inherited and many low-effect quantitative trait loci (QTL) have been mapped in the wheat genome. Synthetic hexaploid wheat (SHW) represents a novel source of FHB resistance derived from Aegilops tauschii and Triticum turgidum that can be transferred into common wheat (T. aestivum). In this study, a panel of 194 spring Synthetic Hexaploid Derived Wheat (SHDW) lines from the International Maize and Wheat Improvement Center (CIMMYT) was evaluated for FHB response under field conditions over three years (2017–2019). A significant phenotypic variation was found for disease incidence, severity, index, number of Fusarium Damaged Kernels (FDKs), and deoxynivalenol (DON) content. Further, 11 accessions displayed < 10 ppm DON in 2017 and 2019. Genotyping of the SHDW panel using a 90 K Single Nucleotide Polymorphism (SNP) chip array revealed 31 K polymorphic SNPs with a minor allele frequency (MAF) > 5%, which were used for a Genome-Wide Association Study (GWAS) of FHB resistance. A total of 52 significant marker-trait associations for FHB resistance were identified. These included 5 for DON content, 13 for the percentage of FDKs, 11 for the FHB index, 3 for disease incidence, and 20 for disease severity. A survey of genes associated with the markers identified 395 candidate genes that may be involved in FHB resistance. Collectively, our results strongly support the view that utilization of synthetic hexaploid wheat in wheat breeding would enhance diversity and introduce new sources of resistance against FHB into the common wheat gene pool. Further, validated SNP markers associated with FHB resistance may facilitate the screening of wheat populations for FHB resistance.
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The physical properties of peat substrates from eight tree nurseries were characterized to determine bulk density, air-filled porosity, saturated hydraulic conductivity, pore effectiveness, relative gas diffusivity and chemical properties. There were significant variations among nurseries both in growth of white spruce [Picea glauca (Moench) Voss] seedlings (1+0) and substrate properties. Shoot dry mass and root collar diameter were negatively correlated with air-filled porosity and saturated hydraulic conductivity, whereas root dry mass was positively correlated with bulk density. Seedling growth increased with increasing substrate bulk density up to ~0.11 g cm−3, above which value conditions may become limiting to plant performance. Our results suggest that there was no growth limitation due to restricted aeration (Ds D0−1 > 0.005 m2 s−1 m−2 s for all substrates except one) and that over-aeration reduced seedling growth under dry irrigation management.
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During more than five centuries of human colonization, 50% of original peatlands in Azores have been destroyed or degraded, mainly due to pasture use, resulting in landscape changes and a loss of ecosystem biodiversity and services. This study aims to identify the most effective measures in the recovery and detect initial responses (2 years). These are the first restoration experiments, carried out in a background of complete absence of strategies adapted for Azores pastured peatlands. In a post-pastured peatland, 24 experiments were implemented, repeated in more degraded (northern) and more natural (southern) areas. The experiments were combinations of restoration measures, with the introduction of Juniperus brevifolia, Calluna vulgaris and Sphagnum palustre and the use of management techniques to control herbaceous cover (alien-rich herbaceous cover competing with Sphagnum or peatland natural shrubs): intensive and extensive grazing and grass cutting. Principal response curves and redundancy analyses were used to assess changes in the flora and the physicochemical parameters. Globally there was an increase in Sphagnum cover and a decrease in herbaceous species. This was accompanied by an increase in organic matter and a decrease in nitrogen. In more natural parcels (large artificially divided areas where different experiments were implemented), the most positive tendencies were associated with the combination of J. brevifolia planting and grass cutting. In more degraded areas, treatments had a lower impact, as the results tended to be similar to the control, pointing to an important natural regenerative succession dynamic that overlapped the response to the implemented treatments.
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Wild bees are declining worldwide, and many species are now threatened with extinction. Decline is caused by a combination of threats, including disease spillover from managed bees that reduces health of wild bees. An increasing number of studies thus aim to characterize bee health. The common approaches, however, require sacrificing tens to hundreds of individual bees per site or species, with reports of several thousand individuals collected per study. Considering the widespread need to assess bee health, this sampling approach is not sustainable, especially for endangered populations or species. Here, we propose a non-destructive method to assess parasite loads of wild-caught bumble bees. The standard protocol consists of net-capturing individual bumble bees and placing them in a 10 cm (diameter) petri dish to collect faeces. Although this approach is frequently used in laboratory settings, it is not in the field, because of the low success in collecting faeces. Placing bumble bees in a previously refrigerated cooler, we significantly improved faecal collection in the field from 76% with the standard protocol to 86% with the cooler protocol. We also successfully identified spores and cells of two common gut parasites Crithidia spp. and Vairimorpha spp. in faecal samples. The efficacy of the cooler protocol, combined to the low-cost and widespread availability of the equipment should promote its use in field studies. Implication for insect conservation: As there are calls to reduce destructive sampling methods in bee research, using the updated cooler protocol will contribute to achieving this goal. This opens future avenues of combining this non-destructive approach to assess bee health with molecular tools.
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Evidence points out that increasing plant productivity associated with greater erect shrub abundance alters soil organic carbon (SOC) stocks in the Arctic. However, the underlying plant economic traits remain poorly examined, which limits our understanding of plant–environment interactions driving tundra carbon cycling. We explored how erect shrub abundance leads to SOC variation in a High Arctic tundra (Bylot Island, Nunavut, Canada), where the only erect shrub, Salix richardsonii, has settled along currently active and abandoned channel zones of alluvial fans. The effects of vegetation and local environmental changes on SOC were evaluated through a paired sampling of soil materials and plant aboveground functional traits associated with plant carbon supply and nutrient demand processes. The occurrence of S. richardsonii, characterized by a tenfold increase in aboveground biomass, induced a 28% increase in SOC compared to adjacent plots dominated by prostrate shrubs and graminoids. Yet, this vegetation effect was solely observed along active channels, where higher SOC was associated with greater leaf and stem biomass. A path analysis showed that shrub leaf area index and total leaf nutrient content best represented plant carbon supply and nutrient demand dynamics, respectively, and jointly regulated SOC variation. This study underscores that vegetation structural changes associated with increasing erect shrub abundance in the Arctic can promote soil organic carbon storage, but that this pattern may be mediated by strong plant–environment interactions. Accounting for changes in functional traits driving plant carbon supply and nitrogen demand proves important for a better mechanistic understanding of how shrubification impacts tundra carbon cycling.
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Main conclusion: Ammonium sulfate is well known to salt out proteins at high concentrations. The study revealed that it can serve to increase by 60% the total number of identified carbonylated proteins by LC–MS/MS. Abstract: Protein carbonylation is a significant post-translational modification associated with reactive oxygen species signaling in animal and plant cells. However, the detection of carbonylated proteins involved in signaling is still challenging, as they only represent a small subset of the proteome in the absence of stress. In this study, we investigated the hypothesis that a prefractionation step with ammonium sulphate will improve the detection of the carbonylated proteins in a plant extract. For this, we extracted total protein from the Arabidopsis thaliana leaves and subjected the extract to stepwise precipitation with ammonium sulfate to 40%, 60%, and 80% saturation. The protein fractions were then analyzed by liquid chromatography–tandem mass spectrometry for protein identification. We found that all the proteins identified in the non-fractionated samples were also found in the prefractionated samples, indicating no loss was incurred during the prefractionation. About 45% more proteins were identified in the fractionated samples compared to the non-fractionated total crude extract. When the prefractionation steps were combined with the enrichment of carbonylated proteins labeled with a fluorescent hydrazide probe, several carbonylated proteins, which were unseen in the non-fractionated samples, became visible in the prefractionated samples. Consistently, the prefractionation method allowed to identify 63% more carbonylated proteins by mass spectrometry compared to the number of carbonylated proteins identified from the total crude extract without prefractionation. These results indicated that the ammonium sulfate-based proteome prefractionation can be used to improve proteome coverage and identification of carbonylated proteins from a complex proteome sample.
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Despite the increased efficiency of sequencing technologies and the development of reduced-representation sequencing (RRS) approaches allowing high-throughput sequencing (HTS) of multiplexed samples, the per-sample genotyping cost remains the most limiting factor in the context of large-scale studies. For example, in the context of genomic selection (GS), breeders need genome-wide markers to predict the breeding value of large cohorts of progenies, requiring the genotyping of thousands candidates. Here, we introduce 3D-GBS, an optimized GBS procedure, to provide an ultra-high-throughput and ultra-low-cost genotyping solution for species with small to medium-sized genome and illustrate its use in soybean. Using a combination of three restriction enzymes (PstI/NsiI/MspI), the portion of the genome that is captured was reduced fourfold (compared to a “standard” ApeKI-based protocol) while reducing the number of markers by only 40%. By better focusing the sequencing effort on limited set of restriction fragments, fourfold more samples can be genotyped at the same minimal depth of coverage. This GBS protocol also resulted in a lower proportion of missing data and provided a more uniform distribution of SNPs across the genome. Moreover, we investigated the optimal number of reads per sample needed to obtain an adequate number of markers for GS and QTL mapping (500–1000 markers per biparental cross). This optimization allows sequencing costs to be decreased by ~ 92% and ~ 86% for GS and QTL mapping studies, respectively, compared to previously published work. Overall, 3D-GBS represents a unique and affordable solution for applications requiring extremely high-throughput genotyping where cost remains the most limiting factor.
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Light regulates important metabolic processes in microalgal cells, which can further impact the metabolism and the accumulation of biomolecules such as lipids, carbohydrates, and proteins. Different characteristics of light have been studied on various strains of the model diatom Phaeodactylum tricornutum, but not on transconjugant cells and information on wild-type strains is still limited. Therefore, we studied the impact of different light characteristics such as spectral quality, light intensity and light shift on the growth, and the composition in lipids and fatty acids of P. tricornutum cells to provide a comprehensive context for future applications. Initially, we tested the impact of spectral quality and light intensity on P. tricornutum transformed with an episomal vector (Ptev), harboring the resistance gene Sh ble. Results indicated that Ptev cells accumulated more biomass and overall lipids in spectral quality Red 1 (R1: 34% > 600 nm > 66%) more effectively as compared to Red 2 (R2: 8% > 600 nm > 92%). It was also detected that cell granularity was higher in R1 as compared to R2. Furthermore, by testing two light intensities 65 μmol·m-2·s-1 and 145 μmol·m-2·s-1 light, it was observed that 145 μmol·m-2·s-1 led to an increase in growth trend, total biomass and lipid content. Combining spectral qualities and light intensities, we show that the lipid accumulation raised by 2.8-fold. Studying the light intensity and spectral quality allowed us to optimize the light conditions to R1 spectral quality and light intensity 145 μmol·m-2·s-1. These initial results showed that red light R1 at 145 μmol·m-2·s-1 was the best condition for biomass and total lipids accumulation in Ptev cells. Next, we further combined these two-light optimizations with a third light characteristics, i.e. light shift, where the cultures were shifted during the early stationary phase to a different light environment. We studied Red light shift (Rs) to investigate how light condition variations impacted P. tricornutum transconjugants Ptev and with an episomal vector containing the reporter gene YFP (PtYFP). We observed that Rs induced growth and fatty acid eicosapentaenoic acid (EPA) in Ptev as compared to PtYFP. Altogether, the study shows that red light shift of R1 at 145 μmol·m-2·s-1 promoted biomass and total lipids accumulation in Ptev and PtYFP cells. The study provides a comprehensive approach to using different light characteristics with the aim to optimize growth and lipids, as well as to fatty acid production.
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Abstract. The main objective of this study was to inventory the abundance and species richness of wild bees and hoverflies in the Lac Saint-Pierre floodplain according to a land-use gradient. In 2019 and 2020, pollinators were sampled using pan-traps in three landscape types: Crop field margins, Perennial hayfields, and Natural habitats. Bee and hoverfly populations were dominated by a few species throughout the study area. Crop field margins contained greater floral availability and attracted more individuals and species of bees than other landscape types. Although hoverflies were not affected by either land-use type or flooding, the abundance and species richness of bees appeared to be reduced when spring flooding lasted longer, suggesting a mortality effect of flooding on their populations. Implications for insect conservation. Our results make a case for the key role of field margins in the conservation of pollinating insects in agricultural landscapes, especially in a floodplain context.
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Developing and implementing improved management practices are necessary to enhance the sustainability of organic cropping systems. This study examined the effects of various organic cropping systems on soil greenhouse gas (GHG) emissions and crop yields in Québec, Canada. Organic cropping systems combining different: (i) crop sequences (barley [Hordeum vulgare L.]-grain corn [Zea mays L.], soybean [Glycine max (L.) Merr.]-spring wheat [Triticum aestivum L.], and grain corn-soybean), (ii) nitrogen (N) sources (poultry manure [PM] and/or a fall-seeded green manure [GM] or no applied N), and (iii) primary tillage intensities (moldboard plough [MP] or chisel plough [CP]) were compared to a perennial forage (PF) and a bare fallow (BF) control. During the 2019 and 2020 snow-free seasons, nitrous oxide (N2O) and methane (CH4) emissions, soil water content, soil temperature, and mineral N concentrations were monitored periodically on a sandy loam soil. The lowest cumulative N2O emissions were found in CP-GM (0.52 ± 0.11 kg N ha-1 in 2019 and 0.47 ± 0.06 kg N ha-1 in 2020), whereas the highest N2O emissions were found in MP-PM in 2019 (3.55 ± 0.72 kg N ha-1) and BF in 2020 (1.44 ± 0.20 kg N ha-1). For the barley-grain corn sequence, the CP-GM treatment generated N2O emissions that were 40–70 % lower and yields that were 33–51 % lower than the MP-PMGM and CP-PMGM systems, which showed equivalent N2O emissions and yields. Yield-scaled N2O emissions were equivalent for all cropping systems. Peak N2O daily fluxes in the PF occurred shortly after cutting in 2020. During both years, CH4 emissions varied from − 0.65 to + 0.18 kg C ha-1 with no detectable differences among cropping systems. The CP-GM cropping system minimized area-scaled N2O emissions without increasing yield-scaled emissions. However, this was a two-year study on a site that was recently converted from conventional agriculture, so a long-term assessment is still necessary to determine whether the benefits associated with these cropping systems change over time.
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