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Accurate genomic sequencing demands high-quality double-stranded RNA (dsRNA). Existing methods for dsRNA extraction from yeast, fungi, and plants primarily rely on cellulose, suitable only for small volume extractions, or the time-consuming lithium chloride precipitation. To streamline the traditional phenol-chloroform-based dsRNA extraction method, the main challenge is the reduction of mitochondrial DNA (mtDNA) and Single Stranded RNA (ssRNA) to no detectable levels after gel electrophoresis. This challenge is successfully addressed through the modified approach described here, involving phenol extraction at low pH, followed by the addition of ammonium sulfate to the aqueous buffer. The dsRNA isolated using this novel method exhibits comparable quality to that obtained through cellulose purification, and it is readily amenable to RT-PCR. Moreover, a single batch of yeast cell RNA isolation requires only 2-3 h of hands-on time, thus simplifying and expediting the process significantly. • Buffers were redesigned from [32,33,35]. • No DNASE, Ribonuclease A or beads were used during the purification. • Simple and inexpensive dsRNA extraction and purification method is described.

This study focused on the development of a stripping process for recovering nitrogen from compost leachate. Several experiments were carried out to identify the optimum conditions of temperature, airflow rate and pH that provide effective recovery of nitrogen from the leachate. The most energy and chemical saving experimental condition were found out to be at 40 °C, an airflow rate of 1 L/min and a pH of 10.5. A stripping efficiency of about 94% was observed upon the use of an air flow rate of 1 L/min and a 71% recovery in sulphuric acid to finally obtain ammonium sulphate. The experiment was performed using a leachate volume of 500 mL, with an NH4+ concentration of 600 mg/L, and during the absorption, the total ammonium was recovered as 214 mg/150 mL of sulphuric acid solution (2% v/v), which makes a total recovery of 71% of initial ammonium concentration. Overall, it is reported that 1.5 g of ammonium sulphate could be extracted from the processing (stripping and absorption) of 1 L of raw leachate.

Grasslands cover up to 40.5% of the world's landmass and store 30% terrestrial carbon (C). Various practices, including mineral fertilization and liming, are used to manage these ecosystems with potential long-term effects on the size and distribution of soil aggregates and inevitably carbon dynamics. The objective of this study was to examine the long-term effects of nitrogen fertilization and liming on soil carbon storage and its dynamics in water-stable aggregates of a semi-arid grassland. Soil samples (0–10 cm) were collected from Ukulinga long-term grassland trial in Pietermaritzburg, South Africa where nitrogen fertilizers have been applied annually and lime every five years for 70 years. Ten treatments were studied: the control (0 kgN/ha and unlimited), lime at 2250 kg/ha (L), ammonium sulphate at 70 kg/ha (AS70) and 211 kg/ha (AS211); ammonium nitrate at 70 kg/ha (AN70) and 211 kg/ha (AN211); AS70 + lime (AS70L); AS211 + lime (AS211L); AN70 + lime (AN70L) and AN211 + lime (AN211L).Nitrogen fertilizers significantly reduced soil pH and increased total soil N. Liming increased soil pH with no effect on total soil N. Lime and lime + N fertilizer treatments had no effect on mean weight diameter (MWD) while separate N application decreased MWD and large macro-aggregates (LMA). Lime only treatment had no effect on water stable aggregate (WSA) fractions. Nitrogen fertilization and liming (separately or in combination) did not affect total C concentration and stocks. Overall, soils had very high total soil organic carbon ranging from 49.7 – 57.6 g/kg across treatments. Nitrogen fertilization decreased organic carbon in LMA in AS70 (1.52%) and AN211 (1.67%) treatments compared to the control (3.40%) which was in concert with increases in C associated with small macro-aggregates (SMA) and micro-aggregates (MiA and SCA). Organic carbon in SMA was 2.67 % (AS70); AS211 (2.62 %); AN70 (2.02 %); AN211 (2.49 %) compared to 1.26 % in the control. Lime + N fertilizer treatments increased C storage in all aggregate fractions compared to N fertilizer only treatments. The lack of response in total SOC to 70 years of N fertilization and liming suggests possible C saturation given the high soil C concentration. Changes in C associated with WSA fractions suggests their importance as diagnostic indicators of N fertilization and liming induced changes in SOC. Findings also show that ammonium-based N fertilization is associated with soil acidification, dispersion of LMA resulting in an increase of microaggregates and C stored in them. Liming can counteracts acidifying and the dispersive effect on NH4+ associated with ammonium-based fertilizers thus restoring macro-aggregation in N fertilized grasslands. These findings suggests that long-term N addition may result in poor soil physical condition and possible stabilization of C in stable fractions.

With a global population of eight billion people, improving the sustainability and nutritional quality of diets has become critical. Mushrooms offer a promising solution because of their nutritional value and ability to be grown from agricultural residues, in line with the circular economy. This study, therefore, focuses on assessing the environmental compatibility of Agaricus bisporus mushroom production in Italy, the world's third largest per capita consumer, by using a Life Cycle Assessment (LCA) and an integrated Water-Energy-Nitrogen-Carbon-Food (WENCF) nexus analysis. The LCA results reveal that for a functional unit of 23,000 kg of the substrate, the production process emits 2.55 × 10 4  kg of CO 2 eq. Sensitivity analysis shows that changing input quantities can reduce environmental impacts by about 5 %. In addition, one scenario evaluates the environmental effects of recycling resources by introducing water and ammonium sulfate from scratch instead of continuous recycling, along with water purification. The study shows that sustainable food production can mitigate resource depletion, climate-altering emissions, and intersectoral competition. Using agro residues for mushroom cultivation and optimizing resource management contribute to environmental sustainability. This approach could not only improve the resilience and efficiency of the food system but could also improve the sustainability of diets. In conclusion, this study highlights the importance of adopting sustainable and circular approaches in mushroom production to address global challenges related to food sustainability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Biogas production generates digested slurry as a byproduct. It can be used as a fertilizer especially after its conversion into digested liquid. A pot based study was conducted in order to evaluate the effect of the application of digested liquid on CH4 and N2O flux, and plant biomass in paddy. Analysis revealed that digested liquid treated soils released more CH4 compared to ammonium sulphate and the control. Ammonium sulphate treated soil emitted the highest N2O whereas digested liquid application decreased its emission significantly. Further, the cumulative emission over 101 d of the experiment was found to be higher for CH4 (16.9 to 29.9 g m−2) compared to N2O (−49.3 to 18.9 mg m−2) for all treatments. Digested liquid application had positive impact on plant variables such as panicle number and weight of panicles. This study suggests that digested liquid application significantly decrease N2O emission and increase CH4 emission possibly due to affecting the availability of organic C in the soil to microbial activity for methanogenesis. Another possibility for enhancing CH4 emission by following biogas digested liquid could be attributed to the increase in plant biomass.

Agricultural sustainability is compromised by nitrogen (N) losses caused by soil microbial activity. Nitrous oxide (N 2 O) is a potent greenhouse gas (GHG) produced as consequence of nitrification and denitrification processes in soils. Nitrification inhibitors (NI) as 3,4-dimethylpyrazole-succinic acid (DMPSA) are useful tools to reduce these N losses from fertilization. The objective of this work was to test the efficiency of DMPSA in two different tillage management systems, conventional tillage (CT) and no-tillage (NT), in a winter wheat crop under Humid Mediterranean conditions. N fertilizer was applied as ammonium sulphate (AS) with or without DMPSA in a single or split application, including an unfertilized treatment. GHG fluxes (N 2 O, CO 2 and CH 4 ) were measured by the closed chamber method. amoA and nosZI genes were quantified by qPCR as indicators of nitrifying and denitrifying populations. Nitrification was inhibited by DMPSA in both CT and NT, while the higher water filled pore space (WFPS) in NT promoted a better efficiency of DMPSA in this system. This higher efficiency might be due to a greater N 2 O reduction to N 2 as result of the nosZI gene induction. Consequently, DMPSA was able to reduce N 2 O emissions down to the unfertilized levels in NT. Provided that NT reduced CO 2 emissions and maintained crop yield compared to CT, the application DMPSA under NT management is a promising strategy to increase agro-systems sustainability under Humid Mediterranean conditions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 The Author(s). Published by Elsevier B.V. All rights reserved.)

Soil fertilisation affects greenhouse gas emissions. The objective of this study was to compare the effect of different fertilisation strategies on N2O, CH4 emissions and on ecosystem respiration (CO2 emissions), during different periods of rice cultivation (rice crop, postharvest period, and seedling) under Mediterranean climate. Emissions were quantified weekly by the photoacoustic technique at two sites. At Site 1 (2011 and 2012), background treatments were 2 doses of chicken manure (CM): 90 and 170kgNH4(+)-Nha(-1) (CM-90, CM-170), urea (U, 150kgNha(-1)) and no-N (control). Fifty kilogram N ha(-1) ammonium sulphate (AS) were topdress applied to all of them. At Site 2 (2012), background treatments were 2 doses of pig slurry (PS): 91 and 152kgNH4(+)-Nha(-1) (PS-91, PS-152) and ammonium sulphate (AS) at 120kgNH4(+)-Nha(-1) and no-N (control). Sixty kilogram NH4(+)-Nha(-1) as AS were topdress applied to AS and PS-91. During seedling, global warming potential (GWP) was ~3.5-17% of that of the whole rice crop for the CM treatments. The postharvest period was a net sink for CH4, and CO2 emissions only increased for the CM-170 treatment (up to 2MgCO2ha(-1)). The GWP of the entire rice crop reached 17Mg CO2-eqha(-1) for U, and was 14 for CM-170, and 37 for CM-90. The application of PS at agronomic doses (~170kgNha(-1)) allowed high yields (~7.4Mgha(-1)), the control of GWP (~6.5MgCO2-eqha(-1)), and a 13% reduction in greenhouse gas intensity (GHGI) to 0.89kgCO2-eqkg(-1) when compared to AS (1.02kgCO2-eqkg(-1))., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Gel electrophoresis, particularly one- (1DE) and two-dimensional electrophoresis (2DE), remain among the most widely used top-down methods for resolving and analysing proteomes. Detection of the resulting protein maps relies on staining (i.e. colloidal coomassie blue (CCB) or SYPRO Ruby (SR), in addition to many others). Fluorescent in-gel protein stains are generally preferred for higher sensitivity, reduced background, and wider dynamic range. Although traditionally used for densitometry, CBB has fluorescent properties. Indeed, infrared detection of CCB stained protein was comparable to SR, with BioSafe (Bio-Rad) and the Neuhoff formulation (NCCB) identified as potentially superior to SR; a minor sensitivity issue encountered in gel-resolved proteomes; might have been due to the unified staining protocol used. Here the staining protocol for both CCB formulations was optimised, yielding improved selectivity without affecting sensitivity; the resulting linear dynamic range was similar for BioSafe and NCCB and somewhat better than SR. 2D gel-based analyses of mouse brain and Arabidopsis thaliana (leaf) proteomes indicated markedly superior spot detection using the NCCB formulation. Thus more sensitive, quantitative in-gel protein analyses can be achieved using NCCB, at a fraction of the cost., (Copyright © 2013 Elsevier B.V. All rights reserved.)