Effects of fresh and field-aged holm-oak biochar on As, Cd and Pb bioaccumulation in different rice growing environments.

Arsenic, Cd, and Pb environmental fate is influenced when the traditional permanent flooding rice production systems are replaced by water-saving and soil conservation practices, urging for additional strategies that avoid their bioaccumulation in rice grain. The aim of this two-years field study was to evaluate the effects of fresh and field-aged biochar on As, Cd, and Pb bioaccumulation, and on As speciation, in rice grain produced in different growing environments (flooding versus sprinkler and conventional tillage versus direct seeding). Biochar produced from holm-oak pruning residues (pyrolysis at 550 °C, 48 h), in a single application (28 Mg ha-1), reduced As bioaccumulation in rice grain in the permanent flooding system to non-quantifiable concentrations (e.g., from 0.178 mg kg-1 to <0.04 mg kg-1, for inorganic-As, respectively), an effect which remained under field-aging conditions, increasing rice commercial value. When adopting sprinkler irrigation, the undesirable increase in Cd bioaccumulation in rice, relatively to the anaerobic system, was counteracted by biochar application, reducing its bioaccumulation in kernels between 32 and 80 %, allowing a simultaneous control of metals and metalloids bioaccumulation in rice. The bioaccumulation of Pb was also prevented with biochar application, with a reduction in its concentration four- to 13-times, in all the management systems, relatively to the non-amended plots, under fresh biochar effects. However, Pb immobilization decreased with biochar field-aging, indicating that the biochar application may have to be repeated to maintain the same beneficial effect. Therefore, the present study shows that the implementation of sprinkler irrigation with holm-oak biochar could reduce the risk of heavy metals(loids) bioaccumulation in rice grains and, thereby, ensuring food safety aspects, particularly under fresh biochar effects.

Combined use of biochar and alternative management systems for imazamox induced pollution control in rice growing environments.

Imazamox (IMZX) is a persistent herbicide having probable risks for non-target organisms in the environment and water contamination. Alternatives to conventional flooding rice production, including biochar amendment, may induce changes in soil properties which can greatly modify the environmental fate of IMZX. This two-year study is the first to evaluate how tillage and irrigation practices, with or without fresh or aged biochar (Bc), that are alternatives to conventional rice production impact IMZX's environmental fate. The treatments were: conventional tillage and flooding irrigation (CTFI), conventional tillage and sprinkler irrigation (CTSI), no-tillage and sprinkler irrigation (NTSI), and the corresponding Bc-amendment treatments (CTFI-Bc, CTSI-Bc, and NTSI-Bc). Fresh and aged Bc amendment decreased IMZX's sorption onto the soil in tillage treatments, with Kf values decreasing 3.7 and 4.2-fold (fresh case) and 1.5 and 2.6-fold (aged case) for CTSI-Bc and CTFI-Bc, respectively. The transition to sprinkler irrigation reduced IMZX persistence. Overall, Bc amendment also reduced chemical persistence with half-life values decreasing 1.6 and 1.5-fold for CTFI and CTSI (fresh year) and 1.1, 1.1, and 1.3-fold for CTFI, CTSI, and NTSI (aged year), respectively. Sprinkler irrigation reduced IMZX leaching by up to a factor of 2.2. The use of Bc as amendment led to a significant decrease in IMZX leaching only under tillage conditions, but notable in particular for the CTFI case where leaching losses were reduced in the fresh year from 80% to 34% and, in the aged year, from 74% to 50%. Hence the change in irrigation from flooding to sprinkler either alone or in combination with the use of Bc (fresh or aged) amendment could be considered an effective way to sharply mitigate IMZX contamination of water in environments where rice is grown, particularly in those managed with tillage.

Medium-Term Effects of Sprinkler Irrigation Combined with a Single Compost Application on Water and Rice Productivity and Food Safety.

Traditional rice (Oryza sativa L.) management (tillage and flooding) is unsustainable due to soil degradation and the large amount of irrigation water used, an issue which is exacerbated in the Mediterranean region. Therefore, there is a need to explore rice management strategies in order to improve water-use efficiency and ensure its sustainability. Thus, field experiments were conducted to determine the medium-term effects of different irrigation and tillage methods combined with a single compost application on water and rice productivity, as well as food safety in a semiarid Mediterranean region. The management systems evaluated were: sprinkler irrigation in combination with no-tillage (SNT), sprinkler irrigation in combination with conventional tillage (ST), which were implemented in 2015, and flooding irrigation in combination with conventional tillage (FT), and their homologues (SNT-C, ST-C, and FT-C) with single compost application in 2015. In reference to rice grain yield, the highest values were observed under ST treatment with 10 307 and 11 625 kg ha-1 in 2018 and 2019 respectively; whereas between FT and SNT there were no significant differences, with 8 140 kg ha-1 as mean value through the study. Nevertheless, sprinkler irrigation allowed saving 55% of the total amount of water applied in reference to flooding irrigation. Furthermore, the highest arsenic concentration in grains was found under FT but it decreased with compost application (FT-C) and especially with sprinkler irrigation, regardless of tillage management systems. However, sprinkler irrigation favors the cadmium uptake by plants, although this process was reduced under SNT in reference to ST, and especially under amended compost treatments. Therefore, our results suggested that a combination of sprinkler irrigation and compost application, regardless of the tillage system, could be an excellent strategy for rice management for the Mediterranean environment in terms of water and crop productivity as well as food safety.

Effects of Combined Use of Olive Mill Waste Compost and Sprinkler Irrigation on GHG Emissions and Net Ecosystem Carbon Budget under Different Tillage Systems.

Traditional rice (Oryza sativa L.) production by flooding is a source of greenhouse gases (GHG), especially methane. The high consumption of water, as well as the chemical and physical degradation caused by these traditional practices in rice soils, is promoting a decrease in rice production in the Mediterranean area. The aim of this study was to monitor GHG emissions and the net ecosystem carbon balance (NECB) from rice produced with sprinkler irrigation techniques and also assess the impact of olive mill waste compost (C-OW) application and tillage on GHG emissions and the NECB. A field experiment for irrigated rice production was implemented by considering four different treatments: (1) tillage (T); (2) no tillage-direct seeding techniques (DS); (3) application of C-OW followed by tillage (TC); and (4) application of C-OW followed by direct seeding (DSC). The C-OW was only applied in the first year at a dose of 80 Mg ha-1. GHG emissions were monitored over three years in these four treatments in order to estimate the direct (first year) and residual (third year) effects of such practices. The application of C-OW caused an increase of 1.85 times the emission of CO2-C in the TC-DSC compared to the T-DS in the first year. It is noteworthy that the TC treatment was the only one that maintained an emission of CO2-C that was 42% higher than T in the third year. Regardless of the treatments and year of the study, negative values for the cumulative CH4 were found, suggesting that under sprinkler irrigation, CH4 oxidation was the dominant process. A decrease in N2O emissions was observed under direct seeding relative to the tillage treatments, although without significant differences. Tillage resulted in an increase in the global warming potential (GWP) of up to 31% with respect to direct seeding management in the third year, as a consequence of the greater carbon oxidation caused by intensive tillage. DS presented a positive NECB in the accumulation of C in the soil; therefore, it provided a greater ecological benefit to the environment. Thus, under Mediterranean conditions, rice production through a sprinkler irrigation system in combination with direct seeding techniques may be a sustainable alternative for rice crops, reducing their GWP and resulting in a lower carbon footprint. However, the use of C-OW as an organic amendment could increase the GHG emissions from rice fields irrigated by sprinklers, especially under tillage conditions.

Environmental fate and efficiency of bispyribac­sodium in rice soils under conventional and alternative production systems affected by fresh and aged biochar amendment.

Irrigation and tillage practice alternatives to conventional flooding production, with or without organic amendments, are attracting great interest to adapt rice cultivation to climate change. However, they can alter the behaviour of pesticides and their efficiency against weeds. A two-year field experiment was conducted to investigate how the environmental fate and the weed control efficiency (WCE) of bispyribac­sodium (BS) were influenced by biochar produced from holm oak prunings (BHO) testing both the fresh and the aged effects. The treatments were: flooding irrigation and tillage (FT), sprinkler irrigation and tillage (ST), sprinkler irrigation and no-tillage (SNT), and the corresponding homologues with BHO addition (FT-BHO, ST-BHO, and SNT-BHO, respectively). Fresh BHO amendment decreased the sorption of BS onto the soil in all treatments, while, after aging, it also decreased sorption in FT-BHO (1.3-fold) but increased it in SNT-BHO and ST-BHO (1.1-fold). BHO addition reduced BS persistence under non-flooding and flooding incubation conditions, except for FT under the former condition for which t1/2 increased ≈1.5-fold in both years. The addition of BHO led to a decrease in BS leaching from 58.3 % and 44.6 % and from 70.4 % and 58.1 % in ST and FT to 50.1 % and 38.3 % and 63.6 % and 50.3 % in the homologue amended soils for the fresh and aged years, respectively. While fresh BHO addition decreased the WCE of BS in SNT-BHO, ST-BHO, and FT-BHO on average by a factor of 1.5, with aged BHO there was only such a decrease (by a factor of 1.4) in FT-BHO. The use of BHO could be effective for reducing water contamination by BS in flooding or sprinkler irrigation rice farming as long as conventional tillage is used. But it may also contribute to greatly reducing the herbicide's efficiency, although with time to allow aging, this reduction would only persist under conventional flooding production.

Combined use of olive mill waste compost and sprinkler irrigation to decrease the risk of As and Cd accumulation in rice grain.

Sprinkler irrigation has been successfully introduced in rice production as an alternative to the traditional flooding system, allowing water savings and the reduction of As accumulation in the grain. However, the same conditions can increase Cd mobility and grain accumulation, an effect that needs to be countered. A 3-year field experiment was set-up in a Mediterranean region (Extremadura, Spain), to evaluate how the application of compost from olive mill waste (single application, 80 t ha-1), influences the accumulation of As and Cd in the grain under different irrigation regimes. Accumulation of As in the grain was always lower in the sprinkler irrigation when compared with the flooding irrigation, reaching a 5-fold difference in the third year. Compost application did not evidence a clear effect on the As accumulation in the rice grain, but highly significant negative correlations (p < 0.001) were obtained between As content in the grain (total, inorganic, and organic) and the humification parameters in the soil, evidencing the importance of using a mature and stable organic amendment to avoid As accumulation in the grain. Cadmium accumulation in the rice grain decreased in each treatment where compost was applied, relatively to the non-treated counterpart (e.g., from 0.080 to <0.010 mg kg-1, in direct seeding with sprinkler irrigation, in the third year). There were no significant differences in the total inorganic As between treatments with or without compost application, but it was possible to observe an increase in the predominance of the organic As over the more toxic inorganic As, when compost was applied, allowing a decrease in the risk associated to As accumulation. Therefore, the aerobic cultivation of rice, with the simultaneous application of an adequate source of organic matter, can be considered a good solution to cope with the risk of accumulation of As and Cd in the rice grain.

Genome Features of a New Double-Stranded RNA Helper Virus (LBCbarr) from Wine Torulaspora delbrueckii Killer Strains.

The killer phenotype of Torulaspora delbrueckii (Td) and Saccharomyces cerevisiae (Sc) is encoded in the genome of medium-size dsRNA viruses (V-M). Killer strains also contain a helper large size (4.6 kb) dsRNA virus (V-LA) which is required for maintenance and replication of V-M. Another large-size (4.6 kb) dsRNA virus (V-LBC), without known helper activity to date, may join V-LA and V-M in the same yeast. T. delbrueckii Kbarr1 killer strain contains the killer virus Mbarr1 in addition to two L viruses, TdV-LAbarr1 and TdV-LBCbarr1. In contrast, the T. delbrueckii Kbarr2 killer strain contains two M killer viruses (Mbarr1 and M1) and a LBC virus (TdV-LBCbarr2), which has helper capability to maintain both M viruses. The genomes of TdV-LBCbarr1 and TdV-LBCbarr2 were characterized by high-throughput sequencing (HTS). Both RNA genomes share sequence identity and similar organization with their ScV-LBC counterparts. They contain all conserved motifs required for translation, packaging, and replication of viral RNA. Their Gag-Pol amino-acid sequences also contain the features required for cap-snatching and RNA polymerase activity. However, some of these motifs and features are similar to those of LA viruses, which may explain that at least TdV-LBCbarr2 has a helper ability to maintain M killer viruses. Newly sequenced ScV-LBC genomes contained the same motifs and features previously found in LBC viruses, with the same genome location and secondary structure. Sequence comparison showed that LBC viruses belong to two clusters related to each species of yeast. No evidence for associated co-evolution of specific LBC with specific M virus was found. The presence of the same M1 virus in S. cerevisiae and T. delbrueckii raises the possibility of cross-species transmission of M viruses.

Analysing the vineyard soil as a natural reservoir for wine yeasts.

The cultivable microorganisms isolated from grapes and soils of two contiguous vineyards were analysed. Two types of management system were tested in each vineyard: conventional tillage (CT), and no-tillage with natural green cover vegetation (NV), both under semi-arid rainfed conditions. The main objective was to gather evidence as to whether the yeasts present in ripe grapes before harvest and those found in spontaneous wine fermentations came from the vineyard soil which could then be regarded as a natural reservoir for these yeasts. Bacteria isolated from the grapes all corresponded to three genera (Pseudomonas, Stenotrophomonas, and Bacillus) that were very abundant in soil samples taken just before grape harvest, indicative of probable contamination of the grapes with soil microorganisms. The amounts of fermentative yeasts in vineyard soil increased significantly during the dates close to harvest. Some yeasts were isolated from soils and spontaneous fermentations (Saccharomyces and Lanchacea), while others were only isolated from fermentations (Hanseniaspora, Metschnikowia, and Pichia) or from soils (Torulaspora). Saccharomyces yeasts were isolated from vineyard soil only after grape harvest, to thereafter become undetectable. The analysis of sterile-must fermentations inoculated with soil samples showed that soil was not the origin of the most abundant fermentative yeasts in spontaneous grape fermentations (Saccharomyces and Hanseniaspora), regardless of the soil management system applied. In contrast, other fermentative wine yeasts such as Lanchacea and Torulaspora seemed to be permanently resident in the vineyard soil, especially in the NV vineyard. Hence, contamination of the grapes with vineyard soil may increase yeast biodiversity during spontaneous fermentation, which could affect the wine's organoleptic quality.

How the environmental fate of clomazone in rice fields is influenced by amendment with olive-mill waste under different regimes of irrigation and tillage.

BACKGROUND: Irrigation and tillage systems alone or in combination with organic amendments can strongly influence soil properties, which in turn may also modify the environmental fate of any pesticides applied. This study was aimed at determining how amendment with composted olive-mill waste (W) influenced leaching, sorption, and persistence of the herbicide clomazone in rice field soils under different tillage and irrigation management practices. The field trial conducted covered 3 years in succession, with six treatments: irrigation by sprinkler and conventional tillage without (ST) or with W application (80 Mg ha-1 ) (STW), irrigation by sprinkler but no tillage (SNT), irrigation by sprinkler but no tillage with W application (SNTW), and continuous flooding irrigation and tillage without (FT) and with W application (FTW). RESULTS: Application of W significantly increased the adsorption of clomazone to soil in the first and third years. In the first year, the persistence of clomazone under aerobic (t1/2  = 33.1-36.3 days) and anaerobic incubation conditions (t1/2  = 3.43-10.8 days) decreased after W application to t1/2 values in the ranges 18.1-29.7 and 3.06-5.44 days, respectively. However, in the third year, although clomazone persistence increased significantly in SNT and ST when W was applied under both incubation conditions, it decreased significantly in FT management under anaerobic incubation conditions. The addition of W led to less leaching of clomazone, particularly for the FT case where the herbicide leaching losses were 2.8 and 2.6 times lower in the first and third years after W addition, respectively. CONCLUSIONS: Using W as an organic amendment could be regarded as an invaluable strategy to reduce water contamination by clomazone in rice production, especially under traditional tillage and flooding management. © 2019 Society of Chemical Industry.

Olive mill sludge may reduce water contamination by 4-chloro-2-methylphenoxyacetic acid (MCPA) in non-flooding but enhance it in flooding rice cropping agroecosystems.

A field experiment covering three years was conducted to evaluate how composted olive mill sludge (OS) influenced MCPA's environmental fate in rice soils under six combinations of tillage and irrigation cultivation techniques: tillage and sprinkler irrigation without (TS) or with (TSOS) the addition of OS (80 Mg ha-1), no-tillage and sprinkler irrigation without (NTS) or with (NTSOS) OS, and tillage and continuous flooding without (TF) or with (TFOS) OS. The measurements made in the first and third years after OS application were taken to constitute the "direct" and "residual" effects, respectively. After OS amendment, Kd (partition coefficients) values in the direct year were lower by factors of 1.1, 1.3, and 1.9 in TSOS, NTSOS, and TFOS, respectively, relative to the corresponding unamended soils, and in the residual year by factors of 1.1 and 1.5 in TSOS and NTOS, but greater by a factor of 1.5 in TFOS, than in the corresponding unamended soils, respectively. The dissipation of MCPA was very fast under both anaerobic (t1/2 = 1.80-5.29 d) and aerobic (t1/2 = 2.23-9.42 d) incubation conditions. The field application of OS led to a decrease in MCPA persistence under both incubation conditions, especially in the TF case. However, while under aerobic conditions the half-life (t1/2) decreased after OS addition in the direct and residual years, under anaerobic condition it only decreased in the direct year. While the application of OS in TS and NTS led to less leaching of MCPA, in TF it led to 1.4 and 1.2 times more leaching losses of the herbicide for the direct and residual years, respectively. Therefore, the use of OS in rice production could be considered an effective strategy for reducing water contamination by MCPA in at least the short- and medium-terms after its application, but only under non-flooding crop management regimes irrespective of the tillage practice implemented.

Environmental fate of bensulfuron-methyl and MCPA in aerobic and anaerobic rice-cropping systems.

Bensulfuron-methyl (BM) and 4-chloro-2-methylphenoxyacetic acid (MCPA) are herbicides widely used in rice agroecosystems, and are commonly found in their environments, especially in water resources. The objective of this study was to evaluate the sorption-desorption, leaching, and dissipation of BM and MCPA under aerobic and anaerobic rice cropping conditions. For this purpose, a three-year field experiment was conducted in SW Spain using four management systems: aerobic with sprinkler irrigation and tillage (ST), sprinkler irrigation and no-tillage (SNT), long-term sprinkler irrigation and no-tillage (SNTLT), and anaerobic with flooding and tillage (FT). At the end of the experiment, the partition coefficients (Kd-values) in ST were (2.7, 3.1, and 3.9) and (1.2, 1.5, and 1.9) times significantly lower than the values in {SNT, SNTLT, and FT} for BM and MCPA, respectively. Greater sorption was related to lower values of soil pH for both herbicides and to higher contents in humic acids for BM and fulvic acids for MCPA. The persistence was much longer for BM (t1/2 = 26.9-52.1 days) than for MCPA (t1/2 = 1.54-21.1 days) in all management systems, and both herbicides' dissipation rates were generally greater under aerobic than under anaerobic conditions. The mobility of MCPA was much greater than that of BM. Compared with SNT and SNTLT, leaching losses of the applied BM were greater by 51% for ST, and of the applied MCPA by 55% and 99% for ST and FT, respectively. Therefore, only aerobic rice production with no-tillage in the short- or long-terms could be considered as alternative management strategies with which to reduce water contamination by BM and MCPA in rice-growing environments.

New Insights into the Genome Organization of Yeast Killer Viruses Based on "Atypical" Killer Strains Characterized by High-Throughput Sequencing.

Viral M-dsRNAs encoding yeast killer toxins share similar genomic organization, but no overall sequence identity. The dsRNA full-length sequences of several known M-viruses either have yet to be completed, or they were shorter than estimated by agarose gel electrophoresis. High-throughput sequencing was used to analyze some M-dsRNAs previously sequenced by traditional techniques, and new dsRNAs from atypical killer strains of Saccharomyces cerevisiae and Torulaspora delbrueckii. All dsRNAs expected to be present in a given yeast strain were reliably detected and sequenced, and the previously-known sequences were confirmed. The few discrepancies between viral variants were mostly located around the central poly(A) region. A continuous sequence of the ScV-M2 genome was obtained for the first time. M1 virus was found for the first time in wine yeasts, coexisting with Mbarr-1 virus in T. delbrueckii. Extra 5'- and 3'-sequences were found in all M-genomes. The presence of repeated short sequences in the non-coding 3'-region of most M-genomes indicates that they have a common phylogenetic origin. High identity between amino acid sequences of killer toxins and some unclassified proteins of yeast, bacteria, and wine grapes suggests that killer viruses recruited some sequences from the genome of these organisms, or vice versa, during evolution.

Behaviour of bentazon as influenced by water and tillage management in rice-growing conditions.

Bentazon is a widely used herbicide in rice agroecosystems that has commonly been found in water resources. To assess how tillage and water regimes affect sorption/desorption, dissipation and leaching of bentazon in Mediterranean rice-growing conditions, field experiments were carried out using tillage and flooding (TF), tillage and sprinkler irrigation (TS), no-tillage and sprinkler irrigation (NTS) and long-term no-tillage and sprinkler irrigation (NTS7). After 3 years, the Kd values in TS were 2.3, 1.6 and 1.7 times lower than the values in NTS7, NTS and TF respectively. Greater sorption of bentazon was related to higher contents in total organic carbon and, although to a lesser extent, in humic acids and dissolved organic carbon. The persistence of bentazon was significantly greater under anaerobic (half-life DT50 = 94.1-135 days) than under aerobic (DT50 = 42.4-91.3 days) incubation conditions for all management regimes. Leaching losses of bentazon were reduced from 78 and 74% in TS and TF to 61 and 62% in NTS7 and NTS respectively. The mid- and long-term implementation of sprinkler irrigation in combination with no-tillage could be considered a management system that is effective at reducing water contamination by bentazon in Mediterranean rice-growing agroecosystems. © 2017 Society of Chemical Industry.

Influence of the dominance of must fermentation by Torulaspora delbrueckii on the malolactic fermentation and organoleptic quality of red table wine.

Torulaspora delbrueckii can improve wine aroma complexity, but its impact on wine quality is still far from being satisfactory at the winery level, mainly because it is easily replaced by S. cerevisiae yeasts during must fermentation. New T. delbrueckii killer strains were selected to overcome this problem. These strains killed S. cerevisiae yeasts and dominated fermentation better than T. delbrueckii non-killer strains when they were single-inoculated into crushed red grape must. All the T. delbrueckii wines, but none of the S. cerevisiae wines, underwent malolactic fermentation. Putative lactic acid bacteria were always found in the T. delbrueckii wines, but none or very few in the S. cerevisiae wines. Malic acid degradation was the greatest in the wines inoculated with the killer strains, and these strains reached the greatest dominance ratios and had the slowest fermentation kinetics. The T. delbrueckii wines had dried-fruit/pastry aromas, but low intensities of fresh-fruit aromas. The aroma differences between the T. delbrueckii and the S. cerevisiae wines can be explained by the differences that were found in the amounts of some fruity aroma compounds such as isoamyl acetate, ethyl hexanoate, ethyl octanoate, and some lactones. This T. delbrueckii effect significantly raised the organoleptic quality scores of full-bodied Cabernet-Sauvignon red wines inoculated with the killer strains. In particular, these wines were judged as having excellent aroma complexity, mouth-feel, and sweetness.

De-oiled two-phase olive mill waste may reduce water contamination by metribuzin.

The impact of de-oiled two-phase olive mill waste (DW) on the behavior of metribuzin in Mediterranean agricultural soils is evaluated, and the effects of the transformation of organic matter from this waste under field conditions are assessed. Four soils were selected and amended in the laboratory with DW at the rates of 2.5% and 5%. One of these soils was also amended in the field with 27 and 54 Mg ha(-1) of DW for 9 years. Significant increases in metribuzin sorption were observed in all the amended soils. In the laboratory, the 5% DW application rate increased the t1/2 values of metribuzin from 22.9, 35.8, 29.1, and 20.0 d for the original soils to 59.2, 51.1, 45.7, and 29.4d, respectively. This was attributable mainly to the inhibitory effect of the amendment on microbial activity. However, the addition of DW transformed naturally under field conditions decreased the persistence down to 3.93 d at the greater application rate. Both amendments (fresh and field-aged DW) significantly reduced the amount of metribuzin leached. This study showed that DW amendment may be an effective and sustainable management practice for controlling groundwater contamination by metribuzin.

A new wine Torulaspora delbrueckii killer strain with broad antifungal activity and its toxin-encoding double-stranded RNA virus.

Wine Torulaspora delbrueckii strains producing a new killer toxin (Kbarr-1) were isolated and selected for wine making. They killed all the previously known Saccharomyces cerevisiae killer strains, in addition to other non-Saccharomyces yeasts. The Kbarr-1 phenotype is encoded by a medium-size 1.7 kb dsRNA, TdV-Mbarr-1, which seems to depend on a large-size 4.6 kb dsRNA virus (TdV-LAbarr) for stable maintenance and replication. The TdV-Mbarr-1 dsRNA was sequenced by new generation sequencing techniques. Its genome structure is similar to those of S. cerevisiae killer M dsRNAs, with a 5'-end coding region followed by an internal A-rich sequence and a 3'-end non-coding region. Mbarr-1 RNA positive strand carries cis acting signals at its 5' and 3' termini for transcription and replication respectively, similar to those RNAs of yeast killer viruses. The ORF at the 5' region codes for a putative preprotoxin with an N-terminal secretion signal, potential Kex2p/Kexlp processing sites, and N-glycosylation sites. No relevant sequence identity was found either between the full sequence of Mbarr-1 dsRNA and other yeast M dsRNAs, or between their respective toxin-encoded proteins. However, a relevant identity of TdV-Mbarr-1 RNA regions to the putative replication and packaging signals of most of the M-virus RNAs suggests that they are all evolutionarily related.

Environmental fate of the herbicide MCPA in agricultural soils amended with fresh and aged de-oiled two-phase olive mill waste.

Olive oil agrifood industry generates large amounts of waste whose recycling as organic amendment represents an alternative to their disposal. The impact of de-oiled two-phase olive mill waste (DW) on the fate of 4-chloro-2-methylphenoxyacetic acid (MCPA) in Mediterranean agricultural soils was evaluated. Furthermore, the effect of the transformation of organic matter from this waste under field conditions was assessed. Four Mediterranean agricultural soils were selected and amended in laboratory with fresh DW and field-aged DW (DW and ADW treatments, respectively). Adsorption capacity increased by factors between 1.18 and 3.59, for the DW-amended soils, and by factor of 4.93, for ADW-amended soil, with respect to unamended soils, when 5% amendment was applied. The DW amendment had inhibitory effect on dehydrogenase activity and slowed herbicide dissipation, whereas the opposite effect was observed in ADW treatments. In the field-amended soil, the amount of MCPA leached was significantly reduced from 56.9% for unamended soil to 15.9% at the 5% rate. However, leaching losses of MCPA increased in the laboratory-amended soils, because of their high water-soluble organic carbon values which could enhance MCPA mobility, especially in the acidic soils. Therefore, the application of DW as organic amendment in Mediterranean agricultural soils could be an important management strategy to reduce MCPA leaching, especially if the organic matter had been previously transformed by ageing processes.

Behavior of MCPA in four intensive cropping soils amended with fresh, composted, and aged olive mill waste.

An evaluation was made of the impact of olive mill waste and its organic matter transformation on the sorption, desorption, leaching, and degradation of the herbicide MCPA when the waste was applied to four Mediterranean soils. The soils were amended in the laboratory with fresh, composted, and field-aged olive mill waste (OW, COW, and AOW treatments, respectively). It was found that the greater the amount of OW applied to the soils, but especially the greater its organic matter maturity, the greater the adsorption of MCPA. Compared with unamended soils, at the 5% rate of application the adsorption capacity increased by between 9.8% and 40%, 148% and 224%, and by 258% for the OW, COW, and AOW amended soils, respectively. The hysteresis coefficients were significantly lower in the OW-amended soils than in AOW or COW-amended soils, indicating that the adsorbed MCPA could be easily desorbed in OW-amended soils if the amendment is not aged or composted. While the OW addition greatly extended the persistence of MCPA, the application of COW enhanced MCPA degradation in all the soils, as corresponded to the increased soil microbial activity indicated by the higher levels of soil dehydrogenase activity. Fresh OW amendment significantly increased the amount of MCPA leached (from 13.7% in the most alkaline soil to 36.7% in the most acidic, at the 5% rate of application), favored by the higher levels of water soluble organic carbon content. However, leaching losses of the herbicide were reduced by up to 39.9% and 55.3% in the COW- and AOW-amended soils at the 5% loading rate, respectively. The use of OW with a high degree of organic matter maturity may be regarded as a potentially useful management practice to reduce MCPA leaching in soils with low organic matter content. The application of fresh OW, however, could well increase the risk of groundwater contamination by this herbicide, especially in acidic soils.

Impact of oiled and de-oiled olive mill waste amendments on the sorption, leaching, and persistence of S-metolachlor in a calcareous clay soil.

Agricultural practices based on periodic inputs of organic amendments are strongly recommended for Mediterranean agro-ecosystems. Such amendments can change the soil's properties and transport characteristics, and hence affect the behaviour and fate of pesticides. S-metolachlor is an herbicide commonly used in intensive crops. The aim of this study was to investigate the influence of fresh oiled (OW) and de-oiled two-phase olive mill waste (DW) amendments on the sorption, leaching, and persistence of the herbicide S-metolachlor in a calcareous clay soil. The soil was amended in the laboratory with OW and DW at the rates of 2.5% and 5% (w/w). Significant increases in S-metolachlor sorption were observed in all amended soils. The addition of OW and DW increased the herbicide half-life from 27 d for the original soil to 41 and 47 d at the higher application rate of OW and DW, respectively. There was a significantly greater retention of the herbicide at the higher OW and DW loading rate. However, whereas the amount of S-metolachlor in the leachate was reduced by increasing the amount of OW, it was unaffected by increasing the amount of DW. The results lend support to the potential of OW and DW amendments as an effective management practice to increase S-metolachlor persistence in soils. This increase does not necessarily ensure decreased leaching of the herbicide but it could also increase the risk of surface water contamination at higher application rate.

Sorption, leaching and persistence of metribuzin in Mediterranean soils amended with olive mill waste of different degrees of organic matter maturity.

Metribuzin is a widely used herbicide, and worldwide is one of the most important contaminants in ground and surface waters. The aim of this study was to evaluate the impact amendment with fresh, composted, and field-aged olive mill waste (OW, COW, and AOW, respectively) had on the behaviour of this herbicide in four typical Mediterranean soils. A batch equilibration method was used to determine metribuzin adsorption-desorption. Leaching experiments were studied in hand-packed soil columns. Half-lives were calculated with incubation studies. Soil dehydrogenase activity (DHA) was also monitored. Metribuzin adsorption in the soils increased not only with increasing amounts of amendment, but especially with increasing degree of organic matter humification. Compared to unamended soils, the adsorption capacity increased by between 81% and 216%, 135% and 193%, and by 363% for the OW, COW, and AOW amended soils, respectively, at a 5% rate of application. The addition of COW enhanced metribuzin degradation in all the soils. In contrast, OW addition increased metribuzin persistence, attributable mainly to the inhibitory effect of this amendment on microbial activity, especially in the acidic soils. The AOW-amended soils, which had the smallest labile fraction of soil organic matter and greatest degree of humification, showed the shortest herbicide persistence. The OW and COW amendments significantly reduced the amount of metribuzin leached. This was especially so in the latter case because of the higher sorption capacity and the faster degradation of the pesticide. The use of OW as organic amendment, especially when it has a high degree of organic matter humification, may be a useful management practice for reducing the risk of groundwater contamination by metribuzin in soils with low organic matter content.