Volatile Fatty Acids in Liquid Swine Manure Can Kill Microsclerotia of Verticillium dahliae

ABSTRACT Liquid swine manure added to acidic soils killed microsclerotia of the wilt fungus Verticillium dahliae. We investigated whether volatile fatty acids (VFAs) in the manure were responsible for this toxicity. The survival of microsclerotia was determined after exposure to various dilutions of manure or its VFA components. Acetic, propionic, and isobutyric acids constituted the major VFAs in the manure, while n-butyric, n-valeric, iso-valeric, and n-caproic acids were present in lesser amounts. Formic acid was not detected. The individual VFAs were more toxic to microsclerotia as the solution pH was decreased, indicating that the protonated forms of the VFAs were toxic (e.g., acetic acid and not acetate). The effective concentration reducing germination of microsclerotia by 95% (EC(95)) for formic and n-caproic acids was approximately 4 mM, the most toxic of the acids tested; for n-valeric, the EC(95) was 9.2 mM, isovaleric was 16.1 mM acids, and acetic, propionic, n-butyric, and isobutyric acids were approximately 30 mM. The toxicity of acetic acid, and likely all the others, was directly related to the duration of exposure. Inhibition of microsclerotia germination followed identical trends in solutions of the manure or in a mixture of VFAs with equivalent concentrations of the individual acids found in the manure. Similarly, germination declined to the same extent in the atmosphere above the manure or the VFA mixture, confirming the toxicity of VFAs to microsclerotia. Thus, under acid conditions, VFAs in liquid swine manure can kill microsclerotia of V. dahliae.     

Survival of soil-borne fungus Phymatotrichopsis omnivora after exposure to volatile fatty acids

Control of plant-pathogenic Phymatotrichopsis omnivora is difficult. In tests of volatile fatty acids (VFAs), mycelia in empty 9-cm Petri dishes with 3 or 6 μL of acetic, butyric, formic or propionic acid did not grow, but 37–67 % grew on potato dextrose agar with 6 μL VFA. Survival of sclerotia was <20 % on water agar with 154 μg/mL of a VFA or 12–40 % in sandy soil with 616 μg/mL. Survivability after 2-h immersion in 17 mmol/L varied greatly (0–100 %) depending on the VFA, but in 33–50 mmol/L, 0 % survived. This fungus was thus susceptible to VFAs.     

Liquid Swine Manure Can Kill Verticillium dahliae Microsclerotia in Soil by Volatile Fatty Acid, Nitrous Acid, and Ammonia Toxicity

ABSTRACT In previous studies, liquid swine manure (LSM) was sometimes shown to reduce Verticillium wilt of potato caused by Verticillium dahliae. We also observed that microsclerotia of this fungus died within 1 day, or between 3 and 6 weeks, after addition of LSM to some acid soils and within 1 week in some alkaline soils. In this study, we demonstrated that a volatile fatty acid (VFA) mixture with an identical concentration of VFAs as that found in an effective LSM reduced germination in an acid soil (pH 5.1) to the same extent as the LSM after 1 day of exposure. Germination was reduced by 45, 75, and 90% in the 10, 20, and 40% ([wt/wt] soil moisture) treatments, respectively, with the latter being equivalent to an application of 80 hl/ha. Addition to this acid soil of 19 LSMs (30% [wt/wt] soil moisture) collected from different producers resulted in complete kill of microsclerotia with 12 manures. Effective manures had a total concentration of nonionized forms of VFAs in soil solution of 2.7 mM or higher. In some acid soils (pH 5.8), addition of LSM (40% [wt/wt] soil moisture) did not kill microsclerotia until 3 to 6 weeks later. Here, a reduction in viability of microsclerotia was attributed to the accumulation of 0.06 mM nitrous acid in the soil solution at 4 weeks. When an LSM was added (40% [wt/wt] soil moisture) to an alkaline soil (pH 7.9) where VFAs are not toxic, microsclerotia germination was reduced by 80% after 1 week. Here the pH increased to 8.9 and the concentration of ammonia reached 30 mM in the soil solution. An ammonium chloride solution having an equivalent concentration of ammonium as the manure was shown to have the same spectrum of toxicity as the manure in assays ranging from pH 7 to 9, both in solutions and above the solutions. At pH 9, the concentration of ammonia reached 18 mM and 100% mortality of microsclerotia occurred. Thus, in acid soils, LSM can kill microsclerotia of V. dahliae by VFA and/or nitrous acid toxicity and in alkaline soils by ammonia toxicity. In order to take advantage of these mechanisms for disease reduction, the manure chemical composition, rate of addition, and soil characteristics need to be determined for each instance of use.     

Effects of oxadiargyl on direct-seeded rice and Echinochloa crus-galli under aerobic and anaerobic conditions

Dose–response experiments were conducted in glasshouse pot experiments to investigate the selectivity of oxadiargyl, a recently introduced herbicide, in direct‐seeded rice under both aerobic and anaerobic conditions. Crop sensitivity to oxadiargyl was comparatively greater for wet‐seeded (anaerobic) than for dry‐seeded rice (aerobic). Likewise, greater efficacy against Echinochloa crus‐galli (L.) was also observed under anaerobic conditions. These results indicate greater activity of oxadiargyl under anaerobic conditions, but that application pre‐sowing with subsequent flooding would reduce selectivity in wet‐seeded rice. The results are discussed in relation to rice production in Mediterranean agriculture.     

Mediation of partial resistance to rice blast through anaerobic induction of ethylene

ABSTRACT The correlation between anaerobic soil conditions and increased resistance to rice blast disease has long been observed without benefit of an adequate explanation. We researched flood depth, dissolved oxygen (DO), and ethylene relative to expression of partial blast resistance in cvs. M-201, Newbonnet, LaGrue, Mars, and Cypress. Cultivar blast index (BI) and flood DO decreased with increasing flood depth. BIs were positively correlated with DO. Total leaf blast lesions were 3.4 and 3.2 times greater in cvs. M-201 and LaGrue growing in a 5.0-mul liter(-1) DO nutrient solution than when growing in a 0.1-mul liter(-1) DO solution. Treatment with 0.25 mM ethephon, which releases ethylene, lowered BIs of Newbonnet, LaGrue, and Cypress growing upland when applied drench, foliar, or foliar-drench. If flooded, BIs of ethephon-treated cultivars were decreased by drench and foliar-drench applications only. BIs of upland plants were unchanged, whereas BIs of analogous flooded plants increased following treatment with 0.31 mM aminoethoxyvinylglycine (AVG), an ethylene biosynthesis inhibitor. We hypothesize that varying anaerobic conditions mediate production of phytohormones, particularly ethylene, which modify expression of inherent partial blast resistance in these rice cultivars.     

A technique to detect Erwinia carotovora ssp. carotovora in lily bulbs1

Erwinia carotovora ssp. carotovora can be detected very effectively in lily bulbs by anaerobic incubation of the bulbs followed by homogenization of the scales and plating of the diluted sap onto crystal-violet pectate medium. This technique, compared with bulb treatments without preliminary anaerobic incubation, also improves the possibility of recovering E.c.carotovora overall at low storage temperatures in the long term. Differences among Lilium cultivars regarding the interaction with E.c.carotovora probably exist; the bacterium multiplied at a higher rate in bulbs of cvs Enchantment, Mont Blanc, Star Gazer and Yellow Blaze. The technique could allow inspection of lily bulbs before planting.     

Survival of strawberry-pathogenic fungi Fusarium oxysporum f. sp. fragariae, Phytophthora cactorum and Verticillium dahliae under anaerobic conditions

The growth and survival of three strawberry pathogens, Fusarium oxysporum f. sp. fragariae (FOF), Phytophthora cactorum, and Verticillium dahliae, were examined in anaerobic (anoxic) conditions at several temperatures (10–40 °C). The growth and survival of these fungi were suppressed by anaerobic conditions in comparison with those in cultured aerobically. Under anaerobic conditions at 22.5 °C, tested isolates of FOF and P. cactorum grew slightly, but V. dahliae did not grow. The three fungi survive for markedly shorter time in the anaerobic conditions compared with the aerobic conditions at all tested temperatures except 40 °C for FOF and P. cactorum. Moreover, survival periods shortened as the cultivation temperature increased. These results demonstrate that anaerobic conditions contribute to eradicating these pathogens during flooding or reductive soil disinfestation.     

Genes related to flooding tolerance during germination and early growth of weedy rice

Evolution of flooding tolerance in weedy rice has occurred in several rice-growing regions, but the genes related to this process and the environmental effects are unknown. The objective of this study was to analyse the expression of genes related to flooding tolerance in response to temperature and flooding during the initial establishment of weedy rice. The experiments were carried out with rice cultivars IRGA 417 and Nipponbare, which are sensitive to flooding, and weedy rice ITJ03 and AV04 genotypes that have high and intermediate tolerance to flooding, respectively. The expression of genes related to reserve mobilisation, anaerobic respiration, escape and quiescence strategies was analysed at periods up to 24 days after sowing. The flooding tolerance of weedy rice genotype ITJ03 was associated with the expression of RAmy3D and OsTPP7 , which are involved in the mobilisation of carbohydrate reserves, ADH1 and ADH2, which participate in anaerobic respiration, and SNRKL1 that triggers rapid elongation of the coleoptile and emergence. Although the genes PDC1, SUS3 and SUB1 are important for flooding tolerance in cultivated rice, their expression was not directly related to flooding tolerance in weedy rice. A temperature of 20°C reduced levels of expression of the RAmy3D, ADH2 and SNRKL1 genes and low temperature had a negative effect on the establishment of weedy rice. Breeding of rice genotypes with tolerance of low temperatures and anaerobic conditions may be a viable strategy to improve the control of weedy rice in paddy fields.     

The role of ferrous ions in the rapid degradation of oxamyl,methomyl and aldicarb in anaerobic soils

The carbamoyloxime pesticides methomyl, oxamyl and aldicarb, together with the oxidation products of aldicarb, are known to break down much more rapidly in certain anaerobic subsoils than in the aerobic topsoils from the same site. Ferrous ions have now been shown to be involved in this reaction. Oxamyl was degraded in aqueous solutions at 30°C containing 250 μg ml^?1 Fe²⁺ with a half-life of about 10 h, independent of pH in the range of 5.65–7.66; the observed products of this reaction were N,N-dimethyl-l-cyanoformamide and methanethiol. These same products, rather than the oximino hydrolysis product observed from degradation in aerobic soils, were rapidly and quantitatively formed from oxamyl in suspensions of anaerobic reduced subsoils (Fe²⁺ concentration 27–41 μg ml^?1 soil water), but oxamyl was rather stable in water-saturated Vredepeel subsoil (Fe²⁺ concentration 0.65 μg ml^?1) in which the redox potential was much higher. Methomyl behaved similarly. The rates of reaction in the suspensions of anaerobic subsoils were greater than expected from the concentrations of Fe²⁺ in the soil water, but most of the Fe²⁺ present in soil was bound to the soil particles by cation exchange and this bound Fe²⁺ may have participated. Breakdown of aldicarb was accelerated both in solutions of Fe²⁺ and in the suspensions of anaerobic reduced subsoils, though the rate enhancement was less than observed with methomyl and oxamyl; 2-methyl-2-methylthiopropionitrile and 2-methyl-2-methylthiopropionaldehyde were the observed products from aldicarb in anaerobic soil but only the former was produced in Fe²⁺ solutions; the corresponding nitriles and aldehydes were also yielded by aldicarb sulphoxide and aldicarb sulphone in the anaerobic, reduced subsoils.     

猪场废水灌溉对土壤-夏玉米系统氮素分布的影响

通过田间小区试验,研究了猪场废水处理工艺中3个阶段出水(原水、厌氧水和仿生态塘水)与地下水1∶5混水和厌氧水不同灌溉量对土壤中矿质氮含量、夏玉米产量以及植株吸氮量的影响。结果表明:厌氧水不同灌溉量对各土层硝态氮和铵态氮含量影响差异较显著,而不同阶段出水混水灌溉对各土层硝态氮和铵态氮含量影响差异不显著;厌氧水不同灌溉量处理下玉米产量呈现中量厌氧水高量厌氧水低量厌氧水的趋势,不同处理阶段出水混水灌溉时,原水与地下水1∶5混水灌溉产量较其他处理高;仿生态塘混水灌溉玉米籽粒粗蛋白含量最高。建议适宜的猪场养殖废水厌氧出水灌水定额为500 m3/hm2,适宜的混水灌溉处理为仿生态塘水与地下水1∶5的配水比例。     

氟虫腈在三种土壤中的降解特性研究

实验室条件下,研究了氟虫腈在东北黑土、江西红壤和太湖水稻土中的降解特性。结果表明,氟虫腈在土壤中降解较慢,其在好气条件下的东北黑土、江西红壤和太湖水稻土中的降解半衰期分别为165、267和42 d,在渍水条件下的3种土壤中的降解半衰期分别为31、173和32 d。氟虫腈在pH 偏中性的太湖水稻土中降解最快;微生物对氟虫腈在土壤中的降解起主要作用;渍水条件有利于氟虫腈的降解,推测降解氟虫腈的微生物主要是厌氧菌属。     

Growth of facultative anaerobic population of human intestinal bacteria with the carbamate pesticide aminocarb

The effect of different concentrations of the carbamate pesticide, aminocarb (Matacil), on the growth and selection of facultative anaerobic bacteria and degradation of the pesticide by human endoflora of the intestinal tract was examined in vitro. Microorganisms were cultured under aerobic or anaerobic conditions, in nutrient broth and mineral media. The intestinal population was more sensitive to 10–1000 μg/ml aminocarb under anaerobic conditions than in aerobic culture; however, spontaneous degradation of aminocarb in media markedly affected the degree of bacterial growth inhibition in prolonged cultures. In addition, the type of culture medium appeared to influence the degree of aminocarb-induced bacterial growth inhibition. A dose of aminocarb inducing 50% growth inhibition was established for different culture conditions: for mineral medium, aminocarb inhibited bacterial growth by 50% at 600 μg/ml under anaerobic and aerobic conditions, whereas less than 50% inhibition was observed even at 1000 μg/ml aminocarb when bacteria were grown in nutrient broth. A selection of bacterial strains occurred in the presence of increasing aminocarb concentrations, which was determined quantitatively and qualitatively by the identification of codominants. A shift in several Escherichia coli biotypes was also observed in cultures with aminocarb, in comparison to control cultures. Bacterial degradation of aminocarb, under anaerobic and aerobic conditions, was determined in a mixed population of the intestinal microflora by high-performance liquid chromatography analysis of culture media. Data showed that aminocarb can be quickly degraded by human intestinal bacteria at relatively high pesticide concentrations. Moreover, other HPLC data suggest rapid spontaneous degradation of aminocarb in neutral and slightly alkaline pH conditions characteristic of the human intestinal tract, which can effectively eliminate the pesticide. Therefore, aminocarb, at the concentrations used, does not seriously affect the bacterial microflora of the human gut.     

Studies on the mechanisms of accumulation of phorate by the free-living nematode Panagrellus redivivus

Accumulation and metabolism of the organophosphate pesticide, phorate by the free-living nematode Panagrellus redivivus was studied under anaerobic conditions and in the presence of carbon monoxide or SKF 525A, and compared with results obtained under normal, aerobic conditions and using heat-killed nematodes. Both phorate hydrolysis and side-chain oxidation were inhibited under anaerobic conditions and with heat-killed nematodes, but only hydrolysis was affected in the presence of carbon monoxide. The much greater phorate accumulation in the nematodes under anaerobic conditions and with carbon monoxide could be explained by this metabolic inhibition. SKF 525A had no significant effect on phorate metabolism or accumulation. The possible mechanisms involved in phorate breakdown by P. redivivus are discussed.     

三唑酰草胺在不同类型土壤中的降解特性

为科学评价除草剂三唑酰草胺在土壤环境中的生态风险,采用室内模拟方法,研究了三唑酰草胺在吉林黑土、江西红土和安徽水稻土中的降解特性。结果表明:三唑酰草胺在土壤中的降解符合一级动力学方程。好氧条件下三唑酰草胺在3种土壤中的降解半衰期分别为86.5、106和91.4 d;厌氧条件下半衰期分别为106、130和127 d;水稻田厌氧条件下半衰期分别为162、219和188 d。研究表明,三唑酰草胺在水稻田厌氧条件下的降解速率明显慢于其他2种试验条件下。     

Anaerobic soil disinfestation is an alternative to soil fumigation for control of some soilborne pathogens in strawberry production

Alternatives to soil fumigation are needed for soilborne disease control. The aim of this study was to test anaerobic soil disinfestation (ASD) as an alternative to soil fumigation for control of critical soilborne pathogens in Californian strawberry production. Controlled environment experiments were conducted at 25 and 15 °C to test different materials as carbon sources for ASD using soil inoculated with Verticillium dahliae. Field trials were conducted in three locations comparing ASD with 20 Mg ha^?1 rice bran (RB) against fumigated and untreated controls, steam, mustard seed meal and fish emulsion. In ASD‐treated soils, temperature and extent of anaerobic conditions were critical for control of V. dahliae, but multiple carbon inputs reduced inoculum by 80–100%. In field trials, ASD with RB provided control of a number of pathogens, and in three of four trials produced marketable fruit yields equivalent to fumigation. Little weed control benefit from ASD was found. ASD with RB also induced changes in the soil microbiome that persisted through the growing season. When equivalent yields were obtained, net returns above harvest and treatment costs with ASD RB were 92–96% of those with bed fumigation based on average prices over the previous 5 years. ASD can be a viable alternative for control of some soilborne pathogens. Growers are adopting ASD in California strawberry production, but research to determine optimal soil temperatures, anaerobicity thresholds and carbon sources for effective control of specific pathogens is needed.     

Rate of transformation of atrazine and bentazone in water-saturated sandy subsoils

When pesticides leach through the soil to the upper groundwater zone, it is important to know whether further transformation occurs before the pumping wells for drinking water are reached. Atrazine and bentazone were incubated (at 10°C) in five water-saturated sandy subsoils (collected at depths between 1·5 and 3·5 m), simulating the conditions in the field. In three subsoils with comparatively low pH and intermediate to high redox potential, atrazine was transformed gradually, to leave 1·9%, 6·2% and 17·5% of the dose after about five years. In one of these subsoils, hydroxy-atrazine was detected; the amount corresponded to half of the dose of atrazine. In one anaerobic subsoil with high pH, the transformation of atrazine was comparatively fast (half-life about 0·15 year). Another anaerobic subsoil, with similar pH and a somewhat higher redox potential, however, showed hardly any transformation. Sterilization of the first anaerobic subsoil had no effect on the rate of transformation. In the course of about five years, bentazone in the first three subsoils was transformed gradually to leave <0·25%, 11% and 25% of the dose. Bentazone transformation in the two subsoils with high pH and low redox potential was very slow, but the presence of oxygen in one of these subsoils speeded up the transformation. © 1998 SCI     

Methods for assessing the susceptibility of potato tubers of different cultivars to rotting by Erwinia carotovora subspecies atroseptica and carotovora

The susceptibility of tubers of different potato cultivars to soft rot by Erwinia carotovora subspp. uroseptica and carotovora was assessed in 3 years by two methods. In one method, whole tubers inoculated at wounds with either bacterium were incubated under anaerobic conditions for 5 or o days at 15°C. In the other method, wounds made in tuber slices were allowed to heal or not, before inoculation with different concentrations of each bacterium and were then incubated under aerobic conditions for 3 days at 15°C. Most cultivars gave consistent reactions in repeated experiments using the same method, but there was some seasonal variation. A few cultivars were consistently susceptible (Klondyke and Manna) or resistant (Drayton) in both methods but others gave completely contrasting results (Record). In both methods and with all cultivars more rotting was caused by subsp. atroseptica than by subsp. carotovora because of the temperature of Incubation.     

Biological Soil Disinfestation (BSD), a new control method for potato brown rot,caused by <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> race 3 biovar 2

The potential of Biological Soil Disinfestation (BSD) to control potato brown rot, caused by Ralstonia solanacearum race 3 biovar 2, was investigated. BSD involves the induction of anaerobic soil conditions by increasing microbial respiration through incorporation of fresh organic amendments (here: grass or potato haulms) and by reducing re-supply of oxygen by covering with airtight plastic sheets. Control treatments were left without cover and amendment, or amended without covering or covered only without amendment. The effect of BSD on survival of R. solanacearum was tested at three different scales: in 1-l glass mesocosms under laboratory conditions, in 1.2-m-diam microplots positioned in an outdoor quarantine field, and in a naturally infested commercial field. Within a few days, anaerobic conditions developed in the BSD-treated soils. In the mesocosm and microplot experiment, anaerobic conditions persisted till the end of the 4-week experimental period. In the field experiment, the period of anaerobiosis was shorter due to birds damaging the plastic cover. In all three experiments, BSD reduced soil populations of R. solanacearum significantly by 92.5% to >99.9% compared to the non-amended and uncovered control treatments. In the field experiment, BSD also resulted in a significant reduction of R. solanacearum survival in potato tubers buried at 15 or 35 cm and in the rapid decomposition of superficially buried potatoes remaining after harvesting, thus destroying an important inoculum reservoir of R. solanacearum. The treatments with grass amendment only or covering with only plastic did not result in anaerobic conditions and did not decrease R. solanacearum populations during the experimental period. PCR-DGGE analyses of 16S-rDNA from soil samples of the various treatments in the mesocosm and microplot experiments revealed that BSD hardly affected bacterial diversity but did result in clear shifts in the composition of the bacterial community. The possible implications of these shifts are discussed. It is concluded that BSD has the potential to strongly decrease soil infestation levels of R. solanacearum and to become an important element in a sustainable and effective management strategy for potato brown rot, especially in areas where the disease is endemic.     

Clostridium bifermentans and C. subterminale are associated with kiwifruit vine decline,known as moria,in Italy

Since 2012, a new pathogenic syndrome has frequently been observed in many areas of kiwifruit cultivation in Italy. The main symptoms include an initial withering of the leaves followed by a total and sudden collapse of plants, mainly occurring during summer. The withered leaves fall and the main and secondary feeder roots appear rotten, sometimes showing a reddish-brown discoloration. The disease, that affects both the green and yellow-fleshed cultivars, has been called kiwifruit vine decline and is locally known as moria. The syndrome has been found consistently associated with soil waterlogging, which frequently occurs either after the traditional agronomical practice of irrigating orchards through surface irrigation or after very heavy rainfall. So far, the role played by bacteria in this syndrome has not been investigated. In the present study, Clostridium spp. were isolated from both rotten roots and soils obtained from Italian kiwifruit orchards affected by the syndrome, indicating for the first time that anaerobic bacteria are able to cause damage to woody crops. C. bifermentans and C. subterminale incited symptoms in kiwifruit in both in vivo and in vitro pathogenicity tests. Soil waterlogging seems to potentially favour colonization of kiwifruit roots by anaerobic bacteria, probably because saturation of the soil can facilitate proliferation and persistence of these bacteria during long periods of the vegetative growth of the crop. The occurrence of anaerobic bacteria does not exclude the possibility that other microorganisms can play additional/synergic role(s) in causing the kiwifruit vine decline.     

The potential for Fusarium oxysporum f. sp. fragariae,cause of fusarium wilt of strawberry,to colonize organic matter in soil and persist through anaerobic soil disinfestation

Fusarium wilt of strawberry, caused by Fusarium oxysporum f. sp. fragariae, is a disease of primary concern for strawberry production in many countries. Crop rotation and anaerobic soil disinfestation (ASD) have gained recent interest for their potential to contribute to management of this disease. Both techniques involve incorporation of organic matter into soil, which may be utilized by strains of Fusarium that are competitive saprophytes. We show that F. oxysporum f. sp. fragariae can colonize strawberry, lettuce, raspberry, and broccoli leaf tissues, which are sources of organic matter generated during crop rotation. This pathogen increased in soil population density during ASD treatments that did not become anaerobic, possibly as a result of growth on the organic amendment. However, significant population decreases were observed after ASD treatment when at least 100,000 cumulative reduced mV hours occurred in a 14-day experiment. Post-ASD abundance of F. oxysporum f. sp. fragariae in soil was negatively correlated with cumulative reduced mV hours. The only treatment that consistently caused disinfestation was exposed to a maximum temperature of 22 °C, which indicates there is potential for developing effective ASD treatments in the cool climates where strawberries are grown. Awareness that F. oxysporum f. sp. fragariae can act as a competitive soil saprophyte should be further investigated for its potential to alter disease outcomes where organic amendments are applied.