The distribution of acetohydroxyacid synthase in soil bacteria

Most bacteria possess the enzyme acetohydroxyacid synthase, which is used to produce branched-chain amino acids. Enteric bacteria contain several isozymes suited to different conditions, but the distribution of acetohydroxyacid synthase in soil bacteria is largely unknown. Growth experiments confirmed that Escherichia coli, Salmonella enterica serotype Typhimurium, and Enterobacter aerogenes contain isozymes of acetohydroxyacid synthase, allowing the bacteria to grow in the presence of valine (which causes feedback inhibition of AHAS I) or the sulfonylurea herbicide triasulfuron (which inhibits AHAS II) although a slight lag phase was observed in growth in the latter case. Several common soil isolates were inhibited by triasulfuron, but Pseudomonas fluorescens and Rhodococcus erythropolis were not inhibited by any combination of triasulfuron and valine. The extent of sulfonylurea-sensitive acetohydroxyacid synthase in soil was revealed when 21 out of 27 isolated bacteria in pure culture were inhibited by triasulfuron, the addition of isoleucine and/or valine reversing the effect in 19 cases. Primers were designed to target the genes encoding the large subunits (ilvB, ilvG and ilvI) of acetohydroxyacid synthase from available sequence data and a ∼355 bp fragment in Bacillus subtilis, Arthrobacter globiformis, E. coli and S. enterica was subsequently amplified. The primers were used to create a small clone library of sequences from an agricultural soil. Phylogenetic analysis revealed significant sequence variation, but all 19 amino acid sequences were most closely related to published large subunit acetohydroxyacid synthase amino acid sequences within several phyla including the Proteobacteria and Actinobacteria. The results suggested the majority of soil microorganisms contain only one functional acetohydroxyacid synthase enzyme sensitive to sulfonylurea herbicides.     

Differential induction of cytochrome P450-mediated triasulfuron metabolism by naphthalic anhydride and triasulfuron.

Cytochrome P450 monooxygenases play paramount roles in the detoxification of herbicides as well as in the synthesis of lignins, flavonoids, and phenolic acids. Biochemical analysis of triasulfuron metabolism in maize (Zea mays) seedlings has demonstrated that the P450(s) responsible for detoxification of this herbicide is induced by naphthalic anhydride (NA), a plant safener, and by triasulfuron, the herbicide itself. Induction studies conducted with seedlings of different ages suggest that two separate response pathways modulate this P-450 activity. Induction by NA is independent of the developmental age of the seedlings up to 6.5 d; induction by triasulfuron is tightly modulated with respect to developmental age in that triasulfuron metabolism can be induced by triasulfuron in young (2.5 d) but not older (6.5 d) seedlings. Induction by NA administered in combination with triasulfuron synergistically enhances triasulfuron metabolism in younger seedlings to levels substantially above that obtained with either herbicide or safener treatment alone. In older seedlings, NA plus triasulfuron treatment induces triasulfuron metabolism to only the level of NA treatment alone, indicating again that the induction cascade responding to triasulfuron is nonfunctional in later development. MnCl2 studies indicate that the triasulfuron insensitivity of older seedlings does not result from a general limitation in the inducibility of this P-450 detoxification system but rather from specific limitations in the triasulfuron-response pathway.     

除草剂对桉树人工林下植物及土壤微生物群落的影响

除草剂在桉树人工林中的应用越来越普遍,但关于除草剂对桉树人工林林下植物和土壤微生物群落的影响知之甚少。通过桉树人工林低剂量高频率(LHF)、中剂量中频率(MMF)、高剂量低频率(HLF)除草剂喷施试验,并与人工除草(MT)为对照,比较分析不同剂量、不同频率除草剂施用对林下植物和土壤微生物群落的影响。结果表明,施用除草剂导致桉树人工林林下植物种类和功能群组成发生显著变化,但并未显著降低林下植物群落物种丰富度和多样性,随除草剂施用频率的降低及恢复时间的增加,物种丰富度及多样性指数呈恢复趋势。除草剂施用也导致土壤养分含量降低。除草剂通过对林下植物群落和土壤养分的负面影响间接影响土壤微生物群落。LHF显著降低藤本植物而显著提高蕨类植物功能群的重要值,从而显著降低了微生物群落、真菌和放线菌的磷脂脂肪酸(PLFA)含量。MMF显著降低木本和藤本植物而显著提高禾草植物功能群的重要值,导致土壤微生物群落和放线菌的PLFA含量显著降低。HLF未显著影响林下植物及土壤微生物群落,但土壤全磷含量显著降低,速效磷含量也大幅下降。施用除草剂显著降低了土壤微生物生物量碳、氮的含量。因此,生产上应减少除草剂的施用,...     

Some continuing uncertainties in knowledge of herbicide behaviour in the soil

Recent developments concerned with the variability in the performance of soil- applied herbicides are reviewed with particular reference to phytotoxicity and to factors controlling persistence. It remains questionable that measurements of adsorption and herbicide concentration in the soil solution can adequately predict the phytoxicity of a herbicide in the soil. This is because such an approach ignores the spatial distribution of the herbicide and the parts of the plant through which a herbicide can be taken up. It also ignores the influence of climatic factors on persistence, uptake and plant response. In a general way persistence and the factors which control it are understood but detailed, reliable, quantitative analyses of the phenomena are not yet available. Much remains to be done before the interactions of the soil microflora with its environment and consequent effects on herbicides can be defined. There are still opportunities to improve the precision of use of soil-applied herbicides based on better comprehension of the processes involved in the expression and duration of phytotoxicity.     

Simazine application inhibits nitrification and changes the ammonia-oxidizing bacterial communities in a fertilized agricultural soil

s-Triazine herbicides are widely used for weed control, and are persistent in soils. Nitrification is an essential process in the global nitrogen cycle in soil, and involves ammonia-oxidizing Bacteria (AOB) and ammonia-oxidizing Archaea (AOA). In this study, we evaluated the effect of the s-triazine herbicide simazine on the nitrification and on the structure of ammonia-oxidizing microbial communities in a fertilized agricultural soil. The effect of simazine on AOB and AOA were studied by PCR-amplification of amoA genes of nitrifying Bacteria and Archaea in soil microcosms and denaturing gradient gel electrophoresis (DGGE) analyses. Simazine [50?μg g(-1) dry weight soil (d.w.s)] completely inhibited the nitrification processes in the fertilized agricultural soil. The inhibition by simazine of ammonia oxidation observed was similar to the reduction of ammonia oxidation by the nitrification inhibitor acetylene. The application of simazine-affected AOB community DGGE patterns in the agricultural soil amended with ammonium, whereas no significant changes in the AOA community were observed. The DGGE analyses strongly suggest that simazine inhibited Nitrosobacteria and specifically Nitrosospira species. In conclusion, our results suggest that the s-triazine herbicide not only inhibits the target susceptible plants but also inhibits the ammonia oxidation and the AOB in fertilized soils.     

Vegetative reproduction and chemical control with post-emergent herbicides of field bindweed (Convolvulus arvensis L.)

It is clearly seen from data that roots of Convolvulus arvensis L. have more and less intensive regenerative period during growing season. The more intensive period is in autumn, because in that time roots culminate nutrients, carbohydrate as starch and sugar. The less intensive regenerative or shoot-growing period is in spring, called "late spring bud dormancy". Experiments were conducted to get more information and further details about the regenerative capacity of roots close to and far from the collar of Convolvulus arvensis L. Root segments closer to collar have an intensive regenerative capacity than those ones further to collar. By data of Bakke et al. (1939) is well known, roots exhumed from deep soil layers are able to create shoots with low intensity. So finally we can exclaim that regenerative capacity is decreasing further to collar. Using mechanical weed control it is sufficient to till the upper layer of soil, but many times. Chemical treatments are most effective in the integrated weed control. It is clearly seen that auxin-type herbicide such as 2,4-D, fluroxipir, MCPA. dicamba give the best result. They gave 95% weed control effect used them separately or in combination with other herbicides. Combination of Banvel 480 S (dicamba) and Logran 75 WG (triasulfuron) introduced 95% weed control effect. Only one time got absolutely 100% weed control effect, in the case of Glyphosate active substance. Caused total plant destruction. Excellent result was given with the application of Pledge 50WP (flumioxazin). Herbicides mentioned above are absolutely allowed to take an important and significant part in chemical plant protection against Convolvulus arvensis L. Other herbicides like Granstar 75DF (tribenuron-methyl), Basis 75DF (rimsulfuron + tifensulfuron-methyl) and Huszár (jodosulfuron-methyl-sodium + mefenpir-diethyl) are not so effective against Convolvulus arvensis L., as compared to the previous ones.     

Effects of glyphosate herbicide on soil and litter macro-arthropods in rainforest: Implications for forest restoration

Summary    Ecological restoration activities, including reforestation, often involve the use of herbicides for the removal of weedy plant cover. Little is known, however, about the effects of herbicides on assemblages of non-target organisms that colonize restored patches. We describe a field experiment to investigate effects of glyphosate herbicide (Roundup® Biactive™) on rainforest-associated soil- and litter-dwelling macro-arthropods. Our experimental protocol differed in two ways from other ecotoxicological studies of herbicides. First, we applied herbicide at a rate considerably greater than the manufacturer's recommended maximum in order to simulate worst-case scenarios that may occur in the practice of forest restoration. Second, our field experiment was carried out under dense canopy cover with sparse understorey vegetation, so that indirect impacts caused by the loss of existing vegetation were eliminated. Paired herbicide-treated and control plots were created within five rainforest remnants on the Maleny plateau of subtropical eastern Australia. Macro-arthropods were collected using litter extraction before, approximately 3 days after, and 3 months after herbicide application. Responses of arthropods were analysed at two levels of taxonomic resolution: 'coarse' arthropods (arthropods sorted to Order/Class), and ant species. Our results suggest that the use of glyphosate herbicide formulated as Roundup® Biactive™ is suitable for the control of unwanted plants in rainforest restoration sites as it appears to have minimal impact on assemblages of soil and litter macro-arthropods or at least those typical of intact rainforest.     

Effects of three herbicides on soil microbial biomass and activity

Three post-emergence herbicides (2,4-D, picloram and glyphosate) were applied to samples of an Alberta agricultural soil at concentrations of 0, 2, 20, and 200 μg g⁻¹. The effects of these chemicals on certain microbial variables was monitored over 27 days. All herbicides caused enhancement of basal respiration but only for 9 days following application, and only for concentrations of 200 μg g⁻¹. Substrate-induced respiration was temporarily depressed by 200 μg g⁻¹ picloram and 2,4-D, and briefly enhanced by 200 μg g⁻¹ glyphosate. It is concluded that because changes in microbial variables only occurred at herbicide concentrations of much higher than that which occurs following field application, the side-effects of these chemicals is probably of little ecological significance.     

Influence of four herbicides on the algal flora of a prairie soil

Summary Four herbicides (2,4-D, trifluralin, MCPA and TCA) were applied at two concentration levels to isolated cores of a grassland loam soil. After herbicide contact times of 1, 5, and 20 days, samples were taken and the algal population estimated both quantitatively and qualitatively using two selective mineral salts media. Thirty one genera of algae were identified as occurring in the soil. Of these, Chlamydomonas, Chlorococcum, Hormidium, Palmella, and Ulothrix proved to be so sensitive to the four herbicides that they were rarely isolated from the cores after treatment. Other algal genera were found to be less sensitive, and the theoretical percentile sensitivity of fifteen genera was calculated. Chlorella, Lyngbya, Nostoc, and Hantzschia were found to be the most resistant algae, having percentile sensitivity to all four herbicides of less than 50%. Some algal genera varied in their sensitivity to each of the herbicides. Scytonema was sensitive to all of the herbicides except 2,4-D, while Tolypothrix showed a greater tolerance to MCPA. In the top cm of the soil, the reduction in cell numbers experienced by many algal genera after herbicide treatment was offset by an increase in the population of Chlorella. Stichococcus, Oscillatoria, and Spongiochloris all exhibited the ability to recover rapidly after a reduction in cell numbers resulting from the application of one of the herbicides. An overall reduction in cell numbers was noted for the algae growing preferentially on a nitrogen-free medium (i.e. potential nitrogen-fixers). re]19760511     

Effects of the herbicides fluometuron and prometryn of Rhizoctonia solani in soil cultures.

Responses of Rhizoctonia solani to herbicides in soil cultures were assessed by measuring soil enzyme activity and other growth-related factors. Both beta-galactosidase (EC 3.2.1.23) and phosphatase (EC 3.1.3.1.3.1.3.2) activities were highly correlated with amounts of mycelium in soil. Both enzyme activities were reduced significantly by either fluometuron or prometryn at 40 microgram/g of soil; the pathogen was more distinctly suppressed by fluometron and showed a stronger tendency to overcome the effects of prometryn with time. Inhibition was also reflected in reduced ultilization of glucose and less CO2-C evolved. Except for an increase in beta-galactosidase activity in the presence of 1 microgram fluometuron, low levels of either herbicide had little effect on the pathogen.     

Study of soil algae

Summary Algal techniques were used to study the soil factors affecting the toxicity of herbicides. It was found that the organic matter adsorbed 18 times more herbicide than clay. The inherent phytotoxicity of different herbicides was tested by these methods and the results obtained compared favourably to those of higher plants. The order of toxicity as tested by algae was: diuron >neburon>monuron>atrazine>simazine>atratone. The prediction of application rates of diuron and simazine by algal methods was tested in the field with wheat as cereal crop. The data obtained testified that the predictions were correct and better than the commercial recommendation. Good chemical control of weeds was achieved by herbicide at the early stage of crop growth. At later stages of crop development the toxicity of the chemical was reduced to insignificance and the crop plants were then capable to compete successfully against the emerging weeds. Thus a biological weed control was obtained. Such combined chemical-biological weed control technique should be regarded as the most desirable practice in agriculture.     

Relative effect of glyphosate on glyphosate-tolerant maize rhizobacterial communities is not altered by soil properties

The rhizobacterial composition varies according to the soil properties. To test if the effect of herbicides on the rhizobacterial communities of genetically modified NK603 glyphosate-tolerant maize varies according to different soil locations, a comparison was made between the effects of glyphosate (Roundup Plus), a post-emergence applied herbicide, and a pre-emergence applied herbicide (GTZ) versus untreated soil. The potential effect was monitored by direct amplification, cloning, and sequencing of the soil DNA encoding 16S rRNA, and high-throughput DNA pyrosequencing of the bacterial DNA coding for the 16S rRNA hypervariable V6 region. The results obtained using three different methods to analyze the herbicide effect on the rhizobacterial communities of genetically modified NK603 maize were comparable to those previously obtained when glyphosate-tolerant maize was grown in soil with different characteristics. Both herbicides decreased the bacterial diversity in the rhizosphere, with Actinobacteria being the taxonomic group most affected. The results suggest that both herbicides affected the structure of the maize rhizobacterial community, but glyphosate was environmentally less aggressive.     

Effect of drought stress,abscisic acid,and abscisic acid analogues on the efficacy of diclofop-methyl and tralkoxydim

The effects of drought stress, abscisic acid (ABA), and four ABA analogues on diclofop-methyl and tralkoxydim efficacy were investigated in oat (Avenu sativa). Drought stress conditions (6% soil moisture content) reduced the efficacy of diclofop-methyl at 350 g ha^–1, but not at 700 g ha^–1. Similarly, tralkoxydim efficacy was reduced by drought stress at 62.5 and 125 g ha^–1, but not at 250 g ha^–1. ABA (100 m), applied as a root drench 2 days before the herbicide, protected oat plants against all rates of diclofop-methyl and against low rates of tralkoxydim. Two ABA analogues protected oat plants from diclofop-methyl injury, whereas two others had no effect. Foliage applications of ABA were much less effective than root applications in protecting against herbicide injury. Protection by ABA and the two active analogues was dependent on the relative time of application with respect to the herbicides. Optimal protection by ABA and analogue I was obtained when they were applied between 2 days before and 1 day after diclofop-methyl application. Analogue IV protected plants when applied between 3 days before and 1 day after diclofop-methyl application. Partial protection against tralkoxydim activity by ABA was observed when it was applied between 1 day before and 1 day after herbicide application. Analogue I did not afford any protection against tralkoxydim, and analogue IV afforded partial protection when applied the same day or 1 day after tralkoxydim. The results indicate that protection against these postemergence herbicides, similar to that conferred by water stress, can be induced by ABA and structural analogues that apparently mimic the action of ABA.Abbreviations SMC soil moisture content - ABA abscisic acid     

Influence of the soil conditioner cellulose xanthate on the activity and persistence of nine acetanilide herbicides

Applications of cellulose xanthate equivalent to 25 kg cellulose/ha increased the numbers of weed seedlings by up to 23% compared with untreated controls. With propachlor and prynachlor, weed control was poorer in the presence of cellulose xanthate and analyses of soil samples demonstrated that the rate of herbicide loss from the soil was enhanced. Although the soil conditioner increased the rate of loss of some other acetanilide herbicides, weed control was not greatly affected. The results suggest that interactions between cellulose xanthate and acetanilide herbicides are only important with those compounds which are normally of very short persistence.     

Native seeds incorporated into activated carbon pods applied concurrently with indaziflam: a new strategy for restoring annual‐invaded communities?

Reestablishing native perennial vegetation in annual grass‐invaded rangelands is critical to restoring ecosystems. Control of exotics, often achieved with preemergent herbicides, is essential for successful restoration of invaded rangelands. Unfortunately, desirable species cannot be seeded simultaneously with preemergent herbicide application due to nontarget damage. To avoid this, seeding is commonly delayed at least 1 year. Delaying seeding increases the likelihood that annual grasses will begin reestablishing and compete with seeded species. Activated carbon (AC) can provide preemergent herbicide protection for seeded species because it adsorbs and deactivates herbicides. Previous studies suggest that a cylindrical herbicide protection pod (HPP), containing AC and seeds, allows desired species to be seeded simultaneously with the application of the preemergent herbicide imazapic. Unfortunately, imazapic is only effective at controlling annual grasses for 1–2 years. Indaziflam is a new preemergent herbicide which exhibits longer soil activity, with which HPPs may be useful. To assess this possibility, we evaluated seeding two native species (Wyoming big sagebrush [Artemisia tridentata Nutt ssp. wyomingensis] and bluebunch wheatgrass [Pseudoroegneria spicata (Pursh) Á. Löve]), both incorporated into HPPs and as bare seed, at four application rates of indaziflam in a grow room study. HPPs protected seeded species at low, mid, and high rates of indaziflam. The abundance and size of plants was greater in HPPs compared to bare seed treatments. These results suggest that HPPs can be used to seed native grasses and shrubs simultaneously with indaziflam application.     

Sensitive bioassays for determining residues of sulfonylurea herbicides in soil and their availability to crop plants

Sulfonylurea herbicides are potent inhibitors of plant growth and are extremely active against a wide spectrum of weeds. They are used at very low rates (10–50 g ai/ha) and cause rapid inhibition of root and shoot growth of young plants. Routine chemical assays for detecting low levels of these compounds are difficult and there is need to develop sensitive bioassay methods for detecting their extremely low residue levels in the soil.This paper describes a simple pot bioassay method with a self watering system using turnip (Brassica rapa) seedlings as test plants for quantitative determination of sulfonylurea herbicides. Results are presented with six of these compounds whose activity was investigated in widely differing substrates. The potential availability to plants was calculated from the dose-response curves in different substrates. The dose-response relationship has been described by a specifically developed computer model. Details are also given of a direct seeded bioassay method with controlled watering system using several test species for detection of sulfonylurea herbicides. The potential uses and practical applications of both techniques are discussed.     

Multi-Residue Analysis of Herbicides in Soil with an UPLC-ESI-MS Method

A rapid and sensitive method was developed for the determination of 51 herbicides in soil by ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC–ESI–MS). Using acetonitrile effectively extracted 22 kinds of triazine and other basic herbicides, and using 90:10 v/v acetonitrile-phosphate buffer (pH = 7.5) effectively extracted another 29 herbicides. The extract has not cleaned up further. Chromatographic separation was achieved within 10 min using gradient elution with acetonitrile–water as a mobile phase for 22 triazine and phenylurea herbicides, and with 5 mM ammonium acetate containing 0.1% formic acid aqueous solution–acetonitrile as a mobile phase for another 29 herbicides. The response was linear over two orders of magnitude with correlation coefficients (r²) higher than 0.99. The limits of quantification for the herbicides varied from 0.2 to 20 µg kg^?1. The intra- and inter-day precisions (relative standard deviation, RSD) were 2.2–9.3% and 5.7–17.1%, respectively. The average recovery varied from 61.6 to 112% with the RSD of 1.6–11.3%. Analyzing 51 soil samples from 17 counties formed the basis of this method. Three herbicide residues were found in four counties. Atrazine residue in soil for 17 counties was found; its content was 0.4–9.8 μg kg^?1. Nicosulfuron residue in soil for two counties was found, with a high up to 133 or 1317 μg kg^?1. Propazine (0.3 and 1.34 μg kg^?1), atratone (2.14 and 3.93 μg kg^?1), and cynanazine (0.34 μg kg^?1) in soils for some counties were also found. The validated method can ensure the rapid multi-class, multi-residue analysis at low μg kg^?1 level for 47 herbicides in soil. The developed method provides an effective analytical basis for controlling herbicide dosage, investigating their distribution and degradation, and evaluating their hazards on the environment and human health.     

No-tillage system applied to the sunflower (hybrid pioneer PR64E83) resistant to the tribenuron-methyl in the conditions from Romania

In the year 2006, the first experience with the NO-TILLAGE system for sunflower crops was organized in Romania, using the Pioneer PR64E83 sunflower hybrid resistant to the tribenuron-methyl herbicide. The experience took place in the Danube river flood plain, on an alluvial soil with a content between 3.1-4.5% and a content of clay of 34-40%. In the conventional system, in the fall of the year 2005, autumn ploughing was carried out, along with harrowing, and in spring the land was tilled two more times, using both the disk and the harrow. Before sowing, the land was prepared by using the combiner-machine. After the sunflower sprouted, three-time mechanical hoeing between the rows, as well as three-time manual hoeing on the sunflower row were performed. In the no-tillage system, no autumn ploughing, nor any other soil work were performed, and the sunflower was sowed directly, by using the Gaspardo seeder. Before sowing, the soil was covered in proportion of 70% by species of weeds of the Cirsum, Sonchus and Polygonum amphibium type. To destroy these species of weeds, 3 days before sowing, the soil was treated with the Roundup herbicide in a dose of 7 litres/ha. After the sunflower sprouted, when the plants had 4-5 leaves, the following sulphonylurea herbicides were applied post-emergently. (1) Express 50SG--which contains tribenuron-methyl (2) Granstar 75DF--which contains 75% tribenuron-methyl (3) Titus 25DF--which contains 25% rimsulfuron (4) Mistral--which contains 40 g/L nicosulfuron (5) Lintur 70WG--which contains 4.1% triasulfuron + 65.9% dicamba (6) Peak 75WG--which contains 75% prosulfuron The Express 50SG and Granstar 75DF herbicides were selective for the sunflower hybrid PR64E83, and the other herbicides although belonging o the same sulphonylurea group, they proved to have a very phytotoxic effect upon the sunflower crop. In the report, selectivity is presented depending on the dose of herbicide, weed control and sunflower production.     

Combining supercritical fluid extraction of soil herbicides with enzyme immunoassay analysis

Supercritical fluid extraction (SFE) of soil herbicides followed by enzyme immunoassay analysis (EIA) is explained in a step-by-step process. Extracted herbicides, include 2,4-D, simazine, atrazine, and alachlor. The herbicide, trifluralin was not successfully analyzed by EIA because of crossreacting metabolites. Problems with SFE, including uneven packing of cells, leaks, uneven flow and clogging, can largely be eliminated as the method parameters are optimized. It was necessary to add modifiers including methanol or acetone to the SF CO₂ to increase the solubility of the analytes. Detection limits of 2.5 ng/g soil for atrazine and alachlor and 15 ng/g soil for simazine and 2,4-D without concentration of the sample were achieved. Recoveries above 80% and relative standard deviations (RSDs) less than 15% for 2,4-D simazine, atrazine and alachlor were achieved. Atrazine and alachlor recoveries were above 90% with RSDs below 10%. Forty soil samples could be extracted and analyzed in an 8-h day.