Pesticide inactivation of peanut glutamate dehydrogenase: biochemical basis of the enzyme's isomerization.

Glutamate dehydrogenase (GDH) isomerizes in response to pesticides and environmental chemicals, but the biochemical basis of the isomerization is not known. Clearer understanding of the isomerization would permit expansion of its utility in the diagnosis of the responses of plant tissues to challenged environments. Peanut plants were treated with different rates of Basagran (3-(1-methylethyl)-1H-2,1,3-benzothiadiazin-4(3H)-one 2,2-dioxide), Bravo 720 (tetrachloroiso-phthalonitrile), and Sevin XLR Plus (1-naphthyl N-methylcarbamate). Free solution isoelectric focusing, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) fractionated the peanut seed GDH to its constituent subunits and degradation polypeptides. After western transfer to nitrocellulose membrane, the GDH subunits and degradation polypeptides were immunodetected with anti-GDH. The pesticide treatments did not induce increased proteolytic activity, but induced about 50% degradation of the GDH, whereas the GDH of the control peanut suffered only about 25% degradation, thus showing that the degradation rate was about double the rate of de novo synthesis in the pesticide treatments. The heavy displacement of the GDH subunit equilibrium toward degradation explains the biochemical basis of the isomerization reaction.     

Analysis of pesticides in nuts by online reversed-phase liquid chromatography-gas chromatography using the through-oven transfer adsorption/desorption interface

A new method to determine pesticide residues in nuts is presented, in which the pesticides are extracted from samples with a small amount of ethyl acetate and anhydrous sodium sulfate. No additional cleanup or concentration steps are necessary. The extract is directly injected into the high-pressure liquid chromatograph, where preseparation of the pesticide residues from other components coextracted from the nuts is carried out using methanol/water as the eluent. The selected liquid chromatography fraction containing the pesticides is automatically transferred to the gas chromatograph using the through-oven transfer adsorption/desorption interface. The calculated limits of detection for each pesticide varied from 0.1 to 61.3 microg/kg. The repeatabilities of the analysis and the overall procedure (extraction and analysis) were satisfactory. No variations in the retention time were observed. The method was applied to the analysis of pistachio nut, peanut, walnut, hazelnut, and sunflower seed.     

Improved uptake models of nonionized pesticides to foliage and seed of crops.

The residual amount of nonionized pesticides incorporated to foliage and stem (foliage) and seed (fruit) of crops via root hairs from the water phase was estimated using the uptake models newly including metabolic parameters by which the amount of intact pesticide remaining in crops was considered with its proportion in a transpiration stream. A new parameter was also introduced for the seed model that accounts for the pesticide loss by adsorption to the inner surface of xylem tissue. Validation of the model was conducted using six pesticides with soybean and spinach plants. The ratio of the predicted concentration of pesticide to the measured one was 0.44-1.49 and 0.57-2.93 with foliage and seed models, respectively, showing that these improved models would be effective as a prediction tool.     

Near-infrared diffuse reflectance spectroscopy for the analysis of poultry manures

The objective was to evaluate near-infrared diffuse reflectance spectroscopy (NIRS) for determining the composition of poultry manures. Poultry manure samples were obtained from a commercial testing laboratory along with conventionally determined analyte values for total N, NH(4)(+)-N, organic N (computed as total minus NH(4)(+)-N), minerals, and moisture. Samples were blended in a food blender and scanned in polyethylene bags on a FOSS-NIRSystems model 6500 scanning monochromator. Initial results indicated that changes had occurred in NH(4)(+)-N contents from the time of analysis, and samples were reanalyzed for NH(4)(+)-N. Using the new analyses, NIRS was able to accurately determine NH(4)(+)-N, organic N, total N, and moisture in 128 poultry manures. Results were more accurate when determinations were made on an "as is" as opposed to dry matter basis. Finally, results indicated that NIRS was not suitable for the determination of minerals (P, K, Ca, etc.) in poultry manures.     

Assessment of pesticide effects on non-target soil microorganisms

Agricultural pesticides should be free of side effects that may cause a lasting disruption of microbial processes that are essential for continued soil fertility. A test program is described here for the assessment of the potential effects of newly-developed pesticides on non-target soil micro-organisms. The pesticides to be tested are applied at the equivalent of their recommended field application concentration and, for a margin of safety, also at the tenfold thereof. The measurement of pesticide effects on the microbial cycling of C, N and S constitute the core of the test program. These measurements are, in some cases, complemented by evaluation of pesticide effects on total and viable microbial numbers, on the abundance of specific components of the soil microbial community, and on the growth rates of selected soil microorganisms. The above test program was applied to evaluate the safety of the herbicides N-(1-ethylpropyl)-3,4-dimethyl-2,6-clinitrobenzeneamine(pendimethalin) l,2-dimethyl-3,5-diphenyl-1H pyrazolium (difenzoquat), S-(4-chlorophenyl)-N,N-diethyldithiocarbamate (thiobencarb), and the fungicides N-[(trichloromethyl) thio]-phthalimide (folpet) and cis-N-[(l,l,2,2-tetrachloroethyl) thio]-4-cyclohexene-1,2-dicarboximide (captafol). No detectable adverse effects were caused by any of the tested herbicides. The fungicides inhibited some of the C cycling activities, but the inhibition was temporary and was associated with their effect on soil fungi.     

Foliar and soil deposition of pesticide sprays in peanuts and their washoff and runoff under simulated worst-case rainfall conditions

There are few studies that relate the timing and amounts of pesticide washoff from plant foliage during rainfall to runoff losses at the edge of the field. We hypothesized that foliar deposits, if washed onto the soil slowly during rainfall, may then undergo less leaching during the period of infiltration that occurs prior to soil saturation and runoff, thus exhibiting larger runoff losses than pesticides on/in the soil at the beginning of rain. We measured the runoff of ethalfluralin, metolachlor, chlorothalonil, and rhodamine WT dye using simulated rainfall on 450 m2 mesoplots planted in peanut. Ethalfluralin was applied preplant incorporated, and metolachlor was applied preemergence on bare soil. Chlorothalonil and rhodamine WT were applied to the peanut canopy at maturity. Rainfall was simulated 24 h after each chemical application (in May and July, 1998, and May and August, 1999) using raindrop sprinklers, applying 5.5 +/- 0.5 cm over a 2 h period to create reasonable worst-case conditions; between 3 and 9 mm of runoff was generated. Volume-weighted average concentrations of chemicals in runoff were 7, 104, 163, and 179 ug L(-1) for ethalfluralin, metolachlor, chlorothalonil, and rhodamine WT, respectively. The total amounts of chemicals lost in the runoff events were 0.04 +/- 0.01, 0.2 +/- 0.1, 0.6 +/- 0.5, and 0.2 +/- 0.1, as percents of amounts applied, respectively. Rhodamine WT formed a vivid red solution on wetting and provided visual clues to the dynamics of chemical washoff/runoff. The washoff from rain-exposed peanut foliage appeared to be complete within a few minutes of the beginning of rainfall, and disappearance of dye from rain-exposed soil surface occurred within the first 10 min of rainfall. However, dye was present in runoff water at near-constant concentrations throughout the 2 h runoff event, indicating that near-constant amounts of chemical remained in the soil extraction zone. These results confirm earlier studies showing that soil incorporation at application significantly reduces runoff losses and that a majority of foliar residues can be washable if rainfall occurs within a few days after application. Runoff losses of foliar-applied pesticides were small relative to washoff amounts but were sensitive to runoff timing relative to washoff.     

Effect of the carbamate pesticide Sevin on the intestinal absorption of some nutrients in the teleost fish,Channa Punctatus

The effect of four concentrations (0.1, 1, 10, and 100 ppm) of Sevin on the rate of uptake of two sugars (glucose and fructose) and an aminoacid (tryptophan) by the intestine of the fresh water snake-head fish,Channa punctatus has been investigated. All four concentrations of the pesticide decreased the rate of transport of the three nutrients. No direct relationship could be established between the concentration of Sevin in the medium and decrease in the rate of transport of nutrients, although, increase in the concentration of Sevin reduced the absorption of nutrients by the intestine to a greater extent. Experiments conductedin vivo andin vitro showed that in both cases the pesticide decreased the brush border enzyme activity.     

Influence of two insecticides, chlorpyrifos and quinalphos, on arginine ammonification and mineralizable nitrogen in two tropical soil types

Effects of seed treatments with chlorpyrifos [5 g of active ingredient (ai) kg(-1) of seed] and quinalphos (6.25 g of ai kg(-1) of seed) and standing crop treatments with chlorpyrifos (800 g of ai ha(-1)) and quinalphos (1000 g of ai ha(-1)) on arginine deamination and mineralizable nitrogen were monitored, in the sandy loam and loamy sand soils of two tropical semiarid fields, for three consecutive crop seasons. The arginine ammonification activity of rhizospheric microbes was inhibited after seed treatment with chlorpyrifos and quinalphos and their principal metabolites, 3,5,6-trichloro-2-pyridinol (TCP) and 3,5,6-trichloro-2-methoxypyridine (TMP) and 2-hydroxyquinoxaline and quinoxaline-2-thiol, respectively. Quinalphos produced transient inhibitions, whereas chlorpyrifos and its metabolites (TCP and TMP) exerted a greater inhibition in both loamy sand and sandy loam soils. Arginine ammonification by nonrhizospheric microbes was stimulated by standing crop treatments with both pesticides. In the loamy sand soil, the parent compounds stimulated rhizospheric N-mineralization, whereas the metabolites were inhibitory. However, nonrhizospheric N-mineralization was inhibited by both chlorpyrifos and quinalphos and stimulated by their metabolites. A higher magnitude of inhibition of arginine deamination in the loamy sand than in the sandy loam soil could be due to greater bioavailability of the pesticides in the former, resulting from lesser sorption of the pesticides due to alkalinity of the soil and its low content of clay and organic carbon. Although both pesticides affected mineralizable nitrogen, seed treatment with quinalphos and standing crop treatment with quinalphos and chlorpyrifos produced the most significant effects. The recommended doses of the pesticides not only efficiently controlled whitegrubs, which increased pod yields, but also left no residues in harvested kernels. They also caused no long-term inhibition of ammonification, which could have been of significant concern during the short crop period in semiarid areas where nitrogen determines plant productivity.     

黄褐土、潮土中不同氮素形态配比对烤烟根际土壤微生物数量的影响

试验采用盆栽控雨栽培和根网袋的方法 ,在两种土壤类型上初步研究了不同氮素形态配比与芝麻饼肥互作对烤烟不同生育时期根际土壤微生物数量的影响。结果表明 :在黄褐土中 ,当施入 40 %饼肥、NO-3 ∶NH+ 4为 1∶1时 ,土壤中真菌、氨化细菌和好气性纤维素分解菌数量最多 ,促进根系的生长发育。在潮土中 ,当施入 40 %饼肥、NO-3 :NH+ 4为 3∶1时 ,土壤中好气性纤维素分解菌、氨化细菌在移栽后 3 8天迅速上升 ,促进了根系对氮素的吸收利用     

Applications of mineral nitrogen increase the yield and content of crude protein in narrow-leaf lupin seeds

Abstract

In a plant pot experiment with non-inoculated seed stock of narrow-leaf lupin (Lupinus angustifolius, L.) of the variety ‘Prima’ we explored the effects of nitrogen compounds applied in the form of NH₄NO₃ on different dates (before sowing, after emergence and during flowering) and at various levels (0–0.5–2–3 g N per pot) on changes in the number of pods per plant, the 1000-seed weight, seed yields and the content of crude protein in seeds. The experiment included four treatments; the first was a control treatment not fertilized with nitrogen. In the other treatments we applied nitrogen either as a single application before sowing or in three split applications. In all the nitrogen-fertilized variants the number of pods per plant, total seed yields per plant and the concentration of crude protein all increased compared to the unfertilized control variant. The highest levels of nitrogen (3 g N per pot) reduced the 1000-seed weight compared to the unfertilized control variant. The total application of 2 g N per pot split into application before sowing (0.5 g N), after emergence (0.5 g N) and during flowering (1 g N) resulted in a highly significant increase in the number of pods per plant (+78%), the 1000-seed weight (+20.1%) and therefore higher seed yields per plant (+139.8%) compared to the control treatment not fertilized with nitrogen. Increasing levels of nitrogen linearly increased the content of crude protein in seeds during harvest from 24.2% to 40.6%, thus increasing the nutritional quality of the seeds.     

不同花生品种氮代谢相关酶活性的研究

为研究不同品种花生各器官中氮代谢酶活性的差异及与籽仁蛋白质含量间的关系,采用田间小区试验,利用高产大花生品种(花育22号)和小花生品种(白沙1016)研究了两品种不同生长发育时期植株各器官中可溶性蛋白质(Pro)含量、硝酸还原酶(NRase)、谷氨酰胺合成酶(GS)和谷氨酸脱氢酶(GDH)活性的变化。结果表明,两粒型花生品种各器官的Protein、NRase、GS、GDH活性变化态势大致相同,但其含量的高低有异;各器官中的各指标含量均以小花生品种白沙1016较高。各器官中NRase活性随生育期的推进呈渐降趋势,籽仁中NRase活性远高于其他营养器官;两粒型品种叶片、茎和根中GS活性的变化趋势均呈单峰曲线,其峰值均出现在结荚期;叶片GDH活性以苗期较高,花针期最低,之后又迅速升高。茎和根中GDH活性均呈V字型曲线变化,根中峰谷滞后于茎,出现在饱果期,而籽仁中GDH活性成熟期略有升高。各器官中蛋白质含量与GS和GDH活性间的相关关系不明显,但与其相应器官中的NRase活性表现显著或极显著相关。     

Identification of an antioxidant,ethyl protocatechuate,in peanut seed testa

The antioxidant activity and identification of the antioxidant component of peanut seed testa were investigated. The antioxidant activity of peanut seed testa was studied in the linoleic acid model system by using the ferric thiocyanate method. Among the five organic solvent extracts, the ethanolic extracts of peanut seed testa (EEPST) produced higher yields and stronger antioxidant activity than other organic solvent extracts. EEPST was separated into 17 fractions on silica gel column chromatography. Fraction 17, which showed the largest yield and significant antioxidant activity, was separated by thin-layer chromatography. Four major antioxidative subfractions were present. Subfraction 17-2 was found to be effective in preventing oxidation of linoleic acid. This subfraction was further fractionated and isolated and characterized by UV, MS, IR, and (1)H NMR techniques. The active compound was identified as ethyl protocatechuate (3,4-dihydroxybenzoic acid ethyl ester).     

不同氮素供应对烟草品质指标的影响

用盆栽试验研究了不同氮形态和不同氮浓度对烟草钾、氯、总糖和烟碱等品质指标的影响。两种氮形态均有助于叶片钾和氯的积累,铵态氮有助于增加氯的积累,硝态氮更有利于钾的积累。铵态氮有利于总糖提高,其比例超过硝态氮时不利于烟碱含量的提高。任何铵/硝比下,氮浓度增加均会降低糖/烟碱比。下部叶对氮的反应不同于上部和中部叶,也是其烟叶品质不同于上部和中部叶的内在原因。     

水稻氨基酸态氮营养效应及其机理研究

在＾１５无菌水培试验条件下水稻氨基酸态氮和铵态氮的营养效应及其氮营养贡献,氨基酸态氮的同化机理等问题作了比较研究。结果表明,等氮量（１０ｍｇ／Ｌ）无菌水培５１天后,甘氨酸态氮（Ｇｌｙ－Ｎ）单施或与硫酸铵态氮（ＮＨ４＾＋－Ｎ）配施处理的干物重、吸氮量均大于硫酸铵态氮单施处理;Ｇｌｙ－Ｎ对水稻干物重及吸氮量的促进作用大于ＮＨ４＾＋－Ｎ;Ｇｌｙ－Ｎ与ＮＨ４＾＋－Ｎ配施条件下,以吸氮总量为基础计算的Ｇｌｙ－Ｎ氮营养贡献率可达５５．６６％。采用Ｇｌｙ－Ｎ培养的籼、粳稻整株谷草转氨酸ＧＯＴ、谷丙转氨酶ＧＰＴ及谷氨酸脱氢酶ＧＤＨ活性均大于ＮＨ４＾＋－Ｎ,且根中ＧＯＴ和ＧＰＴ活性、叶中ＧＤＨ活性明显高于相同部位ＮＨ４＾＋－Ｎ培养的水稻。水稻ＧＯＴ、ＧＰＴ及ＧＤＨ活性受对应底物谷氨酸（Ｇｌｕ）营养的促进。     

Insecticide dissipation after repeated field application to a Northern Thailand Ultisol

Nontarget effects of pesticides may occur when the active ingredients have a long persistence in the environment. The half-lives of six insecticides commonly used in Thai fruit orchards under tropical field conditions were determined. A mixture of endosulfan-alpha and -beta, chlorpyrifos, malathion, dimethoate, and mevinphos was applied five times in 10-day intervals onto an Ultisol (lychee plantation ground-covered with grass vegetation, northwestern Thailand). On days 1, 3, 5, 7, and 10 after each application, composite samples of the topsoil (0-10 cm) were collected and exhaustively extracted. Fitting a first-order model to the datasets revealed rapid initial dissipation [half-lives from 2.2 +/- 0.4 (malathion) to 5.4 +/- 1.3 days (chlorpyrifos)]. Volatilization appeared to be a major process of pesticide dissipation, especially for malathion and mevinphos. Because 8% of the applied endosulfan-alpha and -beta had been converted to the sulfate metabolite within 1 day after the first application, also microbial degradation contributed significantly to pesticide dissipation. Nevertheless, no trend in half-lives over the five application cycles could be observed, indicating that accelerated microbial degradation did not occur for these insecticides following the five applications. Precipitation and soil moisture were key parameters of dissipation, but dissipation processes were too complex to be generalized for all substances studied. Despite their short half-lives, all pesticides except mevinphos accumulated in soil (up to 656%; endosulfan-alpha), and this accumulation correlated significantly with the hydrophobicity of the substances (r = 0.88). This was interpreted as an aging process, and it was concluded that pesticide aging must be considered relevant also in tropical environments, where it has received very limited attention so far.     

Degradation of Sevin by soil microorganisms

Microorganisms capable of transforming the pesticide 1-naphthyl N-methyl-carbamate (Sevin) were isolated from soil. Three isolates were able to accelerate the hydrolysis of Sevin to 1-naphthol. Several unidentified intermediates were separated by thin-layer chromatography and also by following the decomposition of Sevin-methyl-¹⁴C. Since 1-naphthol is a biological as well as a chemical decomposition product of Sevin, its transformation by the isolated microbes was also studied. A fungus, identified as Fusarium solani, altered 1-naphthol rapidly. Whereas one strain of bacterium degraded the hydrolysis product gradually, another strain accumulated it under certain conditions. Mixed cultures of the investigated microbes were more effective in transforming Sevin than pure cultures, and this phenomenon was also observed with 1-naphthol as substrate with the exception of one bacterial strain.     

不同铁制剂与施用方法对矫正花生缺铁黄化症的效果

研究了在盆栽和田间小区试验条件下,叶面和土壤施用铁肥以及土施酸化肥料对纠正花生缺铁黄化症的作用效果。在盆栽条件下,叶面喷施黄腐酸铁(FeFA)、土施硫酸亚铁及酸化肥料都可以提高花生新叶中活性铁以及叶绿素含量,促进花生植株的生长;在田间小区栽培条件下,叶面喷施FeFA可以提高花生的叶绿素含量,尿素与FeFA结合使用的作用效果大于单喷FeFA。土施硫酸亚铁在花生生长前期提高了花生叶绿素含量,后期作用不明显。叶面喷施与土施铁肥都可以增加花生产量,土施铁肥的作用效果不如叶面喷施效果明显,酸化肥料对提高花生叶绿素含量及产量没有明显效果。     

Method for determination of organohalogen pesticide residues in vegetable oil refinery by-products

A method using gel permeation and Florisil column chromatographic cleanup techniques is described for determination of residues of nonpolar organohalogen pesticides and pesticide alteration products in vegetable oils and their refinery by-products. Supplemental Florisil separation and alkali cleanup techniques are used to facilitate determinations. Residues are determined with a 63Ni electron capture gas chromatographic detection system used in conjunction with 3 different gas chromatographic columns. Residue identities are confirmed by gas chromatography-mass spectrometry. Recoveries of 7 organohalogen pesticides, ranging from 90 to 103%, were determined by the supplemental Florisil separation technique to augment previously reported recovery data determined for initial GPC and Florisil cleanup steps. Soybean, peanut, and cottonseed deodorizer distillates and crude and refined oil, as well as additional refinery by-products, were analyzed. Nine to 13 organohalogen residues ranging from 0.5 to 6.3 ppm were determined in the 2 soybean deodorizer distillate samples used to develop and test the method. Identities of residues present at greater than or equal to 0.3 ppm were confirmed by gas chromatography-mass spectrometry. An intralaboratory trial of the method provided additional recovery and residue determination data as follows: Recoveries ranging from 102 to 116% were obtained for 4 pesticides added to peanut oil deodorizer distillate. Residues determined in 1 soybean deodorizer distillate sample supported previously obtained data for this sample.     

施氮水平对不同花生品种产量与品质的影响

为了探讨氮对不同花生品种产量和品质的影响,采用盆栽法开展了不同氮用量对3个不同花生品种产量和品质影响的研究。结果表明,不同花生品种达到最高产量的施氮量不同,白沙1016和鲁花12施氮 112.5 kg/hm2时荚果产量最高,远杂9102施氮 75 kg/hm2时荚果产量最高。相对于不施氮,施氮后花生子仁蛋白质和氨基酸平均含量分别增加了4.9%和15.0%; 施氮375N1125 kg/hm2比不施氮粗脂肪平均含量提高3.7%; 施氮可有效增加子仁苏氨酸、蛋氨酸、异亮氨酸、苯丙氨酸、赖氨酸等必需氨基酸含量; 除脯氨酸外,其它非必需氨基酸含量均显著提高; 与不施氮相比,施氮后子仁硬脂酸、花生酸和山嵛酸平均含量分别提高了13.6%、6.9%和5.5%。3个品种相比,子仁蛋白质、氨基酸和粗脂肪含量差异显著; 氨基酸组分除苏氨酸、色氨酸、甘氨酸含量外,其它差异显著,而脂肪酸组分除十七酸、硬脂酸外,其它差异不显著。总之,供试不同品种花生达到最高产量的施氮量存在明显差异; 施氮可显著提高花生子仁营养品质。