A non-destructive method for testing two components of the behaviour of soil-applied agricultural chemicals over a long period

BACKGROUND: Owing to the complexity of soil composition, accurate predictions of both apoplastic systemicity of lipophilic xenobiotics and their leaching from the soil are made difficult. Therefore, a non‐destructive method to assess directly these two components of the spatial behaviour of soil‐applied phytochemicals is needed. RESULTS: The plant selected was a dwarf tomato, which can exude an abundant apoplastic fluid through large stomata for several months. The feasibility and reliability of the method were assayed using three triazoles exhibiting different log D values. HPLC‐MS analyses indicated that triadimenol (log D = 2.97) was clearly the most mobile compound within the apoplast, especially its diastereoisomer A. Propiconazole (log D = 3.65) and penconazole (log D = 4.64) exhibited a similar low systemicity. The data remained the same when the three fungicides were applied together on the soil. Long time‐course studies (1.5 months) of penconazole behaviour indicated that, in contrast to leaching, which decrease sharply, root‐to‐shoot translocation remained almost unchanged during the whole experiment, in spite of the high lipophilicity of this fungicide. CONCLUSION: This method must contribute to a better knowledge of the behaviour of commercial soil‐applied phytochemicals. It can also be used to screen new xenobiotics within strategies to satisfy environmental requirements. Copyright © 2011 Society of Chemical Industry     

Systemic effects of thiamethoxam and chlorantraniliprole seed treatments on adult Lissorhoptrus oryzophilus (Coleoptera: Curculionidae) in rice

BACKGROUND: Feeding assays using adult rice water weevils and foliage of plants treated as seeds with chlorantraniliprole and thiamethoxam at different rates were conducted to evaluate the systemic adulticidal and feeding effects. Dose–mortality relationships were determined for thiamethoxam seed treatments by combining leaf area lost due to feeding and insecticide residues analyzed by LC/MS/MS. Changes in adulticidal activity of thiamethoxam were also investigated by contrasting adult mortalities at the 5–6‐leaf and tillering stages of rice. RESULTS: Adult weevil mortalities and leaf consumption rates on foliage were affected in thiamethoxam but not in chlorantraniliprole treatments when rice was at the 6–7‐leaf stage. The LD₅₀ for weevils feeding on thiamethoxam‐treated rice at the 2–3‐leaf stage was 447 pg insecticide weevil^?1 (95% CL: 25–830 pg weevil^?1) but was lower (142 pg weevil^?1; 95% CL: 102–180 pg weevil^?1) in experiments with 3–4‐leaf‐stage plants. Mortalities on leaves from 5–6‐leaf‐stage plants were consistently higher than on leaves from tillering plants. Thiamethoxam residues measured by ELISA increased with seed treatment rate and differed between plant stages. CONCLUSION: The LD₅₀ values developed in this study are the first values for leaf‐feeding insects on foliage of plants treated as seeds with thiamethoxam. The attrition of adulticidal activity of thiamethoxam in foliage of older plants may help to explain the reduced effectiveness of seed treatments against rice water larvae that is seen at later stages of rice growth in field studies. The differential activity of these two seed treatments on adults suggests that adult mortality contributes to the field efficacy of thiamethoxam but not to that of chlorantraniliprole. Copyright © 2012 Society of Chemical Industry     

Design, synthesis and structure-activity relationship of novel coumarin derivatives

BACKGROUND: The lead coumarin derivative (E)‐methyl 3‐methoxy‐2‐[2‐(4‐methylcoumarin‐7‐yloxymethyl)phenyl]acrylate was discovered by using an intermediate derivatisation method. To discover new coumarin derivatives with improved activity, a series of substituted coumarins were synthesised and bioassayed. RESULTS: The compounds were identified by ¹H NMR, IR, MS and elemental analysis. Bioassays demonstrated that some of the title compounds exhibited excellent fungicidal activity against cucumber downy mildew at 25 mg L^?1. The relationship between structure and fungicidal activity is reported. CONCLUSION: The present work demonstrates that coumarin derivatives containing methoxyacrylate moieties can be used as possible lead compounds for developing novel fungicides. Copyright © 2011 Society of Chemical Industry     

优化的QuEChERS-超高效液相色谱-串联质谱法检测黄瓜中29种杀菌剂残留

建立了采用QuEChERS-超高效液相色谱-串联质谱（UPLC-MS/MS）同时检测黄瓜中29种杀菌剂残留的分析方法。黄瓜样品经乙腈提取和前处理方法优化,以乙二胺-N-丙基硅烷（PSA）和石墨化碳黑（GCB）为分散固相萃取（dSPE）吸附剂,涡旋提取后过滤膜,采用UPLC-MS/MS检测。结果表明:在0.000 1~0.05 mg/kg范围内,29种杀菌剂的质量浓度与相应的峰面积间呈良好的线性关系,相关系数均大于0.996,定量限（LOQ）在0.000 1~0.01 mg/kg之间;在0.01、0.05和0.5 mg/kg添加水平下,29种杀菌剂的平均回收率均在74%~112%之间,相对标准偏差（RSD）在1.1%~17%之间（n=5）。经实际样品检测验证,该方法快速、简便、可靠、高效,且成本低廉,适用于同时检测黄瓜中29种杀菌剂的残留量。     

Non‐target‐site mechanism of glyphosate resistance in Italian ryegrass (Lolium multiflorum)

In Shizuoka Prefecture, Japan, glyphosate‐resistant Lolium multiflorum is a serious problem on the levees of rice paddies and in wheat fields. The mechanism of resistance of this biotype was analyzed. Based on LD₅₀, the resistant population was 2.8–5.0 times more resistant to glyphosate than the susceptible population. The 5‐enolpyruvyl‐shikimate‐3‐phosphate synthase (EPSPS) gene sequence of the resistant biotype did not show a non‐synonymous substitution at Pro106, and amplification of the gene was not observed in the resistant biotype. The metabolism and translocation of glyphosate were examined 4 days after application through the direct detection of glyphosate and its metabolite aminomethylphosphonic acid (AMPA) using liquid chromatograph‐tandem mass spectrometer (LC‐MS/MS). AMPA was not detected in either biotype in glyphosate‐treated leaves or the other plant parts. The respective absorption rates of the susceptible and resistant biotypes were 37.90 ± 3.63% and 41.09 ± 3.36%, respectively, which were not significantly different. The resistant biotype retained more glyphosate in a glyphosate‐treated leaf (91.36 ± 1.56% of absorbed glyphosate) and less in the untreated parts of shoots (5.90 ± 1.17%) and roots (2.76 ± 0.44%) compared with the susceptible biotype, 79.58 ± 3.73%, 15.77 ± 3.06% and 4.65 ± 0.89%, respectively. The results indicate that the resistance mechanism is neither the acquisition of a metabolic system nor limiting the absorption of glyphosate but limited translocation of the herbicide in the resistant biotype of L. multiflorum in Shizuoka Prefecture.     

The search for biologically active secondary metabolites

Plants provide a rich source of novel biologically active compounds. Biological and chemical screenings are complementary approaches for the rapid detection and isolation of interesting new plant constituents. Biological screening followed by activity-guided fractionation has been used successfully in our laboratories for the discovery of antifungal, larvicidal and molluscicidal compounds. High performance liquid chromatography (HPLC) coupled to UV spectroscopy (LC/UV), mass spectrometry (LC/MS) and nuclear magnetic resonance (LC/NMR) has proved to be highly efficient for the chemical screening of crude plant extracts. In particular LC/MS and LC/MS/MS used with different ionisation techniques such as thermospray (TSP), continuous flow-FAB (CF-FAB) and electrospray (ES) have proved to be very efficient for the early recognition of molluscicidal saponins in Swartzia madagascariensis and Phytolacca dodecandra. The combination of LC/UV/NMR/MS was of great value for the investigation of polyphenols and bitter principles in Gentianaceae species. Among other examples, LC/NMR analysis of the antifungal crude extract of the African plant Swertia calycina is presented. ©1997 SCI     

Degradation kinetics and safety evaluation of tetraconazole and difenoconazole residues in grape

BACKGROUND: Tetraconazole and difenoconazole are triazole fungicides with proven bioefficacy against grapevine powdery mildew disease. In the present work, the authors explored the residue dynamics of these two compounds in grapes and determined their preharvest intervals (PHIs) corresponding to multiple field applications at recommended and double rates considering the most critical use pattern in Indian viticulture. A confirmatory residue analysis method was validated for trace-level determination of both the compounds. RESULTS: Dissipation of both the fungicides followed non-linear two-compartment first + first-order rate kinetics. Tetraconazole and difenoconazole dissipated with PHIs of 12.5 and 25.5 days at recommended rates and of 28.5 and 38.5 days at double application rates respectively. On all the sampling days, the residues were below the maximum permissible intake, indicating consumer safety. The residues in the grape samples drawn from the farms where these two fungicides were applied, maintaining the above PHIs, were below their respective MRLs. CONCLUSION: The rate of degradation of tetraconazole was faster than that of difenoconazole. Thus, the growers will have the choice of using these new chemicals for the management of powdery mildews in succession, difenoconazole at early growth stages, followed by tetraconazole during the last month before harvest. The recommendations of PHIs proved to be effective in minimizing residues in farm grape samples. Thus, this work is of high significance to the grape industry of India, and will support the registration of these new fungicides for effective management of powdery mildews with minimum residue problems.     

N-(2-三氟甲基-4-氯苯基)-2-吡唑酰氨基环己烷基磺酰胺类化合物的合成与杀菌活性

吡唑酰胺类杀菌剂是近年新农药开发的热点。本研究采用EDCI/HOBt酰胺化法合成了14个结构新颖的N-(2-三氟甲基-4-氯苯基)-2-吡唑酰氨基环己烷基磺酰胺类化合物 ( 3a ~ 3n ),其结构均经1H NMR、13C NMR、质谱和元素分析确认,并用X-射线衍射法确定了化合物 3g 的单晶结构和立体构型。菌丝生长速率法试验结果表明,化合物 3a 、 3e 和 3j 对番茄灰霉病菌Botrytis cinerea KZ-9的EC₅₀值分别为4.28、10.08和11.31 mg/L,抑菌活性不及对照药剂啶酰菌胺和腐霉利;但在孢子萌发试验中,目标化合物表现出与对照药剂相近的抑菌活性,在10 mg/L下有7个化合物对灰霉病菌孢子萌发的抑制率超过了85%;在番茄活体盆栽试验中,化合物 3e 在200 mg/L下对番茄叶片及其花上灰霉病菌的防治效果分别为77.5%和65.2%,高于对照药剂啶酰菌胺 (防效分别为59.8%和30.3%),有进一步研究的价值。     

Uptake by roots and translocation to shoots of two morpholine fungicides in barley

Despite being lipophilic, morpholine fungicides are systemic in plants. Such transport may be explicable by their protonation (pKa∽7·5) at the pH of plant compartments to yield the more polar cation. This behaviour might be a useful attribute to be incorporated into other classes of lipophilic pesticides. To understand quantitatively the behaviour of the morpholine fungicides, the uptake by roots and transport to shoots in barley of two such ¹⁴C-labelled compounds, dodemorph and tridemorph, were investigated using bathing solutions of differing pH. At pH 5, uptake and transport were small, but increased by approximately two orders of magnitude at pH 8. Tridemorph, the more lipophilic of the two compounds, was highly accumulated by roots at pH 8 and moderately translocated to shoots. In contrast, dodemorph was translocated to shoots at pH 8 with remarkable efficiency, moving into the xylem across the endodermis at 23 times the efficiency of water, though accumulation in roots was less than that of tridemorph. Behaviour at 24 h was largely similar to that at 48 h for both compounds, indicating that uptake and translocation are equilibrium processes maintained over time. Transport to shoots for each compound was directly proportional to the concentrations accumulated in the roots, except at low pH where partitioning into root solids became proportionately more important with such material not being directly available for transport to the xylem across the endodermis. Uptake and transport of these basic fungicides are explained in terms of their partitioning and of their accumulation in acidic plant compartments by ion trapping as the protonated form; this behaviour is discussed in relation to the pKa and lipophilicity of these compounds. © 1998 Society of Chemical Industry     

Application of coupled liquid chromatography–mass spectrometry in photolysis studies of the herbicide triflusulfuron‐methyl

The photochemical fate of the herbicide triflusulfuron‐methyl in aqueous solution under UV light (λ > 290 nm) was investigated. Nine photoproducts were detected and tentatively identified by LC/MS and LC/MS/MS analysis. The main routes of degradation involve the cleavage and/or contraction of the sulfonylurea bridge with the elimination of sulfur dioxide, which increases the acidity of the reaction medium. On the basis of the results obtained a photodegradation pathway is proposed. © 2001 Society of Chemical Industry     

野艾蒿中双四氢呋喃类木脂素的提取分离及其抑菌活性

为筛选新型植物源杀菌剂,对野艾蒿Artemisia lavandulaefolia DC. 地上组织中的化学成分进行了研究。采用甲醇提取、液-液萃取、柱层析及半制备高效液相色谱等技术从乙酸乙酯浸膏中分离得到6个化合物;利用质谱及核磁共振等波谱技术确定这6个化合物分别是epiashchantin ( 1 )、ashantin ( 2 )、sesartemin ( 3 )、epimagnolin A ( 4 )、epiyangambin ( 5 ) 和diayangambin ( 6 ),均属于双四氢呋喃类木脂素。采用菌丝生长速率法测定了其抑菌活性,结果表明:化合物 1 和 3 对番茄早疫病菌Alternaria solani的菌丝生长表现出明显抑制作用,EC₅₀值分别为14.3和11.0 μg/mL;化合物 3 、 5 和 6 对玉米纹枯病菌Rhizoctonia solani的菌丝生长也表现出抑制作用,EC₅₀值分别为17.4、42.6和54.3 μg/mL。     

Characterization of new photosynthesis-inhibiting imidazolidine derivatives. 1. Interpretation of post-emergence herbicidal performance from photosynthesis-inhibiting activity and systemicity

An attempt was made to account for quantitatively measured herbicidal performance of foliage-applied 5-hydroxy-3-methyl-2–oxo-imidazolidine-1-carboxamide derivatives by their photosynthesis-inhibiting activity and systemicity. Photosynthesis-inhibiting activity was estimated from the increase of chlorophyll fluorescence intensity in Chlorella vulgaris Beijer cells measured by a microplate scanner, and systemicity was also evaluated by computer-aided chlorophyll fluorescence imaging. The highest herbicidal performance was recorded for N-cyclohexyl-5-hydroxy-3-methyl-2-oxo-imidazo～idine-l-carboxamide, a compound with the second strongest photosynthesis-inhibiting activity and intermediate systemicity. Though neither photosynthesis-inhibiting activity nor systemicity showed significant correlation with the actual herbicidal performance in simple regression analyses, a high predictability was found for a multiple regression on both parameters as two independent variables, suggesting that these two factors work cooperatively in the field performance.     

Nested PCR‐RFLP is a high‐speed method to detect fungicide‐resistant Botrytis cinerea at an early growth stage of grapes

BACKGROUND: Grey mould caused by the fungus Botrytis cinerea Pers. ex Fr. is one of the major diseases in grapes. The use of fungicides is a simple strategy to protect grapes against B. cinerea disease. However, phenotypes exhibiting resistance to fungicides have been detected in B. cinerea populations. The variation of fungicide‐resistant B. cinerea isolates renders B. cinerea disease control difficult in grapevine fields. RESULTS: The authors have developed a nested polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) method to detect fungicide‐resistant B. cinerea isolates at an early growth stage of grapes in grapevine fields. The nested PCR‐RFLP method was carried out to detect benzimidazole‐, phenylcarbamate‐ and/or dicarboximide‐resistant B. cinerea isolates from grape berries and leaves at Eichorn–Lorenz growth stage 25 to 29. This method successfully detected fungicide‐resistant B. cinerea isolates at an early growth stage of grapes. In addition, only 8 h was required from tissue sampling to phenotyping of fungicide resistance of the isolates. CONCLUSION: It is proposed that the early diagnosis of fungicide‐resistant B. cinerea isolates would contribute to further improvement of integrated pest management against B. cinerea in grapevine fields, and that the nested PCR‐RFLP method is a high‐speed, sensitive and reliable tool for this purpose. Copyright © 2008 Society of Chemical Industry     

Experiments with some of the newer systemic fungicides on apple and marrow

In field trials against apple scab (caused by Venturia inaequalis) and powdery mildew (caused by Podosphaera leucotricha), improved control of one or other disease has been given by the newer systemic fungicides benomyl, triarimol and the thiophanates. However, in tests on apple seedlings in the greenhouse, biological evidence of translocation from individual deposits was generally limited to movement within the treated leaf. In greenhouse tests with cucumbers and marrows, using Oidium sp., there was again little evidence for movement of toxicant from a treated leaf, although effective disease control was readily obtained by root application of several compounds at low dosage. These results suggest that the systemic properties of the compounds evident when used as soil or seed treatments are of little account when they are applied as foliar sprays.     

N-甲氧基-N- 苯基氨基甲酸甲酯的合成及生物活性研究

为了寻找新的高效的Strobilurin类杀菌剂,以1-取代基-3-三氟甲基吡唑啉-5-酮与N-甲氧基-N-(2-溴甲基苯基)氨基甲酸甲酯反应,合成了系列新型N-甲氧基-N- 苯基氨基甲酸甲酯化合物 (IV1~IV13) ,其结构经IR、1H NMR、LC/MS及元素分析确认。生物活性测定结果表明,该类化合物在50 mg/L下对稻瘟病菌Pyricularia oryzae、黄瓜灰霉病菌Botrytis cinerea、小麦白粉病病菌Erysiphe graminis等具有较高的抑菌活性,如 IV13 对稻瘟病菌P.oryzae 的抑制率达94.0%.     

茚酮肟醚衍生物的合成与生物活性

通过傅－克反应及环化、肟化、成醚等反应,合成了10个未见文献报道的茚酮肟醚类化合物,其结构均经1H NMR、IR、LC/MS及元素分析确证。初步生物活性测试结果表明,所合成的化合物具有明显的杀菌活性或杀虫活性,如在100 mg/L时,化合物 V1 对小麦白粉病菌Erysiphe graminis 的抑制率为 95.6%, V5 对粘虫Mythimna separata的致死率为100%。     

Modelling uptake into roots and subsequent translocation of neutral and ionisable organic compounds

A study on uptake of neutral and dissociating organic compounds from soil solution into roots, and their subsequent translocation, was undertaken using model simulations. The model approach combines the processes of lipophilic sorption, electrochemical interactions, ion trap, advection in xylem and dilution by growth. It needs as input data, apart from plant properties, log K_OW, pK_a and the valency number of the compound, and pH and chemical concentration in the soil solution. Equilibrium and dynamic (steady‐state) models were tested against measured data from several authors, including non‐electrolytes as well as weakly acidic and weakly basic compounds. Deviations from the measured values led to further development of the model approach: sorption in the central cylinder may explain the small transpiration stream concentration factor of lipophilic compounds. For non‐electrolytes, the model predicted uptake and translocation with high accuracy. For acids and bases, the tendency of the results was satisfactory. The dynamic model and the equilibrium approach gave similar results for the root concentration factor. The calculation of the transpiration stream concentration factor was more accurate with the dynamic model, but still gave deviations up to factor of ten or more. The dominating process for monovalent weak electrolytes was found to be the ion trap effect. © 2000 Society of Chemical Industry     

4"-脱氧阿维菌素脲类及硫脲类衍生物的合成与杀虫活性

以构效关系为线索,根据类同合成法和活性官能团拼接策略,以阿维菌素为先导分子,在其4"-位引入硫脲或脲活性结构单元,合成了一系列酰基硫脲（9a~9o）和芳基脲（10a~10i）阿维菌素衍生物,所有目标化合物均经1H NMR、MS和元素分析确证。室内杀虫活性测定结果表明:所有衍生物对朱砂叶螨Tetranychus cinnabarinus和甘蓝蚜Brevicoryne brassicae均表现出不同程度的毒杀活性,大部分化合物对甘蓝蚜的杀虫活性优于阿维菌素B1a,其中化合物9h对朱砂叶螨和甘蓝蚜的LC50值分别为0.100和1.71 μmol/L,表现出很高的毒杀活性,化合物9o对甘蓝蚜的毒性最高,LC50值为1.13 μmol/L。     

新的吡啶杂环硫代磷酸酯类化合物的合成及杀虫活性

为了寻找高活性的新化合物,通过相转移催化反应,在二氯甲烷/水体系中合成了一系列新的吡啶杂环硫代磷酸酯类化合物,其结构均经1H NMR、13C NMR、31P NMR、IR和MS的表征,并通过元素分析确证。室内生物活性测试结果表明,部分化合物表现出较好的杀虫活性,其中化合物 3a 对苜蓿蚜Aphis medicaginis Koch和化合物 3c 对东方粘虫Mythimna separate的 LC50值分别为 1.64和 0.73 mg/L,与对照药剂毒死蜱的活性相当。