Build-up and subsequent decline of residues of quintozene and hexachlorobenzene in lettuce in sequential crops

The build-up and decline of quintozene and hexachlorobenzene residues in protected lettuce, resulting from one to five treatments of the soil (each of 35 g m-2) with a quintozene formulation, have been studied. Residue levels were monitored in each experimental crop during growth until harvest. Residues in the soil at harvest were also determined. Quintozene residues in the harvested lettuce were in the range < 0.02–0.80 mg kg^?1 and those of hexachlorobenzene were in the range < 0.02–0.05 mg kg^?1 (expressed on a fresh weight basis). Residue levels in the soil were 5.4–231 mg kg^?1 for quintozene, and 0.20–5.4 mg kg^?1 for hexachlorobenzene (expressed on a dry weight basis). Under the experimental conditions of the trial there was no significant build-up of quintozene or hexachlorobenzene in harvested lettuce, even after five treatments to the same site.     

建议我国重视五氯硝基苯的应用风险

五氯硝基苯是一种化学性质稳定的有机氯农药,在环境中很难降解,并容易通过食物链富集到生物体内。由于五氯硝基苯及其制备过程中的杂质(六氯苯、二噁英等)对生态环境和人类产生的危害极大,因此多个国家对其采取了禁用或限制使用的监管措施。本文概述了五氯硝基苯的毒性和环境危害及在国内外的应用情况,介绍了五氯硝基苯的替代产品和替代技术,并对其今后的管理和使用提出了建议。     

Inactivation of seed-treatment fungicides by Rhizobium

2-Methoxyethylmercury chloride (MEMC) applied to cowpea seeds with Rhizobium provided little or no control of seedling rot caused by Rhizoctonia solani; similar treatment, but without Rhizobium, gave >40% control of disease. Treating seeds with quintozene and then with Rhizobium rendered the fungicide completely ineffective against R. solani infections; disease control was >70% when seeds were treated with quintozene only, and >60% when seeds were coated first with Rhizobium and then with quintozene; treatment with quintozene+Rhizobium gave about 25% control. Simultaneous treatment with Rhizobium considerably reduced the efficacy of captafol, chloroneb, carbendazim and thiophanate-methyl. Disease control by carboxin, benomyl and thiabendazole remained unaltered in the presence of Rhizobium, but only thiabendazole gave good control. Seeds treated with MEMC and Rhizobium produced seedlings without root nodules; nodulation was also decreased by captafol and by carboxin but six other fungicides had no effect. The implications of Rhizobium-fungicides interactions are discussed in the context of fungicidal control of root diseases.     

实验室农药残留检测的测量不确定度评定——以GB 23200.8—2016测定草莓中4种农药残留为例

自2018年《CNAS-GL006化学分析中不确定度的评估指南》颁布后,不确定度计算方法的需求日益增强。本研究依据GB 23200.8-2016《水果和蔬菜中500种农药及相关化学品残留量的测定气相色谱-质谱法》,采用气相色谱-质谱法,结合内标定量法,对草莓中丙溴磷、亚胺硫磷、五氯硝基苯和氯氟氰菊酯的残留进行了测量不确定度的详细评定,共涉及标准品纯度、储备液配制、工作液配制、称样量和前处理过程5个B类评定分量,以及标准溶液峰面积、样品溶液峰面积和添加回收率3个A类评定分量。结果显示:在0.08 mg/kg添加水平下,丙溴磷、亚胺硫磷、五氯硝基苯和氯氟氰菊酯的测量合成相对标准不确定度依次为3.55%、3.22%、2.20%和3.66%,样品溶液峰面积和添加回收率2个A类不确定度分量对4种农药合成不确定度的贡献较高,其中丙溴磷、亚胺硫磷和氯氟氰菊酯均超过60%,五氯硝基苯超过31%。本研究中4种农药在草莓样品中的测量结果在95%的置信限(k=2)内可表示为:丙溴磷,0.085±0.006 mg/kg;亚胺硫磷,0.084±0.005 mg/kg;五氯硝基苯,0.079±0.003 mg/kg;氯氟氰菊酯,0.082±0.006 mg/kg。不确定度评定结果将最大限度地减少待测物在最大残留限量附近的残留值判定可能存在的争议。     

Genetic analysis of Grapevine leafroll‐associated virus 3 population from Galicia,Spain

Grapevine leafroll‐associated virus 3 (GLRaV‐3; Ampelovirus, Closteroviridae) isolates from Galicia in northwestern Spain were selected to characterize their genetic diversity according to different factors (age, origin, location, variety, etc.). The vines belonged either to local white and red varieties autochthonous to Galicia or to varieties from other Spanish regions but widely used in Galicia. These GLRaV‐3 isolates came from different vineyards in Galicia located in coastal or inner areas. Multiplex RT‐PCR allowed the detection of isolates belonging to groups I, II, III–V and VI. Two genomic regions were studied in the isolates, the HSP70h and the capsid protein, using specific primers that allow the detection of variants from groups I to V. Some possible recombinants could be detected; however, multiple infections with different variants indicated that they were not genuine recombinants. No differences were found in the population structure considering variety or geographical factors. Isolates belonging to four groups were found in the distinct areas surveyed: groups I and II were the most common, followed by groups VI and III, as is the case in the rest of the world. In the same surveys, the presence of insect vectors for GLRaV‐3 was investigated and found lacking in inland areas but present in those with milder climate. Genetic analysis did not support isolation of the GLRaV‐3 isolates in Galicia, suggesting that the uncontrolled exchange of infected vines and/or rootstocks has been a major agent of virus spread.     

Juvenile hormone III, hydroprene and a juvenogen as soldier caste differentiation regulators in three Reticulitermes species: potential of juvenile hormone analogues in termite control

The efficacy of juvenile hormone III (JH III) and two JH-mimicking compounds was compared in laboratory experiments with Reticulitermes lucifugus (Rossi), R. santonensis Feytaud and R. virginicus (Banks). Induction of presoldier, soldier and/or soldier-worker intercaste differentiation was taken as positive response to the treatment. The novel juvenogen, a fatty acid ester of a juvenoid alcohol, induced greatest soldier differentiation in all species tested, followed by hydroprene. JH III was less effective. Representatives of three Reticulitermes species showed similar trends in soldier induction rates. Differences in mortality in treatments of termites of the same species from different colonies with the same compound were observed and evidently were caused by differences in the conditions of respective colonies.     

Resynthesis of multiple resistance to organophosphorus insecticides from strains with factors of resistance isolated from the SKA strain of house flies

Individual factors of resistance to insecticides attributable to chromosomes II, III and ^V of the SKA strain of houseflies (Musca domestica L) were combined in pairs to determine how their presence affects resistance. The re-synthesised strains with resistance factors on chromosomes II and V, and on chromosomes III and V, were tested with several organophosphorus insecticides and DDT. The penetration delaying mechanism Pen on chromosome III, which alone gives little or no resistance, slightly increased the resistance of flies with the microsomal detoxifying factor Ses on chromosome V to diazinon and malaoxon-ethyl (c. × 1.5), but was more effective in increasing resistance to DDT (× 6). There was no effect on the response to other insecticides tested. The combined effect of the mechanisms of resistance on chromosome II (glutathione S-ethyl transferase and phosphatase) and on chromosome V (microsomal detoxication) approximated to the product of the resistance conferred by each of these mechanisms singly, suggesting that the mechanisms of resistance on the two chromosomes act independently. Therefore, most of the strong resistance to organophosphorus insecticides in the SKA strain results from the interaction between delayed penetration (chromosome III) and the factors of resistance on chromosome II, and the independent action of the resistance factors on chromosomes II and V.     

Biological control of Rhizoctonia solani on potatoes by antagonists. 1. Preliminary experiments with Verticillium biguttatum,a sclerotium-inhabiting fungus

A common mycoparasite,Verticillium biguttatum, was found to kill sclerotia ofRhizoctonia solani placed on an inert material (perlite) as well as in soil at 15°C and 20°C, but not at 10°C. Compared with the effectivity ofV. biguttatum, that ofGliocladium roseum, Gliocladium nigrovirens, Hormiactis fimicola andTrichoderma hamatum on sclerotia was only low. In laboratory experiments, treatment of sclerotia-bearing seed potatoes withV. biguttatum reduced disease symptoms in the first stage of growth of the potato plant.V. biguttatum was found to occur on the subterranean part of the potato plant. On untreated plants the surface of the sprouts was colonised byV. biguttatum originating from the soil, presumably partly in response to the presence ofR. solani mycelium. In a preliminary field experiment,Verticillium treatment did not reduce symptoms on the stem. However, there was a marked reduction in sclerotium formation on the newly formed potato tubers. This offers perspectives for a commercial use ofV. biguttatum in the control ofR. solani.     

The influence of two herbicides on the antifungal activity of some fungicides against Pythium butleri and Rhizoctonia solani causing damping-off of cowpea

The herbicides fluchloralin and alachlor applied to soil altered the effectiveness of fungicide treatments to seed and soil for controlling cowpea damping-off. These herbicides also modified the in-vitro toxicity of the fungicides to the mycelial growth of Pythium butleri and Rhizoctonia solani in a nutrient medium. Both herbicides reduced the toxicity of 2-methoxyethylmercury chloride (MEMC) and propamocarb to the growth of P. butleri, and of carbendazim to the growth of R. solani, but enhanced the toxicity of captafol and quintozene to P. butleri and R. solani, respectively. In pot tests, quintozene gave better control of R. solani damping-off in soil treated with fluchloralin or alachlor than in untreated soil, whereas disease control by carbendazim was decreased in similarly treated soils. Both herbicides attenuated the effectiveness of MEMC and captafol to control the damping-off caused by P. butleri; the efficacy of propamocarb was increased by alachlor but was decreased by fluchloralin. The implications of herbicide-fungicide interactions are discussed in the context of fungicidal control of root diseases in herbicide-treated soil.     

Inhibitory effect of isoprothiolane on metabolism of a phosphoramidate by isolates of Pyricularia oryzae cav. in relation to fungicide sensitivity

The inhibitory effect of isoprothiolane(diisopropyl 1,3-dithiolan-2-ylidenemalonate), a fungicide for rice blast control, on the metabolism of dibutyl N-methyl-N-phenylphosphoramidate (BPA) by 20 isolates of Pyricularia oryzae was examined in relation to sensitivity of the isolates to the reference fungicide IBP(S-benzyl diisopropylphosphorothiolate). The isolates were divided into five groups based on the modes of BPA metabolism and the inhibition of BPA metabolism by isoprothiolane. Every isolate in groups I and II, which was either a field isolate or a stock culture, decomposed BPA rapidly and produced both hydroxylated and N-demethylated BPA as metabolites. BPA decomposition by these isolates was strongly inhibited by isoprothiolane, resulting in the decreased production of both metabolites in group I and of the hydroxylated metabolite in group II. These isolates were almost equally sensitive to isoprothiolane. Isolates in groups III, IV, and V were all obtained from selection of the fungus mutants found growing on media containing isoprothiolane. Isolates in group III, derived by plating large numbers of conidia, did not decompose BPA to any extent. Mutants of groups IV and V were obtained from fast-growing sectors on agar containing isoprothiolane. Both these groups decomposed BPA, but isolates belonging to group IV produced copious amount of N-demethylated BPA whereas isolates in group V did not. BPA metabolism by these in vitro mutants in groups III, IV, and V was not inhibited by isoprothiolane. Thus, the inhibitory effect of isoprothiolane on BPA metabolism was correlated with sensitivity of an isolate to isoprothiolane. The inhibitory effect of IBP on BPA metabolism was not always correlated with the sensitivity of an isolate to IBP.     

棉铃虫不同品系Gqα基因的比较及其原核表达与纯化

为了阐明Gqα的DNA序列情况,分别以cDNA和总DNA为模板,对Gqα进行PCR扩增,结果发现,Gqα在DNA水平上没有内含子。为了了解不同棉铃虫品系Gqα的变异情况,以实验室5个棉铃虫品系的中肠总DNA为模板,使用Pyrobest高保真酶对该基因进行扩增,测序后翻译成氨基酸序列发现,田间的耐药性品系只有2个氨基酸的变异,即:第163位的非极性A(丙氨酸)变成了极性T(苏氨酸);第353位的非极性V(缬氨酸)变成了碱性R(精氨酸);实验室内毒蛋白品系只有一个氨基酸的变异,第353位的V变成了R;其他2种实验室品系没有任何氨基酸的变异。个别氨基酸的变异可能与棉铃虫对Bt毒素产生抗性相关。将Gqα编码区克隆到表达载体pET21b中,表达出带有6个组氨酸标签的重组蛋白并对其进行了纯化,为进一步研究其功能奠定了基础。     

Inactivation of sclerotia of Rhizoctonia solani on potato tubers by Verticillium biguttatum,a soil-borne mycoparasite

Experiments in the laboratory and on farms with potato tubers in storage are described in which sclerotia ofRhizoctonia solani were inactivated after inoculation of infected tubers with a suspension of conidia and hyphal fragments ofVerticillium biguttatum. Sclerotia on freshly harvested tubers can be killed in a period of six to eight weeks, provided that (1) a direct contact between sclerotia and conidia ofV. biguttatum is obtained, (2) the temperature during the storage period is at least 15 °C, but preferably closer to 20 °C during the first weeks, and (3) the relative humidity of the air between the tubers is at least 99%. Seed tubers are only certified as export quality if the infection withR. solani, visible as sclerotia on the tubers, is assessed as below a specified incidence. To restore the economical value of tubers with many sclerotia, living sclerotia can be inactivated byV. biguttatum. However, also dead sclerotia have to be removed, as dead and living sclerotia cannot be distinguished visually by inspectors. A satisfactory way to remove dead sclerotia from tubers has not yet been found.     

Specific Behaviour of French Aphanomyces Euteiches Drechs. Populations For Virulence and Aggressiveness on Pea, Related to Isolates from Europe, America and New Zealand

The pathogenic variability of Aphanomyces euteiches on pea was investigated using a collection of 88 pea-infecting isolates from France and 21 isolates from Denmark, Sweden, Norway, USA, Canada and New Zealand. Aggressiveness and virulence were assessed by scoring the root symptoms on a differential set of six pea genotypes. Eleven virulence types were characterised. The virulence type I, previously described as virulent on the whole set, was predominant and included the most aggressive isolates of all geographical origins. The other types were much less prevalent, existing as one to five isolates. Three virulence types (III, IV and V) contained no French isolates. The type III, avirulent on MN313, was composed of American isolates only, and resembled the major group recently described in the USA. A wide range of aggressiveness was found within the virulence type I, and the French isolates appeared globally more aggressive than the foreign isolates. These findings indicate that isolates from the virulence type I should be used as references in breeding programs, and that pea lines PI180693 and 552 may be the most interesting resistance sources to date, despite their only partial resistance.     

柑橘内生细菌YS45的鉴定、抗菌物质分析及其对油菜菌核病的防治作用

 从柑橘枝条中分离到26株对油菜菌核病菌(Sclerotinia sclerotiorum)具有拮抗作用的内生细菌,其中YS45菌株的拮抗活性最强。16S rRNA基因序列分析及形态学和生理生化鉴定结果表明YS45菌株为枯草芽孢杆菌(Bacillus subtilis)。高效液相色谱及质谱分析结果显示其主要的抑菌活性物质为一组fengycin同系物,包括fengycins A、fengycins B和一种稀少fengycin类型化合物。油菜离体叶片接种试验中,YS45菌株发酵液对油菜菌核病的防效在70%以上,与五氯硝基苯相当;田间小区接种试验表明,YS45菌株发酵液对油菜菌核病的防效也在50%以上。     

Molecular Typing and Presence of Genetic Markers Among Strains of Banana Finger-Tip Rot Pathogen, Burkholderia cenocepacia, in Taiwan

ABSTRACT Burkholderia cenocepacia (genomovar III of B. cepacia complex), the causal agent of banana finger-tip rot, is a common plant-associated bacterium but also an important opportunistic pathogen of humans. To better understand the nature of B. cenocepacia from banana, the genetic variation among B. cenocepacia isolates from various banana-growing regions in southern Taiwan was examined. Forty-four serial isolates recovered from diseased banana stigmata from three banana-growing regions during the periods ranging from 2002 to 2004 were investigated. All B. cenocepacia isolates picked from quinate-yeast extract tetracycline-polymyxin semiselective medium could cause onion maceration and were polymerase chain reaction (PCR) positive for bcscV, which is a type III secretion gene present in all members of the B. cepacia complex except B. cepacia (formerly genomovar I). Genetic diversity was assessed using recA PCR restriction fragment length polymorphism, recA nucleotide sequence analysis, and pulsed-field gel electrophoresis assays. The assays revealed the genetic variability among the isolates and also allowed us to trace the relationship among isolates. The isolates all were assigned to genomovar III and consisted of two groups, A and B, which corresponded to recA lineage IIIA and IIIB. The group B strains were separated into B1 and B2 subgroups and the B1 strains were further divided into distinct lineages. The B1 strains were the most frequently detected and occurred in all regions tested. There was no significant difference between strains from each subgroup in the virulence on banana fingers of cv. Cavendish. PCR assays were further used to determine whether B. cenocepacia from banana contained the cable pilus subunit gene (cblA), IS1356, and B. cepacia epidemic strain marker (BCESM), which are DNA markers associated with epidemic B. cepacia clinic strains. The results indicated that cblA and IS1356 were absent but the BCESM was found in all isolates. The present study revealed that banana is a natural reservoir of genetically diversified B. cenocepacia strains.     

浙江省果蔬灰葡萄孢对啶酰菌胺的抗性

以2004—2006年从浙江、江苏等地采集的灰葡萄孢对啶酰菌胺的敏感性基线[EC₅₀ = (1.07 ± 0.11) mg/L]为依据,采用菌丝生长速率法连续监测了浙江省果蔬灰葡萄孢群体对啶酰菌胺的敏感性变化。结果表明:浙江省果蔬灰葡萄孢对啶酰菌胺的抗性发展迅速,2012—2013年和2017—2018年的平均EC₅₀值分别为 (5.23 ± 7.79) 和 (24.30 ± 49.33) mg/L。其中,2012—2013年的抗药性菌株频率为15.3%,且均为低水平抗性 (LR) 菌株;而2017—2018年的抗药性频率上升至53.2%,并出现了7.5%的中等水平抗性 (MR) 菌株和1.3%的高水平抗性 (HR) 菌株。啶酰菌胺抗性菌株的菌丝生长速率、产孢量、产菌核数和致病力与敏感菌株相比均无显著差异。抗药性分子机制研究表明:啶酰菌胺抗性菌株的琥珀酸脱氢酶B亚基 (SDH B) 均发生了点突变,共包括H272R、P225F和N230I 3种类型,其中H272R型突变占88.5%;其SDH A和SDH D均未发生点突变;而SDH C的突变 (G85A + I93V + M158V + V168I) 与对药剂敏感性之间无明显联系。     

Hyaline mutants from Verticillium dahliae, an example of selection and characterization of strains for host–parasite interaction studies

The objective of the present work was to select stable well-characterized strains of Verticillium dahliae that could be used as biological tools in genetic and plant–microbe interaction studies. Hyaline mutants, known for their stability in pathogenicity were chosen for the study. Diversity in pathogenicity was found among hyaline subclones obtained from a defoliating wild-type clone, but not within those from nondefoliating ones. Most subclones from the defoliating clone had parental pathotypes, but one (V7-2) exhibited weak pathogenicity. This subclone (V7-2), together with a highly virulent one (V7-7) deriving from the same defoliating parent clone (7), were selected for further characterization, because of their differences in pathogenicity. When studied on the basis of their growth requirements, the two subclones expressed marked differences. V7-7 grew better than V7-2 over a wider range of temperature conditions. Both subclones grew similarly in media supplemented with NH₄ as nitrogen source, but in those with NO₃, V7-7 grew more vigorously than V7-2 and only the former could grow when NO₂ was used. In spite of these differences, the two subclones were found to belong to the same vegetative compatibility group, confirming their genetic proximity. These results highlight the physiological and genetic complexity inherent in V. dahliae . In view of their characteristics, the clones obtained in this study should prove to be valuable tools in furthering the understanding of genetic and host– V. dahliae interactions.     

Studies on the possible formation of polychloroazobenzenes in quintozene treated soil

Treatment of different soils with quintozene at a level of 1000 parts/10⁶ and incubation for 24 days at 30 °C did not result in the formation of polychloroazobenzenes, although pentachloroaniline was formed. This negative finding is in line with previous work, where it was shown that azobenzenes can only be formed from substituted anilines with at least one free ortho position. Soil activity was demonstrated by incubation of 3,4-dichloroaniline under similar conditions yielding the expected 3,3′,4,4′-tetrachloroazobenzene. Mass fragmentography with a quadrupole mass spectrometer was used for the detection of polychloroazobenzenes. Mass spectra for the synthesised reference compounds are presented.     

Preliminary evaluation of nine fungicides for control ofPhytophthora cactorum andP. citrophthora associated with crown rot in peach trees

Excised twig assay and excised stem inoculation were used to evaluate nine fungicides (metalaxyl, fosetyl-Al, copper hydroxide, copper sulfate, copper oxychloride, captan, quintozene, propamocarb and chlorothalonil) againstPhytophthora cactorum andP. citrophthora associated with crown rot in peach trees. Segments were soaked in fungicide solutions at different concentrations and then inserted vertically intoP. cactorum orP. citrophthora cultures growing on cornmeal agar plus antibiotics, or inoculated by inserting a mycelium-bearing agar plug directly into the cambium. Following incubation, the bark was scraped off and length of necrosis was measured. Metalaxyl was the only fungicide that inhibited canker development on segments at the manufacturer-recommended concentration. Fosetyl-Al, captan, copper hydroxide and copper sulfate inhibited canker development at 3, 4, 4 and 8 gl^-1, respectively. The other fungicides did not affect canker length significantly compared with non-treated twigs, with the exception of propamocarb, which reduced the development ofP. cactorum on excised stems. The tested methods enabled rapid and effective evaluation of a large number of chemicals to prevent crown rot diseases caused byPhytophthora in the laboratory. http://www.phytoparasitica.org posting Dec. 5, 2001.     

Verticillium longisporum and V. dahliae: infection and disease in Brassica napus

Verticillium wilt of oilseed rape ( Brassica napus ) is caused primarily by Verticillium longisporum and has become a serious problem in northern Europe. In order to evaluate whether V. longisporum and V. dahliae differ in their interaction with oilseed rape, phenotypical and molecular assessments were made. Oilseed rape plants for fungal assessments were inoculated with V. longisporum and V. dahliae via root-dipping and samples were taken from roots, stems, leaves, flowers, pods and seeds during plant development. The infection by V. longisporum was found to start mainly in lateral roots and root-hairs, followed by colonization of the xylem vessels and extensive spread in stems and leaves, whereas V. dahliae infected the main roots and remained in the region below the cotyledon node of the plants. Re-isolation studies, together with PCR analysis of samples taken from early growth stages through to fully ripe plants, showed that the onset of flowering was a critical phase for V . longisporum to colonize plants. No seeds infected with V. longisporum were found. Mycelial growth from V. dahliae but not V. longisporum was significantly reduced on media containing tissue from a low glucosinolate B. napus genotype compared with growth on media containing tissue from a high glucosinolate cultivar. The results of this study suggest that V. longisporum favours B. napus as host and that the transition from the vegetative to the generative phase is of importance for the spread of the fungus in oilseed rape plants.