Insensitivity of acetylcholinesterase as a factor in resistance of houseflies to the organophosphate Rabon

Acetylcholinesterase from an organophosphate-resistant strain of housefly exhibited a decrease in sensitivity to Rabon in vitro, to the extent of 206-fold for the soluble enzyme and 38-fold for the particulate. These effects upon the bimolecular reaction constant, k_i, were due to a 573-fold decrease in affinity of the inhibitor for the soluble and 103-fold for the particulate enzyme, coupled with a small increase in reactivity.     

Mechanism of resistance to dichlorvos in adult houseflies

Houseflies (Musca domestica) originating from agricultural buildings in Jalasjarvi (Vaasa province of Finland) were found to be resistant to contact with a variety of organophosphorus insecticides, including dichlorvos. Tolerance to topical application of bromophos, crotoxyphos, and trichlorfon was fairly high, to dimethoate and dichlorvos it was moderate, and to tetrachlorvinphos it was very low. Much higher degrees of resistance were obtained after exposure to surface deposits.Evidence is presented to suggest that the mechanism of resistance in the Jalasjarvi flies is probably based on reduced penetration. With dichlorvos less of the absorbed chemical penetrates the central nervous system (the thoracic ganglionic complex) of the resistant fly than penetrates that of the susceptible fly. No differences were apparent between the strains in the otherwise small rates of degradation of dichlorvos.     

Development of resistance to pyrethroids in field populations of danish houseflies

Houseflies (Musca domestica) on Danish farms have developed high multiresistance to organophosphorus compounds, after successive use of several OPs, mainly dimethoate, in recent years. Topical application tests 1971–73 with flies from many farms showed that the high OP-resistance did not involve resistance to pyrethroids (± the synergist piperonyl butoxide (pb)) above a level of 3–7 x, unless field pressure with synergised pyrethrum (py/pb) or other pyrethroids was applied. In 1971–72 moderate to high, often heterogeneous, pyrethroid resistance was found on a few trial farms treated frequently with pyrethroid aerosols (mainly py/pb) and in 1973 on most of 23 trial farms treated intensively with aerosols (or space spray) containing py/pb, bioresmethrin ± pb, tetramethrin/pb or tetramethrin/resmethrin. The effect of field pressure with these different pyrethroids on development of pyrethroid resistance is summarised and discussed. Maximum resistance ratios, R/S at LD₅₀-LD₉₅, were: py/pb (1:5), 40->100; bioresmethrin, 191–770; bioresmethrin/pb (1:5), 55–133; tetramethrin/pb (1:5), 171->200; tetramethrin/resmethrin (1:5), 78->370. The intensity of selection pressure with pyrethroids is believed to be an important factor. Although py/pb has been widely used as a supplementary fly control on Danish non-trial farms, pyrethroid resistance has only been found on a few of them.     

Some properties of a mechanism delaying penetration of insecticides into houseflies

The penetration of insecticides into houseflies, with and without the factor that delays penetration, was studied by measuring the amounts of the insecticides on the surface of the flies at intervals after treatment. The factor slowed the entry of all the insecticides tested, but its effect depended on the dose, and had little or no effect when the dose applied was big (20 μmUg/fiy). Penetration in all strains, susceptible and resistant, increased but not proportionally to the size of the dose. The rate insecticide was lost from the surface slowed with time and increasing age of the insects. It was faster in males than females, and was always faster in flies without the penetration factor. The differences in penetration between the flies with and without the factor was less with n-dodecane as solvent than with acetone. With all strains, rate of loss from the surface was slowest with dieldrin. Diazinon, parathion and chlorthion-ethyl were lost at about the same rate; diazoxon penetrated fastest.     

Interaction between the factor delaying penetration of insecticides and the desethylation mechanism of resistance in organophosphorus-resistant houseflies

The desethylation (gene a) and penetration delaying (pen) factors of resistance to organophosphorus insecticides isolated by genetical methods from the diazinon-selected SKA strain of houseflies (Musca domestica L.) were inbred into a strain whose resistance was compared with that of flies with single factors of resistance and the SKA strain. Pen alone decreases kill marginally, and gene a alone raises LD₅₀ to 1·4–48 times that of susceptible flies, depending on the insecticide. The two factors together increase resistance greatly to many organophosphorus insecticides (up to 5–10 times or more to diazinon, malathion-ethyl and chlorthion-ethyl than of flies with gene a only) indicating that the two factors interact. Interaction is greater against the thionates than the corresponding phosphates, probably because pen delays entry of thionates more than of the corresponding phosphates. The role in resistance of each factor when hetero-and homo-zygous, and the reasons for interaction, are discussed.     

Genetics of resistance of houseflies (Musca domestica L.) to pyrethroids. I. Knock-down resistance

Knock-down resistance in Musca domestica, which provides cross-resistance between DDT and pyrethroids, has been genetically separated from three different resistant populations. After careful purification of each factor, the cross-over rates between them and the visible mutants brown body and green eye were estimated. This indicated that these factors are probably identical. The influence and implications of the knock-down resistance factor, kdr, on the total resistance of populations that include it are briefly discussed.     

Problems in obtaining a description of the evolution of dimethoate resistance in Danish houseflies (Musca domestica)

Data on the evolution of a polygenically determined resistance to dimethoate, in populations of houseflies on Danish farms, were examined in order to see whether useful information about the processes involved could be obtained. Examination of the variance of tolerances in samples of flies suggested that there was considerable genetic variability for resistance before dimethoate was first used, but provided no other information. Before examining estimates of log LD⁵⁰ (LLD⁵⁰), the relationship between LLD⁵⁰ and the log weight of the flies assayed was investigated in an experiment in Denmark, and the standard error of LLD⁵⁰ estimates was estimated. Examination of LLD⁵⁰ estimates (adjusted for log weight) provided evidence of substantial migration of flies between farms, which produced large and unpredictable changes in tolerance levels on individual farms, and prevented extraction of further useful information from the data. However, it was concluded that even had migration been negligible, the data would not have provided information suitable for the construction of models of the evolution of this polygenically determined resistance. This conclusion is likely to hold for most existing data. Alternative approaches towards obtaining models of the evolution of polygenically determined resistances are suggested.     

Genetics of resistance of pyrethroid-selected houseflies,Musca domestica L.

Four resistance factors were isolated genetically from the NPR strain of houseflies (Musca domestica L.), which resists natural pyrethrins, and were characterised toxicologically. The four factors were : pen, which reduces the rate of penetration of insecticides through the cuticle; kdr-NPR, a general pyrethroid resistance mechanism unaffected by the synergist sesamex; py-ses, a mechanism of resistance to natural pyrethrins that can be suppressed by sesamex; and py-ex, a factor that gives strong resistance to synergised natural pyrethrins and to the new synthetic esters, e.g. resmethrin, but little or none to natural pyrethrins alone.     

Toxicodynamics of malaoxon in house flies

The fate of malaoxon was studied in a susceptible and a resistant strain of house fly following topical application. Sublethal doses were used: 160 pmol for the S-strain (0.17 × LD₅₀) and 1570 pmol for the R-strain (0.1 × LD₅₀). The penetration rates are dose dependent and semilog plots of the external amount vs time show that these rates are not proportional to this external amount. Internal concentrations of malaoxon rapidly increase following administration, reach maximum values between 30 min and 2 hr (depending on dose), and then slowly decrease. The rate of metabolic degradation is highest in the early stage of the intoxication process. A three-compartment pharmacokinetic model is postulated to explain the experimental data quantitatively. The first compartment represents external malaoxon, the other two represent internal parent compound. Statistical analysis shows that the penetration rate is better described with a sum of two exponentials rather than with a single exponential decay. In the model, degradation occurs in the first internal compartment and is assumed to be first order. Malaoxon is distributed between the two internal compartments slowly with first-order kinetics. Parameter estimation with curve-fitting procedures for the internal processes (degradation and exchange) shows that there is not one set of parameter values that can be used for both strains simultaneously. This prompted a study of possible interstrain differences in degradation capacities. It was found that in vitro the R-strain had a fourfold higher oxidative breakdown rate of malaoxon. Taking this difference into account it is possible to explain the two sets of data with one kinetic model, although other alternatives cannot be excluded.     

Evidence of cork barrier formation as a resistance mechanism to berry disease (Colletotrichum coffeanum) in arabica coffee

The histology of resistance to coffee berry disease (CBD) was studied in artificially inoculated berries and hypocotyls of 6-week old seedlings of a number of varieties ofCoffea arabica L. In resistant varieties a phellogen was quickly formed some cell layers below the site of infection and progress of the fungal invasion was effectively blocked by a complete barrier of suberized cells. Such barriers were absent or incompletely developed in CBD susceptible varieties. A highly significant correlation (r=0.87) was found between the frequencies of complete barrier formation in berries and in hypocotyls of young seedlings, while both were also highly correlated to observed mature plant resistance (r>-0.93). Resistance to CBD in arabica coffee may to an important extent be based on the formation of cork barriers. These cork barriers confine the pathogen to the small volume of tissue external to the barrier so that its growth is severely restricted. Such a resistance mechanism is likely to be stable (race-nonspecific). The almost identical response to infection observed in berries and hypocotyls provides further evidence that plants with resistance to CBD can be reliably preselected by the hypocotyl inoculation test.     

Factors affecting the sequential acquisition by Danish houseflies (musca domestica L.) of resistance to organophosphorus insecticides

The prolonged use of dimethoate, introduced into Denmark to control houseflies (Musca domestica L.) that had become resistant to parathion and diazinon, resulted ultimately in dimethoate resistance. Selection with dimethoate led to the disappearance of the hydrolytic phosphatase, a major mechanism of resistance to parathion and diazinon, and its replacement by the acetylcholinesterase AChER with somewhat decreased sensitivity to inhibition by organophosphorus (OP) insecticides. The hydrolytic phosphatase probably disappeared because low substrate turn-over made it ineffective against dimethoxon (O, O-dimethyl S-methylcarbamoylmethyl phosphorothioate, also known as omethoate). which accumulates at higher concentrations than paraoxon (diethyl4-nitrophenyl phosphate) in the haemolymph. Dimethoate selected AChER preferentially because it improved the chances of houseflies surviving against the relatively poor AChE inhibitor dimethoxon, whereas its relatively small insensitivity to OP insecticides, unimportant against good inhibitors such as paraoxon, prevented its selection by parathion.     

桃蚜对噻虫嗪代谢抗性机制研究

对桃蚜进行室内噻虫嗪抗性品系筛选,选育至15代后抗性倍数达到75.6倍。对噻虫嗪敏感品系(THI-S)和抗性品系(THI-R)桃蚜的谷胱甘肽S-转移酶(GSTs)、酸性磷酸酯酶(ACP)、碱性磷酸酯酶(ALP)、羧酸酯酶(CarE)、多功能氧化酶(MFO)O-脱甲基活性进行了比较,结果显示:敏感品系(THI-S)和抗性品系(THI-R)的谷胱甘肽S-转移酶比活力分别为3.127 5和3.215 9,差异不显著,桃蚜抗性品系体内酸性磷酸酯酶、碱性磷酸酯酶、羧酸酯酶和多功能氧化酶O-脱甲基活性均显著高于敏感品系,分别达到了1.57、2.10、6.12、2.03倍。表明桃蚜对噻虫嗪抗性的产生与酸性磷酸酯酶、碱性磷酸酯酶、羧酸酯酶和多功能氧化酶O-脱甲基的活性相关。     

Hydrolysis of paraoxon and malaoxon in three strains of Myzus persicae with different degrees of parathion resistance

Homogenates of three strains of Myzus persicae, A, R, and E, with an LD₅₀ for topically applied parathion of 9, 93, and 263 ng per aphid, showed an in vitro hydrolytic degradation of paraoxon of 2.3, 4.7, and 8.6 pmol/mg aphid/h, respectively. These values represent V_max; K_m was <10^?7M. The three strains showed a malaoxon degradation of 2.4, 11.9, and 18.8 pmol/mg/h at 10^?6M substrate concentration. V_max for R and E was 21 and 27 pmol, respectively and K_m 7 and 4 × 10^?7M. Activity in strain A was too low to estimate these entities. The breakdown product of paraoxon was mainly diethyl phosphoric acid, that of malaoxon mainly dimethyl phosphoric acid. No hydrolysis of the carboxylester groups of malaoxon was found. Hydrolysis of paraoxon and malaoxon was inhibited by isopropyl and n-propyl paraoxon and by the salioxon-analog K₂. The two latter compounds were shown to act as synergists with parathion when added in amounts that caused little mortality when given alone. The hydrolytic enzyme is soluble and retains its activity during incubations of several hours. It is likely that it is responsible for at least part of the resistance. Resistance was maintained without selection over a period of three years. There was no correlation between degree of resistance and carboxylesterase activity of the strains.     

Nicotinoid and pyrethroid insecticide resistance in houseflies (Diptera: Muscidae) collected from Florida dairies

BACKGROUND: The housefly, Musca domestica L., continues to be a major pest of confined livestock operations. Houseflies have developed resistance to most chemical classes, and new chemistries for use in animal agriculture are increasingly slow to emerge. Five adult housefly strains from four Florida dairy farms were evaluated for resistance to four insecticides (beta‐cyfluthrin, permethrin, imidacloprid and nithiazine). RESULTS: Significant levels of tolerance were found in most field strains to all insecticides, and in some cases substantial resistance was apparent (as deduced from comparison with prior published results). At the LC₉₀ level, greater than 20‐fold resistance was found in two of the fly strains for permethrin and one fly strain for imidacloprid. Beta‐cyfluthrin LC₉₀ resistance ratios exceeded tenfold resistance in three fly strains. The relatively underutilized insecticide nithiazine had the lowest resistance ratios; however, fourfold LC₉₀ resistance was observed in one southern Florida fly strain. Farm insecticide use and its impact on resistance selection in Florida housefly populations are discussed. CONCLUSION: Housefly resistance to pyrethroids is widespread in Florida. Imidacloprid resistance is emerging, and tolerance was observed to both imidacloprid and nithiazine. If these insecticides are to retain efficacy, producer use must be restrained. Copyright © 2009 Society of Chemical Industry     

昆虫对Bt杀虫蛋白的抗性机制及治理策略

转Bt基因抗虫作物在害虫综合防治中发挥重要作用，是新一代植物保护产品，其抗虫性能和经济效益已得到普遍肯定。然而由于转基因抗虫植物在整个生育阶段高水平的表达Bt杀虫蛋白，有可能会导致害虫对Bt杀虫蛋白产生抗性，抗性的产生将严重影响Bt植物的应用。田间已经发现小菜蛾Plutella xylostella（Linnaeus）对Bt杀虫剂产生了抗性，实验室人工汰选条件下已经有多种害虫对Bt杀虫蛋白产生了抗性，并对抗性产生的机制进行了研究，目前主要认为害虫产生抗性与杀虫蛋白与中肠细胞上受体蛋白结合能力的改变以及Bt杀虫蛋白在中肠的水解作用的变化有关，但抗性产生的机制还与其它因素有关，估计是多方面因素造成的，因此，抗性机制的产生还不十分清楚。目前已经制订了“高剂量＋庇护所”和多基因策略等一系列有效抗性治理策略，这些策略还需要在深入研究并阐明抗性机制的基础上，进一步完善，以实现Bt抗虫植物在害虫防治中的可持续利用。     

植物抗病毒的可能机制--基因沉默

根据目前RNA介导的植物病毒抗性(RMVR)、转录后基因沉默(PTGS)的研究成果,综述了基因沉默可能是植物抵抗病毒的一种机制,深入研究基因沉默不仅在抗病机制上有重要的理论意义,而且对彻底解决植物病毒问题有较大的潜在实用价值.     

葡萄抗霜霉病机制研究进展

葡萄霜霉病是危害葡萄的重要病害之一,该病害发生时会给葡萄生产造成严重的经济损失。本文综合分析了国内外对葡萄抗霜霉病机制研究的结果,认为葡萄对霜霉病抗性受多种因素影响,主要表现为生理生化物质、叶片形态结构及抗病遗传基因三个方面。由于各种因素对霜霉病病原菌作用关系复杂,目前对于葡萄抗霜霉病的决定性因子还没有明确结论。     

Cuticular lipids of two resistant and a susceptible strain of houseflies

A study was made of the composition of the cuticular lipids of two resistant strains of houseflies (Rutgers and Fc), both of which show a reduced rate of absorption of insecticides as a partial mechanism of resistance and a susceptible strain (CSMA). Total lipids, monoglycerides, diglycerides and sterol esters (except in the Fc strain), sterols, fatty acids and phospholipid phosphorus were higher in resistant strains than in the susceptible strain. Phosphatidyl-ethanolamine and phosphatidyl-choline were major constituents of the phospholipid fractions and were appreciably higher in the resistant strains. Cuticular wax contents did not differ among strains. Incorporation of lipid precursors, [U-¹⁴]acetate and [³²P]orthophosphate, was greater in the cuticle of one or both resistant strains, depending on the lipid component examined.     

Direct measurement of paraquat in leaf protoplasts indicates vacuolar paraquat sequestration as a resistance mechanism in Lolium rigidum

Sequestration of paraquat away from its target site in the chloroplast has been proposed as a mechanism of paraquat resistance. However, no consensus has been reached as to where paraquat is sequestered. This study quantifies paraquat in leaf protoplasts of paraquat resistant (R) and susceptible (S) Lolium rigidum. Intact protoplasts were prepared from plants treated with commercial dose of paraquat for 2 h. Paraquat absorbed by the leaf protoplasts was determined by light absorption of reduced paraquat following concentration and purification using a cation-exchange resin. Leaf protoplasts from treated paraquat resistant plants contained 2- to 3-fold more paraquat than leaf protoplasts isolated from susceptible plants. Since paraquat is not metabolised in L. rigidum and paraquat readily enters chloroplasts of both R and S plants, this greater amount of paraquat in leaf protoplasts of R plant must be kept away from the target site (chloroplast). This result indicates that paraquat resistance in L. rigidum is associated with a cytoplasmic mechanism, most likely a greater rate of vacuolar sequestration.