杀虫剂防治水稻褐飞虱的有效利用率分析

通过综合分析杀虫剂对水稻褐飞虱的毒力测定结果、褐飞虱田间防治指标、杀虫剂田间推荐剂量及杀虫剂经稻田叶面喷施后在田间的分布等文献资料,发现杀虫剂防治褐飞虱的田间推荐剂量的有效利用率不足0.1%。理论上,通过室内毒力测定可获得杀虫剂杀死褐飞虱种群90%个体的致死剂量 (LD₉₀,单位:μg/头),用稻田中褐飞虱的发生量乘以LD₉₀值即为杀死90%田间虫量的杀虫剂有效用量;而实际上杀虫剂的田间推荐剂量却是其有效用量的千倍以上。分析出现这种现象的原因可能有:1) 在褐飞虱为害的水稻孕穗期和扬花期,采用手动喷雾器进行叶面喷雾时,杀虫剂在水稻上的沉积率为34.25%～46.10%,但其中82%以上分布在水稻冠层以上部位,分布在水稻基部茎秆部位的不足2%,只占杀虫剂使用量的0.5%左右;2) 褐飞虱获取致死剂量的杀虫剂后死亡,但杀虫剂和褐飞虱在田间的分布极不均匀,当最低剂量的杀虫剂和最多虫量的褐飞虱出现在同一株水稻上时,该剂量必须能够控制褐飞虱的为害,那么对于有更多杀虫剂和更少褐飞虱的植株而言,就必然造成杀虫剂的浪费,从而降低其有效利用率;3) 约有50%以上的杀虫剂洒落在稻田水中,经田水稀释后的质量浓度远远低于杀死褐飞虱种群10%个体的致死浓度 (LC₁₀值,单位:mg/L),不能有效杀死褐飞虱。作者认为,通过人工智能,将杀虫剂直接喷洒在褐飞虱发生为害的部位,并根据虫量进行变量施药,必将大幅提高杀虫剂防治水稻褐飞虱的有效利用率。     

丽蝇蛹集金小蜂P450基因家族的发现和分子进化分析

丽蝇蛹集金小蜂是一类重要的天敌昆虫,大量地应用于生物防治中。虽然寄生蜂不是化学农药的直接作用靶标,但其在自然环境中可能接触到化学农药。P450基因家族是昆虫产生抗药性的重要原因之一,同时也参与了昆虫激素的合成与降解。因此,研究P450基因家族对充分利用丽蝇蛹集金小蜂,以及保护其免受农药毒杀具有重要的意义。本文利用生物学信息方法从丽蝇蛹集金小蜂基因组筛选到91个P450基因。家族分析表明,丽蝇蛹集金小蜂具有15个P450基因超家族、27个亚家族。对P450基因的正选择和基因转换开展了进一步分析,研究发现5个基因族存在正选择位点,2个基因族存在基因转换,正选择和基因转换之间存在明显的关联性。各个亚家族的选择压力分析表明,CYP9P亚家族受到选择压力最大（ω=118．9355）。通过MEME程序在丽蝇蛹集金小蜂P450基因上识别到5个频率为90％以上的motif,其中“LAE[NH]P”为新发现的motif.     

The effect of insecticide application sequences on the control and insecticide resistance status of the peach-potato aphid, Myzus persicae (Hemiptera:Aphididae), on field crops of potato

Experiments were done on commercial potato crops in the UK to investigate the effect of different insecticide sequences on the control and insecticide resistance status of Myzus persicae (Sulzer). The work was done to provide field validation of similar laboratory studies done in 'field simulators'. To ensure adequate aphid populations and to influence the initial resistance status of the aphid population, cultured M. persicae from a clone of known resistance status (esterase R1, kdr heterozygote, non-MACE (modified acetylcholinesterase)) were inoculated into both experiments. Two-spray programmes starting with lambda-cyhalothrin (a pyrethroid insecticide) gave poor control in comparison with programmes starting with pirimicarb (a carbamate insecticide) or pirimicarb-containing mixtures. This concurred closely with the results obtained from single applications in field simulator studies. Treatment sequences containing pymetrozine (a pyridine azomethine insecticide) were also effective, though slower-acting. This again concurs with field simulator studies. The proportions of aphids carrying different resistance mechanisms were largely unaffected by treatment in these experiments. The implications of these results for field control strategies are discussed.     

The effective dominance of resistance genes in relation to the evolution of resistance

Field selection for a rare insecticide resistance (R) gene can be substantially delayed if R is rendered effectively recessive [if all or most heterozygotes ( R + ) are killed by the field dosages] and if a representative sample (+ +, R +, RR) of the population is permitted to escape exposure. The relationship between heterozygote mortality and escape in influencing selection rates has been considered at various frequencies of R. The cases when mating precedes selection, and when selection precedes mating, have been calculated using computer simulations. It is concluded that when R genes are rare it is advantageous (a) to apply the highest possible doses of insecticide in such a form that all exposed insects pick up an amount sufficient to kill all or most heterozygotes, (b) to apply the insecticide to adults after they have mated, rather than to younger adults or to larvae, and (c) to ensure that the proportion of insects given the opportunity to escape contact is as high as practical considerations allow. Once resistance is obvious (when samples are brought into the laboratory for tests), the R gene frequency will have risen to a level which makes the suggested strategy no longer valuable. Alternative strategies are briefly discussed.     

A novel method for molecular targeting of insecticide resistance in Rhopalosiphum padi L. (Homoptera: Aphididae)

The bird cherry-oat aphid (Rhopalosiphum padi L.) is a devastating cereal pest that develops high resistance to organophosphate and carbamate insecticides. Because acetylcholinesterase (ACE) is the target of carbamate and organophosphate insecticides, the resistance mechanism usually involves mutations occurring in ACE-encoding genes, Ace1 and Ace2. Here, we describe a novel polymerase chain reaction (PCR)-based method for the diagnosis of resistance cases associated with the point mutation F368L in the Ace2 gene. We amplified a 127 bp DNA fragment from Ace2 gene using a modified reverse primer, and digested the amplification product using SmaI endonuclease. This procedure enabled a simple and rapid distinction between resistant and susceptible genotypes for F368L mutation. Subsequently, we screened 152 R. padi samples, and found that F368L mutation occurred at low frequency, in both the homozygous (R/R) and heterozygous (R/S) states. Based upon the results of this study, we believe that molecular diagnosis of insecticide resistance should be generalized to genes and mutations involved in this process, toward an optimal accuracy of insecticide applications.     

Insecticide resistance status of Aedes aegypti (L.) from Colombia

BACKGROUND: To evaluate the insecticide susceptibility status of Aedes aegypti (L.) in Colombia, and as part of the National Network of Insecticide Resistance Surveillance, 12 mosquito populations were assessed for resistance to pyrethroids, organophosphates and DDT. Bioassays were performed using WHO and CDC methodologies. The underlying resistance mechanisms were investigated through biochemical assays and RT‐PCR. RESULTS: All mosquito populations were susceptible to malathion, deltamethrin and cyfluthrin, and highly resistant to DDT and etofenprox. Resistance to lambda‐cyhalothrin, permethrin and fenitrothion ranged from moderate to high in some populations from Chocó and Putumayo states. In Antioquia state, the Santa Fe population was resistant to fenitrothion. Biochemical assays showed high levels of both cytochrome P450 monooxygenases (CYP) and non‐specific esterases (NSE) in some of the fenitrothion‐ and pyrethroid‐resistant populations. All populations showed high levels of glutathione‐S‐transferase (GST) activity. GSTe2 gene was found overexpressed in DDT‐resistant populations compared with Rockefeller susceptible strain. CONCLUSIONS: Differences in insecticide resistance status were observed between insecticides and localities. Although the biochemical assay results suggest that CYP and NSE could play an important role in the pyrethroid and fenitrothion resistance detected, other mechanisms remain to be investigated, including knockdown resistance. Resistance to DDT was high in all populations, and GST activity is probably the main enzymatic mechanism associated with this resistance. The results of this study provide baseline data on insecticide resistance in Colombian A. aegypti populations, and will allow comparison of changes in susceptibility status in this vector over time. Copyright © 2011 Society of Chemical Industry     

A theoretical study of discriminating parameters in metabolic resistance to insecticides

In the case where resistance to an insecticide is associated with increased metabolism of the insecticide, it should not be concluded that the resistance is due only to the increased metabolism (i.e. metabolic hypothesis). Here, we study theoretically the pharmacokinetic consequences of a resistance mechanism due to increased metabolism. We consider two cases: treatment with the initial dose D₀ applied to the susceptible strain and the treatment with the initial dose αD₀, with α>1, applied to the resistant strain. We show the conditions for which the metabolism hypothesis is conceivable. The time τ, from which the mortality of the susceptible strain is significantly higher than that of the resistant strain, is an important parameter in determining the validity of the metabolic hypothesis. The more τ increases, the more the conditions are favourable to this hypothesis. Our work suggests an approach to test the metabolic hypothesis from experimental results. © 1998 SCI     

Effects of temperature on fitness costs, insecticide susceptibility and heat shock protein in insecticide-resistant and -susceptible Plutella xylostella

Effects of high temperature on insecticide susceptibility and fitness were studied in the field population of Plutella xylostella (L.) (Lepidoptera: Plutellidae) (R DBM) and a susceptible field-insectarium population of P. xylostella (S DBM). R DBM displayed 18.3-fold resistance to methamidophos and 74.0-fold resistance to avermectin. The population growth tendency index (I) values were 41.8 (S DBM) and 27.7 (R DBM) at 25 °C, and 1.19 (S DBM) and 0.23 (R DBM) at 33.5 °C. The level of methamidophos resistance in the progenies of R DBM declined sharply when reared at high temperature for one generation. The increase of susceptibility to methamidophos appeared to pass from generation to generation. S DBM displayed higher up-regulation of Hsp70 expression at high temperature than R DBM. It was suggested that low fitness in R DBM caused by high temperature might be involved in the sharp decline of methamidophos resistance under high temperature conditions.     

Temporal and spatial dynamics of insecticide resistance in Myzus persicae (Hemiptera: Aphididae)

Advances in understanding insecticide resistance in the peach-potato aphid, Myzus persicae (Sulzer), at the genotypic, biochemical and molecular levels have led to rapid and precise methods for the detection of several resistance mechanisms (elevated carboxylesterase, modified acetylcholinesterase or MACE, and knockdown resistance or kdr) in individual insects, and for monitoring their frequencies over space and time. This paper summarises the results of two long-term surveys of resistance dynamics in M persicae in England, based on samples collected directly from field and glasshouse crops or from four 12.2-m suction traps. The study showed marked fluctuations in resistance frequencies that probably reflect the counteracting forces of selection imposed by insecticides for aphids possessing more copies of esterase resistance genes, and selection against these forms when insecticide use is relaxed. There is growing evidence that several different resistance mechanisms in M persicae have associated fitness costs. In the case of esterase and MACE, these costs are apparently strong enough to effect a decline in resistance frequency over winter, and a more prolonged decline over successive cropping seasons when aphid numbers are insufficient to trigger intensive chemical applications. Changes in the overall frequency of resistance genotypes may also be influenced by the predominance of year-round parthenogenesis in M persicae in the UK, leading to non-random associations between mechanisms and selection operating on clonal lineages rather than individual genotypes.     

Testing insecticide resistance management strategies: mosaic versus rotations

BACKGROUND: Developing scientifically valid, economically acceptable insecticide resistance management (IRM) programs is critical for sustainable insect management. The diamondback moth, Plutella xylostella (L.), has demonstrated an ability to develop resistance to many different classes of insecticides, including proteins produced by the bacterium Bacillus thuringiensis Berliner (Bt). Recently it has developed resistance to the novel compounds spinosad and indoxacarb. In greenhouse cage experiments, a laboratory‐selected population of P. xylostella resistant to spinosad, indoxacarb and Bt was used to compare population growth and resistance evolution if these three insecticides were rotated or used in a mosaic fashion. RESULTS: The average population density through nine generations was lowest in the treatment in which the insecticide was rotated every generation (R‐1) (x? = 20.7 ± 3.20) compared with the treatment in which the insecticide was rotated every third generation (R‐3) (x? = 41.4 ± 17.6) or where the insecticides were applied as a mosaic (M) (x? = 41.8 ± 6.53). After nine generations, the survival of resistant individuals increased for each insecticide (7.2–73.5%) compared with the population without selection (CK) (0.73–3.1%). Survival on spinosad was significantly lower (23.7%) in the single‐generation rotation than for the other two treatments, both of which exceeded 72%. The calculated survival on all three insecticides treated simultaneously, according to the survival on each insecticide, was 0.26, 0.81 and 1.6% for R‐1, R‐3 and M treatments respectively. CONCLUSION: Results of both population density and resistance development indicated that insecticide rotation every generation was better for IRM than if the insecticide was rotated every third generation or if the three insecticides were applied as a mosaic. Copyright © 2010 Society of Chemical Industry     

与昆虫抗药性相关的乙酰胆碱酯酶基因突变研究进展

有机磷和氨基甲酸酯类杀虫剂的广泛使用导致许多害虫产生了明显的抗药性。害虫对这些杀虫剂产生抗性的一个重要原因是其乙酰胆碱酯酶 (AChE)基因发生突变,从而导致AChE敏感度下降。简要概述了AChE基因发生突变的昆虫种类,介绍了AChE基因突变对其结构与功能的影响、变构AChE的特性、AChE基因突变对适合度的影响以及AChE突变不同组合对抗性的影响。这些突变可为设计新颖的反抗性化合物开辟新的途径。     

Metabolism and absorption of methyl parathion by tobacco budworms resistant or susceptible to organophosphorus insecticides

Factors involved in insecticide resistance were evaluated by using ¹⁴C-labeled methyl parathion and aldrin to compare rates of absorption and metabolism by Heliothis virescens (F.) larvae that were resistant (R) and susceptible (S) to methyl parathion. Tests with third-stage R and S larvae suggested that the rate of insecticide absorption from the cuticular surface was not a major resistance factor. Further evidence for this assumption was demonstrated by the resistance of R larvae to injected and orally administered doses of methyl parathion. Smaller amounts of unmetabolized methyl parathion and aldrin were recovered from S larvae, an indication that differences in metabolism were probably related to the resistance.     

Association between insecticide use and greenhouse whitefly (Trialeurodes vaporariorum westwood) resistance to insecticides in hawaii

Susceptibility to acephate, methomyl, and permethrin was determined with laboratory bioassays for adults of greenhouse whitefly, Trialeurodes vaporariorum Westwood, from 12 to 14 sites in Hawaii. Comparisons at LC₅₀ showed up to 42-fold resistance to acephate, 36-fold resistance to methomyl, and 8-fold resistance to permethrin. Higher levels of resistance to acephate and methomyl than to permethrin are consistent with greater use of organophosphates and carbamates than pyrethroids by growers. Insecticide use varied from 1 to 98 insecticide sprays per site per season. Significant positive associations between LC₅₀ for each insecticide and frequency of application of the same insecticide were found across sites. This finding suggests that local variation in insecticide use was an important cause of variation in susceptibility.     

灰飞虱对杀虫剂抗药性的研究进展

灰飞虱对杀虫剂产生抗药性是其近年来暴发频繁的重要原因。本文综述了国内外关于灰飞虱抗药性的研究成果,包括灰飞虱抗药性的发展、交互抗性、抗性机理、抗性遗传及生物适合度等。田间灰飞虱种群对多种药剂产生了不同程度的抗药性,其中对新烟碱类药剂吡虫啉和昆虫生长调节剂噻嗪酮产生了高水平到极高水平抗性(抗药性倍数分别为44.6～108.8倍和超过200倍),对有机磷类药剂毒死蜱和乙酰甲胺磷(抗药性倍数分别为10～12.6倍和9～13倍)、氨基甲酸酯类药剂甲萘威和残杀威(抗药性倍数分别为29.8～45.3倍和40.1～131.5倍)和拟除虫菊酯类药剂高效氯氰菊酯和溴氰菊酯(抗药性倍数分别为7.8～108.8倍和12～21倍)产生了中等水平到高水平的抗药性,对氟虫腈、阿维菌素和噻虫嗪没有产生抗药性(抗性倍数5倍)。长期大面积使用化学药剂是灰飞虱产生抗药性的重要原因。因此,必须加强灰飞虱的抗性治理,以延缓其抗药性进一步发展。     

Induction of transient ethylene and reduction in severity of tomato bacterial wilt by Pythium oligandrum

Pythium oligandrum (PO) is a mycoparasite on a wide range of fungi and suppresses diseases caused by fungal pathogens when colonizing the rhizosphere. PO and its cell wall proteins (CWPs) have elicitor activity that induces defence responses in plants. The potential of a mycelial homogenate of PO to suppress bacterial diseases was studied in roots of tomato ( Lycopersicon esculentum cv. Micro-Tom) inoculated with Ralstonia solanacearum . PO-treated plants showed enhanced resistance to R. solanacearum and reduction in severity of wilt symptoms. As ethylene often acts as one of the signal molecules for induced resistance, its production following treatment of tomato roots with the mycelial homogenate or CWP of PO was measured. The level of ethylene in PO- and CWP-treated plants was transiently elevated six- to 11-fold at 4–8 h after treatment, followed by high expression of three basic ethylene-inducible defence-related genes ( PR-2b , PR-3b and PR-5b ). Analysis of PR-5b gene expression in the leaves of PO- and CWP-treated plants suggested that PR gene expression was induced systemically. The expression of LeERF2 and LeETR4 , which confer an ethylene-dependent signalling pathway, was also significantly accelerated by such treatments. These results indicate that PO has the potential to control bacterial wilt disease and that CWP may play an important role in the induction of resistance to R. solanacearum accompanying the activation of the ethylene-dependent signalling pathway.     

A review of the interactions between multiple insecticide resistance loci

Insecticide resistance is an ever escalating problem worldwide in many pest populations and numerous cases of insecticide resistance are polygenic. Therefore, it is important to investigate the types of interactions that occur between insecticide resistance loci as this will dictate the level of resistance (and effectiveness of a chemical control strategy). Interactions also play a role in the evolution and/or maintenance of multigenic resistance in the field. Given that a limited number of mechanisms confer resistance, it might be possible to establish general rules for interactions between mechanisms. Several variables might dictate the type of interaction, such as the nature of the resistance mechanisms, genotype, etc. Interactions can be synergistic, antagonistic or additive. Based on this literature review, the most common interaction of multiple homozygous resistance loci is synergistic and additive when loci are heterozygous. When one locus is homozygous and the other locus is heterozygous the most common interaction was synergistic, although very few studies have examined this type of interaction. Possible factors that drive these interactions, exceptions to the trends, and future research needs are discussed.     

29个马铃薯品种(品系)抗晚疫病基因组成与抗病相关性分析

由致病疫霉引起的马铃薯晚疫病是马铃薯生产中最具危害性的病害,明确现有马铃薯品种(品系)中抗病基因组成情况对于指导抗病品种合理布局及抗病育种具有重要意义。本研究利用农杆菌介导的无毒基因瞬时表达技术分析了致病疫霉8个无毒基因在29个马铃薯品种(品系)中的分布情况。结果表明,品种间抗病基因组成各异,平均每个品种含有4个抗病基因。同时,不同抗病基因在品种中的分布频率也不同,抗病基因R1(76%)、Rpi-blb1(66%)和Rpi-blb2(66%)在品种中分布较普遍;而含抗病基因R3b(45%)、R4(38%)和Rpi-vnt1(34%)的品种较少。此外,品种中所含抗性基因的数量和品种室内抗性程度呈显著正相关(r=0.915 6,P=0.029 1)。     

Resistance evolution in Drosophila: the case of CYP6G1

The massive use of DDT as an insecticide between 1940 and 1970 has resulted in the emergence of a resistant population of insects. One of the main metabolic mechanisms developed by resistant insects involves detoxification enzymes such as cytochrome P450s. These enzymes can metabolise the insecticide to render it less toxic and facilitate its elimination from the organism. The P450 Cyp6g1 was identified as the major factor responsible for DDT resistance in Drosophila melanogaster field populations. In this article, we review the data available for this gene since it was associated with resistance in 2002. The knowledge gained on Cyp6g1 allows a better understanding of the evolution of insecticide resistance mechanisms and highlights the major role of transposable elements in evolutionary processes. © 2016 Society of Chemical Industry     

Insecticide resistance management: Relationship to integrated pest management

Insecticide resistance management (IRM) is a component of integrated pest management (IPM) that has the goal of forestalling resistance development to all insecticides. Since the advent of the organochlorine insecticides in the 1940s, an average of about one class of insecticide has been lost every 10 years because of resistance. Effective insecticides are necessary for optimum IPM and are too important and too expensive a resource to be lost so rapidly. By adhering to the principles of IPM and utilizing other IPM components such as biological and cultural control, IRM has the potential of conserving the susceptibility of pests to insecticides. Although initial attempts of IRM appear to be successful, it is imperative that research to undergird IRM be greatly accelerated.     

Characterization of mutations in the membrane-anchored subunits AaSDHC and AaSDHD of succinate dehydrogenase from Alternaria alternata isolates conferring field resistance to the fungicide boscalid

The molecular basis of resistance to the fungicide boscalid in 25 Alternaria alternata field mutants exhibiting resistance to boscalid and previously tested negative for AaSDHB mutations conferring boscalid resistance was investigated by cloning and sequencing the A. alternata SDHC ( AaSDHC ) and SDHD ( AaSDHD ) genes from a boscalid-sensitive isolate. The SDHB and SDHC/SDHD genes encode the iron sulphur and two membrane-anchored subunits of succinate:ubiquinone oxidoreductase (SQR) that constitute the boscalid fungicide molecular targets. The deduced amino acid sequences exhibited low similarities with SDHC and SDHD peptides from other organisms, but residues essential to form the ubiquinone binding site or important in SQR assembly were particularly conserved. Sequence comparisons of the AaSDHC and AaSDHD genes between resistant mutants and wild-type isolates revealed that two highly conserved histidine residues implicated in the heme b ligation and located at codon 134 in AaSDHC (22 mutants) and codon 133 in AaSDHD (two mutants) were replaced by arginine residues (H134R and H133R). In another mutant, a substitution of an aspartate by a glutamic acid occurred at amino acid position 123 (D123E) in AaSDHD. Additional tests revealed that mycelial growth of boscalid-resistant isolates was reduced when isolates were subjected to oxidative stress. The identified mutations were confirmed using PCR-RFLP assays. This is apparently the first report of mutations located in the heme b ligands of the cytochrome II gene associated with carboxamide resistance.