USING FILTER PAPER TEST FOR DETECTING INHIBITORY FREQUENCY OF ORGANOPHOSPHATE TO ESTERASES IN COTTON APHIDS, APHIS GOSSYPII (GLOVER)

Abstract The individual esterase activity which is measured by filter paper test (FPT) method may determine the resistance of cotton aphids Aphis gossypii (Glover) against organophosphorus (OP) insecticides. For testing accurately resistant level caused by different insecticides, we applied FPT method for measuring inhibitory action of methyl-parathion, monocrotophos and omethoate to α-NA esterase of individual cotton aphids, and compared the inhibitory frequencies of these three insecticides to susceptible population (BCA) and resistant population (GCA). Results showed that their inhibitory frequencies of the susceptible population were evidently higher than that of the resistant population. The inhibitory rate of α-NA esterase in F1 generation individual cotton aphids by monocrotophos was low when the cotton aphid population had been treated in advance with monocrotophos, but it got to 75%-90% when the cotton aphid population had been untreated in advance with monocrotophos. Besides, the differences in esterase activity were not obvious between them. In same region when cotton aphids were treated with insecticides the inhibitory frequency of esterases in individual by the insecticides was lower than counterparts in individual cotton aphids which were not treated with insecticides. All these demonstrated that inhibitory frequency of α-NA esterase in individual cotton aphids by OP insecticides could be used as a technique of forecasting pest resistance.     

Electrophoretic studies on populations of Myzjus persicae in Scotland from March to July, 1976

Specimens of Myzus persicae have been collected from field populations in Scotland and scored electrophoretically for their phenotypes at three polymorphic loci. These phenotypes were almost invariably present in individuals in one of a small number of favoured combinations, suggesting that these populations may be oligogeneous, that is, composed of large numbers of copies of a few favoured combinations of alleles. Temporal and spatial sampling at sites in East and West Scotland in 1976 has shown that different allele combination types were present in the two areas. Populations in E. Scotland contained very few specimens belonging to the organophosphorus (OP) resistant clone types (using esterase activity to infer resistance); instead there were large numbers of an OP susceptible type which has not been detected in England, but which came to predominate on cabbage and potato host plants at every site sampled during the period of rapid population increase. In contrast, populations in W. Scotland were characterised by substantial numbers of specimens belonging to OP resistant clone types and by an almost complete absence of this susceptible type. Two other distinguishable OP susceptible types were found to be common to both areas. The W. Scotland populations closely resemble N. England populations while those in E. Scotland appear to be unique to that area. Specimens of OP resistant types were only detected in E. Scotland after the beginning of June and in such relative proportions as to strongly suggest that they were migrants from W. Scotland or N. England.     

Amplification and methylation of an esterase gene associated with insecticide-resistance in greenbugs, Schizaphis graminum (Rondani) (Homoptera: Aphididae)

The greenbug aphid, Schizaphis graminum (Rondani) has developed resistance to organophosphorus insecticides by the over-production of esterases that have been classified as Type I and Type II. The first twenty N-terminal amino acids of the Type I esterase were determined and used to design an oligonucleotide, which in conjunction with an active site primer derived from conserved sequences of other insect esterases and two internal primers specific for esterases from another aphid species resulted in a 0.85 kb genomic DNA fragment from resistant greenbugs. This was extended by 5′ RACE which provided approximately 1.2 kb of the 5′ end of the esterase gene. The 5′ DNA sequence corresponded to 19 of the 20 known amino acids of the Type I esterase, with the last needing only a one base change (probably resulting from a PCR artifact). Furthermore, the sequence showed very close similarity to the amplified E4/FE4 esterase genes of Myzus persicae (Sulzer). A comparison of sequences suggested that the S. graminum gene has introns in the same positions as the first two introns of E4/FE4, with the second intron being considerably larger in S. graminum. Probing of Southern blots with the 0.85 kb esterase fragment showed that the gene encoding the Type I esterase is amplified 4- to 8-fold in resistant S. graminum and that the amplified sequences contain 5-methylcytosine at MspI/HpaII sites, again in agreement with previous findings for M. persicae genes.     

A naturally occurring chromosomal translocation in Myzus persicae (Sulzer)

Populations of M.persicae from glasshouses in Britain were found to have a structurally heterozygous chromosome complement. Biometric data from somatic metaphase chromosomes, and the pairing configurations of chromosomes at spermatocyte meiosis, indicate either a simple or a reciprocal translocation involving autosomes 1 and 3. All except one of the clones started from these populations showed high levels of esterase activity indicative of resistance to organophosphorus insecticides. The British translocated clones were all androcyclic in reproductive character, but of several distinct genotypes. A similar or identical translocation was found in seven out of nine clones started from M.persicae collected in the field in Japan. The translocation was present in Japanese M.persicae of all reproductive categories, being somehow maintained in the sexual breeding system of the aphid. In breeding experiments, translocation heterozygosity was inherited independently from colour and reproductive characters. The translocation was also found in a sample of M.persicae from southern California.     

Aphid populations showing differential levels of virulence on Capsicum accessions

The green peach aphid,Myzus persicae,is one of the most threatening pests in pepper cultivation and growers would benefit from resistant varietices.Previously,we identified two Capsicum acessions as susceptible and three as resistant to M.persicae using an aphid population originating from the Netherlands(NL).Later on we identified an aphid population originating from a diferent gcographical region(Switserland,SW)that was virulent on all tested Capsicum acessions.The objeetive of the current work is to describe in detail diferent aspects of the interaction between two aphid populations and two sclected Capsicum acessions(one that was susceptible[PB2013046]and one that was resistant[PB2013071]to population NL),including biochemical processes involved.Electrical penetration graph(EPG)recordings showed similar feeding activities for both aphid populations on PB2013046.On acession PB2013071 the aphid population sw was able to devote significantly more time to phloem ingestion than population NL.We also studied plant defense response and found that plants of acession PB2013046 could not induce an accumulation of reactive oxygen species and callose formation after infestation with either aphid population.However,plants of PB2013071 induced a stronger defense response after infestation by population NL than after infestation by population SW.Based on these results,population SW of M.persicae seems to have overcome the resistance of PB2013071 that prevented feeding of aphids from NL population.The potential mechanism by which SW population overcomes the resistance is discussed.     

Evaluating reproductive fitness and metabolic costs for insecticide resistance in Myzus persicae from Chile

The development of insecticide resistance in pest insects is an increasing problem for agriculture, forestry and public health. Aphids are ubiquitous herbivorous insects, with approximately 4700 known species, of which less than 5% exploit the agricultural environment successfully. Of these, the peach‐potato aphid Myzus persicae Sulzer is recognized as one of the most important pests worldwide because it has acquired resistance to many insecticides. Although resistance to insecticides provides important benefits for pests in agricultural fields that are treated with insecticides, it may be associated with fitness (or other) costs in environments that are insecticide free. In the present study, the fitness and energy costs that might be experienced by M. persicae in an insecticide‐free environment when carrying at least one insecticide resistance mutation (IRM), or by having an increased production of esterases, are evaluated. The study investigates whether genotypes that have an IRM also have enhanced esterase production, whether there is any metabolic cost associated with insecticide resistance, and whether there are any fitness costs associated with insecticide resistance and metabolic expenditure. The intrinsic rate of increase, standard metabolic rate (i.e. a measure of maintenance costs) and constitutive esterase activity are determined for 30 different multilocus genotypes carrying (or not carrying) at least one of the two most frequent insecticide resistance mutations (MACE and kdr/super‐kdr) that occur in Chile. The results show that genotypes carrying at least one IRM have higher levels of total esterase activity than genotypes without an IRM, that there is no evidence of an energy cost associated with total esterase activity or IRM, and no evidence for a reproductive fitness cost associated with total esterase activity, IRM or metabolic rate. The results agree with previous studies showing linkage disequilibrium between insecticide resistance mechanisms, although they contrast with those of studies that report fitness costs associated with insecticide resistance in Myzus persicae.     

Co‐occurrence of defensive traits in the potato aphid Macrosiphum euphorbiae

1. Insecticide usage selects strongly for resistance in aphid populations, but this could entail fitness costs in other resistance traits. The potato aphid Macrosiphum euphorbiae Thomas exhibits intraspecific variation in susceptibility to parasitism by braconid wasps and provides a suitable species to study the relation between the defensive traits of parasitism and insecticide resistance. 2. Clonal lines (23 in total) of M. euphorbiae were established from aphids collected in 2013 from geographically separate populations in the U.K. Clonal lines belonged to five aphid genotypes, but one genotype predominated (78% of samples), and the facultative endosymbiont Hamiltonella defensa was detected in c. 40% of lines. 3. Total esterase activity in aphid tissues varied significantly between aphid genotypes and collection areas, but there was no clear pattern in relation to H. defensa infection or between collection sites likely to differ in insecticide pressures. 4. Five clonal lines representing low to moderate levels of enzyme activity, which included different aphid genotypes and presence/absence of H. defensa infection, were assayed for their susceptibility to the parasitoid wasp Aphidius ervi Haliday. Aphid mummification varied significantly between aphid genotypes, with low values in one genotype of aphids irrespective of H. defensa presence. 5. The results revealed that aphid lines belonging to the parasitism‐resistant genotype exhibited moderate levels of total esterase activity, indicating a competitve advantage for this genotype of M. euphorbiae when exposed to chemical and biological control factors in agroecosystems.     

抗性品系棉蚜乙酰胆碱酯酶和羧酸酯酶的变异

用浸叶法测定了采自我国不同地区(泰安、莱阳、南京、北京和安阳)的棉蚜品系Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ对久效磷、甲胺磷、抗蚜威和灭多威等杀虫剂的抗性水平,各棉蚜品系对杀虫剂的抗性依次为Ⅴ>Ⅳ>Ⅲ,Ⅱ>Ⅰ。进一步研究表明,Ⅴ和Ⅳ品系棉蚜乙酰胆碱酯酶对杀虫剂的敏感性显著下降,Ⅱ品系次之,Ⅲ和Ⅰ品系接近于敏感品系。Ⅴ和Ⅳ品系乙酰胆碱酯酶的Km值显著下降,表明酶发生了质的变化。不同棉蚜抗性品系的酯酶（全酯酶和羧酸酯酶）活性均显著升高,其中Ⅲ品系的酯酶活力为Ⅱ品系的2倍。Ⅴ品系羧酸酯酶Km值达2460.4 μmol/L,而Ⅳ品系仅为84.4 μmol/L,该两个品系羧酸酯酶发生了质的变化。研究结果表明,不同抗性程度的棉蚜品系均存在代谢抗性和靶标抗性。低抗水平的棉蚜品系,以代谢抗性为主,靶标抗性为辅;中抗水平的棉蚜品系,抑或由于解毒代谢酶的活性显著增强,也可能由于靶标的敏感性显著下降;而高抗水平的棉蚜品系,依赖于代谢抗性和靶标抗性的联合作用。     

Blockage of stylet tips as the mechanism of resistance to virus transmission by Aphis gossypii in melon lines bearing the Vat gene

Aphis gossypii is the main virus vector in muskmelon crops. The melon gene Vat confers resistance to non‐persistent virus transmission by this aphid. The mechanism of this resistance is not well understood, but no relationship has been detected between resistance and the probing behaviour of aphids on resistant plants. Results presented here suggest that temporary blockage of aphid stylet tips preventing virus particle release may explain the resistance conferred by Vat gene. We performed experiments in which viruliferous aphids were allowed to probe different sequences of resistant (Vat‐bearing) and/or susceptible melon plants. The results demonstrated that A. gossypii inoculates Cucumber mosaic virus (CMV) efficiently in susceptible plants having previously probed resistant plants, showing that the resistance mechanism is reversible. Furthermore, the infection rate obtained for susceptible plants was the same (25%) regardless of whether the transmitting aphid had come directly from the CMV source or had subsequently probed on resistant plants. This result suggests that virus is not lost from stylet to plant during probing of resistant plants, supporting the temporary blockage hypothesis. We also found that the ability of Myzus persicae to transmit CMV is noticeably reduced after probing on resistant plants, providing evidence that this aphid species also responds to the presence of the Vat gene. Finally, we also found that in probes immediately after virus acquisition M. persicae inoculates resistant plants with CMV more efficiently than susceptible plants, perhaps because the Vat gene product induces increased salivation by this aphid.     

Intraspecific variation of Myzus persicae on sugar beet (Beta vulgaris)

Differences in inherited resistance among seven sugar-beet stocks had similar effects on Myzus persicae clones representing the range of variation in aphid response to resistant and susceptible sugar beet observed in fifty-eight clones collected between 1969 and 1971. Three sugar-beet stocks were consistently resistant. Statistically significant interactions between beet stocks and aphid clones did not indicate the existence of biotypes with specific abilities to overcome resistance. M. persicae clones differed in their vigour of colonizing sugar beet, irrespective of the differences between beet stocks. The readiness of adult aphids to settle determined the size of aphid population produced and included a component related to the response of the aphid clone to sugar beet as a host, and a component related to the resistance ranking of the beet stock. Breeding sugar beet with resistance to aphids will be simplified, as the results indicate that, at present, differences between aphid biotypes need not be considered a problem.     

Feeding behavior of Myzus persicae on asparagus species susceptible and resistant to Asparagus virus 1

Asparagus virus 1 (AV‐1) infects Asparagus officinalis L. (Asparagaceae) in the field worldwide. However, various wild relatives of A. officinalis are resistant to AV‐1. Here we study the behavior of the green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae), on 19 AV‐1‐resistant wild relatives of A. officinalis. We focus on behavior that is associated with regular cell penetration, relevant for inoculation of AV‐1, and sieve element penetration to check for vector resistance and its potential influence on AV‐1 transmission. Parameters, relevant for the transmission of non‐persistent viruses and host plant acceptance, were obtained by the electrical penetration graph technique. Furthermore, phylloclade architecture of A. officinalis and its wild relatives was examined to study its influence on aphid behavior. Behavior of M. persicae displays many cell penetrations and long ingestion periods on A. officinalis, compared to the generally shorter cell penetrations (reduced potential for virus transmission) and reduced or no ingestion (phloem‐located aphid resistance) on wild relatives. Because effects on aphid behavior are not consistent throughout the group of the tested wild relatives of A. officinalis, with some wild relatives being susceptible to M. persicae, a common genetic background for AV‐1 and aphid resistance appears to be unlikely. However, the reduced potential of virus transmission as well as aphid resistance shown by some wild relatives may be useful for future breeding programs.     

淡色库蚊酯酶等位基因及其在自然种群中的频率分布

酯酶基因扩增所产生的酯酶活性升高是库蚊Culex pipiens对有机磷杀虫剂抗性的主要机理之一。采用分子杂交技术和限制性酶切片段长度多态性(RFLP)分析,已鉴定出多种酯酶等位基因类型。该文通过酯酶基因特异性片段的PCR扩增及扩增片段的酶切片段分析,对淡色库蚊Culex pipiens pallens四种有机磷抗性品系的酯酶等位基因进行分型,并测定分析自然种群中不同酶型的频率分布。研究结果表明：PCR分型方法具有快速、准确的特点。不同的有机磷杀虫剂对酯酶等位基因具有明显的选择作用。双硫磷品系为B₁型;毒死蜱和敌百虫品系为B₂型;马拉硫磷品系为B₁型和B₁/B₂杂合型。不同地区采集的种群表现出不同的酶型频率分布。该文就杀虫剂对酯酶等位基因选择作用及自然种群的酶型频率分布进行了讨论。     

Genetic, biochemical, and behavioral uniformity among populations of Myzus nicotianae and Myzus persicae

Prior to designation as distinct species, an appellation presently in question, the tobacco aphid, Myzus nicotianae Blackman (Homoptera: Aphididae), was classified as a tobacco-feeding form of the green peach aphid, Myzus persicae (Sulzer). In this study, RAPD polymorphisms distinguished members of the Myzus persicae complex (M. persicae and M. nicotianae) from three outgroup Myzus species (M. cerasi (F.), M. hemerocallis Takahashi, and M. varians Davidson). Polymorphisms within the complex did not separate populations on the basis of host association (tobacco versus other host plants) or geographic origin (collections from the United States, Europe, and Japan). Similarly, while GC-MS analysis of cuticular hydrocarbon profiles revealed both developmental and inter-populational differences within the M. persicae complex, it did not separate populations of tobacco feeding aphids from those collected off non-tobacco hosts. Finally, with the exception of their responses to a choice between lettuce and collards, the host preference behavior of a green peach aphid population, a red tobacco aphid population, and a green tobacco aphid population was indistinguishable in host preference experiments. These results add to a growing body of evidence suggesting M. nicotianae and M. persicae are conspecific.     

Aspects of insecticide resistance in the melon and cotton aphid, Aphis gossypii (Hemiptera: Aphididae)

Strains of the aphid Aphis gossypii, taken from cucumber in UK glasshouses were tested for resistance to pirimicarb, diazinon and heptenophos. Compared to a susceptible strain, resistance factors to diazinon of up to 659 were found in populations collected between 1987 and 1989. Compared to the same susceptible strain, diazinon-resistant populations also showed a reduced response to heptenophos. All field populations tested against pirimicarb were resistant. Host plant preference tests showed that all strains (including stock cultures) tested could readily breed on cotton. However strains originally collected from chrysanthemum did not transfer to cucumber and likewise strains from cucumber would not colonise chrysanthemum. Polyacrylamide gel electrophoresis of four of the aphid strains showed differences in migration rates and intensity of staining of esterase bands. Intensity of staining was correlated with the degree of diazinon resistance.     

Glasshouse tests and field selection for heritable resistance to Myzus persicae in sugar beet

A technique is described to assess and select in the field for resistance to Myzus persicae in sugar beet. Lines developed by this method were aphid resistant when compared with commercial cultivars. The resistance was inherited by progeny lines. Variation between beet lines in glasshouse tests suggested segregation of a number of resistance genes with varying effects. Integration of selection for aphid-resistance and agronomic performance during beet breeding is discussed.     

Possible roles of esterase, glutathione S-transferase, and superoxide dismutase activities in understanding aphid–cereal interactions

Activities of the detoxification enzymes esterase, glutathione S‐transferase, and of superoxide dismutase in aphids and aphid‐infested cereal leaves were assayed using polyacrylamide gel electrophoresis and a spectrophotometer to elucidate the enzymatic mechanisms of aphid resistance in cereal plants. A chlorosis‐eliciting Russian wheat aphid, Diuraphis noxia (Mordvilko), and non‐chlorosis‐eliciting bird cherry‐oat aphid, Rhopalosiphum padi (L.), and four cereals were used in this study. The four cereal genotypes were ‘Arapahoe’ (susceptible) and ‘Halt’ (resistant) wheat (Triticum aestivum L.), ‘Morex’ (susceptible) barley (Hordeum vulgare L.), and ‘Border’ (resistant) oat (Avena sativa L.). Esterase isozymes differed between the two aphid species, although glutathione S‐transferase and superoxide dismutase did not. Esterase, glutathione S‐transferase, and superoxide dismutase activities in either aphid species were not affected by the level of resistance of a cereal to D. noxia. The assays of cereal leaf samples showed that D. noxia feeding elicited an increase in esterase activity in all four cereal genotypes, although R. padi feeding did not. The increase of esterase activity in cereals, however, was not correlated to aphid resistance in the cereals. The time‐series assays of aphid‐infested cereal leaves showed that D. noxia‐infested Morex barley had a significant increase in esterase activity on all sampling dates (3, 6, and 9 days) in comparison with either uninfested or R. padi‐infested barley. No difference in glutathione S‐transferase activity was detected among either aphid infestations or sampling dates. The electrophoretic assays, however, revealed that aphid feeding elicited a significant increase in superoxide dismutase activity, which served as the control of glutathione S‐transferase activity assays. The increase in esterase and superoxide dismutase activities suggested that D. noxia feeding imposes not only toxic, but also oxidative stresses on the cereals. The ramification of using these enzyme activity data to understand the etiology of D. noxia‐elicited chlorosis is discussed.     

杀虫剂及不同地区寄主植物对棉蚜酯酶的影响

通过滤纸酯酶反应法比较 4种有机磷杀虫剂对北京和山东高密地区取食不同寄主植物的棉蚜β-乙酸萘酯 (β -NA)酯酶的抑制作用。结果表明高密地区不同寄主植物上的棉蚜 β -NA酯酶活力高的个体多于北京地区 ,同时辛硫磷对高密地区不同寄主植物上的棉蚜 β -NA酯酶的抑制作用亦高于北京地区 ,而久效磷和DDVP对两地区不同寄主植物上的棉蚜种群中 β-NA酯酶的抑制作用相似。     

Laboratory selection for resistance to sulfoxaflor and fitness costs in the green peach aphid Myzus persicae

Sulfoxaflor is a newly released fourth-generation neonicotinoid insecticide for management of sap-feeding pests that have developed resistance to established insecticide groups. The risk of resistance developing to this pesticide in target pests is unclear. We selected a strain of the green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae), for resistance to sulfoxaflor in the laboratory, which showed 199-fold resistance after 45 generations compared to the starting population. Life table analysis showed that the resistant strain had a fitness of 0.83 compared to the susceptible strain. Adult longevity of the resistant strain was reduced by 9.55% compared to the susceptible strain. The period when adults of the resistant strain produced offspring was reduced by 17.19%, while the mean fecundity of the resistant strain was reduced by 15%. These findings suggest that M. persicae can develop a high level of resistance to sulfoxaflor, but fitness costs may result in a recovery of sensitivity when field populations are no longer exposed to sulfoxaflor.     

Abscisic acid deficiency increases defence responses against Myzus persicae in Arabidopsis

Comparison of Arabidopsis thaliana (Arabidopsis) gene expression induced by Myzus persicae (green peach aphid) feeding, aphid saliva infiltration and abscisic acid (ABA) treatment showed a significant positive correlation. In particular, ABA‐regulated genes are over‐represented among genes that are induced by M. persicae saliva infiltration into Arabidopsis leaves. This suggests that the induction of ABA‐related gene expression could be an important component of the Arabidopsis–aphid interaction. Consistent with this hypothesis, M. persicae populations induced ABA production in wild‐type plants. Furthermore, aphid populations were smaller on Arabidopsis aba1‐1 mutants, which cannot synthesize ABA, and showed a significant preference for wild‐type plants compared with the mutant. Total free amino acids, which play an important role in aphid nutrition, were not altered in the aba1‐1 mutant line, but the levels of isoleucine (Ile) and tryptophan (Trp) were differentially affected by aphids in wild‐type and mutant plants. Recently, indole glucosinolates have been shown to promote aphid resistance in Arabidopsis. In this study, 4‐methoxyindol‐3‐ylmethylglucosinolate was more abundant in the aba1‐1 mutant than in wild‐type Arabidopsis, suggesting that the induction of ABA signals that decrease the accumulation of defence compounds may be beneficial for aphids.     

Antibiotic resistance in potato cultivars to the aphid Myzus persicae

Commercially-grown potato cultivars were assessed for antibiotic resistance to Myzus persicae in a culture room at 19°C with a 2°C range and a 16 h photoperiod. The resistant rankings obtained were consistent for the following aphid metrics: maturation time, teneral adult weight, embryo complement and the production of nymphs. Intrinsic rates of increase (r_m) correlated with the above parameters of aphid growth and reproduction but this was not always the case for nymphal survival rate. The cultivars Ulster Tarn, Record and Maris Piper were relatively resistant to M. persicae and cultivars Desirée and King Edward were the most susceptible. When the aphid clone or the culture plants were changed there was no significant effect on the relative resistance of the potato cultivars under test. The relationship between the number of embryos in teneral adults and the production of nymphs by aphids was examined by grouping aphids in weight batches. There was a strong relationship between these two variables.