Early detection of resistance to tebufenozide in field populations of Cydia pomonella L.: methods and mechanisms

Abstract: Four populations of codling moth Cydia pomonella L. were collected as overwintering larvae from apple orchards with different pesticide pressure (S. Michele, Roncafort, Revò and Vervò) in the Trento province (northern Italy). Mortality rate caused by a predetermined discriminating concentration of tebufenozide topically applied on overwintering larvae was evaluated. Neonate F1 progeny of the same populations were assayed for susceptibility to tebufenozide by feeding them on thinning apples treated with an appropriate discriminating dose of the insecticide. The activities of the main enzyme systems involved in the detoxification of insecticides were also evaluated in each population and related to their susceptibility to tebufenozide. The topical test detected a significant loss in susceptibility to tebufenozide in two populations, S. Michele and Roncafort, while all the overwintering larvae collected in the orchards of Revò and Vervò died when treated topically with the discriminating concentration. The apple‐dipping test performed on the neonate larvae showed a highly significant reduction in the susceptibility of the two populations of S. Michele and Roncafort. A less significant reduction in mortality rate was found in the Revò population; however, no statistical difference was found between the Vervò population and the susceptible reference. None of the four field populations significantly differed from the susceptible strain for Glutathione‐S‐transferase and esterase activity. A significantly higher frequency of individuals of the S. Michele and Roncafort populations exhibited a higher mixed function oxidase activity than the susceptible strain. The small resistance ratio values found for the two populations together with the low frequency of individuals exibiting enhanced enzymatic activity, reveals that the selection process was still at the early stage. Because of its efficiency in early detection of resistance to tebufenozide, topical application on diapausing larvae can thus be considered an appropriate, simple and robust tool for implementing resistance monitoring programmes for tebufenozide.     

Pro-active management of beet armyworm (Lepidoptera: Noctuidae) resistance to tebufenozide and methoxyfenozide: baseline monitoring,risk assessment,and isolation of resistance

Susceptibility to tebufenozide and methoxyfenozide of beet armyworm [Spodoptera exigua (Hübner)] from the southern United States and Thailand was determined through exposure of first and third instars to dipped cotton leaves. Among the field populations evaluated, tebufenozide LC50 values for first and third instars, respectively, ranged from 0.377 to 4.41 and 4.37-46.6 microg (AI) /ml of solution. Methoxyfenozide LC50 values for first and third instars of field populations ranged from 0.058 to 0.487 and 0.601-3.83 microg (AI)/ml of solution. A Thailand field strain exhibiting reduced susceptibility to both compounds was subjected to intense laboratory selection for three nonconsecutive generations. At the LC50 and LC90, selected Thailand strains were 45-68 times and 150-1,500 times less susceptible to tebufenozide and 340-320 times and 120-67 times less susceptible to methoxyfenozide as first and third instars, respectively, when compared with the laboratory reference strain. Among the U.S. field populations evaluated, ones from Belle Glade, FL, and Florence, SC, were generally the most susceptible and ones from Maricopa and Parker, AZ, were the least susceptible. Selection of the Thailand field strain with tebufenozide reduced susceptibility to both compounds, and selection of Thailand strains previously pressured with either compound further reduced susceptibility to both, suggesting at least some commonality of resistance mechanism. Characterization of this resistance will provide information that will be helpful for pro-active management of resistance for this valuable group of insecticides.     

Baseline monitoring of codling moth (Lepidoptera: Tortricidae) larval response to benzoylhydrazine insecticides

A diet-incorporation larval bioassay was developed to measure the response of codling moth, Cydia pomonella (L.), to the benzoylhydrazine insecticides tebufenozide and methoxyfenozide. The bioassay tested neonates and third, fourth, and fifth instars from a laboratory colony and neonates and fourth instars from a pooled population collected from five certified-organic apple orchards. Bioassays were scored after 6 and 14 d. No differences between the laboratory and field population were found for either insecticide. Significant differences were found in the response of third and fifth instars between the 6 and 14 d bioassays, primarily due to a high proportion of moribund larvae in the shorter assay. Larval age had a significant effect in bioassays and was more pronounced in 6- versus 14-d tests. Fifth instars were significantly less susceptible to both insecticides than other stages, while responses of third and fourth instars were similar. The response of neonates was significantly different from third and fourth instars to tebufenozide but not with methoxyfenozide in the 14-d test. Field bioassays excluded the use of fifth instars and were scored after 14 d. LC50s estimated for 18 field-collected populations varied five- and ninefold for tebufenozide and methoxyfenozide, respectively. The responses of all but six field-collected populations were significantly different from the laboratory strain. Five of these six populations were collected from orchards with no history of organophosphate insecticide use. The LC50 for methoxyfenozide of one field-collected population reared in the laboratory for three generations declined fourfold, but was still significantly different from the laboratory population. These data suggest that transforming current codling moth management programs in Washington from a reliance on organophosphate insecticides to benzoylhydrazines may be difficult.     

Baseline susceptibility of Planococcus ficus (Hemiptera: Pseudococcidae) from California to select insecticides

Between 2006 and 2008, 20 populations of Planococcus ficus (Signoret), from Coachella and San Joaquin Valleys of California were measured in the laboratory for susceptibility to buprofezin, chlorpyrifos, dimethoate, methomyl, and imidacloprid. Toxicity was assessed using a petri dish bioassay technique for contact insecticides and by a systemic uptake technique for imidacloprid. Mixed life stages were tested for susceptibility to all insecticides except for buprofezin, which was measured against early and late instars (first, second, and third). Dose-response regression lines from the mortality data established LC50 and LC99 values by both techniques. Responses of populations from the two geographical locations to all five insecticides varied, in some cases significantly. Variations in susceptibility to each insecticide among sample sites showed a sevenfold difference for buprofezin, 11-fold to chlorpyrifos, ninefold to dimethoate, 24-fold to methomyl, and 8.5-fold to imidacloprid. In spite of susceptibility differences between populations, baseline toxicity data revealed that all five insecticides were quite effective based on low LC50s. Chlorpyrifos was the most toxic compound to Planococcus ficus populations as shown by lowest LC50s. Buprofezin was toxic to all immature stages but was more potent to first instars. The highest LC99 estimated by probit analysis of the bioassay data of all 20 populations for each compound was selected as a candidate discriminating dose for use in future resistance monitoring efforts. Establishment of baseline data and development of resistance monitoring tools such as bioassay methods and discriminating doses are essential elements of a sustainable management program for Planococcus ficus.     

Allele frequency of resistance to Bacillus thuringiensis Cry1ab corn in Louisiana populations of sugarcane borer (Lepidoptera: Crambidae)

Transgenic Bacillus thuringiensis (Bt) corn, Zea mays L., has been widely used to manage a corn borer complex in the mid-southern region of the United States. The sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), has become a dominant cornstalk boring species in some areas of this region, especially in Louisiana. Therefore, management of sugarcane borer resistance to Bt corn is critical to ensure the long-term sustainability of Bt corn for the region. This study screened 280 two-parent family-lines of sugarcane borer from four geographical populations in Louisiana during 2005 to determine whether Bt resistance allele frequency in sugarcane borer is sufficiently low to meet the rare resistance assumption of the current "high dose/refuge" resistance management strategy for Bt corn. These sugarcane borer family-lines were examined for Bt resistance by using novel F2 screening procedures. No major Bt resistance alleles were detected in these four populations. The estimated frequency of major Bt resistance alleles was < 0.0027, with a 95% probability and a detection power of 94%. The estimated minor resistance allele frequency was 0.0063, with a 95% CI of 0.0025-0.0117. During a previous study, a major Bt resistance allele was detected in one individual from 213 family-lines of another Louisiana population of sugarcane borer. Combining these data with the current screen, the frequency of major Bt resistance alleles across the five populations was 0.001, with a 95% credibility interval of 0.0001-0.0028 and a detection power of 95%. Major Bt resistance allele frequencies in Louisiana sugarcane borer populations seem to be low, and they should support the rare resistance allele requirement of the high dose/refuge strategy.     

Resistance and cross-resistance to four insecticides in populations of obliquebanded leafroller (Lepidoptera: Tortricidae)

Populations of obliquebanded leafroller, Choristoneura rosaceana (Harris), were collected from organic and conventionally managed orchards located in the Okanagan and Similkameen Valleys of British Columbia Neonate F1 progeny were assayed for resistance to azinphosmethyl, tebufenozide, methoxyfenozide, and indoxacarb using a leaf disk bioassay. Significant differences in resistance levels among populations were observed for all four insecticides. Insects collected from organic sites were more susceptible to all insecticides than were insects collected from conventional sites. Resistance to the benzoylhydrazine insect growth regulators tebufenozide and methoxyfenozide was highly correlated with resistance to azinphosmethyl across populations, indicating cross-resistance between these compounds. The highest levels of resistance were observed with indoxacarb, but resistance levels to indoxacarb did not correlate with those for azinphosmethyl. Dose-response regression lines for tebufenozide were parallel across populations, suggesting that the resistance mechanism(s) were quantitatively, but not qualitatively, different. Cross-resistance between azinphosmethyl and benzoylhydrazine insecticides indicates that a resistance management strategy for obliquebanded leafroller involving the rotation of these materials is not likely to be successful.     

Resistance Monitoring and Enzyme Activity in Field-collected Populations of the Spiraea Aphid,Aphis citricola van der Goot

Susceptibility of the spiraea aphid (Aphis citricola van der Goot) to several classes of insecticides revealed regional variation. Four field-collected A. citricola populations have developed resistance to several classes of insecticides, including pyrethroids, organophosphates, and carbamates. Of the 15 insecticides tested four insecticides (deltamethrin, clopyrifos, omethoate and carbaryl) showed the relative resistance ratio (RRR) values of 10 or above, against four field-collected populations, whereas the other 11 insecticides had RRR values above 2. There were no significant differences in esterase activities among the field-collected populations. Esterase activity was generally homogeneous in the Yeasan-collected population, but was more heterogeneous in the populations collected from Andong, Kunwi and Suwon. Glutathione S-transferase (GST) activity in the Andong-and Yeasan-collected populations was higher than that in the other populations. More heterogeneous pattern of esterase activity and higher activity of GST in field populations of A. citricola would be due to multiple resistance mechanism. Fortunately, the levels of resistance in four field-collected populations examined are still increasing. Therefore, careful selective use of insecticides through both continued resistance monitoring and resistance mechanism study can be warranted to extend the effective life of insecticides for controlling A. citricola that was already resistant to various insecticides.     

Indirect selection for increased susceptibility to permethrin in diamondback moth (Lepidoptera: Plutellidae)

We have been exploring the behavioral response ot insect pests to heterogeneous distribution of toxins (low dose with refugia), and its genetic correlation with physiological tolerance to these toxins. A field-collected population of diamondback moth, Plutella xylostellu (L.) (Lepidoptera Plutellidae), from Celeryville, OH, was selected with permethrin to determine whether low heterogeneous doses could lead to increased susceptibility to permethrin by selecting indirectly on behavior. Two replicates of each of three selection regimes: uniform high concentration hypothesized to result in increased physiological tolerance, heterogeneous low concentration hypothesized to result in increased susceptibility through indirect selection on behavior, and a control with no exposure to permethrin, were maintained in 1-in3 cages in a greenhouse, for 33 generations. All life stages of the diamondback moth were exposed to the selection regimes, and new generations were started with a random selection of pupae from the previous generation. Lines selected with uniform high concentrations developed 76-fold levels of resistance to permethrin by the 17th generation, with little changes thereafter. For generations 1-20, lines selected with heterogeneous low concentrations remained slightly lower in LC50 but not significantly different from the unselected control lines. Based on confidence intervals from probit analyses, the LC50 of the lines selected with heterogeneous low concentration, however, were significantly lower than those of the control lines in generations 21-33. Our results demonstrate that selection on behavioral responses can result in greater susceptibility than no selection at all, despite exposure to the toxin and ample genetic variation and potential for increased physiological tolerance. The implications of our findings, which are based on selection scenarios that could take place in field situations, are that behavioral responses can prevent and even decrease the levels of resistance in insect populations, an important result with respect to resistance and resistance management.     

Resistance to hexythiazox in Brevipalpus phoenicis (Acari: Tenuipalpidae) from Brazilian citrus

The objective of this study was to collect baseline information for implementing an acaricide resistance management program of Brevipalpus phoenicis (Geijskes) to hexythiazox in Brazilian citrus groves. The egg susceptibility of B. phoenicis to hexythiazox was measured by a direct contact bioassay. The estimated LC50 for the S strain was 0.89 mg hexythiazox L(-1) of water (95% FL 0.75-1.03). After hexythiazox selection of a field-collected population associated with intense hexythiazox use, a resistance ratio greater than 10,000-fold was detected. Results from a survey revealed a great variability in the frequency of resistance in populations of B. phoenicis collected from citrus groves located in the State of S?o Paulo. No relationship was observed between the intensity of hexythiazox use and the frequency of resistance. Studies on dynamics of resistance showed that the resistance of B. phoenicis to hexythiazox is stable under laboratory conditions. Therefore, there is an urgent need to implement resistance management of B. phoenicis to hexythiazox in order to prolong its effective use in Brazilian citrus groves.     

Selection, egg viability, and fecundity of the sugarcane borer (Lepidoptera: Crambidae) with tebufenozide.

Two separate attempts to select the sugarcane borer, Diatraea saccharalis (F.), for resistance to tebufenozide were unsuccessful. Both selected colonies were lost after the fourth generation due to a lack of oviposition. Differences were not detected in fecundity or percent egg viability for 5-d-old third instars exposed to concentrations (EC5, EC15, and EC30) of tebufenozide for 7 d. Decreases (P < or = 0.01) in mean female pupal weights were detected in larvae exposed to EC15 and EC30 concentrations. An ovicidal impact using serial dilutions of tebufenozide (10, 100, and 200 ppm) also was detected. Percent viability was reduced from 98% for untreated eggs to 61% for eggs dipped in 10 ppm and below 6% for eggs dipped in > or = 100 ppm. Eggs treated with 200 ppm did not hatch. Though some embryonic development was observed on eggs treated with the high concentrations (100 and 200 ppm), sclerotization of head capsule was not apparent. The ovicidal property of tebufenozide may enhance its effectiveness in controlling populations of the D. saccharalis on an area-wide basis. Fecundity and egg viability were affected in later generations of selection; however, separate studies assessing individuals that were exposed to sublethal concentration (EC5, EC15, and EC30) of tebufenozide as third instars for 7 d in one generation did not detect differences.     

Frequencies of pyrethroid resistance-associated mutations of Vssc1 and CYP6D1 in field populations of Musca domestica L. in Turkey

House flies were collected from 16 different provinces in the Aegean and Mediterranean regions of Turkey, and the frequencies of pyrethroid resistance-associated mutations in Vssc1 and CYP6D1 in these field-collected populations were studied. Although there is no organized resistance management program for house fly control in Turkey, it is known that different groups of insecticides, including pyrethroids, are used. The frequencies of both Vssc1 and CYP6D1 alleles were weighted toward the susceptibles, with Vssc1-susceptible alleles having higher frequencies in both regions (0.75 in Aegean and 0.69 in Mediterranean populations) than CYP6D1-susceptible alleles (0.65 in Aegean and 0.56 in Mediterranean populations). The frequencies of kdr-his alleles were higher than the frequencies of kdr alleles in these populations. While the frequencies of kdr-his alleles were close to each other in the Aegean (0.23) and Mediterranean (0.17) populations, the frequencies of kdr alleles remarkably differed in these two regions, with values of 0.02 and 0.14, respectively. In contrast to Europe, Asia, and the U.S.A., no super-kdr allele was detected in the samples from both regions. We identified six and eight different Vssc1+CYP6D1 genotype classes in the Aegean and Mediterranean regions, respectively. The three most common genotype classes in the regions were susceptible Vssc1 with heterozygous CYP6D1v1 (29%), sus/kdr-his1 with heterozygous CYP6D1v1 (23%), and susceptible Vssc1 with CYP6D1 (22%). The total frequencies of these three most common genotype classes (approximately 75%) obtained in our study were very close to the value obtained in Florida in a previous study, which was related by the similarity of temperature patterns between Florida and the corresponding regions of Turkey. This may reflect the lack of overwintering fitness cost associated with resistance alleles in both climates.     

Baseline susceptibility of western corn rootworm (Coleoptera: Chrysomelidae) to clothianidin

Abstract:  Western corn rootworm, Diabrotica virgifera virgifera LeConte, neonate susceptibility to clothianidin, a contact and systemic neonicotinoid insecticide, was determined from both laboratory and field-collected populations. Neonates were exposed to filter paper treated with increasing clothianidin concentrations and mortality was evaluated after 24 h. Additionally, two populations were exposed to an artificial diet which was surface treated with clothianidin. Although larvae were five- to six-fold more sensitive to treated diet, results with treated filter paper were more reliable in terms of control mortality and required much less manipulation of rootworm larvae. Therefore, initial baseline comparisons were conducted using the filter paper assays. The variation among populations exposed to treated filter paper was generally low, 4.4-fold among laboratory populations tested; however, there was a 14.5-fold difference in susceptibility among all populations tested. In general, clothianidin was very toxic to rootworm neonates, with LC₅₀ values ranging from 1.5 to 21.9 ng/cm². These results indicate the practicability and sensitivity of the paper filter disc assay to establish baseline susceptibility levels, which is an essential first step in resistance management. A baseline response provides a reference for tracking shifts in susceptibility following commercialization of a control agent so that early changes in susceptibility can be detected.     

Phenotypic- and Genotypic-Resistance Detection for Adaptive Resistance Management in Tetranychus urticae Koch

Rapid resistance detection is necessary for the adaptive management of acaricide-resistant populations of Tetranychus urticae. Detection of phenotypic and genotypic resistance was conducted by employing residual contact vial bioassay (RCV) and quantitative sequencing (QS) methods, respectively. RCV was useful for detecting the acaricide resistance levels of T. urticae, particularly for on-site resistance detection; however, it was only applicable for rapid-acting acaricides (12 out of 19 tested acaricides). QS was effective for determining the frequencies of resistance alleles on a population basis, which corresponded to 12 nonsynonymous point mutations associated with target-site resistance to five types of acaricides [organophosphates (monocrotophos, pirimiphos-methyl, dimethoate and chlorpyrifos), pyrethroids (fenpropathrin and bifenthrin), abamectin, bifenazate and etoxazole]. Most field-collected mites exhibited high levels of multiple resistance, as determined by RCV and QS data, suggesting the seriousness of their current acaricide resistance status in rose cultivation areas in Korea. The correlation analyses revealed moderate to high levels of positive relationships between the resistance allele frequencies and the actual resistance levels in only five of the acaricides evaluated, which limits the general application of allele frequency as a direct indicator for estimating actual resistance levels. Nevertheless, the resistance allele frequency data alone allowed for the evaluation of the genetic resistance potential and background of test mite populations. The combined use of RCV and QS provides basic information on resistance levels, which is essential for choosing appropriate acaricides for the management of resistant T. urticae.     

Insecticide susceptibility of Cnaphalocrocis medinalis (Lepidoptera: Pyralidae) in China

Insecticide control is the major measure for suppression of Cnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae) damage, and a few insecticides used for long time have proved to fail to control this pest in China. Several new chemicals have been introduced for control of C. medinalis. However, there was no baseline susceptibility data of C. medinalis to insecticides used or will be in use. In this study, a seedling dipping method was developed for bioassay of insecticide susceptibility of C. medinalis. Dose responses of C. medinalis to 11 insecticides were tested. Interpopulation sensitivity to insecticides was compared. Based on LC50 values, C. medinalis was most susceptible to antibiotic insecticides (abamectin, emamectin benzoate, and spinosad) and least sensitive to monosultap and a Bacillus thuringiensis (Bt) product. Chlorantraniliprole and insect growth regulator (IGR) insecticides (tebufenozide and hexaflumuron) exhibited great efficacy against C. medinalis. No susceptibility difference was observed for antibiotic insecticide and IGR insecticides among three populations. Narrow variation in tolerant level was detected for organophosphates insecticides, chlorantraniliprole, monosultap, and Bt. The results in this study provided baseline susceptibility data of C. medinalis to 11 insecticides and also offered useful information for choice of alternative insecticide and for integrated resistance management of C. medinalis.     

Carbaryl resistance in populations of grape berry moth (Lepidoptera: Tortricidae) in New York and Pennsylvania

We collected grape berry moth, Endopiza viteana (Clemens) (from cultivated and wild Vitis along Lake Erie in Pennsylvania and New York), and measured carbaryl susceptibility in first instars. A model of susceptibility was based on the concentration-mortality curve of laboratory-maintained colonies originating from wild Vitis with no prior history of carbaryl exposure, and a noncommercial vineyard with modest previous exposure to carbaryl. We estimated LC50 and LC90 for susceptible grape berry moth larvae at 45.4 and 2319 microg/ml, respectively. Bioassays on field-collected larvae from commercial vineyards in both states, where grape growers were abiding by current pest management guidelines for carbaryl use, revealed carbaryl resistance ratios from 7 to 71 at the LC50 level. With the loss or restriction of alternative chemical control tactics in the Food Quality Protection Act era, resistance management programs for grape berry moth should be immediately developed and implemented to regain the efficacy of this once effective insecticide and other related chemical compounds.     

Predicting economic losses from the continued spread of the Mexican rice borer (Lepidoptera: Crambidae)

The Mexican rice borer, Eoreuma loftini (Dyar) (Lepidoptera: Crambidae), is an invasive species that originated from Mexico, and it is threatening to cause major economic losses to sugarcane, Saccharum spp., and rice, Oryza sativa L., industries in Louisiana. The insect is expected to reach sugarcane and rice production areas in Louisiana by 2008, and infest all of Louisiana sugarcane and rice industries by 2035. When all sugarcane in Louisiana becomes infested, annual yield losses of $220 million would be expected for a cultivar of comparable susceptibility to LCP 85-384 (assuming this cultivar is planted on 100% of the production area). This also assumes the use of the current practice of rainfed production and one application of insecticide, which is presently used by farmers in Louisiana. Irrigation with 30 cm of water is predicted to reduce estimated losses by 29%, whereas four applications of a biorational insecticide such as tebufenozide are expected to reduce the loss in revenue by 53%. The use of the resistant 'HoCP 85-845' would reduce the projected loss in revenue by 24%. Combining all three management tactics on sugarcane, anticipated net loss in revenue would decrease by 66%. The rice industry in Louisiana is projected to suffer from a loss in revenue of $45 million when the entire state is infested. A 77% reduction in loss in revenue is expected with one application of lambda-cyhalothrin. A quarantine on east Texas sugarcane is estimated to save the Louisiana industry between $1.1 billion and $3.2 billion (depending on management) during the time needed for the insect to fully invade the state's sugarcane and rice producing area by natural migration rather than by accidental introduction. The rapid deployment of appropriate management tactics will have a key role in reducing the anticipated economic impact of E. loftini once it becomes a pest in Louisiana sugarcane and rice.     

Trend of temephos resistance in Aedes (Stegomyia) mosquitoes in Thailand during 2003-2005

The diagnostic dose for temephos susceptibility test was established based on Aedes aegypti, the susceptible Bora (French Polynesia) strain, for practical and routine use. The diagnostic dose was subsequently used to evaluate the susceptibility/resistance status in F1 progenies of field-collected samples from Bangkok and various parts of Thailand. It appeared that Ae. aegypti mosquitoes of one collection site each in Bangkok, Nakhon Sawan (northcentral), and Nakhon Ratchasrima (northeast) were resistant to temephos, with mortality ranging from 50.5 to 71.4%. Moreover, there was a trend of resistance to temephos among Ae. aegypti populations of all studied districts of Nakorn Ratchasima and most areas of Nakhon Sawan, of which those in one area were susceptible. However, various levels of temephos susceptibility were found in Bangkok populations, including resistance and incipient resistance. In Chonburi Province (eastern), all mosquitoes were susceptible to temephos with an indication of tolerance in one sample. Additionally, mosquitoes from Songkhla (south), Chiang Rai (north), Kanchanaburi (west), and Chanthaburi (east) remained susceptible to temephos during the sample collecting period. Bioassay tests on Aedes albopictus populations collected in this study from Nakhon Sawan, Nakorn Ratchasima, Songkhla, and Kanchanaburi revealed high susceptibility to temephos. Although the use of temephos seems to be potentially effective in many areas of the country, a noticeable trend of resistance indicated that alternative vector control methods should be periodically applied.     

Establishment of Cry9C susceptibility baselines for European corn borer and southwestern corn borer (Lepidoptera: Crambidae)

In 1997 and 1998, Cry9C susceptibility baselines were established for field-collected populations of European corn borer, Osrinia nubilalis (Hubner), and southwestern corn borer, Diatraea grandiosella Dyar. Bioassay of neonate European corn borer larvae of 16 colonies collected from the midwestern United States indicated LC50 values ranging from 13.2 to 65.1 ng of Cry9C protein per square centimeter. Neonate European corn borer LC50 values ranged from 46.5 to 214 ng/cm2. Neonate larvae of three colonies of southwestern corn borer collected from the southern and southwestern United States exhibited LC50 values from 16.9 to 39.9 ng of Cry9C protein per square centimeter. Southwestern corn borer neonate LC90 confidence limit values ranged from 40.3 to 157 ng of Cry9C protein per centimeter. The most sensitive southwestern corn borer colony was collected from the Mississippi delta exhibiting an LC50 value of 22.6 ng of Cry9C per cm2 and also displayed the widest LC0 confidence limits of 40.3-94.8 ng of Cry9C per cm2. Geographic baseline susceptibility data establishes the natural genetic variation and provides the foundation for future testing of insect populations exposed to increased use of Bacillus thuringiensis-based crops. Insect resistance management and stewardship of Cry9C will rely upon baseline data for the validation of discriminating dose assays for European corn borer and southwestern corn borer.     

Monitoring and characterization of diamondback moth (Lepidoptera: Plutellidae) resistance to spinosad

Fourteen populations of the diamondback moth, Plutella xylostella (L.), were collected from fields of crucifer vegetables in the United States, Mexico, and Thailand in 1999 and 2000 for susceptibility tests with spinosad. Most populations were susceptible to spinosad and similar to earlier baseline values, but populations from Thailand and Hawaii showed high levels of tolerance. A statewide survey in Hawaii in 2000 and 2001 indicated resistance problems on several islands. One colony collected in October 2000 from Pearl City, HI, was subjected to further selection pressure, using spinosad in the laboratory, and then was used as the resistant strain (Pearl-Sel) for other tests. Spray tests using the recommended field rates of spinosad on potted broccoli plants in the greenhouse confirmed that field control failures due to resistance were possible in the areas of these collections. Analysis of probit lines from F1 reciprocal crosses between the Pearl-Sel and S strain indicated that resistance to spinosad was inherited autosomally and was incompletely recessive. A direct test of monogenic inheritance based on the F1 x Pearl-Sel backcrosses suggested that resistance to spinosad was probably controlled by one locus. The synergists S,S,S-tributyl phosphorotrithioate and piperonyl butoxide did not enhance the toxicity of spinosad to the resistant colony, indicating metabolic mediated detoxification was probably not responsible for the spinosad resistance. Two field colonies in Hawaii that were resistant to spinosad were not cross-resistant to emamectin benzoate or indoxacarb. Resistance developed in Hawaii due to the continuous cultivation of crucifers in which as many as 50 applications of spinosad per year may have been made to a common population of P. xylostella in sequential plantings, although each grower might have used the labeled restrictions for resistance management. Resistance management strategies will need to address such cropping and pest management practices.     

甜菜夜蛾抗药性研究现状

就有关甜菜夜蛾Spodoptera exigua(Hbner)对常用杀虫剂的抗药性现状作综述。甜菜夜蛾对拟除虫菊酯类、有机磷类、氨基甲酸酯类杀虫剂已产生了较高水平的抗药性,其中山东泰安抗性种群对氯氟氰菊酯的抗性高达2445.5倍;对多杀菌素等生物杀虫剂产生了中低水平的抗性;对昆虫生长调节剂如虫酰肼的敏感性也有所降低,但昆虫生长调节剂依然是比较理想的防治药剂。对交互抗性及抗性治理也作了阐述。