Investigation of resistance mechanisms to fipronil in diamondback moth (Lepidoptera: Plutellidae)

The SZ-F strain of Plutella xylostella (L.) (Lepidoptera: Plutellidae) was derived from a field-collected strain (SZ) by 20 generations of continuous selection with fipronil. The selection resulted in 300-fold increase in resistance to fipronil, and 3.5- and 6.5-fold cross-resistance to dieldrin and endosulfan, respectively, in the SZ-F strain compared with the unselected SZ strain. Analysis of detoxification enzyme activities and bioassay with synergists indicated that metabolic mechanisms are not important to fipronil resistance of the SZ-F strain and that the fipronil resistance is most likely attributed to target site insensitivity. The genomic DNA fragments flanking the second membrane-spanning region of Rdl gamma-aminobutyric acid (GABA) receptor gene from P. xylostella, PxRdl, were cloned and sequenced. A single allele of the PxRdl gene (encoding A302 or allele PxRdl-Ala) was found in both the Roth (susceptible) and SZ strains. In addition to the wild-type allele PxRdl-Ala, the resistant SZ-F strain carried a mutant PxRdl allele with the conserved amino acid replacement A302(GGC)--> S302(TCC) (allele PxRdl-Ser). The mutant PxRdl-Ser allele frequency in the SZ-F strain was 30%. After treatment of 20 mg/l fipronil on the SZ-F strain, the PxRdl-Ser allele frequency in the survivors increased to 57%. High frequency of the PxRdl-Ala allele remaining in the resistant SZ-F strain suggested that the A302S mutation in the PxRdl gene is partially associated with fipronil resistance and that other mutation(s) in the PxRdl gene or other Rdl-like subunit(s) may contribute to fipronil resistance.     

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.     

Fipronil resistance in the diamondback moth (Lepidoptera: Plutellidae): inheritance and number of genes involved

Bioassays (at generation 1, G1) using fipronil, spinosad, indoxacarb, and Bacillus thuringiensis toxins Cry1Ac and Cry1Ca with a newly collected field population of Plutella xylostella (L.) from farmers fields in the Cameron Highlands, Malaysia, indicated a resistance ratio of approximately 400-, 1,170-, 330-, 2,840-, and 1,410-fold, respectively, compared with a laboratory-susceptible population of P. xylostella (ROTH). At G3, the field-derived population was divided into two subpopulations, one was selected (G3 to G7) with fipronil (fip-SEL), whereas the second was left unselected (UNSEL). Bioassays at G8 found that selection with fipronil gave a resistance ratio of approximately 490 compared with UNSEL and approximately 770 compared with ROTH. The resistance ratio for fipronil, spinosad, indoxacarb, Cry1Ac, and Cry1Ca in the UNSEL population declined significantly by G8. Logit regression analysis of F1 reciprocal crosses between fip-SEL (at G8) and UNSEL indicated that resistance to fipronil in the fip-SEL population was inherited as an autosomal, incompletely recessive (D(LC) = 0.37) trait. At the highest dose of fipronil tested, resistance was completely recessive, whereas at the lowest dose it was incompletely recessive. A direct test of monogenic inheritance based on a backcross of F1 progeny with fip-SEL suggested that resistance to fipronil was controlled by a single locus. The fip-SEL population at G8 showed little change in its response to spinosad and indoxacarb compared with G1, whereas its susceptibility to Cry1Ac and Cry1Ca increased markedly over the selection period. This suggests that there may be some low level of cross-resistance between fipronil, spinosad, and indoxacarb.     

Functional and genetic characteristics of Chlorantraniliprole resistance in the diamondback moth,Plutella xylostella (Lepidoptera: Plutellidae)

The diamondback moth (Plutella xylostella) is a globally distributed and important economic pest, and it has developed resistance to all conventional insecticide classes used in the field. Chlorantraniliprole is a new chemical class of insecticide that acts as a conformation‐sensitive activator of the insect ryanodine receptor (RyR). In the present study, a field strain (16.3‐fold resistance to chlorantraniliprole) was collected in Korea and lab‐selected with chlorantraniliprole for more than one year. The resulting strain presented 2,157‐fold resistance to chlorantraniliprole. A point mutation (G4946E) in the RyR gene was observed at a high frequency in the resistant strain. Enzyme assays indicated that glutathione S‐transferase (GST) and P450 activity in the resistant strain were 2.4‐ and 1.96‐times higher than that of the susceptible strain, respectively. The expression of the RyR, GST (sigma, omega, and zeta) and CYP321E1 gene was higher in the resistant strain than in the susceptible strain. The F₁ progeny resulting from reciprocal crosses did not reveal maternal effects or a diamide‐susceptible phenotype, which suggests an autosomal nearly recessive mode of inheritance. In addition, we surveyed the susceptibility to 13 insecticides (3 diamides, 2 synthetic pyrethroids, 2 spinosyns, 1 organophosphate, 1 oxadiazine, 1 avermectin, and 3 others) in the chlorantraniliprole‐resistant strain. The resistant strain exhibited high cross‐resistance to flubendiamide (5,910 fold) and showed no cross‐resistance to spinetoram, spinosad, indoxacarb, and metaflumizone. These results can serve as an important basis for guiding the use of insecticides in the field.     

Manipulating the attractiveness and suitability of hosts for diamondback moth (Lepidoptera: Plutellidae)

Ovipositional preference and larval survival of the diamondback moth, Plutella xylostella (L.), were compared among cabbage, Brassica oleracea L. variety capitata; glossy collards, Brassica oleracea L. variety acephala; and yellow rocket, Barbarea vulgaris (R. Br.) variety arcuata in different treatments of planting density, host plant age, intercropping, and water stress in 2003 and 2004. P. xylostella laid nearly twice as many eggs per plant in the high planting densities of glossy collards and yellow rocket than in the standard planting densities. Ovipositional preference was positively correlated with plant age in cabbage, glossy collards, and yellow rocket. Larval survival on cabbage was 1.9 times higher on 6-wk than on 12-wk-old plants, whereas larval survival on collards was 12.1 times higher on the younger plants. No larvae survived on either 6- or 12-wk-old yellow rocket plants. Intercropping cabbage with either tomato, Lycopersicon esculentum Mill., or fava bean, Vicia fava L., did not reduce the number of eggs laid on cabbage. No significant differences in oviposition were found between water-stressed and well-irrigated host plants treatments. Yet, P. xylostella larval survival on water-stressed cabbage was 2.1 times lower than on well-irrigated cabbage plants. Based on our findings, the effectiveness of trap crops of glossy collards and yellow rocket could be enhanced by integrating the use of higher planting densities in the trap crop than in the main crop and seeding of the trap crop earlier than the main crop.     

Evaluating trap crops for diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)

Potential trap crops for the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), were evaluated through a series of ovipositional preference and larval survival experiments in outdoor screenhouses in 2002 and 2003. Hosts examined as trap crops were glossy and waxy collards, Brassica oleracea L. variety acephala; Indian mustard, Brassica juncea (L.) Czern; and yellow rocket, Barbarea vulgaris (R. Br.) variety arcuata. More eggs were laid on the potential trap crops, with the exception of waxy collards, than on cabbage. When P. xylostella was offered multiple hosts at the same time, numbers of eggs laid on glossy collards, Indian mustard, and yellow rocket were 3, 18, and 12 times greater than on cabbage, respectively. Similarly, when P. xylostella was offered a single trap crop host and cabbage, numbers of eggs laid on glossy collards, Indian mustard, and yellow rocket were 300, 19, and 110 times greater than on cabbage, respectively. Our studies suggest differences in oviposition between the potential trap crops and cabbage were likely due to host volatiles, leaf morphology and color, or a combination of these factors, rather than to total leaf areas, leaf shape, or plant architecture. Two-choice tests with a Y-tube olfactometer indicated that plant volatiles were major factors in P. xylostella host preference. The percentage larval survival from egg to pupation was 22.2% on cabbage, 18.9% on waxy collards, and 24.4% on Indian mustard, whereas survival was significantly lower on glossy collards (6.7%) and yellow rocket (0%). Based on our tests, it seems that yellow rocket may be the best candidate for use as a trap crop for P. xylostella because it is highly attractive for oviposition, but larvae do not survive on it.     

Monitoring of diamondback moth (Lepidoptera: Plutellidae) resistance to spinosad, indoxacarb, and emamectin benzoate

Six to nine populations of the diamondback moth, Plutella xylostella (L.), were collected annually from fields of crucifer vegetables in the United States and Mexico from 2001 to 2004 for baseline susceptibility tests and resistance monitoring to spinosad, indoxacarb, and emamectin benzoate. A discriminating concentration for resistance monitoring to indoxacarb and emamectin benzoate was determined based on baseline data in 2001 and was used in the diagnostic assay for each population in 2002-2004 together with a discriminating concentration for spinosad determined previously. Most populations were susceptible to all three insecticides, but a population from Hawaii in 2003 showed high levels of resistance to indoxacarb. Instances of resistance to spinosad occurred in Hawaii (2000), Georgia (2001), and California (2002) as a consequence of a few years of extensive applications in each region. The collaborative monitoring program between university and industry scientists we discuss in this article has provided useful information to both parties as well as growers who use the products. These studies provide a baseline for developing a more effective resistance management program for diamondback moth.     

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.     

羧酸酯酶介导的小菜蛾对氟虫腈的抗性

【目的】羧酸酯酶（carboxylesterases, CarEs）是昆虫重要的解毒代谢酶之一,可以介导靶标昆虫对多种杀虫剂的代谢抗性。本研究检测了羧酸酯酶对小菜蛾Plutella xylostella 抗药性的介导功能,旨在阐明羧酸酯酶在小菜蛾代谢解毒中的生理生化和分子机理。【方法】采用点滴法测定氟虫腈对小菜蛾敏感种群和抗氟虫腈种群的毒力,以及羧酸酯酶抑制剂磷酸三苯酯（triphenyl phosphate, TPP）对氟虫腈的增效作用;以LC₃₀和LC₅₀浓度的氟虫腈处理抗性小菜蛾,测定药剂处理后CarEs酶活性的变化;利用qRT-PCR技术分析Pxae22和Pxae31两个基因在小菜蛾不同发育阶段、组织和种群的表达模式;利用dsRNA干扰Pxae22和Pxae31后观察基因的表达变化和小菜蛾3龄幼虫对药剂敏感性的变化。【结果】TPP可以削弱小菜蛾3龄幼虫对氟虫腈的抗性,增效倍数约为6倍;使用较低剂量（LC₃₀和LC₅₀）氟虫腈处理小菜蛾3龄幼虫后,处理组CarEs比活力明显高于对照,提示氟虫腈对小菜蛾CarEs活性具有诱导作用。对羧酸酯酶基因Pxae22和Pxae31在小菜蛾不同发育阶段、4龄幼虫不同组织和不同种群3龄幼虫中的表达模式分析发现,这两个基因在小菜蛾4龄幼虫中的表达量最高;在4龄幼虫中以中肠组织中的表达量较高,头、表皮、脂肪体中的表达量很低; 抗性种群中的表达量显著高于敏感种群。通过干扰 Pxae22和 Pxae31后的qRT-PCR验证,两个基因的表达量均显著降低,进一步的氟虫腈毒力测定发现,干扰P xae22和 Pxae31后的小菜蛾3龄幼虫对氟虫腈的敏感性分别增加了1.63倍和1.73倍。【结论】羧酸酯酶在小菜蛾对氟虫腈解毒代谢中具有重要作用;Pxae22和Pxae31是小菜蛾的两个抗性相关基因,其表达水平的变化直接影响小菜蛾对氟虫腈的敏感性。     

Management of diamondback moth,Plutella xylostella (Lepidoptera: Plutellidae) by mating disruption

Abstract Field trials were conducted in China in 2008 and 2009 to evaluate the efficacy of mating disruption (MD) on diamondback moth, Plutella xylostella, in cabbage, Brassica oleracea var. capitata. Effectiveness was positively correlated with the MD dispenser density in the field. A density of 167 MD dispensers per ha produced an average population decrease of about 50% compared to the conventional‐practice field. Significant fewer males were captured in pheromone‐treated and conventional‐practice fields than in the blank control field, but the difference was not significant between the pheromone‐treated and conventional‐practice fields. In addition, fewer eggs and larvae were observed in pheromone‐treated fields. Our results suggest mating disruption coupled with minimal insecticidal supplements is a promising solution for resistance management and control of diamondback moth infestation.     

Using yellow rocket as a trap crop for diamondback moth (Lepidoptera: Plutellidae)

Yellow rocket, Barbarea vulgaris (R. Br.) variety arcuata, was evaluated as a trap crop for diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), in cabbage, Brassica oleracea L. variety capitata, in 2003 and 2004. In 2003, the numbers of P. xylostella larvae found in field plots of cabbage alone were 5.2-11.3 times higher than those on cabbage plants in plots that included cabbage and several rows of yellow rocket. In an outdoor experiment in screenhouses, P. xylostella oviposition on cabbage was compared among six treatments that varied in the percentage of yellow rocket in relation to cabbage (0, 4, 8, 16, 24, and 32% of the plants were yellow rocket). Results indicated that the percentage of eggs laid on cabbage decreased as the percentage of yellow rocket in the treatment increased, but this decrease was not significant beyond 20% of the plants being yellow rocket. In 2004, the numbers of P. xylostella larvae in field plots of cabbage alone were 1.6-2.4 and 1.7-2.8 times higher than numbers in treatments with 10 and 20% trap crop, respectively. Sticky trap and sweep net captures of P. xylostella adults indicated that within-field dispersal was reduced by the presence of yellow rocket and aggregation occurred around yellow rocket plants. Our study suggests that using yellow rocket as a trap crop may reduce P. xylostella infestations in cabbage fields, and this possibility is discussed in the context of general crop and insect pest management practices in crucifers.     

Oviposition site selection by the diamondback moth,Plutella xylostella (L.) (Lepidoptera: Plutellidae)

AlthoughPlutella xylostella (L.) is a worldwide pest of cruciferous crops, relatively little is known about its oviposition behaviour. This study was undertaken to provide necessary information about mechanisms involved inP. xylostella host selection. Four oviposition behaviours were described. Moths were given artificial substrates treated with water, sinigrin, orBrassica napus (cv. Westar) squashes, combined withB. napus volatiles and/or grooves in the substrate. No eggs were deposited in the absence of olfactory and gustatory stimuli. Moths given gustatory but not olfactory stimuli deposited similar numbers of eggs but spent significantly more time performing olfactory-related behaviours. Conversely, moths given olfactory but not gustatory stimuli did not oviposit.SSubstrate grooves did not influence egg numbers but appeared to influence egg location. The order in which oviposition behaviours occurs and the relative importance of stimuli type are discussed.     

Examination of the F2 screen for rare resistance alleles to Bacillus thuringiensis toxins in the diamondback moth (Lepidoptera: Plutellidae)

A synthetic laboratory population of the diamondback moth, Plutella xylostella (L.), was used to test the F2 screen developed for detecting the frequency of rare resistance alleles to Cry1Ac and Cry1C toxins of Bacillus thuringiensis (Bt). Of the 120 single-pair matings set up, 106 produced enough F2 families for screening of Cry1Ac or Cry1C resistance alleles using both transgenic broccoli and an artificial diet overlay assay with a diagnostic dose. When using Bt broccoli plants as the F2 screen method, only one F2 family was detected for Cry1Ac resistance and no family was detected for Cry1C resistance. Six families were detected for either Cry1Ac or Cry1C resistance using the diet assay. The survivors in the diagnostic diet assay were crossed with the resistant individuals to confirm their resistance genotypes. Four F2 families were confirmed to contain one copy of an allele resistant to Cry1Ac in the original single-pairs and four other F2 families contained an allele resistant to Cry1C. Our results suggest that using transgenic plants expressing a high level of a Bt toxin in an F2 screen may underestimate the frequency of resistance alleles with high false negatives, or fail to detect true resistance alleles. The diagnostic diet assay was a better F2 screen method to detect alleles, especially for the Cry1Ac resistance with monogenic inheritance in the diamondback moth. The estimated probabilities of false positives and false negatives were 33 and 1%, respectively, for detecting Cry1Ac resistance at the allele frequency of 0.012 using the diagnostic diet assay. Careful validation of the screening method for each insect-crop system is necessary before the F2 screen can be used to detect rare Bt resistance alleles in field populations.     

微小花蝽对小菜蛾捕食控制的能力

为了明确微小花蝽以小菜蛾卵做为饲料的适宜性及其对小菜蛾的控制能力,在室内以桃蚜作为参比猎物,研究捕食小菜蛾卵对微小花蝽生长发育、繁殖的影响,并结合捕食功能反应评价微小花蝽对小菜蛾的捕食能力.结果表明: 微小花蝽取食小菜蛾卵能够完成世代发育,并能够正常繁育后代.取食小菜蛾卵时,微小花蝽雌、雄若虫的历期(♀: 12.3 d,♂: 12.2 d)、成虫体长[♀: (2.13±0.01) mm,♂: (1.91±0.00) mm]、体宽[♀: (0.87±0.01) mm,♂: (0.71±0.01) mm]、单雌产卵量(12.7±1.1)、产卵前期[(5.1±0.6) d]和产卵期[(3.7±0.4) d]均与桃蚜处理组无显著差异;雌、雄成虫寿命[♀: (10.7±1.4) d,♂: (9.1±1.3) d]显著长于桃蚜处理组[♀: (8.5±0.5) d,♂: (6.4±0.3) d];若虫存活率[(65.0±6.8)%]不及桃蚜处理组[(80.0±8.2)%],且雌性比例偏低.微小花蝽对小菜蛾的捕食功能反应符合HollingⅡ型方程.微小花蝽1～5龄若虫对小菜蛾卵的日均最大捕食量(N_max)分别为7.5、16.3、23.3、29.1和38.7粒;雌、雄成虫的日均最大捕食量分别为39.0和26.9粒;5龄若虫对小菜蛾低龄幼虫的日均最大捕食量为41.3头;雌、雄成虫的日均最大捕食量分别为40.8和23.9头.单头雌、雄微小花蝽一生中最多可捕食小菜蛾卵(711.3±58.1)和(535.4±30.6)粒,小菜蛾低龄幼虫(371.9±52.0)和(253.9±32.3)头.微小花蝽以小菜蛾卵饲养可行,且对小菜蛾具有良好的控制作用.     

Infectivity studies of a new baculovirus isolate for the control of the diamondback moth (Plutellidae: Lepidoptera)

This study describes a new baculovirus isolate recovered from infected larvae of the diamondback moth, Plutella xylostella (L.), and identified as a multiple nucleopolyhedrovirus (MNPV). The plaque purified isolate designated as PxMNPVCL3 was found to be pathogenic to P. xylostella, Heliothis virescens (F.), Trichoplusia ni (Hübner), H. subflexa (Guenée), Helicoverpa zea (Boddie), Spodoptera exigua (Hübner), and S. frugiperda (J. E. Smith) larvae in decreasing order of susceptibility. The LC50 for diamondback moth, the most susceptible, was 6 occlusion bodies (OB)/cm2, whereas the most resistant species, namely S. frugiperda, was 577 OB/cm2. PxMNPVCL3 was more pathogenic to diamondback moth by 3-4 log cycles as compared with 2 broad-spectrum baculoviruses, namely Autographa california (alfalfa looper) MNPV and Anagrapha falcifera (celery looper) MNPV. The 3 baculoviruses were compared with each other and characterized by restriction endonuclease (REN) analysis, hybridization, and neutralization tests. Fragmentation profiles generated by REN showed that the 3 baculoviruses shared some fragments in common. Hybridization studies employing digoxigenin labeled PxMNPVCL3 DNA as a probe revealed the close but distinct relationship of these 3 viruses. Neutralization tests confirmed the hybridization studies, namely that the 3 viruses although genetically similar are distinguishable from each other.     

The effects of local selection versus dispersal on insecticide resistance patterns: longitudinal evidence from diamondback moth (Plutella xylostella (Lepidoptera: Plutellidae)) in Australia evolving resistance to pyrethroids

When strong directional selection acts on a trait, the spatial distribution of phenotypes may reflect effects of selection, as well as the spread of favoured genotypes by gene flow. Here we investigate the relative impact of these factors by assessing resistance to synthetic pyrethroids in a 12-year study of diamondback moth, Plutella xylostella, from southern Australia. We estimated resistance levels in populations from brassicaceous weeds, canola, forage crops and vegetables. Differences in resistance among local populations sampled repeatedly were stable over several years. Levels were lowest in samples from weeds and highest in vegetables. Resistance in canola samples increased over time as insecticide use increased. There was no evidence that selection in one area influenced resistance in adjacent areas. Microsatellite variation from 13 populations showed a low level of genetic variation among populations, with an AMOVA indicating that population only accounted for 0.25% of the molecular variation. This compared to an estimate of 13.8% of variation accounted for by the resistance trait. Results suggest that local selection rather than gene flow of resistance alleles dictated variation in resistance across populations. Therefore, regional resistance management strategies may not limit resistance evolution.     

小菜蛾鞣化激素基因克隆及其功能分析

鞣化激素是调节昆虫表皮骨化和翅膀发育的一种神经激素, 尽管已经在许多不同种昆虫上克隆了鞣化激素基因, 但是关于小菜蛾 Plutella xylostella鞣化激素及其基因的研究至今未见报道。本研究克隆了两个小菜蛾鞣化激素基因Pxbursα和Pxbursβ （GenBank 登录号分别为KF498645和KF498646）全长cDNA, 其序列长度分别为537 bp和360 bp, 与已报道的其他昆虫的鞣化激素氨基酸序列一致性分别为51%~68% 和37%~57%。实时定量PCR分析发现Pxbursα和Pxbursβ均在蛹期表达量高, 而在幼虫期和成虫期的表达量低。以Pxbursα部分序列的双链RNA（dsRNA）饲喂小菜蛾4龄末期幼虫, 发现蛹期Pxbursα的表达受到了显著抑制, 小菜蛾的发育停滞在蛹期而无法正常羽化, 并最终死亡。由此推测, 小菜蛾鞣化激素基因在蛹期的大量表达对其生长发育和羽化具有重要的作用。     

四种农药对小菜蛾体温的影响

【目的】本研究旨在为阐明小菜蛾Plutellaxylostella体温在其防治中的应用价值提供资料。【方法】在不同人工气候箱内温度(环境温度)下,测定小菜蛾2, 3和4龄幼虫的体温,建立各龄幼虫体温(y)与环境温度(x)的关系方程;同时测定了不同环境温度下不同浓度阿维菌素、毒死蜱、氟虫腈和高效氯氰菊酯分别处理后小菜蛾3龄幼虫在不同处理时间的体温。【结果】小菜蛾2, 3和 4龄幼虫体温(y)与环境温度(x)关系方程分别为y＝0.95x+1.19(r=0.9463), y＝0.95x+1.18(r=0.9988),以及y＝0.93x+1.45(r=0.9989),等温点分别为22.16℃,21.40℃和21.41℃。在气候箱温度设定为15℃或40℃时,4种农药都对小菜蛾3龄幼虫体温无影响;而其他温度条件下,农药处理都可能改变小菜蛾3龄幼虫体温。对于阿维菌素,25℃下 2, 4和8 mg/L处理12 h,2和4 mg/L处理24 h, 0.5, 2, 4和8 mg/L处理36 h以及0.5, 1, 2和8 mg/L处理48 h时3龄幼虫体温均显著高于对照, 8 mg/L阿维菌素处理24 h时3龄幼虫体温显著低于对照;30℃下0.5 mg/L处理24 h及1 mg/L处理36 h 3龄幼虫体温显著低于对照,1 mg/L处理48 h和各浓度处理60 h时3龄幼虫体温均显著高于对照;35℃下只有1和8 mg/L处理48 h时3龄幼虫体温显著低于对照。对于毒死蜱,20℃下50, 200和800 mg/L处理24 h, 100, 400和800 mg/L处理36 h时3龄幼虫体温都显著低于对照;25℃下100和200 mg/L处理12h, 800 mg/L处理24 h, 100, 200和800 mg/L处理60 h时3龄幼虫体温均显著低于对照,而50, 100, 200和400 mg/L处理24 h, 100和200 mg/L处理36 h及100和400 mg/L处理48 h时3龄幼虫体温均显著高于对照;30℃下只有800 mg/L处理24 h时3龄幼虫体温显著低于对照,50, 100, 200和800 mg/L处理60 h时3龄幼虫体温显著高于对照。对于氟虫腈,20℃下只有0.5 mg/L处理36 h时3龄幼虫体温显著低于对照;25℃下 4 mg/L处理12 h和各浓度处理60 h时3龄幼虫体温显著低于对照,0.5 mg/L处理24 h以及0.25, 1和2mg/L处理48 h时3龄幼虫体温均显著高于对照;30℃下 0.25和0.5 mg/L处理12 h, 0.25和2 mg/L处理24h, 4 mg/L处理48 h以及2 mg/L处理60 h时3龄幼虫体温显著低于对照;35℃下只有0.25和0.5 mg/L处理60 h时3龄幼虫体温显著高于对照。对于高效氯氰菊酯,20℃下2和8 g/L处理36 h, 4和8 g/L处理48 h时3龄幼虫体温显著高于对照;25℃下 2, 4和8 g/L处理12 h时3龄幼虫体温均显著低于对照, 0.5, 4和8g/L处理24 h, 1, 4和 8 g/L处理36 h以及1, 2和4 g/L处理60 h时3龄幼虫体温均显著高于对照;30℃下 0.5和1 g/L浓度处理12 h, 0.5, 1, 4和8 g/L处理24 h以及1, 2和8 g/L处理60 h时3龄幼虫体温都显著低于对照。【结论】小菜蛾幼虫自律性体温调节能力低;阿维菌素、毒死蜱、氟虫腈或高效氯氰菊酯处理影响小菜蛾3龄幼虫的体温,影响形式随农药种类和浓度,环境温度及处理时间不同而不同。本研究拓宽了农药毒理学及害虫防治研究内容。     

光周期对小菜蛾羽化节律的影响

【目的】本研究旨在探究光周期对小菜蛾Plutella xylostella(Linn.)羽化节律的调控作用,为小菜蛾的季节性种群动态预测提供理论基础。【方法】在自然光周期和逆转光周期条件下,观察小菜蛾每日的羽化情况。【结果】在自然光周期和逆转光周期条件下,小菜蛾蛹在24 h均可以羽化。正常光周期下,羽化行为表现为近似抛物线形昼夜节律,雌成虫通常在黑暗阶段羽化（18:00-6:00）,而雄成虫大多在光照阶段羽化（6:00-18:00）。在逆转光周期下,小菜蛾蛹的羽化无明显规律。在正常光周期下,雌成虫的羽化频率在光期和暗期没有显著差异,但是,相比于暗期,雄成虫在光期羽化率较高;在逆转光周期后,雌成虫在暗期（6:00-18:00）的羽化数量比光期（18:00-6:00）高,雄成虫暗期的羽化数量比光期低。【结论】光周期逆转后小菜蛾雌成虫和雄成虫的羽化节律明显改变,这将有助于阐明害虫光周期变化与生物钟之间的关系。