虫酰肼及其衍生物0593对家蚕的毒性及作用机理

为明确虫酰肼衍生物0593对家蚕Bombyx mori的毒性,本研究采用食下毒叶法测定了虫酰肼及其衍生物0593对家蚕的毒性,观察了亚致死浓度下对家蚕生长发育的影响,并测定了虫酰肼及其衍生物0593对家蚕幼虫体内保护酶的影响,对虫酰肼0593的作用机理进行了初步探索。结果表明：虫酰肼及其衍生物0593对2龄家蚕96 h的LC₅₀值分别为1.2863和0.3364 mg·L^-1,属高毒级药剂;虫酰肼及其衍生物0593在亚致死剂量下对家蚕的生长发育有明显的不利性,可使幼虫历期缩短0.5～2 d;处理组眠期体重、全茧量、蛹重和化蛹率与对照相比均显著降低;对4龄幼虫体内多酚氧化酶和几丁质酶也有较明显影响,虫酰肼及其衍生物0593处理家蚕,处理后6 h对体内多酚氧化酶有明显激活作用,12 h后表现出显著的抑制作用;虫酰肼及其衍生物0593对家蚕体内几丁质酶均有激活作用。0593对保护酶的影响较虫酰肼明显。结果提示虫酰肼及其衍生物0593对家蚕毒性高,对其生长发育和保护酶类均有不利性,不适合在桑园及其周边农田使用。     

甜菜夜蛾对甲氧虫酰肼抗药性选育及其抗性生化机理初探

在室内用甲氧虫酰肼对甜菜夜蛾Spodoptera exigua(Hübner)幼虫进行抗药性选育,经过13代汰选,甜菜夜蛾抗性种群(Met-R)对甲氧虫酰肼的抗性倍数为4.19倍;交互抗性测定发现,Met-R甜菜夜蛾对毒死蜱、高效氯氰菊酯、虫螨腈、虫酰肼、甲胺基阿维菌素苯甲酸盐等药剂的交互抗性比值在1.05～2.07之间,不存在交互抗性。离体酶活性测定结果表明,Met-R甜菜夜蛾5龄幼虫中肠谷胱甘肽S-转移酶比活力是相对敏感种群(SS)甜菜夜蛾的2.44倍,存在显著性差异,说明谷胱甘肽S-转移酶活性增强可能与甜菜夜蛾对甲氧虫酰肼产生抗药性有关;但Met-R甜菜夜蛾全酯酶催化活性与SS的无显著性差异,说明其抗药性可能与酯酶无关。     

虫酰肼和甲维盐对甜玉米草地贪夜蛾的生物活性及田间防效

为了研究虫酰肼和甲氨基阿维菌素苯甲酸盐(甲维盐)对新入侵我国的草地贪夜蛾毒杀活性并为田间应用提供依据,采用浸叶法进行了室内毒力测定,采用喷雾法进行盆栽试验和田间小区试验。毒力测定表明虫酰肼对草地贪夜蛾2龄、3龄幼虫48 h LC_(50)分别为0.30 mg/L和0.26 mg/L,甲维盐对草地贪夜蛾2龄、3龄幼虫48 h LC_(50)分别为0.005 mg/L和0.002 mg/L,其LC_(50)活性是虫酰肼的60～128倍。盆栽试验表明,20%虫酰肼SC在20 mg/L、5%甲维盐ME 1mg/L喷雾处理后7 d后防效均为100%,在甜玉米田间小区试验最高防效均可达到90%以上。本研究结果表明,虫酰肼可推荐用于草地贪夜蛾的防治并可与甲维盐轮换使用或复配使用。     

虫酰肼对甜菜夜蛾汰选种群生长发育及生殖的影响

研究虫酰肼对甜菜夜蛾汰选种群生长发育及生殖的影响。结果显示:经虫酰肼处理后,甜菜夜蛾SS种群和RR种群卵的孵化抑制中浓度分别为179.38和160.71mg/kg,孵化幼虫的存活抑制中浓度分别为18.64和30.53mg/kg,虫酰肼对甜菜夜蛾卵的孵化率影响较小,而对孵化后幼虫的存活率影响较大。处理初孵幼虫6d后,2种群幼虫存活率及化蛹率随处理浓度的增加而降低,幼虫历期最高饲毒浓度与最低饲毒浓度相比分别延长了2.71d和6.19d,同一浓度下,RR种群幼虫历期比SS种群缩短了0.94～2.59d。处理甜菜夜蛾3龄幼虫12h后,随虫酰肼浓度的增加,2种群化蛹率逐渐降低,雌雄蛹重均有递减的趋势,雌蛹比例随浓度的增加变化较小,但SS种群雌蛹比例≤50.00%,而RR种群雌蛹比例≥50.00%,雌雄成虫羽化率总体上依次递减,处理浓度大于1.00mg/kg时虫酰肼对甜菜夜蛾成虫羽化的影响较大。     

甜菜夜蛾虫酰肼抗性品系选育及相对适合度研究

本试验在室内对甜菜夜蛾Spodoptera exigua(Hübner)武汉敏感品系进行了8代群体汰选后分别于9、10、12代采用了单对汰选。结果表明虫酰肼处理后各单对品系后代幼虫存活率发生明显分离,F9(s1)、F10(s2)和F12(s3)代幼虫平均存活率分别为48.34%、11.72%和2.37%,筛选后分别为84.00%、83.33%和7.29%。经过12代汰选后,汰选品系抗性倍数为敏感品系的5.24倍,且单对汰选世代对虫酰肼的抗性发展较快,其抗性倍数分别是群体汰选的1.27、1.67和1.38倍。比较单对汰选世代与群体汰选世代某些生物学特性,显示单对汰选世代并不存在生长发育和生殖不利性,表明在虫酰肼群体汰选中穿插几代单对汰选可以加速抗性品系的选育。汰选品系相对于敏感品系具有0.31的适合度,表明甜菜夜蛾对虫酰肼产生抗性后存在适合度缺陷。     

Inhibitory effects of tebufenozide on dopa decarboxylase and tyrosine hydroxylase in 5th instar larvae of Spodoptera exigua (Hiibner) ( Lepidoptera: Noctuidae)

虫酰肼模拟昆虫蜕皮激素的作用干扰新表皮的形成.为了探讨虫酰肼对昆虫新表皮形成的影响是否与抑制表皮形成相关酶的活性有关,本研究应用高效液相色谱-荧光检测法(HPLC-RP),测定了甜菜夜蛾Spodoptera exigua5龄幼虫用虫酰肼处理不同时间(24,48和72 h)后多巴脱羧酶和酪氨酸羟化酶的活性.结果表明:用LCu( 28.41 μmol/L)和LC33 (85.23 μmol/L)两个亚致死剂量的虫酰肼处理5龄幼虫后,多巴脱羧酶和酪氨酸羟化酶的活性均受到明显抑制,高浓度的抑制作用大于低浓度的抑制作用.随着处理时间的延长,同一剂量的抑制作用逐渐增强.进一步测定虫酰肼处理24,48和72 h后5龄幼虫血淋巴、脂肪体、中肠、表皮和头部的多巴脱羧酶和酪氨酸羟化酶的活性,可看出虫酰肼对幼虫不同组织的多巴脱羧酶和酪氨酸羟化酶的活性也具有相似的抑制作用.结果提示,虫酰肼对甜菜夜蛾幼虫多巴脱羧酶和酪氨酸羟化酶活性具有明显抑制作用,幼虫新表皮形成受阻可能与虫酰肼抑制多巴脱羧酶和酪氨酸羟化酶的活性有关.     

甜菜夜蛾对虫酰肼的抗性选育、风险评估及交互抗性

虫酰肼是目前防治甜菜夜蛾Spodoptera exigua（Hübner）的一种主要杀虫剂,为评估其抗性风险,在室内进行了抗性筛选和交互抗性的研究。采用饲料感染法,在甜菜夜蛾饲养75代期间用虫酰肼筛选62代,与起始种群相比抗性上升39.2倍,与室内敏感品系相比抗性上升141.3倍。在筛选的早、中、后期,现实遗传力h²分别为0.1075（F₀～F₂₅）、0.2780（F₂₆～F₅₀）和0.0538（F₅₁～F₇₅）,整个筛选62代现实遗传力为0.1556。抗性种群筛选43次（F55）后停止用药筛选,饲养21代后,与敏感品系相比,抗性水平由63.5倍下降到21倍,抗性下降3倍。说明甜菜夜蛾具有对虫酰肼产生抗性的风险,且抗性衰退缓慢,短期内很难恢复到敏感水平。交互抗性测定结果表明,上述室内选育的抗性品系对甲氧虫酰肼具有71.4倍的高水平交互抗性,对阿维菌素具有13.1倍的中等水平交互抗性,对甲维盐、茚虫威和呋喃虫酰肼分别具有7.0、8.4和4.7倍的低水平交互抗性,但对溴虫腈交互抗性不明显（1.9倍）。结果提示： 间断交替使用虫酰肼可以延缓抗性的发展,但除溴虫腈外,虫酰肼和其他几种新型杀虫剂之间的轮用可能不是甜菜夜蛾抗性治理的有效策略。     

虫酰肼对甜菜夜蛾多巴脱羧酶和酪氨酸羟化酶的抑制作用

虫酰肼模拟昆虫蜕皮激素的作用干扰新表皮的形成。为了探讨虫酰肼对昆虫新表皮形成的影响是否与抑制表皮形成相关酶的活性有关, 本研究应用高效液相色谱-荧光检测法（HPLC-RP）, 测定了甜菜夜蛾Spodoptera exigua 5龄幼虫用虫酰肼处理不同时间（24, 48和72 h）后多巴脱羧酶和酪氨酸羟化酶的活性。结果表明： 用LC₁₁ （28.41 μmol/L）和LC₃₃ （85.23 μmol/L）两个亚致死剂量的虫酰肼处理5龄幼虫后, 多巴脱羧酶和酪氨酸羟化酶的活性均受到明显抑制, 高浓度的抑制作用大于低浓度的抑制作用。随着处理时间的延长, 同一剂量的抑制作用逐渐增强。进一步测定虫酰肼处理24, 48和72 h后5龄幼虫血淋巴、 脂肪体、 中肠、 表皮和头部的多巴脱羧酶和酪氨酸羟化酶的活性, 可看出虫酰肼对幼虫不同组织的多巴脱羧酶和酪氨酸羟化酶的活性也具有相似的抑制作用。结果提示, 虫酰肼对甜菜夜蛾幼虫多巴脱羧酶和酪氨酸羟化酶活性具有明显抑制作用, 幼虫新表皮形成受阻可能与虫酰肼抑制多巴脱羧酶和酪氨酸羟化酶的活性有关。     

甜菜夜蛾对氯氟氰菊酯抗性的表皮穿透机理

用三种方法测定了采自南京江浦的甜菜夜蛾Spodoptera exigua对氯氟氰菊酯的抗性。结果表明,抗性水平的次序为3龄幼虫点滴法（5 499.5倍）和5龄幼虫点滴法（3 973.2倍）＞3龄幼虫浸叶法（1 041.6倍）＞5龄幼虫叶片夹毒法（24.7倍）,因此该品系触杀毒力的抗性水平至少为胃毒毒力LD50的160倍。用¹⁴C标记氯氟氰菊酯测定甜菜夜蛾抗性和敏感品系5龄幼虫表皮穿透率结果表明,处理后8h,抗性品系5龄幼虫的表皮穿透率仅为敏感品系幼虫表皮穿透率的55.5%。证实表皮穿透率的降低是产生抗性的一个重要机理。     

甲氧虫酰肼对棉铃虫生长发育的亚致死效应研究

以一个对甲氧虫酰肼具有18.5倍抗性的棉铃虫Helicoverpa armigera(Hübner)种群(R)和其同源对照种群(S)为对象,研究了甲氧虫酰肼对棉铃虫生长发育的亚致死效应。结果表明,甲氧虫酰肼亚致死剂量(LC25)处理R和S种群的3龄初幼虫,对S种群幼虫的生长抑制率(37.30%)显著大于R种群(21.20%)。与空白对照处理相比,致死剂量处理R和S种群后,均表现出生长发育延缓,蛹期延长,化蛹率、羽化率、卵孵化率降低,且对S种群的影响更明显。研究结果表明亚致死剂量的甲氧虫酰肼对棉铃虫存在一定的不利影响,但对甲氧虫酰肼具有一定抗性的棉铃虫种群能够减轻这种不利影响。     

A Comparison of Growth and Development of Three Major Agricultural Insect Pests Infected with Heliothis virescens ascovirus 3h (HvAV-3h)

Ascoviruses are double-stranded DNA viruses that are pathogenic to lepidopteran hosts, particularly noctuid larvae. Infection of a larva is characterized by retarded growth, reduced feeding and yellowish body color. In this paper, we reported the growth and development of three major agricultural noctuid insect pests, Helicoverpa armigera (Hübner), Spodoptera exigua (Hübner) and Spodoptera litura (Fabricius), infected with Heliothis virescens ascovirus 3h (HvAV-3h). Using 10-fold serial dilutions (0 to 7) of HvAV-3h-containing hemolymph to infect S. litura larvae, we found no significant difference in larval mortalities from 0 to 10³-fold dilutions; however, significant differences were observed at 10⁴-fold dilution and above. Using a 10-fold dilution of HvAV-3h-containing hemolymph to infect H. armigera, S. exigua and S. litura larvae, we found that the growth and development were significantly affected. All infected larvae could not pupate; the survival times of treated H. armigera, S. litura and S. exigua larvae were significantly longer than untreated control larvae. Body weight showed significant difference between treated and untreated control group from day 1 after inoculation in H. armigera and S. exigua, but day 2 in S. litura. Additionally, food intake also showed significant difference between treated and untreated control group from day 2 after inoculation in H. armigera and S. litura, but day 3 in S. exigua.     

Monitoring of four major lepidopteran pests in Korean cornfields and management of Helicoverpa armigera

Lepidopterans such as Helicoverpa armigera are emerging pests of corn in Korea, causing huge yield losses and deteriorating the quality of the corn crop. We monitored four major lepidopteran pests in major corn growing areas of Korea by employing sex pheromone traps from 2012 to 2015 to understand population dynamics. H. armigera, Ostrinia furnacalis, Spodoptera litura, Spodoptera exigua and Mythimna separata were collected from April to October. Among these species, H. armigera was found to be the main pest based on crop damage intensity (80–90% of the total damage). H. armigera was generally observed during the earing stage of corn and would migrate to other fields during September to October for overwintering. We also tested select insecticides against larvae of H. armigera in the laboratory. The most effective insecticide was indoxacarb, which was applied at specific times to manage H. armigera in cornfields. Optimal timing of spraying was estimated at about 70 days after planting (earing season) to control for H. armigera. Additionally, two species of parasitoid (Therion circumflexum and Ophioninae sp.) were identified from H. armigera. However, more extensive surveys are needed to organize a control program based on natural enemies.     

Biocontrol Potential of Steinernema thermophilum and Its Symbiont Xenorhabdus indica Against Lepidopteran Pests: Virulence to Egg and Larval Stages

Under laboratory conditions, the biocontrol potential of Steinernema thermophilum was tested against eggs and larval stages of two important lepidopteran insect pests, Helicoverpa armigera and Spodoptera litura (polyphagous pests), as well as Galleria mellonella (used as a model host). In terms of host susceptibility of lepidopteran larvae to S. thermophilum, based on the LC₅₀ 36 hr after treatment, G. mellonella (LC₅₀ = 16.28 IJ/larva) was found to be more susceptible than S. litura (LC₅₀ = 85 IJ/larva), whereas neither host was found to be significantly different from H. armigera (LC₅₀ = 54.68 IJ/larva). In addition to virulence to the larval stages, ovicidal activity up to 84% was observed at 200 IJ/50 and 100 eggs of H. armigera and S. litura, respectively. To our knowledge this is the first report of entomopathogenic nematode pathogenicity to lepidopteran eggs. Production of infective juvenile (IJ) nematodes/insect larva was also measured and found to be positively correlated with rate of IJ for H. armigera (r = 0.990), S. litura (r = 0.892), as well as G. mellonella (r = 0.834). Both Phase I and Phase II of symbiotic bacteria Xenorhabdus indica were tested separately against neonates of H. armigera and S. litura by feeding assays and found to be virulent to the target pests; phase variation did not affect the level of virulence. Thus S. thermophilum as well as the nematode’s symbiotic bacteria applied separately have the potential to be developed as biocontrol agents for key lepidopteran pests.     

Seasonal parasitism and biological characteristics of Habrobracon hebetor (Hymenoptera: Braconidae) – a potential larval ectoparasitoid of Helicoverpa armigera (Lepidoptera: Noctuidae) in a chickpea ecosystem

Seasonal parasitism of Habrobracon hebetor (Say) on Helicoverpa armigera (Hübner) in chickpea was studied for three consecutive years. Parasitism by H. hebetor on larvae of H. armigera reached 12.3%. The parasitoid maintained reproductive activity on H. armigera from February to April coinciding with pod formation and maturation stages of the crop. In laboratory assays, we investigated the suitability of larval instars of H. armigera to the parasitoid H. hebetor. This parasitoid attacked third to sixth instars, though fourth and fifth instar larvae were found most suitable with 100% parasitism and development to adults. Parasitoid developmental time was longest in fifth instar (9.1 days) compared to other instars (8.1–8.9 days). Fifth instar larvae resulted in highest numbers of cocoons and adult emergence. In addition, suitability of seven lepidopteran species to H. hebetor was investigated. Corcyra cephalonica, Galleria mellonella and H. armigera were the most suitable hosts with 100% parasitism and development to adults. It was followed by Maruca vitrata and Autographa nigrisigna with 60–76.7% and 40–70% parasitism and parasitoid developmental success, respectively. Though there was 23.3% parasitism, there was no parasitoid development in Spodoptera litura. No parasitism was recorded in Spilarctia obliqua. Development of H. hebetor was most rapid in C. cephalonica (8.7 days), and longest in G. mellonella (9.3 days). Parasitoids that developed on these hosts resulted in highest numbers of cocoons and adult emergence. The parasitoid could be exploited for the biological control of H. armigera in a chickpea ecosystem.     

A single semi‐synthetic diet with improved antimicrobial activity for mass rearing of lepidopteran insect pests of cotton and maize

Helicoverpa armigera (Hübner), Earias vittella (Fabricius), Spodoptera litura (Fabricius), Spodoptera exigua (Hübner) (all Lepidoptera: Noctuidae), Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), and Chilo partellus (Swinhoe) (Lepidoptera: Crambidae) are the major pests of cotton and maize. Mass‐rearing of these insects under controlled conditions is necessary to obtain the numbers needed to conduct bioassays to screen insecticides, proteins, and other compounds, as tools for insect pest management. We present a diet suitable for rearing the six lepidopteran pests (five cotton and one maize pest). We further show that this diet is on par with or superior to the published diet recipes for each of the insect species, which were studied for three generations. We also discuss the advantages of antimicrobials other than formalin for keeping microbial growth under check. A combination of antimicrobial solution and benomyl provided effective control and suppressed the growth of microbes for a longer period than a formalin‐containing diet. A common diet for six pests provide opportunities for automation of diet preparation in addition to improved throughput and consistency in the process, while eliminating diet‐batch related errors.     

Consumption and utilization of a soyflour-wheat germ diet by larvae of the tobacco budworm parasitized by the tachinidEucelatoria sp

The number ( \(\bar X\) =2.4) ofEucelatoria sp. maggots that completed development in 4th- or 5th-instar larvae of the tobacco budworm (TBW),Heliothis virescens (F.), was significantly greater (P<0.05) than the number ( \(\bar X\) =1.2) that completed development in 3rd-instar larvae. Maggot development time decreased with increasing number of maggots per host larva. It also decreased with advancing larval instars. The range was 6.9±1.1 days in early 3rd-instar TBW larvae and 5.0±0.8 days in early 5th-instar TBW larvae. Unparasitized 3rd- or 4th-instar TBW larvae consumed significantly more food than did similar aged larvae parasitized byEucelatoria sp., but larvae parasitized during the early 5th-instar consumed more food than did similar aged unparasitized larvae. Consumption by 4th- or 5th-instar larvae increased significantly as maggot densities increased from 1 to 3 per host larva, but decreased at a density of 4 or more maggots per host larva. Although body weight gain and consumption were both significantly reduced 48 and 120 h after parasitization of late 3rd-instar larvae (6 days old), the approximate digestibility (AD) value was significantly greater for parasitized than for unparasitized larvae. Unparasitized larvae were more efficient in converting digested food to body substance (ECD) than parasitized larvae, but the efficiency in conversion of ingested food to body substance (ECI) was similar for both parasitized and unparasitized larvae.     

Selective Feeding of <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> (Hübner) and <Emphasis Type="Italic">Spodoptera litura</Emphasis> (Fabricius) on Meridic Diet with <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> Toxins

Laboratory experiments were conducted to evaluate the behavior of Helicoverpa armigera (Hübner) and Spodoptera litura (Fabricius) larvae on meridic diet with different concentrations of Bt spray formulation Delfin or isolated Cry1Ac protein or the foliage and bolls from transgenic cotton, Bollgard hybrid RCH-317 Bt. Both insect species selectively fed on nontreated diet compared with the diet treated with Delfin. While H. armigera exhibited concentration response with Cry1Ac, this protein did not affect S. litura larvae. In general Helicoverpa selected diet with low concentrations (EC₂₀ and EC₅₀ levels) of Cry1Ac compared with higher concentrations of Cry1Ac. In order to develop appropriate management strategies, a thorough understanding of the behavioral mechanisms leading to the responses of insects to the proteins in transgenic varieties is required. Thus, based on results of the insects fed individually on the leaf discs or bolls from transgenic cotton plants alone or under choice situation with meridic diet revealed that H. armigera larvae preferred meridic diet to transgenic leaves or bolls expressing Cry1Ac protein. H. armigera larvae preferred meridic diet to plant material; more than 70% larvae were seen on the meridic diet, and average larval weight gain was in the range of 121.7–130.5 mg. However, in case of S. litura the larvae showed no significant discrimination between meridic diet and the leaf discs. In fact more than 60% larvae preferred leaf discs for feeding, though Cry1Ac expression in leaf discs was in the range of 0.9–2.18 μg/g. Thus differences in behavioral response could potentially impact the level of efficacy of crop cultivars that have been genetically engineered to produce these proteins.     

甜菜夜蛾抗高效氯氟氰菊酯近等基因系的构建

将甜菜夜蛾Spodoptera exigua抗性种群雄成虫与敏感种群雌成虫杂交,杂交后代再自交后,用高效氯氟氰菊酯杀死敏感和部分杂合个体的剂量(50 μg/mL)处理4龄幼虫,对1天后存活的幼虫再用250 μg/mL汰选,将存活幼虫发育成的雄成虫与敏感种群的雌成虫回交,再经自交和选育,如此循环6次,获得了甜菜夜蛾抗高效氯氟氰菊酯近等基因系。对该基因系构建过程中各代幼虫的抗性水平和相关生物学特性进行了监测和比较,结果表明获得的抗高效氯氟氰菊酯近等基因系抗性种群的抗药性仍保持较高水平;酯酶同工酶的电泳分析表明抗高效氯氟氰菊酯近等基因系抗性种群与敏感种群图谱相近,而与亲本抗性种群之间存在明显差异。