草地贪夜蛾与斜纹夜蛾解毒相关基因的比较分析

草地贪夜蛾Spodoptera frugiperda近期在中国为害猖獗,由于繁殖速度快,迁飞能力强,在本土呈现爆发趋势,严重为害了我国农作物。与本地近缘物种斜纹夜蛾Spodoptera litura相比,草地贪夜蛾更偏好玉米、水稻、小麦等禾本科农作物,且对多种化学杀虫剂及转基因Bt玉米产生抗性。寄主适应性以及杀虫剂抗性与解毒代谢相关蛋白密切相关。因此,本研究对这两种夜蛾科害虫的解毒代谢相关蛋白——细胞色素P450、谷胱甘肽转移酶(GST)及ABC转运蛋白进行了全基因组水平系统的搜集和数目比较,构建系统发育树并对P450和GST部分基因扩张分支进行氨基酸差异位点分析。结果显示,在草地贪夜蛾中共鉴定出213个P450基因、58个GST基因、102个ABC基因,其中P450基因与GST基因数目远远多于斜纹夜蛾(116,37),而ABC基因数目与斜纹夜蛾(99)接近。系统发育树分析表明,草地贪夜蛾P450在CYP6、CYP9以及CYP4功能簇,GST在部分进化分支上都发生了显著基因扩张,发生显著扩张的基因中有数个氨基酸突变,其中一些突变被预测可能影响蛋白质功能。但出乎意料的是,ABC亚家族B和E在斜纹夜蛾聚集成簇并发生了显著的基因扩张现象。以上结果暗示入侵种草地贪夜蛾和本地近缘物种斜纹夜蛾在抗性方面可能发展出各自独特的应对机制。本研究为解析草地贪夜蛾解毒抗性特征机制提供基因数据,为草地贪夜蛾的生物防治及抗性研究提供参考依据。     

草地贪夜蛾与斜纹夜蛾嗅觉味觉相关基因的比较分析

草地贪夜蛾Spodoptera frugiperda,近期侵入中国并迅速扩张。与其近缘物种斜纹夜蛾Spodoptera litura不同,草地贪夜蛾主要偏好玉米、水稻、小麦等禾本科农作物,暴食危害重。此外两种害虫在交配、产卵频率等繁殖特征上也存在差异。这些差异可能与嗅觉、味觉相关蛋白的分歧密切相关。本研究对两种夜蛾科害虫的嗅觉、味觉相关基因进行了系统的鉴定及系统发育分析,并对部分嗅觉结合蛋白进行了选择信号与氨基酸差异位点分析。分别在草地贪夜蛾中鉴定到261个嗅觉相关基因,数目明显大于斜纹夜蛾(179个);鉴定到233个味觉相关基因,数目略大于斜纹夜蛾(217个)。嗅觉相关基因中气味结合蛋白(OBP)基因数目差异最为明显。系统发育分析显示该基因存在夜蛾科特异的分支,且该分支中草地贪夜蛾中基因数目远高于斜纹夜蛾;发现位于I类触角结合蛋白(ABPI)分支的一类OBP成员在草地贪夜蛾中拷贝数增加,其中两个基因拷贝受到正选择,且在草地贪夜蛾中特有的氨基酸替换还可能影响蛋白质功能。本研究为草地贪夜蛾食性特征的遗传机制解析提供线索,为草地贪夜蛾的生物防治提供参考依据和基因素材。     

斜纹夜蛾（Spodoptera litura）的胚胎发育

本文利用实验胚胎学方法详细研究了斜纹夜蛾胚胎发育的全过程。结果表明：在２４±１℃的恒温条件下,仅需８７小时即可孵化出幼虫,同时证明了马氏管起源于外胚层。     

Interaction of Xestia c-nigrum granulovirus with peritrophic matrix and Spodoptera litura nucleopolyhedrovirus in Spodoptera litura

Xestia c-nigrum granulovirus (XcGV) was tested for its ability to increase Spodoptera litura nucleopolyhedrovirus (SINPV) infection in larvae of S. litura (F.). The interaction of XcGV with peritrophic matrix and SINPV in S. litura also was studied to account for the synergism. In dose-response bioassays with a constant XcGV concentration of 5-mg/ ml capsules and SINPV concentration that varied from 10(3) to 10(7) polyhedral inclusion bodies (PIB) per larva, XcGV increased the virulence of SINPV infection in fifth instars of S. litura. The lethal concentration of 50% individuals (LC50) of SINPV combined with XcGV was 3.35 x 10(5)PIB/ml, which was significantly lower than that of SINPV alone (2.17 x 10(6)). Compared with 10(7) PIB/ml SINPV alone, the lethal time of 50% individuals (LT50) of 10(7) PIB/ml SINPV combined with XcGV was not significantly shortened. In addition, no significant improvement in the activity and killing speed of SINPV progeny was noted after propagation with XcGV, indicating that native characters of SINPV associated with viral potency were not altered by XcGV. Investigation via environmental scanning electronic microscopy showed that the peritrophic matrix (PM) of S. litura exposed to XcGV or XcGV enhancin, or the combination treatment, was markedly disrupted. The outer surface of the PM was loose, or ruptured, which potentially facilitated the passage of virions through the PM. Therefore, it is reasonable to conclude that the synergy between XcGV and SINPV was closely associated with the disruption of the PM in S. litura.     

Significance of penetration and metabolism on topical toxicity of diflubenzuron in Spodoptera littoralis and Spodoptera exigua

Toxicity assays indicated that when diflubenzuron (DFB) was topically applied on larvae of the Egyptian cotton leafworm, Spodoptera littoralis (Boisduval), and the beet armyworm, Spodoptera exigua (Hübner), estimated LD₅₀ values were similar, reaching 0.47 and 0.44 ng/larva, respectively. This study evaluated the importance of the rate of uptake and excretion, and of enzymatic metabolism in building up an insecticidal toxicity after topical application on the insect cuticle in both species. In general, penetration of DFB in S. littoralis was about 2-fold higher than in S. exigua, whereas metabolic breakdown was of minor importance in S. littoralis as compared with S. exigua. The current data indicate the importance of cuticle penetration and discuss that extrapolation from oral feeding toxicity results to topical toxicity data should be evaluated with caution, especially for insect growth regulators used in integrated pest management.     

CO2浓度升高对草地贪夜蛾与斜纹夜蛾生长发育及繁殖的影响

为明确大气CO2浓度升高对草地贪夜蛾Spodoptera frugiperda及其本地近缘种斜纹夜蛾Spodoptera litura生长发育和繁殖的影响。本研究在正常二氧化碳浓度(400 μL/L)和倍增二氧化碳浓度(800 μL/L)下,用寄主植物小麦饲养草地贪夜蛾和斜纹夜蛾,研究CO2浓度升高对草地贪夜蛾及斜纹夜蛾发育历期和繁殖力的直接影响(高CO2浓度直接影响试虫)、间接影响(高CO2浓度通过影响小麦间接影响试虫)及综合影响(小麦和试虫同时受高CO2浓度影响)。结果表明,CO2浓度升高对草地贪夜蛾各虫态的发育历期均有较显著影响,完成整个世代的时间显著缩短,其中间接影响组幼虫期较对照组显著延长0.7 d,而蛹期和成虫期寿命分别显著缩短1.5 d和4.9 d;综合影响组幼虫期较对照组显著延长0.8 d,而蛹期和成虫期寿命分别显著缩短1.2 d和5.7 d;直接影响组草地贪夜蛾蛹期显著缩短2.7 d;CO2浓度升高对斜纹夜蛾各虫态发育历期影响不显著,仅直接影响组的各虫态发育历期缩短较为明显。在CO2浓度升高条件下,草地贪夜蛾直接影响组单雌产卵量显著增加1 220.48粒,综合影响组略有减少;而斜纹夜蛾单雌产卵量在高CO2浓度下均呈现下降趋势,其中综合影响显著减少818.20粒。说明CO2浓度升高加快了草地贪夜蛾的种群增长速率,使其完成整个世代的时间显著降低;但对本地近缘种斜纹夜蛾的影响不显著。因此,在未来CO2浓度升高的情况下,入侵种草地贪夜蛾的种群增长速率将大于其本地近缘种斜纹夜蛾。     

草地贪夜蛾核型多角体病毒对我国草地贪夜蛾不同地理种群毒力的比较分析

草地贪夜蛾Spodoptera frugiperda(J. E. Smith)入侵我国多个地区,逐渐形成地理种群。在草地贪夜蛾核型多角体病毒Spodoptera frugiperda multiple nucleopolyhedrovirus(SfMNPV)生物农药的生产过程中发现,不同地理来源的宿主对SfMNPV的敏感性和产量存在明显差异,显著影响了SfMNPV的生产效率。为解析其敏感性差异及其产生的原因,本研究首先对云南德宏、广东广州、广西钦州、西藏林芝4个地区草地贪夜蛾种群基因型进行了鉴定,采用生物测定法测试SfMNPV对4龄幼虫的口服毒力,然后,通过向幼虫体内注射草地贪夜蛾核多角体病毒出芽型病毒粒子(budded virions, BVs)的方式,越过口服感染中肠的过程,分析敏感性差异发生的阶段。最后比较了高敏感和低敏感种群中肠肠液pH,并基于16S rDNA测序测定了肠道菌群组成。结果表明,广西种群属于纯合玉米型,其余种群为带有水稻型COI标记的杂合玉米型。广西种群对SfMNPV的口服敏感性最低,西藏种群的敏感性最高,但两者注射BV后死亡率差异无统计学意义,暗示病毒敏感性差异...     

溴虫氟苯双酰胺对草地贪夜蛾的亚致死效应

为明确溴虫氟苯双酰胺对草地贪夜蛾Spodoptera frugiperda的亚致死效应,采用点滴法测定溴虫氟苯双酰胺对草地贪夜蛾3龄幼虫的室内毒力及亚致死剂量溴虫氟苯双酰胺(LD₁₅和LD₃₀)对草地贪夜蛾生物学特性的影响,同时测定了亚致死剂量(LD₁₅、LD₃₀)和半致死剂量(LD₅₀)溴虫氟苯双酰胺对草地贪夜蛾3龄幼虫体内解毒酶(GST、EST、MFO)、保护酶(SOD、POD、CAT)活性的影响。结果表明,溴虫氟苯双酰胺对草地贪夜蛾3龄幼虫具有良好的触杀活性,LD₅₀值为0.50μg/g。亚致死剂量(LD₁₅和LD₃₀)溴虫氟苯双酰胺处理草地贪夜蛾3龄幼虫后,幼虫存活率显著下降,较对照分别下降了6.7%和32.88%;LD₃₀处理的成虫畸形率(8.33%)较对照(1.75%)显著升高,产卵前期(3.31 d)较对照(2.88 d)显著延长。溴虫氟苯双酰胺3个不同剂量(LD_15...     

甜菜夜蛾发生规律及其防治研究

甜菜夜蛾在福建漳州龙海每年发生9－10年,3月中下旬开始发生,4－10月份是该虫为害农作物的严重时期,其中8－9月份为全年发生危害的高峰期,通过田间药效试验,评价了不同类型的12种杀虫剂对甜菜夜蛾的防治效果,结果表明,除尽,菜喜以及米满对田间甜菜夜蛾具有较好的防效,锐颈特,杀虫单,敌杀死等药剂对田间甜菜夜蛾几乎失去防效。     

斜纹夜蛾天敌作用的评价

通过组建斜纹夜蛾第4代和第8代自然种群生命表,运用排除作用控制指数分析了生物因子对斜纹夜蛾种群的自然控制作用。结果表明,低龄(1～3龄)幼虫的捕食性天敌是影响斜纹夜蛾种群数量动态的重要因子。对第4代和第8代种群的排除作用控制指数分别为13.904和12.946.如果没有捕食性天敌的作用,下代种群数量将分别增长到当代的15.1206和74.678倍。病原微生物是影响第4代斜纹夜蛾种群数量的另一重要因子,其排除作用控制指数为2.4726.     

Intermediates of stigmasterol metabolism in Spodoptera littoralis

Stigmasterol-24,28-epoxide, 22E-stigmasta-5,22,24(28E)-trien-3 beta-ol, and 22E-cholesta-5,22,24-trien-3 beta-ol were identified as normal metabolites of [3H]stigmasterol in Spodoptera littoralis larvae. Relative concentrations of all three of these metabolites increased when a diazasterol inhibitor was fed in combination with stigmasterol in the artificial diet. Identification of these sterols as intermediates in the conversion of stigmasterol to cholesterol in this insect indicates that intermediates analogous to fucosterol and fucosterol-24,28-epoxide in the conversion of sitosterol to cholesterol are produced in the metabolism of stigmasterol. This is the first published identification of stigmasterol-24,28-epoxide and 22E-stigmasta-5,22,24(28E)-trien-3 beta-ol as intermediates in this pathway in an insect.     

Electron microscope study on the replication of Autographa nuclear polyhedrosis virus and Spodoptera nuclear polyhedrosis virus in Spodoptera exigua

A comparative study of Spodoptera nuclear polyhedrosis virus (NPV) and Autographa NPV replication in Spodoptera exigua revealed some cytopathologic differences. Infection with Spodoptera NPV was accompanied by electron-dense intranuclear granules. Autographa infection within the midgut led to secretion within the lumens of the goblet cells of paracrystalline arrays of small, round particles, 9.5 nm in diameter. Autographa virus was also observed in various stages of possible replication within the cytoplasm.     

红彩瑞猎蝽对草地贪夜蛾幼虫的控害能力

为明确红彩瑞猎蝽Rhynocoris fuscipes Fabricius成虫对草地贪夜蛾Spodoptera frugiperda J. E. Smith幼虫的控害潜能,室内条件下研究了不同饥饿时间处理红彩瑞猎蝽对草地贪夜蛾3龄幼虫的捕食功能反应、自身密度干扰效应和搜寻效应。结果表明,饥饿0 h、24 h、48 h、72 h和96 h条件下,红彩瑞猎蝽雌、雄成虫对草地贪夜蛾3龄幼虫的功能反应符合Holling Ⅱ型功能反应方程;饥饿程度对红彩瑞猎蝽24 h内捕食量影响明显,饥饿48 h和72 h处理的红彩瑞猎蝽对草地贪夜蛾具有更旺盛的捕食欲,其捕食速度曲线分别为v=0.9047-0.0592t+0.009t2和v=1.1512-0.0702t+0.0011t2,对草地贪夜蛾的捕食量随着时间的变化模型分别为Na=20/（1+e1.968-0.79t)和Na=20/（1+e1.689-0.93t）。不同饥饿程度红彩瑞猎蝽搜寻效应随猎物密度的增加而逐渐降低,其个体间竞争和干扰作用与天敌自身密度变化呈正比,符合Hassell-Varley模型。结果表明,红彩瑞猎蝽成虫对草地贪夜蛾幼虫有较好的防治潜力。     

斜纹夜蛾天敌作用的评价

通过组建斜纹夜蛾第４代和第８代自然种群生命表,运用排除作用控制指数分析了生物因子对斜纹夜蛾种群的自然控制作用．结果表明,低龄（１～３龄）幼虫的捕食性天敌是影响斜纹夜蛾种群数量动态的重要因子．对第４代和第８代种群的排除作用控制指数分别为１３．９０４和１２．９４６．如果没有捕食性天敌的作用,下代种群数量将分别增长到当代的１５．１２０６和７４．６７８倍．病原微生物是影响第４代斜纹夜蛾种群数量的另一重要因子,其排除作用控制指数为２．４７２６．     

甜菜夜蛾幼虫饲养技术的改进

利用一次性塑料杯饲养甜菜夜蛾SpodopteraexiguaH櫣bner,一次性接入初孵幼虫,直至化蛹不再转移,也不再更换饲料。与已报道的方法相比,该方法操作简单,省工省时,可以提高工作效率。在各项饲养指标中,除平均蛹重略微偏低之外,其余指标均与已报道的数据基本一致。试验证明,采用该方法可以满足室内甜菜夜蛾的继代繁殖和批量饲养。     

草地贪夜蛾生殖行为及其昼夜节律研究

草地贪夜蛾Spodoptera frugiperda是当前跨国迁飞性重大害虫,云南是其重要虫源地及其向内地迁移的重要通道。生殖行为是性选择和物种进化的重要基础,基于生殖行为的害虫监测和防治策略也有重要价值。本研究模拟自然条件下可能出现的择偶情况,采用永久配对和每日更换配偶两种处理,对草地贪夜蛾成虫一生(约10 d)的生殖行为和节律进行了连续观测。结果表明,该蛾在羽化当日(0日龄)即出现召唤和求偶行为但无交配发生,在1日龄开始出现交配行为并在2日龄开始产卵,从2日龄到6日龄期间一直保持了较高的生殖活动,之后开始下降。雄虫求偶与雌虫召唤在节律上有较高的同步性,交配高峰多出现在召唤和求偶高峰之后。每日更换配偶与永久配对相比,雌蛾表现出较高的求偶频率和持续时间,较高交配比率和频率。雌虫在遇到新配偶或与新配偶交配后,会延缓产卵或下调产卵速率。从性选择理论来看,这些行为策略将有助于雌虫获得物质或基因利益,值得进一步探究。交配次数、召唤时长与雌虫寿命之间相关性不显著,但前5 d产卵速率与雌虫寿命间存在显著负相关,即早期产卵量较高的雌虫寿命较短,符合早期繁殖-后期存活基因拮抗假说。     

Predation behavior of Podisus nigrispinus on Spodoptera eridania

The behavior and effective predation time can affect the prey death in pest biological control programs. This work studied the Podisus nigrispinus (Dallas, 1851) (Heteroptera: Pentatomidae) behavior on Spodoptera eridania (Cramer, 1782) (Lepidoptera: Noctuidae) caterpillars, and its implications in case of prey escape. The preference bioassay (B1) aimed to verify the caterpillars body region (anterior: head and thorax; median and posterior) preferred by the predator and its implication in prey mortality. The predation duration bioassay considered the following effective predation times: 1, 2, 4, 8, 16, 32, 64 and 128 min; the caterpillars were removed after each predation time, to simulate prey escape, and the dead were counted until the seventh day. This experiment was performed in two ways: with randomly selected and not repeated predators (B2); and with the same predators in successive times (B3). The predator preferred to attack the caterpillars anterior region. The caterpillars mortality increased with increasing effective predation time. The mortality was 90% after 64 min under B2. This value was estimated for 16 min under B3. The P. nigrispinus prefers to attack the caterpillars anterior region and mortality of S. eridania caterpillars was favored in predators that have suffered predation interruption.     

甜菜夜蛾的饲养方法介绍

介绍一种甜菜夜蛾的室内饲养方法 ,采用本方法累代饲养甜菜夜蛾 8代 ,平均蛹重达 1 1 2 .9mg ,平均化蛹率 97% ,平均羽化率为 91 .3% ,室内饲养结果表明 ,该饲养方法可用于室内大量饲养甜菜夜蛾     

Molecular and phylogenetic characterization of Spodoptera litura granulovirus

Baculovirus, Spodoptera litura granulovirus (SlGV) was isolated from the infected S. litura larvae, and was characterized. The granule of SlGV was ovoidal shape with an approximate size of 240∼340 nm× 140∼180 nm. Each granule contained one single rod-shape virion with a mean size of 180∼200 nm×20∼40 nm. Restriction endonuclease fragment analysis estimated that the total genome size of SlGV is about 115 kb. Necleotide sequence analysis of the granulin gene showed that the gene encodes 249 amino acids with a predicted molecular mass of 29 kDa. When the phylogenic relationship was analyzed using the nucleotide sequence of the granulin gene, SlGV was most closely related to Trichoplusia ni granulovirus (TnGV) and Xestia c-nigrum granulovirus (XcGV) which belong to Type I granulovirus.