斜纹夜蛾发生期预测预报的探讨——有效积温预测法

根据斜纹夜蛾发育起点温度和有效积温,探讨了斜纹夜蛾发生期预测方法。结果表明,田间斜纹夜蛾发生高峰期与预测的日期基本吻合。     

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。     

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。     

几种虫生真菌对斜纹夜蛾的致病性

用布氏白僵菌、球孢白僵菌、玫烟色拟青霉、绿僵菌和莱氏野村菌5种真菌的固体培养物,对斜纹夜蛾2、3龄幼虫进行了毒力测定.结果表明：布氏白僵菌和莱氏野村菌两种菌株对斜纹夜蛾幼虫有明显的致病效果,对2龄幼虫的致死中时(LT₅₀)分别为2.95 d和4.10 d,累计校正死亡率分别为100%和95.2%;对3龄幼虫的致病力低于2龄,致死中时(LT₅₀)分别为19.67 d和19.63 d,累计校正死亡率分别为56.6%和52.2%.玫烟色拟青霉、球孢白僵菌两菌株也有一定的致病力,对2龄幼虫的致死中时(LT₅₀)分别为4.89 d 和6.34 d,累计校正死亡率分别为85.7%和71.4%.     

斜纹夜蛾性信息素诱捕器田间应用技术

考查斜纹夜蛾Spodoptera litura(Fabricius)性信息素诱捕器放置高度、间隔距离以及气象因子对性信息素诱集效果的影响。结果表明,放置高度为1m时,性信息素诱捕器易引起斜纹夜蛾的反应,明显优于0.5m和1.5m的诱集效果。诱捕器放置的间隔距离,以25m为诱集效果最佳,与10,15,20及30m的具有显著差异。夜间风向对诱捕器的诱蛾效果具有明显影响,在3个成一列放置的诱捕器中,诱集量最大的是放在上风口位置的诱捕器;同时,与夜间风向平行设置的诱捕器,其诱集量显著高于与夜间风向垂直设置诱捕器的诱集量。     

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。     

斜纹夜蛾空间分布图式的研究

斜纹夜蛾幼虫在不同作物上的空间分布特征研究表明：幼虫在低龄阶段高度聚集,聚集的集团是成虫产卵的行为造成。在高龄阶段,聚集度下降,幼虫不断扩散,扩散的原因是密度制约因素引起。在白菜上,幼虫在4龄前为聚集分布,5龄以后为均匀分布,棉花和向日葵上各龄幼虫均为聚集分布。根据种群空间分布图式,导出了数据代换公式。     

Characterization of the active fragments of Spodoptera litura Lebocin-1

Insects can produce various antimicrobial peptides (AMPs) upon immune stimulation. One class of AMPs are characterized by their high proline content in certain fragments. They are generally called proline-rich antimicrobial peptides (PrAMPs). We previously reported the characterization of Spodoptera litura lebocin-1 (SlLeb-1), a PrAMP proprotein. Preliminary studies with synthetic polypeptides showed that among the four deductive active fragments, the C-terminal fragment SlLeb-1 (124-158) showed strong antibacterial activities. Here, we further characterized the antibacterial and antifungal activities of 124-158 and its four subfragments: 124-155, 124-149, 127-158, and 135-158. Only 124-158 and 127-158 could agglutinate bacteria, while 124-158 and four subfragments all could agglutinate Beauveria bassiana spores. Confocal microscopy showed that fluorescent peptides were located on the microbial surface. Fragment 135-158 lost activity completely against Escherichia coli and Staphylococcus aureus, and partially against Bacillus subtilis. Only 124-149 showed low activity against Serratia marcescens. Negative staining, transmission, and scanning electron microscopy of 124-158 treated bacteria showed different morphologies. Flow cytometry analysis of S. aureus showed that 124-158 and four subfragments changed bacterial subpopulations and caused an increase of DNA content. These results indicate that active fragments of SlLeb-1 may have diverse antimicrobial effects against different microbes. This study may provide an insight into the development of novel antimicrobial agents.     

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.     

印楝素乳油对斜纹夜蛾的生物活性及田间防效研究

在室内测定了印楝素乳油对斜纹夜蛾(Spodopteralitura)的生物活性,并进行了田间防治试验.结果表明,印楝素乳油对斜纹夜蛾具有多种生物活性,主要表现在对幼虫的拒食作用、生长发育的抑制作用和一定的毒杀活性,以及对成虫产卵的忌避作用.经印楝素处理后,幼虫平均发育历期延长、幼虫体重、蛹重、化蛹率和蛹羽化率显著低于对照,印楝素乳油12μl·L^-1浓度处理3龄幼虫5和7d后的体重分别是对照组的36.8%和14.5%,而幼虫平均发育历期比对照组延长6d左右,化蛹率和蛹羽化率分别是27.79%和37.23%,显著低于对照组85.66%和82.11%.印楝素对低龄幼虫有一定的毒杀活性,而对高龄幼虫活性则很低.田间防治试验表明,印楝素乳油对斜纹夜蛾种群有良好的控制作用,施药7d后其防治效果要优于化学农药10%除尽悬浮剂.     

Flight activity of three Spodoptera spp., Spodoptera litura, S. exigua and S. depravata, measured by flight actograph

Flight activities of three Spodoptera species were measured by the aid of flight actograph: S. litura and S. exuiga being regarded as long‐distance migratory insects, and S. depravata being non‐migratory and diapause‐inducible species. In all species tested, flight activities were observed only in scotophase, males showed far higher activities than females, being several times higher at the time of maximum flight activity, which was observed within 2 days after adult eclosion. Total flight activity in males was highest in S. litura, some being flyable even 12 days after eclosion, followed by S. exigua being one‐third compared to the former species, while in S. depravata flight activity was nearly half of that of the second species and most ceased to fly within a week after eclosion. There occurred species‐specific daily rhythms in flight activity during respective scotophase. In S. litura, both females and males exhibited a peak of flight activity shortly after light‐off and exhibited the second flight activity in late scotophase, the females slightly but the males more actively compared to early scotophase. In S. exigua, both sexes did not respond to light‐off, did not show a peak of flight activity in early scotophase, whereas males, but not females prominently increased activity toward the end of scotophase. In S. depravata, both sexes exhibited a peak of flight activity in early scotophase, and the males revived flight activity, being maximum shortly before light‐on, but the females did not show a clear rhythm in flight activity. These features observed in flight activity were discussed in relation with migratory capability.     

Biological and genetic characterization of a Pakistani isolate of Spodoptera litura nucleopolyhedrovirus

Spodoptera litura is an emerging insect pest in a wide range of crops worldwide. The insect is difficult to control because of resistance development to synthetic insecticides and emerging resistance to Bacillus thuringiensis toxins. Therefore, there is a need to develop biological control agents, preferably from an indigenous source to avoid risks associated with the importation of exotic natural antagonists. A Pakistani isolate of S. litura nucleopolyhedrovirus (SpltNPV, Baculoviridae), SpltNPV-Pak-BNG, was obtained from the field and characterized biologically and genetically, and compared to a SpltNPV reference isolate, SpltNPV-G1, thought to be of Chinese origin. The dose–mortality response (LD₅₀) of SpltNPV-Pak-BNG was not significantly different from that of the reference isolate SpltNPV-G1, but the time-to-death (LT₅₀) was significantly shorter for SpltNPV-Pak-BNG than for SpltNPV-G1. DNA restriction enzyme profiling indicated that SpltNPV-Pak-BNG and SpltNPV-G1 are different viruses. Sequence analysis of ‘ORF24’, specific for SpltNPV (and S. littoralis NPV as ORF21), and the conserved baculovirus core genes polyhedrin, DNApol, pif-2 and lef-8 confirmed that this was indeed the case and that SpltNPV-Pak-BNG is a genuine SpltNPV variant, whereas the SpltNPV-G1 isolate we used is, in fact, a SpliNPV variant, renamed to SpliNPV-G1. The newly isolated SpltNPV-Pak-BNG has the potential for development as a biocontrol agent of S. litura in Pakistan.     

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．     

杀菌剂对斜纹夜蛾SL细胞系和幼虫的生物活性

以MTT法筛选了19种杀菌剂对斜纹夜蛾Spodoptera litura SL细胞的毒杀活性,并以该方法研究了三唑类杀菌剂对SL细胞的毒力。结果表明: 福美双、烯唑醇、己唑醇、氟硅唑、苯霜灵、苯醚甲环唑、戊唑醇和腈菌唑等杀菌剂对SL细胞具有优异的毒杀活性。三唑类杀菌剂腈菌唑、烯唑醇、己唑醇和戊唑醇处理SL细胞48 h后,LC₅₀值分别为21.94 μg/mL、 23.80 μg/mL、 33.16 μg/mL和47.63 μg/mL。以考马斯亮蓝G₂₅₀法研究了腈菌唑对SL细胞中蛋白质含量和乳酸脱氢酶(LDH)漏出率的影响, 20 μg/mL腈菌唑处理12 h、24 h、48 h和72 h后,SL细胞中蛋白质含量分别降低8.55%、25.95%、42.95%和67.05%;处理24 h和48 h后,SL细胞的LDH漏出率分别为30.66%和32.05%。以浸叶喂食法处理斜纹夜蛾3龄幼虫,三唑类杀菌剂可显著抑制试虫体重增长。以0.5 μg/头、 1.0 μg/头和2.0 μg/头剂量的腈菌唑注射处理72 h后,斜纹夜蛾4龄幼虫血细胞数量分别降低12.31%、 25.96%和25.73%;腈菌唑注射处理48 h和72 h后,对斜纹夜蛾幼虫的LD₅₀值分别为1.59 μg/头和1.53 μg/头。结果显示以离体培养细胞为对象,从现有杀菌剂中寻找新的杀虫剂先导化合物具有良好的研究潜力。     

Specific Incorporation of L-Glutamine into Volicitin in the Regurgitant of Spodoptera litura

Volicitin, [N-(17-hydroxylinolenoyl)-L-glutamine], was identified as an elicitor of plant volatiles from a Spodoptera exigua regurgitant. It has been proposed that gut microbes synthesize volicitin from glutamine, a predominant amino acid component in the insect gut. However, we found that glutamine was not a major component in the regurgitant of Spodoptera litura, although L-glutamine was exclusively incorporated into volicitin by S. litura fed on diets enriched with various amino acids. This selectivity of glutamine as a substrate was not due to a dominant occurrence in the insect gut.     

日本三株斜纹夜蛾核型多角体病毒的增殖特性及其多角体蛋白基因的序列分析

利用斜纹夜蛾Spodoptera litura培养细胞,对近年来在日本本州、九州和四国等地发现并筛选出的对斜纹夜蛾幼虫具有强烈杀虫活性的3株斜纹夜蛾核型多角体病毒(SpltMNPV)（K-3、G1-2和G10-3）进行了生物学活性和分子生物学的初步研究,克隆了多角体蛋白基因,并进行了序列分析和比较。结果表明:（1）SpltMNPV日本分离株K-3、G1-2和G10-3分别具有不同的特征性酶切图谱,分别属于3种基因型（A型、B型和C型）; （2）3个分离株的芽生型病毒（budded virus）产生能力和多角体产生能力有差异,免疫印迹分析表明,多角体蛋白的分子量也不同;（3）日本株SpltMNPV核型多角体蛋白结构基因由747个核苷酸编码序列（编码249个氨基酸）组成,其序列与中国株SpltMNPV的同源性为98.9%,与其他6种核型多角体病毒有较高的同源性（61.7%～74.2%）,但其5′端侧翼序列(nt-1～-100)与AcMNPV和BmNPV相比差异显著,在对该基因表达调控起决定性作用的8个高度保守核苷酸序列中（nt-44～-51）有2处发生自然突变。     

斜纹夜蛾药剂防治综合决策的研究

根据不同药剂对斜纹夜蛾的防治效果、防治工本费、对天敌的影响、防治后挽回的产量损失等资料,以净收益最大作为最优防治目标,运用优化理论和多目标决策方法,对11种药剂防治措施进行了综合分析。结果表明,在11种药剂防治措施中,以每667m^2施1．8％阿维菌素乳油16．7ml最优,其次是1．8％阿维菌素乳油25ml,20％氰戊菊酯乳油25ml,2．5％功夫乳油16．7ml,2．5％功夫乳油25ml,20％氰戊菊酯乳油50ml,4．5％高效氯氰菊酯乳油50ml,4．5％高效氯氰菊酯乳油25ml,50％辛硫磷乳油50ml,10％除尽悬浮剂16．7ml,10％除尽悬浮剂25ml,为合理地选择农药和用量提供了科学依据。     

斜纹夜蛾Cecropin D成熟肽的原核表达及活性检测

采用RT-PCR方法从斜纹夜蛾Spodoptera litura脂肪体组织中扩增得到了Cecropin D成熟肽基因序列,分析发现Cecropin D成熟肽与斜纹夜蛾Cecropin B之间存在2个氨基酸残基的差异。将获得的基因序列连接入原核表达载体pGEX-4T-1,并在原核细胞中实现了该蛋白的融合表达。SDS-PAGE结果表明,诱导后的宿主菌比未诱导菌中多出了一条融合蛋白表达带,诱导后1 h就可以检测到该蛋白,从诱导后1 h到5 h该蛋白在表达量上没有明显的差异。生长曲线显示在IPTG诱导后宿主菌的生长受到明显的抑制,纯化后的蛋白对细菌具有一定的抑制作用。     

斜纹夜蛾抗性种群中酶抑制剂对杀虫剂的增效作用（英文）

采用浸液生测法研究了斜纹夜蛾Spodoptera litura巴基斯坦抗性种群中酶抑制剂［胡椒基丁醚（PBO）和脱叶膦（DEF)］对丙溴磷、灭多威、硫双灭多威、氯氰菊酯、氯氟氰菊酯、联苯菊酯、茚虫威和多杀菌素等杀虫剂的增效作用。结果表明：PPO和DEF对氨基甲酸酯杀虫剂灭多威和硫双灭多威均具有增效作用,但对有机磷杀虫剂丙溴磷不具有增效作用。两种抑制剂对氯氰菊酯均产生增效作用,但对联苯菊酯没有增效作用。PPO 和DEF增加了氯氟氰菊酯对Multan种群的毒性,但没有增加其对Mailsi种群的毒性。DEF对多杀菌素具有增效作用,但PBO对其没有增效作用。PBO和DEF对氨基甲酸酯杀虫剂、拟除虫菊酯杀虫剂、茚虫威和多杀菌素具有明显的增效作用,这说明细胞色素P450单加氧酶和酯酶的解毒作用至少部分参与了斜纹夜蛾对这些杀虫剂的抗性过程。不过,两种增效剂对杀虫剂增效作用范围有限,暗示对于斜纹夜蛾巴基斯坦种群而言,其他的机制（如靶位点不敏感、表皮穿透作用降低）可能是更重要的抗性机制。