甜菜夜蛾核型多角体病毒对甜菜夜蛾幼虫的毒力测定

室内毒力测定表明,在一定范围内,不同浓度甜菜夜蛾核型多角体病毒(SeNPV)感染甜菜夜蛾2龄幼虫后,浓度越高,甜菜夜蛾死亡率越高,开始死亡时间越早,全程感染至死亡的天数越短,致死中时(LT50)为2.44~5.60 d,致死中浓度LC50=7.11×104PIB/m l。以剂量为2×105PIB/m l的SeNPV样品感染甜菜夜蛾1~4龄幼虫,低龄幼虫的死亡高峰期为4~5 d,全程期为6~7 d;3龄以上幼虫死亡高峰期为7 d左右,全程期为10~12 d。     

Laboratory-based flight performance of the fall armyworm,Spodoptera frugiperda

In late 2018, the fall armyworm(FAW) Spodoptera frugiperda Smith(Lepidoptera: Noctuidae) made its arrival in China and its populations have since proliferated across most of eastern Asia. While S. frugiperda exhibits a considerable dispersal capacity and engages in long-distance migration, there's only scant information on the species' flight capability. Here, we empirically assessed S. frugiperda flight activity under varying climatic conditions using a flight mill. More specifically, under laboratory conditions, FAW exhibited superior flight performance at 20–25°C and 60–90% relative humidity(RH). When quantifying flight performance over five consecutive nights(i.e., 10 h/night), all flight parameters initially increased and then gradually dropped and FAW adults attained a total flight distance, duration and velocity equal to 63.73 km(48.42–94.12 km)(median, quartile range), 24.12 h(20.87–27.73 h) and 2.73 km h~(–1)(2.13–3.33 km h~(–1)), respectively. Our work constitutes a first comprehensive assessment of S. frugiperda flight performance and provides baseline information for future efforts to forecast spatiotemporal changes in its geographical distribution, movement patterns and invasion trajectories. Such can ultimately permit a timely and targeted deployment of area-wide pest management measures against this newly-invasive pest in China and across eastern Asia.     

甜菜夜蛾防治指标的测定

通过室内测定甜菜夜蛾对空心菜、上海青、菠菜的取食量 ,进行田间模拟剪叶 ,确定虫量与产量损失率的关系 ,并根据当地生产水平、产品价格、防治成本及药剂防治效果等 ,分别制定了我省常规产量下空心菜、上海青及菠菜的防治指标 [分别为 12 60 0 0、65 5 0 0 0和 2 640 0 0头· hm-2 (3龄幼虫 ) ].此外 ,还提供了在不同产量、不同经济系数下的防治指标 ,以指导生产上防治     

草地贪夜蛾的综合防治

草地贪夜蛾Spodoptera frugiperda (J. E. Smith)属于入侵性农业害虫,其幼虫可以大量啃食玉米、高粱、水稻等多种农作物。草地贪夜蛾具有迁飞能力强、突发性、多产性、暴食性、杂食性、隐蔽性、钻蛀性、钻土性、夜出性、趋嫩性和独占性等特性,防治难度极大。对于入侵的暴发性害虫来说,化学防治依旧是目前能够在较短时间内控制其在我国大规模暴发的主要方法。虽然有很多昆虫都是草地贪夜蛾的天敌,但基本上没有足够用来防止作物受伤的措施。目前的防治措施主要包括物理防治、化学防治、生物防治,其中生物防治又可具体为植物源农药、性诱剂、生防菌、寄生蜂、寄生性天敌等。     

斜纹夜蛾的人工饲养技术

介绍了斜纹夜蛾的人工饲养技术,结果表明:室内饲养的斜纹夜蛾平均蛹重达473 mg,幼虫成活率、羽化率均在95%以上。该饲养方法可用于室内大量饲养斜纹夜蛾,但为减少霉菌的污染,应在人工饲料中加入适量的甲醛。     

斜纹夜蛾寄主植物名录

在田间调查和查阅国内外文献的基础上,整理出斜纹夜蛾寄主植物名录。该虫寄主涉及到蕨类植物、裸子植物、双子叶植物、单子叶植物,共计109科389种(包括变种)。     

Fitness of fall armyworm,Spodoptera frugiperda to three solanaceous vegetables

The fall armyworm(FAW), Spodoptera frugiperda Smith(Lepidoptera: Noctuidae) is an important agricultural pest that invaded China in the middle of December 2018. As a polyphagous pest, FAW is identified as a serious threat to agricultural production and food security in China. Pepper(Capsicum annuum L.), tomato(Solanum lycopersicum Mill.) and eggplant(Solanum melongena L.) are three of dominant solanaceous vegetables of this country. To our knowledge, the effects of these plants on the performances of FAW have not been well studied. In this study we assessed the fitness of this pest to these three plants. Results showed that FAW can complete its life cycle when fed with tomato and pepper, but not on eggplant. The population parameters of FAW fed with maize(Zea mays L.) and the three solanaceous vegetables were compared using the age-stage, two-sex life table method. Developmental duration was significantly different in the larval stage, but not in the pupae stage. FAW fed with pepper had the longest pre-adult period(41.73 d) and the lightest pupal weight(0.1134 g); the survival rate was lower than FAW fed with tomato. Significant differences were observed in the mean fecundity of female, with the highest(943.95 eggs) laid by FAW fed with tomato. FAW had the shortest mean generation time(T), the highest intrinsic rate of increase(r) and finite rate of increase(λ) on maize, and the highest net reproductive rate(R_0) on tomato. Overall, FAW fitness on the three solanaceous vegetables was: tomatopeppereggplant. This study provides the foundation for further assessment of FAW risk to solanaceous vegetables and for establishing corresponding control strategies in China.     

甜菜夜蛾双重氧化酶基因SeDuox的克隆、表达及功能分析

【目的】昆虫双重氧化酶（dual oxidase,Duox）介导产生活性氧（reactive oxygen species,ROS）是调节昆虫肠道微生物动态平衡的重要免疫机制。本研究通过分析甜菜夜蛾（Spodoptera exigua）双重氧化酶基因（SeDuox）的序列特征与表达模式,RNAi技术探究SeDuox基因沉默对甜菜夜蛾肠道细菌载量的影响,分析SeDuox对苏云金杆菌（Bacillus thuringiensis,Bt）的免疫响应,从而明确SeDuox在昆虫肠道免疫调控中的作用,为甜菜夜蛾的综合防控提供新的思路和作用靶标。【方法】利用反转录PCR（RT-PCR）技术克隆SeDuox,序列特征进行生物信息学分析,采用ClustalX和MEGA_X软件构建系统发育树;克隆SeDuox膜外91—1 767 bp编码基因SeDuox-OM,利用Bac to Bac表达系统构建Bacmid-SeDuox-OM,脂质体转染法转染昆虫细胞Sf9,表达SeDuox-OM膜外蛋白;采用实时荧光定量PCR（RT-qPCR）方法分析甜菜夜蛾不同组织（围食膜、血淋巴、脂肪体、马氏管、中肠和表皮）和不同发育时期（卵、1—5龄幼虫、雌蛹、雄蛹）SeDuox的表达水平;利用RNAi技术进行功能分析,向4龄幼虫注射SeDuox的dsRNA,以注射绿色荧光蛋白（green fluorescent protein,GFP）的dsRNA作为对照,注射48 h和72 h后检测基因沉默效果及肠道细菌载量的变化;向甜菜夜蛾幼虫饲喂Bt,RT-qPCR方法分析SeDuox及相关免疫基因的诱导表达情况。【结果】SeDuox序列全长为4 497 bp,编码1 498个氨基酸,预测蛋白质的相对分子量为171.62 kD。SMART分析结构域发现SeDuox含有过氧化物酶结构域（peroxidase）、烟酰胺腺嘌呤二核苷酸结构域（NAD）、N-端钙离子结合域（EFH）、铁还原酶结构域（Ferric）和黄素腺嘌呤二核苷酸结构域（FAD）,与斜纹夜蛾（Spodoptera litura）、草地贪夜蛾（Spodoptera frugiperda）等昆虫的Duox结构域类似。TMHMM分析显示有7个跨膜区,跨膜片段SeDuox-OM 在昆虫细胞中成功表达约80 kD的重组蛋白;RT-qPCR分析结果显示SeDuox在甜菜夜蛾4龄幼虫不同组织中均有表达,其中围食膜和中肠表达量较高;在不同发育时期均有表达,但在卵期表达量最高。与注射dsGFP相比,注射 dsSeDuox 的甜菜夜蛾幼虫在48 h和72 h时,SeDuox表达量分别下调了62.08%和74.94%,肠道微生物载量显著升高。甜菜夜蛾幼虫取食Bt GS36 48 h时,SeDuox表达量显著增加,随着侵染时间的延长,其表达量呈现出先升高后降低再趋于稳定的特点。【结论】甜菜夜蛾SeDuox具有保守的结构域,在中肠和围食膜中高表达,SeDuox在宿主肠道免疫调控中起重要作用,可能与抗菌肽基因协同作用抵御Bt的侵染。     

几种天敌对斜纹夜蛾幼虫的捕食作用

测定了几种主要捕食性天敌捕食斜纹夜蛾1－2龄幼虫的功能反应，结果表明，草间小黑蛛雌成蛛、拟水狼蛛雌成蛛和叉角厉蝽2龄若虫对斜纹夜蛾1－2龄幼虫均有较强的捕食能力，其理论最大捕食量分别为6.12次、26.00头和10.11头。在斜纹夜蛾幼虫的6个龄期中，最适合于叉角厉蝽雌成虫捕食的龄期为4龄，其理论最大捕食量高达10.59头。     

草地贪夜蛾肠道细菌分离鉴定与功能初探

【目的】明确草地贪夜蛾幼虫肠道细菌的种类和功能,以诠释肠道细菌对草地贪夜蛾寄主植物适应性的影响,为揭示草地贪夜蛾的寄主适应机制及进一步预测其寄主谱扩张趋势提供依据。【方法】采用传统的微生物分离纯化方法对草地贪夜蛾肠道细菌进行分离纯化,对分离获得的细菌菌株进行16S rRNA序列同源性分析鉴定;利用筛选培养基对产生纤维素酶、木聚糖酶、果胶酶及对苯酚有代谢能力的菌株进行初筛,并进一步用DNS法测定相关菌株产纤维素酶、木聚糖酶和果胶酶的酶活性,用含苯酚的无机盐培养基培养菌株,检测菌株的苯酚降解效率。【结果】共分离获得45株细菌菌株,经同源序列比对分析,45株细菌菌株分属于3门5属8种,分别是厚壁菌门（Firmicutes）的葡萄球菌属（Staphylococcus）和芽孢杆菌属（Bacillus）,变形菌门（Proteobacteria）的克雷伯氏菌属（Klebsiella）和不动杆菌属（Acinetobacter）,放线菌门（Acinobacteria）的短杆菌属（Curtobacterium）,其中克雷伯氏菌属的丰度最高。45株菌株中有产纤维素酶菌株11株,酶活力最高的是变栖克雷伯氏菌菌株K3,为0.105±0.007 U/mL;产木聚糖酶菌株10株,酶活力最高的是枯草芽孢杆菌菌株B9,为1.090±0.468 U/mL;产果胶酶菌株5株,酶活力最高的是变栖克雷伯氏菌菌株K27,为0.193±0.047 U/mL;降解苯酚的菌株9株,降解速率最高的是沙福芽孢杆菌菌株B8,为（0.347±0.042）%。【结论】草地贪夜蛾幼虫肠道细菌的产酶菌株多样性较高,推测这是导致草地贪夜蛾寄主谱广,对寄主为害严重的原因之一。     

核型多角体病毒对斜纹夜蛾幼虫致病性的研究

利用从日本引进的斜纹夜蛾核型多角体病毒(SlNPV)研究了对斜纹夜蛾幼虫的致病性。结果表明,随着龄期增加,致病性减弱,LC50值升高。在15~30℃范围内,饲育温度越高,致病性越强,LC50最低。     

Cold hardiness of the invasive fall armyworm,Spodoptera frugiperda in China

Fall armyworm, Spodoptera frugiperda(J. E. Smith, 1797), a crop pest native to tropical and subtropical regions of America, has invaded and spread into most regions in China, posing a severe threat to China's agriculture. The cold hardiness directly determines its geographic distribution through adapting to winter temperatures of different regions. Here, we measured supercooling points and lethal time(LT) at low temperatures of S. frugiperda. The supercooling points for developmental stages in increasing order were: adults(–15.05°C)pupae(–13.25°C)prepupae(–10.50°C)larvae(–9.03°C). Among eggs and 1 st to 4 th instar larvae, the 99% lethal time(LT_(99)) was the highest for 4 th instar larvae, with 99% of larvae dying after 18.59 d at 2°C, 58.72 d at 7°C, and 66.28 d at 13°C. LT_(99) was the lowest for eggs with LT_(99) of 5.33 d at 2°C, 9.28 d at 7°C, and 12.97 d at 13°C. This study provides an understanding of overwintering regions of S. frugiperda in China, which will be helpful for population forecasting and management.     

甜菜夜蛾人工饲料筛选

为工厂化生产甜菜夜蛾卵,对几种甜菜夜蛾人工饲料的饲养效果进行了比较分析。结果表明:3号饲料具有产卵量大、成本低廉和制作简单等优点,可作为工厂化生产甜菜夜蛾卵的人工饲料,为规模化生产赤眼蜂提供理论和技术保障。     

蜕皮激素类似物美满防治甜菜夜蛾田间药效试验

蜕皮激素类似物美满(methoxyfenozide)对甜菜夜蛾(Spodoptera exigua Hubner)有突出的防治效果,田间使用24%美满悬浮剂3 000倍、6 000倍,药后1天、3天、7天防治效果分别达到93.42%,97.05%,98.39%和82.61%,88.87%,89.38%,与10%除尽(chloffenapyr)1 500倍相当.美满与乐斯本(chlorpyrifos)混配使用,也可以达到理想的防治效果.且能节约用药成本,在作物生长前期可适当使用.仿生的蜕皮激素类似物是当前无公害蔬菜生产中值得推荐的好农药.     

光敏剂对斜纹夜蛾细胞的光活化杀虫活性、单线态氧及稳定性

[目的]提高光敏剂的光稳定性,延长光敏剂在环境中的半衰期。[方法]该研究以典型光敏剂α-三联噻吩为模板,利用生物等排原理,以苯环代替噻吩环,合成了光敏剂2,5-二苯基噻吩和2,5-二乙炔苯基噻吩,并研究了所合成的光敏剂的对斜纹夜蛾细胞的光活化毒力、单线态氧及光稳定性。[结果]光敏剂2,5-二苯基噻吩对斜纹夜蛾细胞的24和48 h的抑制中浓度IC50值分别为0.22和0.16μg/ml,2,5-二乙炔苯基噻吩斜纹夜蛾细胞的24和48 h的抑制中浓度 IC50值分别为0.06和0.04μg/ml;在紫外光照下,光敏剂2,5-二苯基噻吩和2,5-二乙炔苯基噻吩产生的1O2能力分别为1.0475和1.5294μg/mmol;2,5-二苯基噻吩和2,5-二乙炔苯基噻吩在甲醇中降解动力学方程分别为 Ct=5.2271e-0.061t和 Ct=5.0842e-0.0973t,其半衰期分别为111.79和7.12 h。[结论]所合成的光敏剂2,5-二苯基噻吩具有良好的光活化激发产生单线态氧的能力,对斜纹夜蛾SL细胞具有良好的光活化毒杀作用,克服了光活化杀虫剂见光易分解等不足,因此2,5-二苯基噻吩具有开发成光活化杀虫剂的潜力。     

唐古特瑞香对斜纹夜蛾的拒食活性研究

报道了青藏高原特有植物唐古特瑞香的杀虫作用。利用甲醇、丙酮、乙酸乙酯、氯仿、苯、石油醚６种极性不同的溶剂对唐古特瑞香进行了连续抽提,其中甲醇提取率最大为２２．３％,石油醚的提取率最低为２．２１％,唐古特瑞香的甲醇、丙酮、乙酸乙酯、氯仿、苯抽提物对斜纹夜蛾３龄幼虫的ＡＦＣ５０值分别为以干粉计１０８．７２、１１１．６９、１０２．１４、７１．２５、６５．５１ｇ／Ｌ,石油醚抽提物在以干粉主地５００ｇ／Ｌ的     

光敏剂对斜纹夜蛾细胞的光活化杀虫活性、单线态氧及稳定性(英文)

[目的]提高光敏剂的光稳定性,延长光敏剂在环境中的半衰期。[方法]该研究以典型光敏剂α-三联噻吩为模板,利用生物等排原理,以苯环代替噻吩环,合成了光敏剂2,5-二苯基噻吩和2,5-二乙炔苯基噻吩,并研究了所合成的光敏剂的对斜纹夜蛾细胞的光活化毒力、单线态氧及光稳定性。[结果]光敏剂2,5-二苯基噻吩对斜纹夜蛾细胞的24和48 h的抑制中浓度IC50值分别为0.22和0.16μg/ml,2,5-二乙炔苯基噻吩斜纹夜蛾细胞的24和48 h的抑制中浓度IC50值分别为0.06和0.04μg/ml;在紫外光照下,光敏剂2,5-二苯基噻吩和2,5-二乙炔苯基噻吩产生的1O2能力分别为1.047 5和1.529 4μg/mmol;2,5-二苯基噻吩和2,5-二乙炔苯基噻吩在甲醇中降解动力学方程分别为Ct=5.227 1e-0.006 1t和Ct=5.084 2e-0.0973t,其半衰期分别为111.79和7.12 h。[结论]所合成的光敏剂2,5-二苯基噻吩具有良好的光活化激发产生单线态氧的能力,对斜纹夜蛾SL细胞具有良好的光活化毒杀作用,克服了光活化杀虫剂见光易分解等不足,因此2,5-二苯基噻吩具有开发成光活化杀虫剂的潜力。     

甜菜夜蛾UAP的克隆、时空表达及RNAi研究

【目的】在昆虫中,几丁质的合成对于其生长发育至关重要,虽然几丁质合成通路在真菌中的研究比较深入,但是在昆虫中,目前大多数的研究都集中在海藻糖酶和几丁质合成酶方面,而对于通路上其他基因的研究却非常少。论文旨在阐明昆虫几丁质合成通路中的UDP-N-乙酰氨基葡萄糖焦磷酸化酶（UAP）对甜菜夜蛾（Spodoptera exigua）几丁质合成酶基因和害虫存活的影响。【方法】以甜菜夜蛾5龄第1天幼虫的cDNA为模板,通过简并引物扩增出甜菜夜蛾UAP基因（SeUAP）的中间片段,然后利用特异性引物和RACE技术扩增出3′和5′ race序列,拼接得到cDNA全长。随后提取甜菜夜蛾各组织和各龄期的总RNA,并通过RT-PCR方法分析SeUAP在各个组织和龄期的表达情况。最后通过向甜菜夜蛾5龄第1天的幼虫注射dsUAP观察其生长状况,检测RNAi的效率以及对几丁质合成通路下游基因的影响。【结果】获得全长为2 098 bp的SeUAP cDNA,编码一个491个氨基酸残基的蛋白,与双翅目昆虫致倦库蚊、埃及伊蚊、黑腹果蝇和冈比亚按蚊的UAP亲缘关系较近。该基因在表皮和卵巢中大量表达,在气管和中肠中的表达次之,在马氏管中表达微弱,在脂肪体中则基本检测不到其转录本的存在。该基因在甜菜夜蛾生长发育的各个阶段均有表达,在卵的整个阶段、L22（幼虫2龄第2天）、L31、L42、L51、P0、P5、P7以及A3这些天中的表达量较高,其余天数表达量较低,其表达的高峰主要出现在几丁质需求量较高的时期。RNAi结果表明,通过向甜菜夜蛾5龄第1天的幼虫注射5 µg dsRNA,导致部分甜菜夜蛾出现蛹期畸形及死亡,并出现两种畸形现象：（1）虫体腹部蛹壳已基本形成并已硬化,但头部蛹壳形成有障碍,化蛹时无法突破旧表皮;（2）虫体头部蛹壳形成正常,也能正常突破旧表皮,但蜕皮过程仅到此为止,无法正常进行下去。定量PCR结果显示SeUAP的沉默对几丁质合成酶A基因（SeCHSA）下调不明显,而对几丁质合成酶B（SeCHSB）的表达有明显下调作用。【结论】UAP可以影响几丁质合成通路的下游基因并对甜菜夜蛾幼虫-蛹的生长发育起一定作用。     

斜纹夜蛾感受器超微形态特征

采用扫描电镜技术,系统研究了斜纹夜蛾触角、跗节和产卵器的感受器形态结构特征,发现触角感受器主要包括毛形、锥形、刺形、耳形和腔锥感受器.雌、雄虫触角锥形感受器形态特征区别明显,雄蛾锥形感受器长约10～15 μm,基部直径约4～5μm,显著比雌虫锥形感受器长、粗,雄蛾锥形感受器顶端有一小锥,雌蛾没有.雌成虫和雄成虫前足、中足、后足跗节均分布有毛形和刺形感受器,但数量明显少于触角同类感受器数量.雌成虫产卵器上主要为长毛形和短毛形感受器,这些感受器可能与其感受环境信息、产卵场所选择等行为有关.

Abstract：

The ultrastructures of the sensilla on the antennae, the tarsi and the ovipositor of Spodoptera litura were observed with scanning electron-microscopy. The antennae of both male and female moths were found to contain five kinds of antennal sensilla, namely, sensillum trichodeum, sensillum basiconicum,sensillum chaeticum, ear-shaped sensillum and sensillum coeloconicum. The characters of sensillums basiconicum on the antennae were different between the male and the female. The length was 10-15 μm, the diameter at the base was approximately 4-5 μm for the male antennae, which were significantly longer and bigger than the female antennae, and the male moths had onesmall bradawl at the bass of the antennae while the female moths did not. The tarsi of both male and female moths contained two kinds of sensilla:sensillum trichodeum and sensillum chaeticum; however, their numbers were significant less than those on the antennae. The long sensillun trichodeum and short sensillun trichodeum were mainly distributed on the ovipositor. These results provided basis for exploring the molecular mechanism of oviposition deterrent.     

Flight activity promotes reproductive processes in the fall armyworm,Spodoptera frugiperda

The fall armyworm(FAW), Spodoptera frugiperda(Lepidoptera: Noctuidae), has invaded many countries in Africa and Asia since 2016, posing a major threat to world food security. Long-distance migration and strong reproductive ability form the biological basis of its rapid population expansion, but the relationship between the flight and reproduction of FAW has not been studied in depth. Here, an empirical assessment of this relationship in an invasive FAW population in China found that 1–3-day-old adults which had undergone 10-h tethered flights had a significantly shorter pre-oviposition period and greater oviposition synchronization, but did not show any differences in fecundity, oviposition period, mating percentage or other reproductive variables. Further studies on moths after 1.25–15-h tethered flights indicated that the reproductive process of adults could be fully triggered by flight activity longer than 2.5 h. Dissection of the reproductive organs also showed that tethered flight promoted ovarian and testicular development of FAW. These results show that appropriate moth flight activity significantly speeds up the reproductive process of FAW, which increases our knowledge on its migratory biology in relation to regional outbreaks.