褐飞虱长翅型和短翅型雌雄成虫体内磁性物质定位和定量检测

【目的】地磁定向是昆虫远距离迁飞定向的重要机制之一。本研究以迁飞性害虫褐飞虱Nilaparvata lugens长翅型和短翅型雌雄成虫为研究对象,系统开展虫体内磁性物质定量研究。【方法】本研究利用MPMS-7型超导量子磁强计检测褐飞虱长翅型和短翅型雌雄成虫体内的磁性物质,并将经普鲁士蓝染色后的虫体超薄石蜡切片于JEM-2100型透射电子显微镜下观察磁性物质的分布状况,最后利用原子发射光谱法对虫体内的磁性物质进行定量分析与比较。【结果】超导量子磁强计检测发现,仅在褐飞虱长翅型和短翅型雌雄成虫腹部的磁滞曲线有明显闭合现象,证明该部位存在磁性物质;经透射电镜观察发现,普鲁士蓝沉淀主要呈点簇状存在于虫体腹部,证明该部位存在铁磁性物质;通过等离子发射光谱检测发现,同一性别长翅型成虫体内铁离子含量显著高于短翅型成虫,且相同翅型雄成虫体内铁离子含量显著高于雌成虫。【结论】褐飞虱长翅型和短翅型雌雄成虫腹部普遍存在铁磁性物质,且不同翅型和性别间磁性物质含量差异显著。     

褐飞虱flightin的原核表达及其差异表达检测

flightin最早发现于果蝇Drosophila melanogaster的间接飞行肌中,并且定位于粗肌丝。这种蛋白对维持肌节的结构和功能起到了重要的作用,但其在具有长短翅型分化的褐飞虱Nilaparvata lugens Stl的不同翅型间差异并不清楚。本研究以长翅型雌虫褐飞虱cDNA为模板,通过PCR扩增得到褐飞虱flightin基因ORF全长,将其连接到表达载体pGEX-6P-1中以与谷胱甘肽S-转移酶(GST)融合表达。将表达载体转入大肠杆菌表达株Rosseta,在不同温度、不同浓度IPTG的条件下诱导表达flightin,得到了最优表达条件,获得了高水平可溶性表达。在用GST抗体进行Western blotting验证GST-flightin融合重组蛋白表达的正确性后,我们通过GST柱纯化了的GST-flightin,进而用纯化后的蛋白免疫新西兰兔制备了高特异性的多克隆抗体。最后,我们用制备的多克隆抗体检测了长、短翅型雌成虫和不同发育阶段的褐飞虱体内flightin的表达差异。结果显示,flightin仅在长翅型成虫中表达,在短翅型雌成虫中未检测到其明显表达,而且flightin只在成虫期表达。本研究为进一步研究褐飞虱的flightin与其它蛋白互作、翅肌发育和翅型分化打下了基础。     

高温对稻褐飞虱发育与生殖的影响

通过高温处理及交配产卵实验,探讨了高温对稻褐飞虱Nilaparvata lugens (Stal)发育与生殖的影响。研究了不同高温条件对褐飞虱若虫发育历期、产卵量、产卵前期、寿命等生物学特性的影响。34℃以上高温导致褐飞虱若虫发育历期延长。高温处理4龄若虫使羽化后的雌成虫产卵量减少。高温处理不同日龄雌成虫也致使其产卵量减少,其中以1日龄短翅型、3日龄长翅型的产卵量影响最大。高温处理后褐飞虱寿命缩短。高温恒温处理,对短翅型雌成虫的产卵前期影响不大,但能延长长翅型雌成虫的产卵前期;而高温变温处理致使短、长翅型雌成虫的产卵前期均延长。高温变温对褐飞虱生殖的影响程度大于高温恒温。高温处理组雌、雄成虫与对照组相应的成虫交配试验表明,高温对雌性的影响大于雄性。实验初步确定34℃为对褐飞虱发育与生殖产生影响的临界温度。     

三种杀虫剂对褐飞虱海藻糖含量和海藻糖酶活性的影响

为了解农药处理导致褐飞虱Nilaparvata lugens (Stål）飞行能力增强的生理机制, 本文采用蒽酮法和酶促反应终止法, 研究了吡虫啉、 三唑磷和溴氰菊酯3种杀虫剂亚致死剂量对褐飞虱3龄、 5龄若虫及长、 短翅型雌雄成虫体内海藻糖含量和海藻糖酶活性的影响。结果表明： 杀虫剂处理的褐飞虱3龄若虫海藻糖含量和海藻糖酶活性与对照相比没有显著差异（P>0.05）。40 mg/L三唑磷处理的褐飞虱5龄若虫体内海藻糖含量显著低于对照（P<0.05）, 比对照降低了24%; 而20和40 mg/L三唑磷处理的褐飞虱5龄若虫海藻糖酶活性显著高于对照（P<0.05）, 分别比对照高出了100%和129%。10 mg/L吡虫啉, 20 和40 mg/L三唑磷以及3和6 mg/L溴氰菊酯处理的褐飞虱短翅雌成虫和雄成虫体内海藻糖含量显著低于对照（P<0.05）, 雌成虫体内海藻糖含量比对照分别降低了36%, 53%, 67%, 58%和69%, 雄成虫体内海藻糖含量比对照分别降低了59%, 71%, 65%, 70%和77%; 而40 mg/L三唑磷以及3和6 mg/L溴氰菊酯处理的褐飞虱短翅型雌成虫和雄成虫体内海藻糖酶活性显著高于对照（P<0.05）, 雌成虫体内海藻糖酶活性比对照分别高出了124%, 100%和88%, 雄成虫体内海藻糖酶活性比对照分别高出了146%, 132%和118%。10 mg/L吡虫啉, 40 mg/L三唑磷和3 mg/L溴氰菊酯处理的褐飞虱长翅型雌成虫和雄成虫海藻糖含量显著低于对照（P<0.05）, 雌成虫海藻糖含量比对照分别降低了44%, 34%和37%, 雄成虫体内海藻糖含量比对照降低了48%, 54%和43%; 而5和10 mg/L吡虫啉处理的长翅型雌成虫和雄成虫海藻糖酶活性显著高于对照（P<0.05）, 雌成虫体内海藻糖酶活性比对照分别高出了317%和300%, 雄成虫体内海藻糖酶活性比对照分别高出了170%和97%。这些结果说明这3种杀虫剂亚致死剂量处理可以增强褐飞虱体内海藻糖酶活性, 并导致海藻糖含量下降。本研究结果对深入阐明农药诱导褐飞虱再猖獗及杀虫剂处理增强其飞行能力的生理机制具有一定的科学价值。     

近零磁场对褐飞虱翅型分化、趋光性及飞行能力的影响

【目的】地磁场(geomagnetic field,GMF)不是恒定不变的,而是随时间和空间时刻变化的。目前,动物对磁场变化的响应研究主要集中于迁徙性动物地磁定向导航中的磁感受方面,而迁徙性动物迁出地和迁入地之间地磁场强度差异对动物生理和行为潜在的磁场效应尚不明确。【方法】迁飞性昆虫褐飞虱Nilaparvata lugens试虫采自江苏省农业科学院试验田。本文采用亥姆霍兹线圈室内模拟近零磁场(near-zero magnetic field,NZMF)和GMF,调查了褐飞虱成虫在NZMF和GMF下的翅型分化、趋光性和飞行能力。【结果】结果表明,与GMF相比,NZMF显著提高了褐飞虱雄成虫短翅型比例(6. 4%)(P <0. 05),但对雌成虫长翅型比例影响不显著(P> 0. 05);对于长翅雌成虫,NZMF显著提高了其2日龄成虫的上灯比例(55%)(P <0. 05),但显著降低了其4日龄成虫的上灯比例(22%)(P <0. 05),对趋光性的影响总体呈现出随着龄期先增强后减弱的效应。NZMF对长翅雄成虫趋光性的影响也呈现出相同的效应,但对各日龄成虫的影响不显著(P> 0. 05); NZMF显著缩短了2日龄长翅雄成虫的飞行时间(46%)(P <0. 05),并显著提高了长翅雌成虫(65%)和长翅雄成虫(101%)的飞行速度(P <0. 05)。此外,GMF对照组的褐飞虱长翅雄成虫飞行速度显著低于长翅雌成虫(96%),而NZMF处理组中二者无显著差异。【结论】结果说明,近零磁场可提高褐飞虱成虫短翅比例,对长翅成虫趋光性的影响呈现出基于龄期增长先增强后减弱的效应,并在未影响长翅成虫飞行距离的情况下,改变了其飞行策略,即提高飞行速度,同时缩短飞行时间。     

病原线虫对褐飞虱寄生状况调查

褐飞虱Nilaparvata lugens在浙北稻区常年发生严重,在中晚稻后期常暴发成灾.病原线虫是褐飞虱短翅型成虫的主要寄生性天敌之一.由于短翅型成虫数量与下一代发生和为害动态关系密切,故研究线虫对该虫主害代前一代短翅型成虫的寄生情况,有助于剖析褐飞虱成灾的规律,也可为增强线虫这一重要生物因子对该虫的控制作用提供依据.作者调查了浙江安吉1985～1998年线虫对褐飞虱主害代前一代短翅型成虫的寄生情况,讨论了线虫对该虫主害代的控制作用.结果如下.     

稻株营养物质与褐飞虱翅型分化的关系

褐飞虱(Nilaparvaia lugens Stl)是我国水稻的主要害虫之一,特别是短翅成虫繁殖力强、危害大。稻株中的营养物质和褐飞虱性别分化、翅型分化及趋性的关系与防治对策直接有关。本文研究水稻各生育期被褐飞虱取食的部位营养物质的种类和含量与褐飞虱翅型分化的关系,找出主导因素,为开展褐飞虱的综合防治提供资料。     

外源激素对褐飞虱翅型分化的影响

该文报道外源蜕皮激素(ED)和保幼激素类似物(JHA)对褐飞虱Nilaparvata lugens (Stal)翅型分化及体内酯酶活性、可溶性糖和游离氨基酸含量等的影响。褐飞虱在3、4龄若虫期经JHA处理后,所羽化出的成虫短翅型比例明显升高;而在5龄若虫期处理,所羽化出成虫的短翅型比例无明显变化。ED处理能增加3、4龄若虫体内酯酶活性、可溶性糖及游离氨基酸含量,而JHA处理则能有效地抑制ED的这些生物活性,这可能是JHA处理后褐飞虱成虫短翅型比例提高的生理生化基础。     

褐飞虱的迁人和翅型分化规律的研究

褐飞虱Nilaparvata lugens(Stal)属迁飞性水稻害虫,其成虫有短翅和长翅两种翅型。 褐飞虱迁入雌虫均未经过交配,其短翅型由一对显性等位基因控制。翅型分化同时受到遗传和内分泌系统的协调控制,外界条件如密度和寄主等因子通过内分泌系统来影响翅型的分化。褐飞虱翅型分化的敏感龄期雌虫为1～3龄,而雄虫为1-5龄;不同若虫蜜度处理对褐飞虱成虫的前翅形成有一定的影响。分蘖期水稻饲养的褐飞虱短翅型比例明显高于孕穗期水稻饲养的褐飞虱。不同密度下各生物型间的翅型分化差异不显著。迁入地的浙江各种群属温带型,其雌虫短翅率低且与密度呈显著负相关;菲律宾热带种群雌虫在不同密度下均为短翅型,而雄虫的短翅率随密度增加而上升。广西种群接近热带型,其雌虫短翅率高但不随密度而变化。试验各种群的雄虫在中等密度甚至高密度时其短翅率出现最高。     

水稻挥发性次生物质对褐飞虱寄主定向及生长的影响

从褐飞虱的抗虫与感虫水稻品种植株中提取挥发性次生物质,分析其对褐飞虱的定向选择、生长发育及成虫触角电位反应影响.结果表明,感虫品种TN1挥发性次生物质对褐飞虱生物型Ⅱ若虫的取食及生长发育未表现不良作用,对其定向选择有引诱作用;而抗虫品种IR36的挥发性物质对褐飞虱为负作用,表现为褐飞虱取食添加挥发性物质的人工饲料后,其死亡率均增加,生长发育受阻,且随着浓度的增加,不良影响加大.触角电位试验结果表明褐飞虱的电位反应总的趋势是对相同浓度的TN1挥发性物质的触角电位反应值比IR36大,且触角电位值基本都随着挥发性物质浓度的升高而增大;短翅型成虫电位值强于长翅型成虫,雄性成虫的电位值强于雌性成虫.挥发性物质气质联用检测分析结果发现抗虫与感虫水稻中挥发性次生物质的化学组成及含量相差较大,从抗感两个品种的挥发性物质中共检到44种已知化合物,部分化合物对褐飞虱有引诱作用.     

灰飞虱长、短翅型品系对高、低温的适应能力差异

【目的】灰飞虱Laodelphax striatellus种群存在长翅和短翅个体,其翅型分化受遗传和环境条件的影响。本研究旨在明确经连续定向选育的灰飞虱长、短翅型品系对高、低温的适应能力差异。【方法】在室内分别长期定向筛选63和65代建立灰飞虱长、短翅型品系,并在南京田间采集灰飞虱自然种群;分别在25℃, 30℃和35℃恒温条件下测定上述灰飞虱长、短翅型品系和自然种群的繁殖力、存活率和发育历期,测定若虫在5℃和-20℃下的存活率以及若虫和成虫的过冷却点,通过品系间存活率、发育历期、生殖力和过冷却点的比较分析,确定灰飞虱长、短翅型品系对高温和低温的耐受能力。【结果】在25℃, 30℃和35℃下,灰飞虱自然种群雌成虫的繁殖力均显著高于室内筛选后的长、短翅型品系,而长、短翅型品系雌成虫之间的繁殖力均无显著差异;长、短翅型品系的繁殖力在25℃和30℃之间均无显著差异,但自然种群在25℃下的繁殖力显著高于30℃条件下的繁殖力;35℃下长、短翅型品系基本无若虫产生。25℃下长翅型品系、短翅型品系和自然种群若虫的存活率无显著差异,但在30℃下,短翅型品系若虫的存活率显著低于自然种群,而与长翅型品系差异不显著;35℃下长、短翅型品系的若虫最长只能存活到3龄,且两品系间无显著差异,而自然种群可存活到5龄。25℃下,短翅型品系若虫的发育历期显著短于自然种群,而与长翅型品系无差异;但是在30℃下,短翅型品系若虫历期显著长于长翅型品系和自然种群。长翅型品系、短翅型品系和自然种群灰飞虱3-4龄若虫在5℃低温下的存活天数无显著差异,并且长翅和短翅型品系在-20℃极端低温下暴露10 min后其死亡率也无显著差异且均显著高于自然种群。3个品系(种群)成虫的过冷却点无显著差异,但5龄若虫的过冷却点表现为短翅型品系显著高于自然种群,而与长翅型品系无显著差异。【结论】经连续定向选育的灰飞虱长、短翅型品系对低温和高温的适应力基本相当,且低于自然种群。在高温条件下灰飞虱短翅型品系的发育速率低于长翅型品系。     

丽斗蟋翅二型雌虫飞行肌和卵巢发育间的资源分配差异

丽斗蟋Velarifictorus ornatus具有明显的翅二型现象, 长翅型与短翅型雌虫的卵巢和飞行肌存在着生理权衡。本研究分别应用蒽酮比色法、 硫代磷酸香草醛法、 考马斯亮蓝染液对羽化后10 d内两型雌虫飞行肌与卵巢内糖原、 总脂及蛋白质含量进行了定量分析。结果表明： 成虫羽化后10 d内, 两型雌虫体重无明显差异(P>0.05), 但短翅型雌虫怀卵量明显多于长翅型雌虫, 而人工脱翅能够促进长翅型雌虫怀卵量增加(P<0.05)。短翅型雌虫飞行肌内蛋白质、 糖原及总脂含量在成虫羽化后10 d内无明显变化, 但长翅型雌虫飞行肌内蛋白质在成虫羽化后3 d时达到最大值564.4±87.5 μg/♀, 糖原与总脂含量分别于羽化后第5天达到最大值85.2±21.7 μg/♀和5 284.7±1 267.4 μg/♀。然后开始下降, 各实验处理天数内, 长翅型雌虫飞行肌内蛋白质、 糖原及总脂含量都显著多于短翅型雌虫(P<0.05)。相反, 各处理天数内, 短翅型雌虫卵巢内蛋白质、 糖原及总脂含量则明显多于长翅型雌虫(P<0.05), 同时虫龄对蛋白质、 糖原及总脂在两型雌虫飞行肌与卵巢内分配也产生明显影响(P<0.05)。人工脱翅能够促进长翅型雌虫卵巢内蛋白质、 糖原及总脂含量增加, 同时诱导飞行肌内蛋白质、 糖原及总脂含量降低, 其中总脂含量在脱翅后10 d时降为2 394.9±1 461.8 μg/♀, 只有最大值的一半, 而与短翅型雌虫相似(P>0.05), 表明总脂为丽斗蟋飞行的主要能源物质。外用保幼激素Ⅲ能够促进长翅型雌虫卵巢内蛋白质、 糖原及总脂含量增加(P<0.05), 但对飞行肌内三者含量无明显影响(P>0.05), 外用早熟素Ⅰ对短翅型雌虫卵巢内蛋白质、 糖原及总脂含量亦无明显影响(P>0.05)。上述结果表明, 丽斗蟋长翅型雌虫首先将获得的资源用于发育飞行所需的飞行肌, 短翅型雌虫则首先将所获得的资源用于发育繁殖所需的卵巢, 但长翅型雌虫飞行肌与卵巢间的资源分配方式受保幼激素的影响。     

Seasonal wing length dimorphism in a tropical seed bug: ecological significance of the short-winged form

Summary Short-winged forms of the bug, Jadera aeola, were discovered during the late dry season at a study site on Barro Colorado Island, Panama. They were smaller than long-winged adults not only in elytron and hindwing sizes but also in head, thorax, rostrum and hind femur dimentions. When newly emerged adults were kept under natural photothermal conditions, short-winged females started ovipositing earlier, produced more eggs, and died earlier than did long-winged females. This was partly due to the fact that long-winged adults usually enter diapause after adult energence (Tanaka et al. 1987b). Egg size was another factor contributing to the high egg production in the short-winged form; it was significantly smaller in the short-winged form than that in the long-winged form. Wing form was apparently environmentally induced because all off-spring of short-winged adults became long-winged. No short-winged form was obtained in the laboratory when nymphs were reared on different species of seeds at different photoperiods. Although the factor(s) responsible for determination of the wing form remains unknown, the ecological significance of this short-winged form seems clear, i.e. exploitation of the food resource left in the late dry season. Wing length dimorphism in J. aeola could be a response to unpredictable availability of the food resource determined by seed production of the host plants (Sapindaceae) and by the timing of the onset of a wet season.     

Relative contribution of genetic and environmental factors to determination of wing morphs of the brown planthopper Nilaparvata lugens

Wing dimorphism is a fascinating feature of the ability of insects to adapt to environments. The brown planthopper (BPH) Nilaparvata lugens, a serious pest of rice, can switch between the long- and short-winged morphs. It has been known that environmental factors can affect the wing morph of BPH. However, it is still unclear whether the effect of environment is dependent on BPH genetic backgrounds or not. In the present study, we established the pure-bred lineages of short- and long-winged BPHs via multigenerational selection, and we found that survival and fecundity were similar between these 2 lineages. Wing morphs of the pure-bred lineages were almost fully dependent on genetics, but independent of the environmental factors, nymphal density and rice plant stage, 2 key factors affecting BPH wing morphs. In the unselected BPH population, short- and long-winged morphs were produced depending on those 2 environmental factors, indicating the contribution of environment to wing morph. In the wing-selected lineages, 4 developmental regulated genes of wing, NlInR1, NlInR2, NlAkt, and NlFoxo were expressed stably in the short-winged adults, but almost silenced in the long-winged adults. However, all these genes were expressed normally with a similar level in both the short- and long-winged adults in an unselected population except NlFoxo. The pure-bred lineages of long- and short-winged morphs exhibited different expression patterns of wing development-regulated genes, suggesting the genetic determination of wing morphs. Effects of environmental factors on wing morphs occurred only in the genetic mix population.     

Genetic variation in developmental times among four pure lines exhibiting specific wing form and body color in the brown planthopper, <Emphasis Type="Italic">Nilaparvata lugens</Emphasis> (Hemiptera: Auchenorrhyncha)

Developmental times were compared among four pure lines of wing-dimorphic Nilaparvata lugens generated by successive selections for the following designated characteristics: blackish short-winged (BS), blackish long-winged (BL), yellowish brown short-winged (YS) and yellowish brown long-winged (YL). When the nymphs were reared on rice seedlings at low density, the developmental time was in the order of BS = BL < YS = YL, while with an increase in rearing density, the time lengthened markedly in long-winged lines but only slightly in short-winged lines, rearranging the order to BS < BL = YS < YL. Among F₁ progeny of BS × YL, the developmental time in short-winged phenotypes was the same as those of BS lines at any of the densities tested, but the long-winged phenotypes emerged as adults at intermediate time between BS and YL lines at high density. This method of inheritance of developmental time and the differences in the duration observed among the four pure lines suggest that the genes regulating wing form and body color expression are also involved in the regulation of developmental time.     

Diversities in morphological characteristics and seed germination behavior in fruits ofSalsola komarovii Iljin

Two different types of dispersal units (called fruits in this study) were observed inSalsola komarovii Iljin. One is a fruit which has dark brown lignified tepals with long wings and a green seed and falls easily from the mother plant (long-winged type). Another has light brown lignified tepals with short wings and a yellow seed and attaches tightly to the mother plant (short-winged type). This difference of fruit type appeared independently from maturity of fruit. Seeds in the short-winged fruits were in dormancy for a longer period of time than those in the long-winged fruits. The germination rate was significantly higher in the seeds of long-winged fruits. The dormancy in seeds of the long-winged fruits was effectively terminated by reducing the temperature but the effect of chilling was very weak in seeds of the short-winged fruits. It was concluded from these observations that there exists a dimorphism in the fruits of, or a heterocarpy in,S. komarovii. The plants grown under water stress produced mostly short-winged fruits and those grown under well-watered conditions bore fruits of both types. Exogenously applied abscisic acid (ABA) tended to produce the short-winged fruits, suggesting that the heterocarpy was, at least partly, regulated by ABA.     

Developmental control of ultrasound sensitivity by a juvenile hormone analog in crickets (Teleogryllus oceanicus)

Cricket ears are sensitive to ultrasound as well as to lower, cricket-like sound frequencies. Ultrasound stimuli evoke negative phonotaxis in flying crickets, a behavior that has been interpreted as a defensive response against predation by echolocating bats. A recent study on a wing-dimorphic species, Gryllus texensis, showed that short-winged individuals, which are incapable of flight, are less sensitive to ultrasound, but not to lower sound frequencies, than their long-winged counterparts. The developmental decision to develop as a long- or short-winged individual is made during the last two larval instars, and there is some evidence suggesting that juvenile hormone (JH) has an instructive role, such that high levels of JH result in short-winged individuals. We show that treatment of last-instar larvae of a monomorphic long-winged species, Teleogryllus oceanicus, with a JH analog causes a decrease in sensitivity to ultrasound, but not to the lower sound frequency used for intraspecific communication.     

Effects of wing-pad removal and corpus allatum implantation on development of wings, flight muscles, and related structures in the striped ground cricket, Allonemobius fasciatus

ABSTRACT. Removal of a hindwing pad from late instar nymphs of the striped ground cricket, Allonemobius fasciatus DeGeer (Gryllidae) inhibits the formation of long-winged adults. All nymphs become short-winged when their hindwing pad is removed during the sixth (penultimate) instar or during the first 2 days of the seventh instar. When a hindwing pad is removed on day 4 of the seventh instar or later, there is no effect on wing development and at least 50% of the crickets emerge as long-winged adults as in controls.
The period sensitive for removal of a hindwing pad varies among different structures, e.g. elytra, hindwings, flight muscles, and the acrotergite of the first abdominal segment. As a result, some intermediate forms are produced when a hindwing pad is removed in the middle of the seventh instar.
Implantation of corpora allata (CA) into late instar nymphs also inhibits formation of long-winged adults. When CA are implanted on day 2 or 3 of the seventh instar, some of the resulting adults have abnormally short elytra. Implantation of CA at an earlier stage produces supernumerary (eighth instar) nymphs.