First Record of Steinernema kraussei (Rhabditida: Steinernematidae) from Turkey and Its Virulence against Agrotis segetum (Lepidoptera: Noctuidae)

During a survey of entomopathogenic nematodes (EPNs) in the eastern Black Sea region of Turkey in 2009–2012, a steinernematid species was recorded and isolated using the Galleria-baiting method. The isolate was identified as Steinernema kraussei based on its morphological and molecular properties. The analysis of the ITS rDNA sequence placed the Turkish population of S. kraussei in the “feltiae-kraussei” group in the clade that contains different isolates of the species. This is the first record of S. kraussei from Turkey. The efficacy of S. kraussei was tested on Agrotis segetum (Lepidoptera: Noctuidea) larvae at different densities (100, 300, and 500 infective juveniles (IJs) g⁻¹ dry sand ) in laboratory conditions at 25 °C. The highest mortality (98%) was obtained with 500 IJs g⁻¹ dry sand within 7 d after inoculation. Our results indicate that the new isolate is a highly promising biological control agent against A. segetum, one of the most serious soil pests of agricultural area and fruits worldwide.     

小地老虎食诱剂糖醋酒液配方筛选及发酵增效作用

【目的】为筛选和规范诱集小地老虎Agrotis ipsilon成虫的最佳糖醋酒液配方及最佳发酵时间,确定发酵液中的挥发物成分。【方法】优选以单一纯物质糖、醋、酒和水进行混配的配方A, B, C和D, 其配比分别为蔗糖(g)∶乙酸(mL)∶无水乙醇(mL)∶纯水(mL)(m/v/v/v)=3∶1∶3∶80, 3∶1∶3∶160, 3∶1∶6∶80和1∶1∶3∶80及常见配方E［白糖(g)∶白醋(mL)∶白酒(mL)∶自来水(mL)= 6∶3∶1∶10(m/v/v/v)］,分别并采用Y型嗅觉仪及GC-MS测试和分析小地老虎成虫对发酵1~15 d后的糖醋酒液的趋性及发酵液中的挥发物成分。【结果】趋性试验结果表明,糖醋酒液B配方对小地老虎诱集效果显著优于A, C, D和E配方,C配方次之。小地老虎成虫对B和C这两种配方发酵8 d发酵液的选择反应率均显著高于对未发酵及发酵5 d和7 d外的其他发酵时间发酵液的选择反应率。挥发物成分组成分析结果显示,糖醋酒液B配方发酵8 d发酵液的挥发物共有41种化合物,主要包括17种烃类化合物、8种醛类化合物、2种酮类化合物、4种醇类化合物、5种酯类化合物和5种醚类化合物。而且发酵8 d的糖醋酒液B配方与其未发酵以及发酵4 d和发酵12 d的糖醋酒液B配方之间的挥发物成分差异较大。【结论】优选并明确单一物质糖、醋、酒和水进行混配的小地老虎食诱剂配方(3∶1∶3∶160, m/v/v/v),其糖醋酒液通过发酵可以产生增效作用,最佳发酵时间为8 d。     

Agrotis segetum雄蛾触角叶性信息素反应神经元对电刺激触角神经的反应

为探讨电刺激Agrotis segetum雄蛾触角神经是否可以作为MGC中神经元的识别手段,采用细胞内电生理记录方法,共记录34个对性信息素有反应的MGC神经元,并测试了其中12个神经元对性信息素刺激的反应,22个神经元对性信息素刺激和电刺激的反应。结果表明,MGC神经元对性信息素及电刺激的反应模式基本一致,为一种双相反应模式。两种刺激方式均能诱导出兴奋反应,电刺激得到的兴奋反应比由信息素刺激引起的要短;MGC神经元对两种刺激的超极化反应（抑制反应）幅度影响没有显著性差别,在电刺激实验的22个神经元上,超极化反应幅度和抑制时间都与神经元本身放电频率有一定的相关性。超极化反应是在LN参与下一定的神经回路对刺激所产生的反应而形成的。这提示两种刺激所作用的神经回路应是一致的,但从整个实验过程记录到的神经元情况来看,还须进一步结合形态学实验来验证电刺激触角神经作为MGC神经元的识别手段。     

小地老虎变态期脂肪体变化及保幼激素类似物的影响

本文对小地老虎Agrotis ypsilon(Rottemberg)从四龄幼虫开始经预蛹和蛹的变态期及羽化为成虫后的脂肪体出现的超微结构变化,蛋白质含量的变动,以及蛋白质颗粒的形成和消失过程,进行了系统观察和组织化学分析。结果表明:(1)在幼虫期的后期,脂肪体扩大成宽带状,细胞体积增大的同时出现双核和多核。进入预蛹期,细胞内开始出现嗜碱性“蛋白质颗粒”,血细胞吞噬部分脂肪体细胞。蛹龄一天时,脂肪体转变成块状,细胞内充满大型蛋白质颗粒。在蛹龄5—10天内,“幼虫脂肪体”逐步崩解,围膜及细胞膜消失。至蛹龄12天时转变为预成虫,脂肪体细胞重新出现,并以气管分支为中心聚合成花朵状圆球体,再组成串状“成虫脂肪体”,仍充满蛋白质颗粒。幼虫期发达的粗面内质网和线粒体,至预蛹期则衰变成几种类型的蛋白质颗粒。(2)六龄幼虫在1—5日龄期间,每克脂肪体的蛋白质含量稳定在7.1—8.4毫克之间,随后逐步升高,至预蛹期达16.3毫克的峰值。蛹初期,雄蛹和雌蛹的含量分别增高到预蛹期的1.63和2.4倍。但在蛹龄2—8天内迅速下降到六龄幼虫期的水平。至蛹龄9—13天时间(包括预成虫),含量又突然猛增,雌蛹尤为显著。蛹期脂肪体细胞充满着的几种蛋白质颗粒,在羽化为成虫后的24小时内全部消失。在六龄幼虫期用保幼激素类似物ZR-515(20微克)作体壁处理,可使幼虫期延长4天,并使九日龄幼虫的脂肪体仍保持幼虫型状态。     

小地老虎β-微管蛋白基因cDNA序列的克隆、序列分析和表达检测

β-微管蛋白在昆虫生长发育、信号传导、抗药性等方面具有重要作用.本研究以小地老虎Agrotis ypsilon(Rottemberg)3龄幼虫为材料,利用RT-PCR、cDNA末端快速扩增技术(RACE)克隆得到小地老虎β-微管蛋白基因的cDNA序列,命名为AgTubB(GenBank登录号:JN029962),并检测...     

Molecular characterization of Agrotis segetum nucleopolyhedrovirus from Poland

The turnip moth, Agrotis segetum (Lepidoptera, Noctuidae), is an important pest insect in Europe, Asia, and Africa. We have genetically characterized and classified a nucleopolyhedrovirus isolated from A. segetum larvae in Poland (AgseNPV-P). The restriction pattern of AgseNPV-P was distinct from an isolate from England/France (AgseNPV-UK and AgseNPV-F). Sequence analysis of three conserved baculovirus genes, polyhedrin, lef-8 and pif-2, revealed that AgseNPV-P differs substantially from the already described NPVs isolated from A. segetum and possibly represents a new NPV species. Phylogenetic analysis placed AgseNPV-P among group II NPVs and showed the closest relationship to Agrotis ipsilon (Agip) NPV and Spodoptera exigua (Se) MNPV.     

混合感染后杆状病毒间的增效作用——病毒蛋白及核酸的初步分析

AsNPV+HasNPV、AsNPV+HaNPV、AsNPV+PsNPV分别感染烟青虫、棉铃虫和粘虫幼虫,对分离到的核型多角体病毒(AsNPV+HasNPV)-Helicoverpa assulta、(AsNPV+HaNPV)- H.Armigera和(AsNPV+PsNPV)-Pseudaletia separata,经电镜观察,多角体蛋白及病毒粒子蛋白SDS-PAGE电泳,病毒核酸的限制性内切酶酶解分析等研究,证明各病毒的多角体形态不规则,大小差异极大,病毒粒子为杆状,(AsNPV+HasNPV)-H.Assulta 和(AsNPV+HaNPV)-H.Armigcra病毒粒子有单粒和多粒包埋类型, (AsNPV+PsNPV)-P.Separata为多粒包埋型。各病毒的多角体蛋白基本上只有一种多肽,分子量为25 000道尔顿左右。(AsNPV+HasNPV)-H.Assulta、(AsNPV+HaNPV)-H.armigera和(AsNPV+PsNPV)-P.Separata的病毒粒子分别有10、14、5条多肽,分子量大小在13 500～98 000,13 000～88 000,18 500～52 000道尔顿之间。病毒核酸经EcoRI,HindIII,HindIII+BamHI酶解,其DNA的酶切位点,大小及DNA的总分子量与AsNPV和原寄主的Has-NPV,HaNPV和PsNPVDNA的酶切图谱存在一定差异,混合病毒侵染昆虫后新复制的病毒核酸发生一定的变化,从而导致病毒蛋白和病毒形态的变化。混合感染后AsNPV对Has-NPV、HaNPV和PsNPV的侵染有明显的增效作用,其机理有待深入研究。     

A Comparison of the Efficacy of Nuclear Polyhedrosis and Granulosis Viruses in Spray and Bait Formulations for the Control of Agrotis segetum (Lepidoptera: Noctuidae) in Maize

Agrotis segetum nuclear polyhedrosis virus (AsNPV) and granulosis virus (AsGV), propagated in laboratory cultures of A. segetum in England and A. ipsilon in Spain, respectively, were applied to plots of maize plants at the one‐ to four‐leaf stage of growth. Plots were arranged in a 6 x 6 Latin square design and infested with second‐instar A. segetum larvae (the common cutworm). Each virus was applied in separate treatments by two application methods; as an aqueous spray containing 0.1% Agral as a wetting agent, and as a bran bait. The NPV was applied at a rate of 4 X 10¹² polyhedra/ha, and the GV at 4 X 10¹³ granules/ha. Soil and plants were sampled for larvae on three occasions following virus treatment: 24 h, 4 days and 11 days. The larvae were reared on diet in the laboratory, until death or pupation, to examine the rate and level of viral infection. Infection data showed 87.5% and 91% NPV infection and 12.5% and 55% GV infection in spray and bait treatments, respectively, in larvae sampled 24 h after treatment. In larvae sampled 4 days after treatment, the results were 78% and 100% NPV infection, and 13% and 6% GV infection. A total of only six larvae were retrieved on day 11. In both treatments larvae infected with AsNPV died significantly more rapidly and at an earlier instar than those infected with AsGV, indicating that AsNPV appears to have better potential as a control agent for A. segetum.     

小地老虎及粘虫幼虫对灭幼脲表现自然耐药力的体壁结构和生化基础

小地老虎Agrotis ypsilon rottem.幼虫对灭幼脲具有一定的自然耐药力。本文以粘虫Mythimna separata(Walker)作为敏感性虫种与之进行比较,实验结果表明,灭幼脲对两种试虫的室内毒力相差4倍左右,引起差异的原因,在体壁结构方面主要在于:(1)小地老虎幼虫的表皮层较粘虫的厚4.2倍左右;(2)上表皮不是匀质结构,依靠少数蜡道与体表沟通;(3)几丁质片层内的孔道数较少,仅及粘虫的1/4。由此构成了表皮对疏水性的灭幼脲表现抗穿透的性能。小地老虎幼虫体壁还含有较强的生化防卫体系,灭幼脲对多功能氧化酶、芳基酰胺酶有明显激活效应,这两种酶都是灭幼脲的降解酶。由此认为,小地老虎幼虫对灭幼脲所表现的自然耐药力,是由体壁的抗穿透性能以及由灭幼脲所激活的适应酶所造成。