茶尺蠖核型多角体病毒研究

本文报道茶尺蠖核型多角体病毒(EoNPV)基本特性及应用价值研究结果。EoNPV属昆虫杆状病毒属A亚组,单粒包埋型。其病毒粒子平均大小62×220nm,具有特异的肽谱、DNA内切酶谱和血清学反应特性。在动物机体及离体细胞中均未显示对非宿主昆虫和人畜的致病能力和遗传诱变效应 室内毒力测定及田间防治试验结果显示EoNPV对其宿主昆虫茶尺蠖有较强的毒力和较好的防效。茶尺蠖人工饲养及EoNPV人工增殖技术的建立,及具有光保护作用的EoNPV杀虫剂剂型的成功筛选,为EoNPV实际应用的可行性提供了依据。EoNPV作为茶尺蠖的生物控制剂,应用于茶园是有效、安全和可行的。     

土荆芥精油对灰茶尺蠖幼虫的熏蒸活性及对其体内两种酶活力的影响

采用皿内测定法,研究了土荆芥精油对灰茶尺蠖的熏蒸活性及对其体内2种解毒酶活性的影响。结果表明,随着土荆芥精油剂量的增加和处理时间的延长,灰茶尺蠖幼虫的死亡率也相应升高;LC_(50)随着处理时间的延长,逐渐降低;通过测定土荆芥精油对灰茶尺蠖幼虫体内解毒酶活力变化表明,其精油对灰茶尺蠖幼虫体内的乙酰胆碱酯酶呈明显的抑制作用,对羧酸酯酶呈激活后抑制的作用。     

两近缘种茶尺蠖的抗菌肽基因在抵抗EoNPV感染过程中的表达差异

小茶尺蠖Ectropis obliqua Prout和灰茶尺蛾Ectropis grisescens Warren是危害茶园的两近缘种害虫。用小茶尺蠖核型多角体病毒E.obliqua nucleopolyhedrovirus(Eo NPV)喂饲两近缘种茶尺蠖,10 d后小茶尺蠖的死亡率为100%,而灰茶尺蛾的死亡率仅为35.5%。以感染病毒的两近缘种3龄幼虫的cDNA为模板,采用RT-PCR技术克隆获得了gloverin、attacin和moricin 3个抗菌肽基因,通过邻位连接法构建系统发育树聚类分析显示小茶尺蠖和灰茶尺蛾的Gloverin、Attacin和Moricin蛋白均分别归为一个分支;进一步采用qPCR检测发现,病毒处理后gloverin、attacin和moricin 3个基因在两近缘种茶尺蠖体内均表达上调,gloverin基因在12 h达到最大值,moricin基因在48 h达到最大值,且gloverin和moricin基因在灰茶尺蛾中表达量显著高于小茶尺蠖,而attacin基因在病毒侵入后的不同时期在两近缘种茶尺蠖体内表达量是变化的,推测gloverin、moricin和attacin基因可能与病毒的侵入有关。这一研究结果为进一步揭示抗菌肽基因在昆虫与病毒互作中的作用打下了基础。     

茶尺蠖绒茧蜂放蜂试验

茶尺蠖(茶尺蛾)Ectropis obliqua hypulina Wehrli是我国华东地区的一种主要茶树害虫。单寄生于其幼虫体内的茶尺蠖绒茧蜂Apanteles sp.是目前已知的最主要的天敌。为了在此虫防治上开辟一条新的途径,减少用药,减少农药残留和环境污染,在几年来茶尺蠖幼虫期寄生天敌自然寄生率考查、茶尺蠖绒茧蜂生物学特性研究以及     

性引诱剂对茶尺蠖防治效果研究

为探索咸宁茶区茶尺蠖发生规律及性引诱剂对茶尺蠖的诱杀效果,本研究对咸宁茶区茶尺蠖发生期及发生量进行了监测,同时,研究并分析了性引诱剂对茶尺蠖的诱杀效果,为茶园害虫绿色防控提供了科学的依据。     

适用于高温季节的茶尺蠖核型多角体病毒制剂的筛选

生物测定表明,直接使用茶尺蠖核型多角体病毒(EoNPV)病死虫匀浆稀释液,抗紫外线能力最强,提纯病毒加苋菜红食品染料次之。苋菜红食品染料的保护作用极显著强于胭脂红食品染料以及黑色素等,可作为保护剂供生产实际应用。杀灭菊酯和硫酸铜对EoNPV具有一定的增效作用。两者各自与该病毒混用后,在34℃下实际杀虫率分别为理论杀虫率的1.65和1.35倍。天皇星、灭幼脲Ⅲ、灭扫利、乐果、CpNPV及克螨特则对此病毒无增效作用。正交筛选的结果表明,34℃下9种EoNPV制剂中以EoNPV-Ⅰ乳剂为最佳,EoNPV-Ⅱ、Ⅶ乳剂和EoNPV-Ⅳ水剂次之,对三龄初幼虫的杀虫率由单用EoNPV的4.76%分别提高到89.04%和71.02—75.11%,可供生产上试用。     

不同地理种群茶尺蠖适合度差异研究

为了明确不同地理种群茶尺蠖种群分化特征,对室内条件下饲养的茶尺蠖发育和生殖力进行研究.结果表明:不同地理种群间茶尺蠖卵和幼虫的发育历期具有显著差异,即偏低纬度茶尺蠖种群的发育历期长于偏高纬度种群,而蛹和成虫的发育历期差异不明显;茶尺蠖各种群的净生殖率(R0)、内禀增长率(Rm)和周限增长率(λ)均为武汉种群最大,而种群加倍时间(Dt)则以武汉种群最小,故利于该种群茶尺蠖的发展;同一种群中不同世代茶尺蠖间的幼虫发育历期和平均世代周期随着世代的推移有增加的趋势,但差异不显著;不同地理种群茶尺蠖间存活率和生殖力随着在实验条件下的不断饲养,差异会逐渐缩小.     

两株茶尺蠖核型多角体病毒毒株对灰茶尺蠖的毒力及基因组差异

为明确茶尺蠖核型多角体病毒（Ectropis obliqua nucleopolyhedrovirus,EcobNPV）毒株对灰茶尺蠖E.grisescens的毒力差异及其机制,以前期筛选获得的高效毒株EcobNPV-QF4和原始毒株EcobNPV-QV为研究对象,采用PCR扩增技术对其进行分子鉴定,通过叶盘法毒力生测试验测定毒力差异,并用定量PCR技术比较病毒拷贝数及基因组测序方法分析基因组差异。结果表明,EcobNPV-QV经分子鉴定存在3个基因型,而EcobNPV-QF4则存在2个基因型,2株毒株的基因型存在差异;EcobNPV-QV和EcobNPV-QF4对灰茶尺蠖的幼虫毒力分别为4.26×10⁶ PIB/头和9.72×10⁷ PIB/头,EcobNPV-QF4对灰茶尺蠖的毒力是EcobNPV-QV的22.8倍;饲毒0~48 h之间EcobNPV-QF4在灰茶尺蠖体内的增殖拷贝均显著高于EcobNPV-QV,EcobNPV-QF4比EcobNPV-QV具有更快的繁殖能力;比较2株毒株的基因组,EcobNPV-QF4比EcobNPV-QV长315 bp,与EcobNPV-QV相比EcobNPV-QF4基因组的hr1~hr3区域出现倒位,其中核苷酸还原酶小亚基基因在EcobNPV-QF4中重复1次,3个同源重复区的回文序列存在差异但2株毒株仍为α杆状病毒亚家族II亲缘种分离株。表明2株毒株的基因组在同源重复区的序列及回文个数间存在差异,推测这可能与导致其对灰茶尺蠖毒力差异有关。     

茶尺蠖和灰茶尺蠖幼虫两种共有寄生蜂的鉴定

茶尺蠖Ectropis obliqua(Prout)及其近缘种灰茶尺蠖E.grisescens Warren是我国茶树上最为重要的两种食叶性害虫,茶尺蠖绒茧蜂Apanteles sp.和单白绵绒茧蜂Apanteles sp.是茶尺蠖Ectropis spp.幼虫期最重要的两种寄生性天敌昆虫。为了明确这两种绒茧蜂的物种名称及形态差别,同时探明这两种绒茧蜂是否是茶尺蠖和灰茶尺蠖的共有寄生蜂,本研究采集了我国8省22个地方的茶尺蠖或灰茶尺蠖幼虫,通过室内饲养获得两种绒茧蜂的标本,对标本进行形态鉴定;同时将两种绒茧蜂的COI基因序列与Bold数据库中已知物种进行比对。结果表明,从野生茶尺蠖和灰茶尺蠖幼虫中均饲养出之前报道的茶尺蠖绒茧蜂Apanteles sp.和单白绵绒茧蜂Apanteles sp.两种寄生蜂;形态鉴定和分子比对结果表明,这两种广义绒茧蜂Apanteles spp.实为尺蠖原绒茧蜂Protapanteles immunis(Haliday,1834)和单白绵副绒茧蜂Parapanteles hyposidrae(Wilkinson,1928)。本研究明确了40多年前国内报道的茶尺蠖幼虫期的两种绒茧蜂Apanteles spp.的物种名称和主要形态特征,确定了这两种绒茧蜂是茶尺蠖和灰茶尺蠖的共有寄生蜂,同时报道了一个中国新记录种尺蠖原绒茧蜂。研究结果为今后正确判定本地优势寄生蜂物种进而对茶尺蠖和灰茶尺蠖进行生物防治提供科学依据。     

茶尺蠖性信息素的田间使用技术及防治效果

为明确茶尺蠖Ectropis obliqua性信息素的田间使用技术,在田间设置茶尺蠖性信息素诱捕器的5、10和20 m挂放密度处理区及其对照区,并对不同处理区和对照区中诱捕器诱集的茶尺蠖成虫和幼虫数量以及同一挂放密度下距性信息素诱捕器不同距离的茶尺蠖幼虫数量进行调查,确定性信息素诱捕器的最佳挂放间距;在此基础上确定性信息素诱捕器对茶尺蠖的田间防治效果,并利用性信息素诱捕器对茶尺蠖成虫的田间动态进行监测。结果表明:在任何一个挂放密度下,处理区诱捕器诱集的茶尺蠖成虫数量均显著高于对照区诱捕器,但在不同挂放密度的小区之间单个诱捕器诱集的成虫数量差异不显著;当茶尺蠖幼虫重度发生(1 361.57头/m~2)时,10 m间隔挂放1套性信息素诱捕器可显著降低茶尺蠖幼虫的发生数量;当茶尺蠖幼虫轻度发生(38.40头/m~2)时,间距为20 m挂放一套性信息素诱捕器时,茶尺蠖的校正防治效果可达88.44%;茶尺蠖性信息素诱捕器在6月上旬、8月上旬、8月下旬至9月初、9月下旬分别有一个明显的诱捕高峰期,与田间茶尺蠖的发生高峰期基本一致。表明茶尺蠖性信息素诱捕器可作为大量诱捕和虫情监测的手段在茶园中应用。     

混合病毒在不同宿主中交替繁殖 对病毒毒力的影响

研究了混合病毒在不同宿主中交替繁殖对病毒毒力的影响,选用对甜菜夜蛾Spodoptera exigua (Hüber)较专化的苜蓿银纹夜蛾核多角体病毒(AcNPV)和对斜纹夜蛾Spodoptera litura (Fabricius) 较专化的斜纹夜蛾核多角体病毒(SlNPV),按多角体数量1∶ 1混合后分别连续感染或交替感染甜菜夜蛾和斜纹夜蛾,比较各代病毒对两种夜蛾的活性。测定结果表明:SlNPV中国株和AcNPV混合,起始感染甜菜夜蛾,再在斜纹夜蛾和甜菜夜蛾中交替繁殖后,F4代斜纹夜蛾病毒(由斜纹夜蛾中收集的病毒)对3龄甜菜夜蛾幼虫的致死率极显著高于F2代斜纹夜蛾病毒,F3代甜菜夜蛾病毒(由甜菜夜蛾中收集的病毒)对斜纹夜蛾的致死率也显著高于F1代甜菜夜蛾病毒;混合病毒起始感染斜纹夜蛾,再经甜菜夜蛾和斜纹夜蛾交替繁殖后,斜纹夜蛾病毒对甜菜夜蛾的毒力以及甜菜夜蛾病毒对斜纹夜蛾的毒力也都显著提高。交替传代后的病毒比同代次连续传代后的病毒对非宿主的毒力显著提高,并且它们对原宿主的毒力未有减弱。AcNPV和SlNPV日本株混合病毒在甜菜夜蛾和斜纹夜蛾中交替繁殖后也表现出对非宿主毒力的提高。表明混合病毒在两种不同原宿主中交替繁殖比其在同种宿主中连续繁殖后对非繁殖原宿主的毒力显著提高。     

复合型苜蓿银纹夜蛾核型多角体病毒制剂

复合型苜蓿银纹夜蛾核型多角体病毒制剂主要由苜蓿银纹夜蛾核型多角体病毒和斜纹夜蛾核型多角体病毒两株病毒毒株和苏云金杆菌按一定比例复合而成,经单一型,复合型病毒制剂毒力测定比较,获得增效比值为５．１－９．５的最佳配比,并以甜菜夜蛾为宿扩大增殖形成雏型产品。     

三种强启动子和egt基因缺陷在重组核型多角体病毒杀虫剂研发中的意义

核型多角体病毒 (nuclearpolyhedrosisvirus ,NPV)是鳞翅目昆虫的重要病原微生物。采用构建重组高效表达外源毒蛋白的 p10、ph、ocu基因的强启动子 ,以提高野生型NPV的毒力 ;探讨删除杆状病毒中的蜕皮甾体尿苷二磷酸葡糖转移酶 (ecdysteroidUDP-glucosyltraansferase ,EGT)基因对改良病毒杀虫剂的杀虫效果 ,为进一步完善重组NPV杀虫剂的研发和安全生产提供了理论基础     

The effects of azadirachtin and nucleopolyhedrovirus on midgut enzymatic profile of Spodoptera litura Fab. (Lepidoptera: Noctuidae)

The effects of azadirachtin (AZA) and nucleopolyhedrovirus (NPV) on midgut enzyme activity in Spodoptera litura Fabricius (Lepidoptera: Noctuidae) (Tobacco cutworm) were evaluated. Gut enzyme activities were decreased by AZA and NPV individually and in combination. When S. litura larvae were fed a diet of castor leaves treated with AZA and NPV in bioassays, gut enzyme—acid phosphatases, alkaline phosphatases, adenosine triphosphatases, and lactate dehydrogenase—activities were decreased. There were statistically significant differences (P ? 0.05) in enzyme activities between combined and individual treatment. A synergistic effect of botanical insecticides and virus was found when combined in low doses. These effects are most pronounced in early instars. Maximum weight loss (59-72%) occurred, when AZA and NPV were combined.     

棉铃虫核多角体病毒的生产方法及其在生物防治中的应用

本文是报道棉铃虫Heliothis armigera核多角体病毒(NPV)的生产方法及其在棉田应用的试验研究结果。NPV用棉铃虫幼虫生产。从虫卵接种开始,用人工饲料饲养棉铃虫幼虫,到四龄用病毒感染,六天后收集病死虫,病毒致死率80～85%,单虫含量4～6×10~9PIB。由棉饼粉、粗棉油等配制成诱饵加低剂量西维因作病毒增效剂与NPV混用,显著提高病毒的防治效果。四年田间试验表明,NPV杀虫剂防治棉铃虫的效果相当于或优于常用高效化学农药。1979年开展了大面积飞机微量喷雾PVN杀虫剂的试验,也取得了比较好的防治效果。     

昆虫病毒在害虫防治上的应用及其对寄生蜂的影响

本文综述了昆虫病毒，包括核多角体病毒（NPV）、昆虫痘病毒（EPV）和颗粒体病毒（GV），在防治农林害虫中的应用及其对寄主寄生蜂影响的研究进展，同时也介绍了昆虫杆状病毒诱导细胞凋亡及基因工程研究的近况。     

The impact of co-infection by a nucleopolyhedrovirus and the endoparasitoid Microplitis pallidipes Szepligeti on the protective enzymes in hemolymph of Spodoptera exigua

We determined the effects of parasitism by Microplitis pallidipes and/or nucleopolyhedrovirus (NPV) infection on protective enzymes in the hemolymph of Spodoptera exigua larvae. We found that concentrations of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) were constantly higher within the five observation days after treatment in parasitized/virus-infected than healthy (control) larvae. In parasitized + infected larvae, enzyme levels reduced from day 1 compared to parasitized larvae. Compared to enzyme levels in virus-infected larvae, in parasitized + infected larvae, CAT concentrations were generally lower, while SOD concentrations were significantly lower in the first two days and POD concentrations were lower in the first two days, but higher in following three days. We concluded that joint and separate parasitism and NPV infection promoted the activities of protective enzymes. Our findings also revealed that NPV infection inhibited the activities of protective enzymes induced by parasitism of M. pallidipes and that the parasitism inhibited the CAT activity induced by NPV infection.     

Improving the efficacy of nuclear polyhedrosis virus and Bacillus thuringiensis against Helicoverpa spp. with ultra-violet light protected petroleum spray oils on cotton crops in Australia

Nuclear polyhedrosis virus (NPV) and Bacillus thuringiensis (Bt) are the most commonly used biopesticides for the control of Helicoverpa spp. larvae on cotton crops in Australia. The performance of NPV and Bt against Helicoverpa spp. larvae on cotton crops, is inconsistent and at times totally unsatisfactory against high densities of Helicoverpa spp. larvae. We determined the effect of mixing petroleum spray oils, containing ultra-violet light absorbing compounds, with NPV and Bt for efficacy against Helicoverpa spp. larvae, levels of cotton plant damage, and persistence of efficacy. The study showed that the efficacy and persistence of NPV and Bt were increased when mixed with petroleum spray oil (PSO?–?Canopy®) at the rate of 2% (v/v). In the field experiments, mixing NPV with 1 and 2% (v/v) PSO, increased Helicoverpa spp. mortality from 25.9 to 31.5 and 44.8%, respectively. Similarly, the mortality caused by Bt, when mixed with 1 and 2% (v/v) PSO, was increased from 31.5 to 36.0 and 48.2%, respectively. In addition, 1 and 2% PSO mixtures with NPV increased persistence of efficacy from 1.1 to 1.6 and 2.5 days, respectively, whilst persistence of Bt was increased from 1.5 to 1.8 and 2.6 days, respectively. In another study using potted cotton plants, in which the plants were left outdoors throughout the study, the average NPV induced mortality of first instar Helicoverpa larvae was increased from 20.9% to 35.9 and 43.4% by 1 and 2% (v/v) PSO, respectively. Persistence of NPV efficacy was enhanced by 2 and 3.1 times by 1 and 2% (v/v) PSO, respectively. Similarly, Bt induced mortality of Helicoverpa larvae was increased by 1 and 2% PSO from 68.1 to 78.8 and 83.2%, respectively, and the persistence of Bt efficacy was enhanced 1.3?–?2.0 times, respectively. In a mesh house study, young cotton plants, treated with a PSO/biopesticide mixture, suffered less leaf damage than cotton plants treated with the biopesticides alone. In conclusion, the results of this study showed synergies from the combined use of UV protected PSO and NPV or Bt, against Helicoverpa spp. larvae on cotton. Such a biopesticide-PSO combinations could be a useful tool for IPM program in cotton.     

Economic and financial model to the mass-rearing of Macrolophus pygmaeus (Rambur) (Heteroptera: Miridae), a biological control agent against the tomato moth Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) in protected culture

BACKGROUND

Tuta absoluta (Meyrick) is a major pest of tomato produced in glasshouses and open field, causing severe damages to crops, reducing the quality of tomato fruits. The current maintenance of the pest populations below the economic threshold is not achieved by natural and classical control, thus requiring the continuous application of biological control agents (BCAs), under an augmentative or inoculative approach. The present study aims to develop an economic and financial model to evaluate the commercial viability of a continuous mass production of Macrolophus pygmaeus (Rambur), a BCA commonly used against the tomato moth, Tuta absoluta, in protected culture. The estimations for our model were based on two approaches: the farm-level impact analysis and the benefit–cost analysis.

RESULTS

The results of the farm-level analysis show that the adoption of a more sustainable biological control approach is profitable for farmers and the benefit–cost analysis provides evidence that the investment on a new factory dedicated to the mass rearing of M. pygmaeus to control tomato moth populations generates a positive net present value (NPV) of 7.2 million euros, corresponding to an internal rate of return (IRR) of 28.4% per year.

CONCLUSION

Our results are in line with (i) the more recent European Commission proposals for a new Regulation on sustainable use of plant protection products, which includes the reduction of 50% the use and risk of chemical pesticides by 2030 and (ii) most of the existing literature which conclude that new projects on BCA production are worth investments. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

棉铃虫C型凝集素抑制多角体病毒体内增殖

为探究C型凝集素（C-type lectin）在病毒传播中的可能作用,采用半定量RT-PCR法检测了棉铃虫Helicoverpa armigera不同C型凝集素在核型多角体病毒（nuclearpolyhedrosis virus,NPV）感染后的表达变化,并通过real-time qPCR和RNA干扰技术研究了NPV在棉铃虫体内的增殖状况及对棉铃虫致死率的影响。结果显示,在感染NPV后,棉铃虫血细胞C型凝集素Ha-lectin1和Ha-lectin5基因表达变化明显,其中Ha-lectin1在血细胞和脂肪体中的表达显著上调;注射dsRNA后,Ha-lectin1基因相对于actin的表达量均值由0.19下降到0.04,下降幅度达79%,显示Ha-lectin1基因被成功干扰。随着Ha-lectin1基因的沉默,病毒滴度由0.0015快速升高到1.95,棉铃虫的死亡率也随之增加到97.7%。表明C型凝集素能够抑制NPV在棉铃虫体内的增殖。