茶尺蠖生物防治进展

茶尺蠖Ectropis obliqua Prout是茶园中的重要害虫,对它的生物防治研究国内已做了较多的工作.本文从茶尺蠖的研究基础-大量饲养和它的病原性天敌、捕食性天敌、寄生性天敌及信息素等方面对茶尺蠖的生物防治研究作了综述.     

美国白蛾天敌研究进展

美国白蛾Hyphantria cunea(Drury)是国家林业和草原局的重点防控对象,也是我国重大外来林业检疫性有害生物.现阶段美国白蛾在我国的种群密度持续增加和扩散,缺乏有效天敌的控制应该是重要的原因之一.本文综述了国内、外美国白蛾的捕食性和寄生性天敌的种类.捕食性天敌主要包括昆虫、蜘蛛、两栖类和鸟类,整理出国外报道的捕食性天敌名录119种,国内捕食性天敌名录29种;寄生性天敌主要包括寄生蜂和寄生蝇类,整理出国外报道的寄生性天敌名录47种,国内寄生性天敌名录53种.本文回顾了我国在美国白蛾天敌利用方面取得的阶段成果,并针对将来天敌复合体的应用和原产地天敌的引进提出了展望.     

烟草害虫天敌及其自然控制作用

介绍了云南省烟草主要害虫天敌资源和天敌昆虫的自然控制作用。在云南省烟田内 ,控制烟草害虫发生的生物因子大致可归纳为寄生性和捕食性两大类。充分发挥自然因素的生态调控作用 ,对保护利用天敌、维护烟田生态平衡、提高害虫综合治理效果、保护生态环境都有重要意义     

转双抗虫基因741杨节肢动物群落营养结构及生态位变化

转双抗虫基因741杨（简称转基因741杨）节肢动物群落中,基位物种的植食性昆虫丰富度显著降低,但中性节肢动物丰富度却明显增加.高抗和中抗的节肢动物群落中位物种和顶位物种较之对照有所增多.转基因741杨节肢动物群落的害虫功能团,其优势状况,与对照相比有所改变：天敌优势度高于对照,中性节肢动物丰富度增加,并在天敌-害虫的营养链中起着重要的调控作用.鳞翅目害虫的空间生态位宽度最小,其它各功能类群的生态位宽度较大;捕食性和寄生性天敌与鳞翅目害虫的生态位重叠均较小,而与腐生和游逛种类的生态位重叠较大;各类害虫之间、捕食性天敌与寄生性天敌之间亦存在激烈竞争.转基因741杨对寄生性天敌和捕食性天敌在利用时间资源上有正作用.各种功能类群的时-空二维生态位宽度和生态位重叠均不如单维生态位宽度和生态位重叠值大,但抗性株系天敌类群对环境的适应性优于对照.     

小菜蛾的天敌类群及其利用现状

本文介绍了小菜蛾Plutella xylostella的天敌类群包括寄生性天敌、捕食性天敌和病原微生物、原虫、线虫等共约97种,并对利用天敌进行生物防治的现状作了概述.     

浙江茶树黑刺粉虱的发生危害及天敌的研究

本文报道了浙江省茶树害虫黑刺粉虱天敌的调查结果,共发现天敌昆虫27种,其中捕食性天敌19种,寄生性天敌8种;捕食性蜘蛛9种;寄生性真菌7种.     

潍坊地区大袋蛾天敌研究

1991—1993年在潍坊地区,共收集到大袋蛾天敌11种,其中寄生蝇类3种,捕食性动物6种,病毒和白僵菌各1种。在幼虫期,优势天敌类群是奇蝇和病原微生物,可杀死35.82％的幼虫。蛹、成虫和卵期未发现寄生性天敌,研究表明,寄生性天敌对控制大袋蛾种群数量消长有重要作用。     

桔小实蝇生物防治研究进展

本文综述了国内外桔小实蝇的生物防治研究进展,概括了国内外对桔小实蝇起主要控制作用的主要天敌种类,论述了桔小实蝇寄生性天敌、捕食性天敌的利用;病原微生物中真菌、线虫、共生菌等的利用;对桔小实蝇具有引诱作用的水解蛋白、化学物质和植物次生物利用;桔小实蝇不育技术的应用等方面,并讨论了今后持续控制桔小实蝇中生物防治方面的发展趋势,以期为今后桔小实蝇持续控制提供生物防治方面的参考.     

麦田天敌种群动态及主要天敌对麦蚜捕食效能的研究

查明麦田害虫天敌有98种,其中捕食性天敌80种,寄生性天敌18种,对天敌种群动态作了系统观察,分析了影响天敌种群数量的环境因素,对大灰食蚜蝇,黑带食蚜蝇,七星瓢虫捕食麦蚜效能进行了探讨。     

害虫的生物防除

昆虫天敌对于昆虫的发生常有抑制的作用,科学工作者根据这一原理,利用害虫的天敌去防治害虫,其方法称为害虫生物防除法. 昆虫的天敌很多,包括病原微生物(病毒、细菌、真菌和原生动物)线虫、壁虱目动物,昆虫(捕食性昆虫及寄生性昆虫)和脊椎动物.除此以外,还可包括一些高等植物.昆虫的天敌,虽然有上述的几大类,但并非全部都能够用来防治害虫,利用得最普遍的是捕食性昆虫和寄生性昆虫,其次是病原微生物和脊椎动物,至于线虫和壁虱目动物,只是偶有利用.     

Mixed-genotype infections of Trichoplusia ni larvae with Autographa californica multicapsid nucleopolyhedrovirus: Speed of action and persistence of a recombinant in serial passage

Fast-acting recombinant baculoviruses have potential for improved insect pest suppression. However, the ecological impact of using such viruses must be given careful consideration. One strategy for mitigating risks might be simultaneous release of a wild-type baculovirus, so as to facilitate rapid displacement of the recombinant baculovirus by a wild-type. However, at what ratio must the two baculoviruses be released? An optimum release ratio must ensure both fast action, and the eventual competitive displacement of the recombinant virus and fixation of the wild-type baculovirus in the insect population. Here we challenged Trichoplusia ni larvae with different ratios of wild-type Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and a derived recombinant, vEGTDEL, which has the endogenous egt gene (coding for ecdysteroid UDP-glucosyltransferase) deleted. Time to death increased with the proportion wild-type virus in the inoculum mixture, although a 1:10 ratio (wild-type: recombinant) resulted in equally rapid insecticidal action as vEGTDEL alone. Five serial passages of three different occlusion body (OB) mixtures of the two viruses were also performed. OBs from 10 larval cadavers were pooled and used to initiate the following passage. Although the wild-type baculovirus was maintained over five passages, it did not go to fixation in most replicates of the serial passage experiment (SPE), and there was no good evidence for selection against the recombinant. Long-term maintenance of a recombinant in serial passage suggests an ecosystem safety risk. We conclude that for assessing ecological impact of recombinant viruses, SPEs in single and multiple larvae are relevant because of potential modulating effects at the between-host level.     

构建重组杆状病毒的几种新方法

昆虫杆状病毒作为高效的表达载体,现已广泛地用于各种外源基因的表达.但是,用传统的方法构建重组杆状病毒,存在着重组率低,纯化难及耗时长等缺点,围绕如何快速、简便、高效地构建重组杆状病毒,近几年来人们进行了一些重大的改进,包括使病毒DNA线状化以提高重组病毒的比例;在体外进行重组;同源重组和重组病毒的纯化与筛选在酵母和大肠杆菌中一次完成;使重组病毒可以形成多角体等,从而从根本上改变了传统方法中的不足;文章着重介绍了这几种新的改进方法.     

In vitro and in vivo gene delivery by recombinant baculoviruses

Although recombinant baculovirus vectors can be an efficient tool for gene transfer into mammalian cells in vitro, gene transduction in vivo has been hampered by the inactivation of baculoviruses by serum complement. Recombinant baculoviruses possessing excess envelope protein gp64 or other viral envelope proteins on the virion surface deliver foreign genes into a variety of mammalian cell lines more efficiently than the unmodified baculovirus. In this study, we examined the efficiency of gene transfer both in vitro and in vivo by recombinant baculoviruses possessing envelope proteins derived from either vesicular stomatitis virus (VSVG) or rabies virus. These recombinant viruses efficiently transferred reporter genes into neural cell lines, primary rat neural cells, and primary mouse osteal cells in vitro. The VSVG-modified baculovirus exhibited greater resistance to inactivation by animal sera than the unmodified baculovirus. A synthetic inhibitor of the complement activation pathway circumvented the serum inactivation of the unmodified baculovirus. Furthermore, the VSVG-modified baculovirus could transduce a reporter gene into the cerebral cortex and testis of mice by direct inoculation in vivo. These results suggest the possible use of the recombinant baculovirus vectors in combination with the administration of complement inhibitors for in vivo gene therapy.     

Evaluation of viral contamination in a baculovirus expression system

Insect expression systems based on baculovirus are widely used for generating recombinant proteins. Here, the infectivity of baculoviruses under the physiological stresses of ‘freeze–thaw’ and sonication and the baculoviral contamination of recombinant proteins after protein purification were evaluated. Our findings suggest that Nonidet P‐40 (NP‐40) treatment of baculoviruses completely abolishes their infectivity and that recombinant proteins purified with affinity beads do not include infectious baculoviruses. We therefore suggest that baculovirus is completely inactivated by NP‐40 treatment and that recombinant proteins are unlikely to be contaminated with infectious baculoviruses after their affinity purification.

     

Use of baculoviruses as biological insecticides

Naturally occurring baculoviruses can be used to control a wide range of insect pests. Most baculoviruses are used as biopesticides, that is, they are sprayed onto high-density pest populations in a manner akin to the use of synthetic chemical pesticides. However, other strategies that use the biological features of the viruses are also possible and should increase as we expand our knowledge of baculovirus ecology. In order to develop a baculovirus control program, several areas need to be studied before progressing to large scale field studies and commercialization. These range from laboratory efficacy testing and the development of production systems to detailed study of pest behavior and the development of appropriate application strategies.     

A Baculovirus Expression System for Insect Cells

The review considers the biology of baculoviruses, construction of transfer vectors for the baculovirus expression system (BES), selection of recombinant baculoviruses, approaches to expression of multimeric proteins, and BES potentialities and prospects.     

A baculovirus expression system for insect cells

The review considers the biology of baculoviruses, construction of transfer vectors for the baculovirus expression system, selection of recombinant baculoviruses, approaches to expression of multimeric proteins, and the potentialities and prospects of the system.     

Virion proteomics of large DNA viruses

Large DNA viruses normally have complex structures with many of protein components derived from both viral and host origins. The development in proteomics, especially mass spectrometry identification techniques provide powerful tools for analyzing large viruses. In this review, we have summarized the recent achievements on proteomic studies of large DNA viruses, such as herpesvirus, poxvirus, nimavirus and baculoviruse. The proteomics of baculovirus occlusion-derived virions (ODV) were emphasized. Different mass spectrometry techniques used on ,carious baculoviruses were introduced, and the identified structurally associated proteins of baculoviruses are summarized.     

Ligand-directed gene targeting to mammalian cells by pseudotype baculoviruses

The baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) can infect a variety of mammalian cells, as well as insect cells, facilitating its use as a viral vector for gene delivery into mammalian cells. Glycoprotein gp64, a major component of the budded AcMNPV envelope, is involved in viral entry into cells by receptor-mediated endocytosis and subsequent membrane fusion. We examined the potential production of pseudotype baculovirus particles transiently carrying ligands of interest in place of gp64 as a method of ligand-directed gene delivery into target cells. During amplification of a gp64-null pseudotype baculovirus carrying a green fluorescent protein gene in gp64-expressing insect cells, however, we observed the high-frequency appearance of a replication-competent virus incorporating the gp64 gene into the viral genome. To avoid generation of replication-competent revertants, we prepared pseudotype baculoviruses by transfection with recombinant bacmids without further amplification in the gp64-expressing cells. We constructed gp64-null recombinant bacmids carrying cDNAs encoding either vesicular stomatitis virus G protein (VSVG) or measles virus receptors (CD46 or SLAM). The VSVG pseudotype baculovirus efficiently transduced a reporter gene into a variety of mammalian cell lines, while CD46 and SLAM pseudotype baculoviruses allowed ligand-receptor-directed reporter gene transduction into target cells expressing measles virus envelope glycoproteins. Gene transduction mediated by the pseudotype baculoviruses could be inhibited by pretreatment with specific antibodies. These results indicate the possible application of pseudotype baculoviruses in ligand-directed gene delivery into target cells.     

杆状病毒用于哺乳动物细胞快速高效表达外源基因的研究

现已发现杆状病毒可进入某些培养的哺乳动物细胞,这提示可将杆状病毒作为一种对哺乳动物细胞的新型基因转移载体。对杆状病毒转移载体的改造及对哺乳动物细胞的基因转移方式进行了进一步的研究。以绿色荧光蛋白基因为报告基因,利用Bac-to-Bac系统构建了分别含有正向和反向CMV启动子表达盒的两种重组杆状病毒。可观察到CMV启动子在Sf9细胞中可启动报告基因的表达,但表达效率较低。用重组杆状病毒感染后Sf9细胞的培养上清直接与HepG2细胞作用,以流式细胞术检测基因转移效率及荧光表达强度,发现这两种病毒在相同的感染复数下对HepG2细胞具有相似的基因转移及表达效率。同时,利用流式细胞术进一步研究了直接使用重组杆状病毒感染4d后Sf9细胞的培养上清对哺乳动物细胞进行基因转移的方法。通过对HepG2细胞的实验结果显示,将带毒Sf9细胞培养上清(1.2×10⁷PFU/mL)用哺乳动物细胞培养基1倍稀释后,37℃下孵育靶细胞12h(moi=50),可达到较高的基因转移及表达效率,同时不会对细胞造成明显损伤。将重组杆状病毒与脂质体和逆转录病毒这两种系统对HepG2及CV1细胞的基因转移效率进行了比较,结果发现在同样未经浓缩等特殊处理的条件下重组杆状病毒对这两种细胞的基因转移效率是最高的。因此可以认为,经过适当改造后的Bac-to-Bac重组杆状病毒系统可作为一种对哺乳动物细胞简便高效的基因转移表达载体。