草地贪夜蛾核型多角体病毒对我国草地贪夜蛾不同地理种群毒力的比较分析

草地贪夜蛾Spodoptera frugiperda（J. E. Smith）入侵我国多个地区,逐渐形成地理种群。在草地贪夜蛾核型多角体病毒Spodoptera frugiperda multiple nucleopolyhedrovirus（SfMNPV）生物农药的生产过程中发现,不同地理来源的宿主对SfMNPV的敏感性和产量存在明显差异,显著影响了SfMNPV的生产效率。为解析其敏感性差异及其产生的原因,本研究首先对云南德宏、广东广州、广西钦州、西藏林芝4个地区草地贪夜蛾种群基因型进行了鉴定,采用生物测定法测试SfMNPV对4龄幼虫的口服毒力,然后,通过向幼虫体内注射草地贪夜蛾核多角体病毒出芽型病毒粒子（budded virions,BVs）的方式,越过口服感染中肠的过程,分析敏感性差异发生的阶段。最后比较了高敏感和低敏感种群中肠肠液pH,并基于16S rDNA测序测定了肠道菌群组成。结果表明,广西种群属于纯合玉米型,其余种群为带有水稻型COI标记的杂合玉米型。广西种群对SfMNPV的口服敏感性最低,西藏种群的敏感性最高,但两者注射BV后死亡率差异无统计学意义,暗示病毒敏感性差异发生在口服感染阶段。广西种群中肠pH略低于西藏种群,并且较于西藏种群,广西种群相肠道乳杆菌Lactobacillus丰度高。本文结果表明,肠道微环境的差异可能是不同地理种群草地贪夜蛾对SfMNPV口服敏感性产生差异的原因。     

虫螨腈对草地贪夜蛾幼虫的室内毒力及田间防效

草地贪夜蛾Spodoptera frugiperda(J. E. Smith),是一种广泛存在于美洲的杂食性农业害虫。据报道2019年1月草地贪夜蛾入侵我国云南,截止2019年6月6日,已在全国18个省份的884个县(市、区)监测到发生为害,发生面积342万亩,已经对我国粮食安全造成重大威胁。在2019年5月,在广东省广州市花都区儒林村玉米田开展喷雾和喇叭口点施虫螨腈对草地贪夜蛾幼虫的防治试验以及室内毒力测定试验。试验结果表明,虫螨腈对草地贪夜蛾2龄幼虫具有良好的毒力活性;综合考虑在草地贪夜蛾1～3龄幼虫期,用药量67.5 g.a.i/hm~2、药液量675 L/hm~2常规喷雾效果更佳;在草地贪夜蛾4～6龄幼虫期,用药量22.5 g.a.i/hm~2、药液量225 L/hm~2喇叭口点施效果更佳,且喇叭口点施可以定位定向定量地直接到达草地贪夜蛾为害部位,可减少2/3的农药使用量,达到农药减量增效的目的。该研究结果为拟定草地贪夜蛾应急防控技术方案提供技术支撑和理论依据。     

4种昆虫病原线虫对草地贪夜蛾的致死作用

【目的】探讨昆虫病原线虫对草地贪夜蛾幼虫的侵染效果。【方法】在室内条件下采用生测试验法,测定了小卷蛾斯氏线虫AⅡ、长尾斯氏线虫X-7、夜蛾斯氏线虫SN和嗜菌异小杆线虫H06等4种昆虫病原线虫对草地贪夜蛾2龄和5龄幼虫的致死作用。【结果】小卷蛾斯氏线虫AⅡ与草地贪夜蛾数量比为50∶1时,36 h后草地贪夜蛾2龄和5龄幼虫的死亡率分别为92%和100%。长尾斯氏线虫X-7与草地贪夜蛾数量比为50∶1时,36 h后草地贪夜蛾2龄幼虫的死亡率为80%;在数量比30∶1时,36 h后草地贪夜蛾5龄幼虫死亡率为100%。夜蛾斯氏线虫SN与草地贪夜蛾数量比为400∶1时,36 h后草地贪夜蛾2龄和5龄幼虫的死亡率分别为88%和85%。嗜菌异小杆线虫H06与草地贪夜蛾在400∶1时,36 h后草地贪夜蛾2龄和5龄幼虫的死亡率分别为32%和67.5%。【结论】小卷蛾斯氏线虫AⅡ具有较好的草地贪夜蛾生物防治潜质,其次是长尾斯氏线虫X-7。     

稠合杂环化合物唑虫酯对草地贪夜蛾的活性研究

【目的】草地贪夜蛾Spodoptera frugiperda原产于美洲,是一种对农作物造成严重危害的昆虫,被国际农业和生物科学中心CABI评为世界十大植物害虫。目前已入侵我国14个省区,对我国玉米等种植业造成严重威胁,为做好草地贪夜蛾的防控工作,及时控制其为害。【方法】采用浸叶法测定了唑虫酯对草地贪夜蛾幼虫的室内毒力,并进行了田间药效试验。【结果】唑虫酯对草地贪夜蛾幼虫具有优异的活性,处理后1、2、3d,对2龄幼虫的LC50值分别为5.51、1.95、0.91mg/L;对3龄幼虫的LC50值分别为7.50、3.92、1.94mg/L;对4龄幼虫的LC50值分别为13.79、10.30、8.78mg/L。田间药效结果表明,玉米喇叭口期,喷雾处理后3d,唑虫酯对1-3龄幼虫的防效达到90%以上,对4-6龄幼虫的防效在50%以下。药液喇叭口点施处理后3 d,唑虫酯对草地贪夜蛾的防效在93.65%以上,施药后7 d,防效在98.78%以上。【结论】药剂对低龄幼虫杀灭效果好,高龄幼虫具有钻进性,不容易防控。应结合虫情监测和田间监测情况,把握住1-3龄幼虫防控关键时期。     

室内测定6种化学杀虫剂对草地贪夜蛾幼虫的毒力

2019年草地贪夜蛾Spodoptera frugiperda入侵我国多个省份和地区,对我国农业生产造成重大影响。化学防治是应对入侵害虫的主要措施,因此,本研究测定了6种不同作用机制杀虫剂对草地贪夜蛾初孵和3龄幼虫的生物活性。结果表明,对初孵幼虫具有较高毒力作用的为氯虫苯甲酰胺、多杀菌素、茚虫威、阿维菌素和甲氧虫酰肼;对3龄幼虫具有较高毒力作用的为氯虫苯甲酰胺、多杀菌素和茚虫威;与初孵幼虫相比,阿维菌素、茚虫威、多杀菌素和甲氧虫酰肼的LC 50分别提高了78.06、1.70、11.73和23.09倍。本研究筛选出了对草地贪夜蛾初孵幼虫和3龄幼虫杀虫效果较好的药剂,证明了低龄幼虫对部分农药抵抗力较弱,而3龄后的幼虫抵抗力显著增加,为田间草地贪夜蛾的化学防治提供了科学的用药指导。     

绿色杀虫剂对草地贪夜蛾杀虫活性比较测定

为了减缓草地贪夜蛾Spodoptera frugiperda对化学农药抗性的增加,减少传统化学农药的使用量,提高草地贪夜蛾的防控效果。本研究测定了15种生物源、矿物源和昆虫激素类等绿色杀虫剂对草地贪夜蛾卵和2龄幼虫的室内毒杀效果。结果表明,灭幼脲、虱螨脲、短稳杆菌和苏云金杆菌是替代传统化学农药防控草地贪夜蛾的首选农药,其中尤以低毒、易降解的昆虫激素类农药灭幼脲和活体微生物农药短稳杆菌最佳,在实际生产中有较好的推广应用价值。而植物源农药藜芦碱、鱼藤酮、印楝素、苦参碱、除虫菊素、茶皂素以及矿物源农药矿物油对该虫控制效果相对较差,不建议单独大面积推广使用。     

草地贪夜蛾高致病力绿僵菌菌株的筛选与鉴定

为获取对草地贪夜蛾Spodoptera frugiperda具有高致病力的生防真菌，从福建省不同地区分离得到8株寄主为鳞翅目和半翅目幼虫僵虫的绿僵菌Metarhizium，采用浸渍法测定了其对草地贪夜蛾2龄幼虫和蛹的致病力，并根据形态学和分子生物学方法对高致病力菌株进行种类鉴定。结果表明，8株绿僵菌菌株对草地贪夜蛾2龄幼虫和蛹均表现出不同程度的致病力，其中菌株FJMR2和FJXY7表现出较强的致病力。在5×107个/mL孢子浓度下，FJMR2和FJXY7对草地贪夜蛾2龄幼虫的致死率分别为88.76%和82.13%，对蛹的致死率分别为86.57%和84.00%；对草地贪夜蛾2龄幼虫的LT50分别为4.81 d和4.93 d，对蛹的LT50分别为4.94 d和4.83 d。经鉴定菌株FJMR2和FJXY7均为莱氏绿僵菌Metarhizium rileyi。本研究获得2株对草地贪夜蛾2龄幼虫和蛹具有高致病力的莱氏绿僵菌菌株，在草地贪夜蛾的生物防治中具有较大应用潜力。     

大草蛉对草地贪夜蛾捕食潜能研究

草地贪夜蛾Spodoptera frugiperda JE Smith于2018年底至2019年初入侵我国后,迅速扩散,并在半年内对西南地区的玉米生产造成为害。田间调查和观测研究表明,田间虽然可见该害虫的天敌,但由于天敌种群数量低,无法形成有效控害的模式。为研究大草蛉对草地贪夜蛾的控害潜能,本研究进行了捕食功能反应试验。结果表明,大草蛉成虫对草地贪夜蛾卵以及大草蛉3龄幼虫对草地贪夜蛾1龄幼虫的捕食效应均能够很好的拟合HollingⅡ功能反应模型。其中,大草蛉成虫对贪夜蛾卵的理论日最大捕食量、瞬时攻击率和处理时间分别为1115.56头、1.004和0.0009 d。大草蛉幼虫对贪夜蛾低龄幼虫的理论日最大捕食量、瞬时攻击率和处理时间分别为358头、1.074和0.003 d。结果表明大草蛉具备对草地贪夜蛾卵和低龄幼虫进行有效控害的潜力,为应用天敌昆虫防治草地贪夜蛾提供了理论依据。     

草地贪夜蛾海藻糖合成酶基因的克隆、时空表达谱及对温度胁迫的响应

【目的】本研究旨在通过克隆草地贪夜蛾Spodoptera frugiperda的海藻糖合成酶(trehalose-6-phosphate synthase)基因SfTPS,分析其在草地贪夜蛾不同发育阶段、不同组织中的表达水平及不同温度胁迫时5龄幼虫中的相对表达量,为进一步探究TPS在草地贪夜蛾生长发育及抗逆应激反应中的功能奠定基础。【方法】运用RT-PCR技术克隆草地贪夜蛾SfTPS的全长编码区,并进行生物信息学分析。运用RT-qPCR技术检测SfTPS在草地贪夜蛾不同发育阶段(卵、1-6龄幼虫、蛹和成虫)、5龄幼虫不同组织(体壁、中肠和脂肪体)中和经短期(2, 4和8 h)高(35℃)低温(10℃)胁迫后5龄幼虫中的表达变化。【结果】克隆获得2 571 bp的草地贪夜蛾TPS cDNA序列,命名为SfTPS(GenBank登录号： MT920672),全长开放阅读框(ORF)长2 481 bp,编码的826个氨基酸具有TPS和TPP两个保守结构域。同源比对和系统进化分析表明,昆虫TPS蛋白具有较高的保守性,SfTPS与斜纹夜蛾S. litura的TPS亲缘关系最近,序列一致性达到99.15%。SfTPS中α-螺旋、β-折叠和无规则卷曲占比分别为38.14%, 12.23%和48.55%;SfTPS的三级结构为同源二聚体。RT-qPCR结果表明,SfTPS在草地贪夜蛾卵期和1-5龄幼虫期低表达,在6龄幼虫期、蛹期和成虫期高表达,且草地贪夜蛾变态前后SfTPS的表达量变化较大。组织分布结果显示,SfTPS在5龄幼虫脂肪体中表达量最高。草地贪夜蛾5龄幼虫经2~8 h低温(10℃)和高温(35℃)胁迫后,SfTPS的相对表达量显著高于对照(25℃),分别为对照的4.43~9.34和2.50~6.03倍。【结论】SfTPS基因在草地贪夜蛾生长发育过程及抵御高低温度胁迫中可能具有重要作用。     

四氯虫酰胺玉米全株喷雾和喇叭口点施防治草地贪夜蛾的药效评价

评价四氯虫酰胺玉米全株喷雾和喇叭口点施处理对草地贪夜蛾的活性与防效,为草地贪夜蛾综合防控提供技术支持。采用叶片喷雾法,测定了对草地贪夜蛾2龄幼虫毒力;以40%辛硫磷乳油为对照药剂,采用喷雾和喇叭口点施两种方式田间施用10%四氯虫酰胺悬浮剂,药前1 d和药后1 d、3 d、7 d,调查挂牌标记玉米上草地贪夜蛾活虫数,计算防治效果。结果表明:四氯虫酰胺对草地贪夜蛾2龄幼虫具有良好的毒力;10%四氯虫酰胺悬浮剂制剂用量600 g/hm 2喷雾处理后1 d、3 d、7 d对草地贪夜蛾的防效分别为75.90%、91.91%和92.02%,制剂用量300 g/hm 2喇叭口点施处理后1 d、3 d、7 d对草地贪夜蛾的防效分别为82.28%、89.52%和91.23%,40%辛硫磷乳油制剂用量1500 g/hm 2处理,两种施药方式防效均不佳。四氯虫酰胺对草地贪夜蛾具有良好的防治效果,喇叭口点施方式节省药剂可达50%以上,在防治草地贪夜蛾等害虫方面具有广阔的应用前景。     

Linearization of baculovirus DNA enhances the recovery of recombinant virus expression vectors.

Engineered derivatives of Autographa californica multiple nucleocapsid nuclear polyhedrosis virus (AcMNPV) possessing a unique restriction site provide a source of viral DNA that can be linearized by digestion with a specific endonuclease. Circular or linearized DNA from two such viruses were compared in terms of their infectivity and recombinogenic activities. The linear forms were 15- to 150-fold less infectious than the corresponding circular forms, when transfected into Spodoptera frugiperda cells using the calcium phosphate method. Linear viral DNA was, however, proficient at recombination on co-transfection with an appropriate transfer vector. Up to 30% of the progeny viruses were recombinant, a 10-fold higher fraction of recombinants than was obtained from co-transfections with circular AcMNPV DNA. The isolation of a recombinant baculovirus expression vector from any of the AcMNPV transfer vectors currently in use can thus be facilitated by linearization of the viral DNA at the appropriate location.     

Improving baculovirus resistance to UV inactivation: increased virulence resulting from expression of a DNA repair enzyme

The use of baculoviruses as biological control agents is hampered by their susceptibility to inactivation by ultraviolet (UV) light. In an attempt to reduce UV inactivation, an algal virus pyrimidine dimer-specific glycosylase, cv-PDG, was expressed in the baculovirus Autographa californica M nucleopolyhedrovirus (AcMNPV), and the infectivity of recombinant viruses expressing cv-PDG was measured after exposure to UV light. Expression of cv-PDG resulted in a 3-fold decrease in inactivation of budded virus by UV as measured by plaque assay in Spodoptera frugiperda Sf21 cells. However, occluded viruses expressing cv-PDG were not more resistant to UV inactivation than wild type AcMNPV when fed to either S. frugiperda or Trichoplusia ni neonate larvae. Surprisingly, however, viruses expressing cv-PDG showed a significant decrease in both the dose of occluded virus required for oral lethality and the time required for lethality compared to control virus, but these effects were only seen in S. frugiperda and not in T. ni larvae.     

Cellular VPS4 is required for efficient entry and egress of budded virions of Autographa californica multiple nucleopolyhedrovirus

Membrane budding is essential for the egress of many enveloped viruses, and this process shares similarities with the biogenesis of multivesicular bodies (MVBs). In eukaryotic cells, the budding of intraluminal vesicles (IVLs) is mediated by the endosomal sorting complex required for transport (ESCRT) machinery and some viruses require ESCRT machinery components or functions to bud from host cells. Baculoviruses, such as Autographa californica multiple nucleopolyhedrovirus (AcMNPV), enter host cells by clathrin-mediated endocytosis. Viral DNA replication and nucleocapsid assembly occur within the nucleus. Some progeny nucleocapsids are subsequently trafficked to, and bud from, the plasma membrane, forming budded virions (BV). To determine whether the host ESCRT machinery is important or necessary for AcMNPV replication, we cloned a cDNA of Spodoptera frugiperda VPS4, a key regulator for disassembly and recycling of ESCRT III. We then examined viral infection and budding in the presence of wild-type (WT) or dominant negative (DN) forms of VPS4. First, we used a viral complementation system, in combination with fluorescent tags, to examine the effects of transiently expressed WT or DN VPS4 on viral entry. We found that dominant negative VPS4 substantially inhibited virus entry. Entering virus was observed within aberrant compartments containing the DN VPS4 protein. We next used recombinant bacmids expressing WT or DN VPS4 proteins to examine virus egress. We found that production of infectious AcMNPV BV was substantially reduced by expression of DN VPS4 but not by WT VPS4. Together, these results indicate that a functional VPS4 is necessary for efficient AcMNPV BV entry into, and egress from, insect cells.     

In Vitro Translation of Autographa californica Nuclear Polyhedrosis Virus Early and Late mRNAs

A preliminary translational map of the Autographa californica genome was constructed. Eighteen viral DNA restriction fragments were either purified from agarose gels or obtained from pBR322 recombinant DNA plasmids to locate specific gene products. The DNAs were immobilized on nitrocellulose filters and used to select viral mRNAs isolated from RNA obtained from the cytoplasm of infected Spodoptera frugiperda cells at 21 h postinfection. The fragment-specific mRNAs were translated in vitro in the presence of l-[(3)H]leucine by using a rabbit reticulocyte lysate system and analyzed on sodium dodecyl sulfate-polyacrylamide gels. The approximate locations of 19 A. californica nuclear polyhedrosis virus (AcMNPV) gene products were mapped. The genes for mRNAs present late in viral infection were mapped to DNA fragments that represent nearly the entire genome. The molecular weights of many of these proteins were similar to those present in purified AcMNPV extracellular virus and to proteins being made in infected cells at 18 to 21 h postinfection. Cytoplasmic RNA was isolated at 4 h postinfection from infected cells, a time early in the viral infection cycle, and hybridized to AcMNPV DNA immobilized on nitrocellulose filters. AcMNPV-specific early RNA was translated in vitro into at least six polypeptides, the most abundant having a molecular weight of 39,000. Viral polypeptides were detected in cells pulse-labeled with l-[(3)H]leucine at 3 to 6 h postinfection, with molecular weights similar to those of polypeptides made in vitro from early AcMNPV mRNA.     

Baculovirus infection induces a DNA damage response that is required for efficient viral replication

Several mammalian viruses have been shown to induce a cellular DNA damage response during replication, and in some cases, this response is required for optimal virus replication. However, nothing is known about whether a DNA damage response is stimulated by DNA viruses in invertebrates. Cell cycle arrest and apoptosis are two of the downstream effects of the DNA damage response, and both are stimulated by baculovirus infection, suggesting a possible relationship between baculoviruses and the DNA damage response. In the study described in this report, we found that replication of the baculovirus Autographa californica M nucleopolyhedrovirus (AcMNPV) in the cell line Sf9, derived from the lepidopteran insect Spodoptera frugiperda, stimulated a DNA damage response, as indicated by an increased abundance of the S. frugiperda P53 protein (SfP53) and phosphorylation of the histone variant protein H2AX. Stimulation of the DNA damage response was dependent on viral DNA replication. Inhibition of the DNA damage response prevented both the increase in SfP53 accumulation and H2AX phosphorylation and also caused a 10- to 100-fold reduction in virus production, along with decreased viral DNA replication and late gene expression. However, silencing of Sfp53 expression by RNA interference did not significantly affect AcMNPV replication or induction of apoptosis by a mutant of AcMNPV lacking the antiapoptotic gene p35, indicating that these processes are not dependent on SfP53 in Sf9 cells.     

Role of early and late replication events in induction of apoptosis by baculoviruses.

Autographa californica nuclear polyhedrosis virus (AcMNPV) mutants that lack the apoptotic suppressor gene p35 cause apoptosis in Spodoptera frugiperda SF21 cells. To identify a viral signal(s) that induces programmed cell death, we first defined the timing of apoptotic events during infection. Activation of a P35-inhibitable caspase, intracellular fragmentation of host and AcMNPV DNA, and cell membrane blebbing coincided with the initiation of viral DNA synthesis between 9 and 12 h after infection and thus suggested that apoptotic signaling begins at or before this time. Virus entry was required since binding of budded virus to host cell receptors alone was insufficient to induce apoptosis. To therefore determine the contribution of early and late replication events to apoptotic signaling, we used the AcMNPV mutant ts8 with a temperature-sensitive lesion in the putative helicase gene p143. At the nonpermissive temperature at which viral DNA synthesis was conditionally blocked, ts8 caused extensive apoptosis of the SF21 cell line p3576D, which dominantly interferes with anti-apoptotic function of viral P35. Confirming that apoptosis can be induced in the absence of normal viral DNA synthesis, parental SF21 cells also underwent apoptosis when infected with a ts8 p35 deletion mutant at the nonpermissive temperature. However, maximum levels of ts8 p35 deletion mutant-induced apoptosis required a temperature-sensitive event(s) that included the initiation of viral DNA synthesis. Collectively, these data suggested that baculovirus-induced apoptosis can be triggered by distinct early (pre-DNA synthesis) and late replicative events, including viral DNA synthesis or late gene expression.     

The deletion of the pif gene improves the biosafety of the baculovirus-based technologies

Our goal was to improve the biosafety of baculovirus-based technologies by deleting the pif (per os infectivity factor) gene from baculovirus expression vectors. Such a deletion would block transmission in nature without disturbing protein production. A pif deletion mutant of Autographa californica multiplecapsid nucleopolyhedrovirus (AcMNPV) was constructed and its infectivity to two host species was tested by oral or intrahemocoelic inoculation. Virus replication after oral inoculation was monitored using PCR. Oral inoculations with a mixture of the wild type and the pif deletion viruses were carried out. The pif deletion blocked oral infection but it did not hamper infectivity in cell culture. The blockage took place early after inoculation and could not be overcome by mixed inoculations with the wild type. The cat gene was inserted under the control of the polyhedrin promoter in the deletion mutant and the wild type CAT yield was measured in Spodoptera frugiperda insect cells (Sf9) infected with either recombinant. The pif deletion did not hamper CAT production. This deletion significantly improved CAT yields early in the infection. Hence, expression vectors lacking pif may produce higher quality protein. The pif deletion is a simple measure that dramatically reduces the chances of virus spread or gene transfer in nature.     

Semipermissive Replication of a Nuclear Polyhedrosis Virus of Autographa californica in a Gypsy Moth Cell Line

Several gypsy moth cell lines have been previously described as nonpermissive for the multiple-embedded nuclear polyhedrosis virus of Autographa californica (AcMNPV). In this report, we demonstrate the semipermissive infection of a gypsy moth cell line, IPLB-LD-652Y, with AcMNPV. IPLB-LD-652Y cells infected with AcMNPV produced classic cytopathic effects but failed to yield infectious progeny virus. Results of experiments employing DNA-DNA dot hybridization suggested that AcMNPV DNA synthesis was initiated from 8 to 12 h postinfection (p.i.), continued at a maximum rate from 12 to 20 h p.i., and declined from 20 to 36 h p.i. The rate of AcMNPV DNA synthesis approximated that observed in the permissive TN-368 cell line. AcMNPV-infected IPLB-LD-652Y cells, pulse-labeled with [(35)S]methionine at various time intervals p.i. and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, revealed four virus-induced proteins, one novel to the semipermissive system and three early alpha proteins, synthesized from 1 to 20 h p.i. Thereafter, both host and viral protein synthesis was completely suppressed. These results suggest that AcMNPV adsorbed, penetrated, and initiated limited macromolecular synthesis in the semipermissive gypsy moth cell line. However, the infection cycle was restricted during the early phase of AcMNPV replication.     

AcMNPV as a model for baculovirus DNA replication

Baculoviruses were first identified as insect-specific pathogens, and it was this specificity that lead to their use as safe, target specific biological pesticides. For the past 30 years, AcMNPV has served as the subject of intense basic molecular research into the baculovirus infectious cycle including the interaction of the virus with a continuous insect cell line derived from Spodoptera frugiperda. The studies on baculoviruese have led to an in-depth understanding of the physical organization of the viral genomes including many complete genomic sequences, the time course of gene expression, and the application of this basic research to the use of baculoviruses not only as insecticides, but also as a universal eukaryotic protein expression system, and a potential vector in gene therapy. A great deal has also been discovered about the viral genes required for the replication of the baculovirus genome, while much remains to be learned about the mechanism of viral DNA replication. This report outlines the current knowledge of the factors involved in baculovirus DNA replication, using data on AcMNPV as a model for most members of the Baculoviridae.