甜菜夜蛾核多角体病毒可湿性粉剂生产技术的研究

报道了甜菜夜蛾核多角体病毒可湿性粉剂的生产关键技术,包括甜菜夜蛾的饲养,多角体病毒的增殖,可湿性粉剂的生产,产品质量检测等。     

Optical brightener M2R destroys the peritrophic membrane of Spodoptera exigua (Lepidoptera: Noctuidae) larvae

Observations under an environmental scanning electron microscope showed that the peritrophic membrane (PM) of Spodoptera exigua (Hübner) was smooth without pores, but there were many pores and slits in the PM of larvae fed with 10 g L(-1) optical brightener Calcofluor white M2R. After incubation of S. exigua PM in vitro with 10 g L(-1) M2R, a significant amount of proteins was released from the PM structure. M2R disrupted the structure of larval PM, resulting in greatly increased permeability and increased larval susceptibility to Syngrapha falcifera multiple nucleopolyhedrovirus infection. Continuous feeding of larvae on a diet treated with a 10 g L(-1) M2R solution significantly retarded larval development and resulted in high mortality. The destructive effect of M2R on PM was transient and reversible, depending on the presence of M2R within the midgut.     

茶尺蠖生物学特性及防治技术研究现状

综述了茶尺蠖[Ectropis oblique(Prout)]的为害特点和发生规律,阐述了近年来基于农业防治、物理防治、生物防治、化学防治等防治方法的研究进展,并展望了今后的研究重点。     

Combined effect of nucleopolyhedrovirus and Microplitis pallidipes for the control of the beet armyworm, Spodoptera exigua

BACKGROUND: Nucleopolyhedrovirus (SeNPV) and Microplitis pallidipes are important biological control agents of Spodoptera exigua populations. The interactions between these agents and their combined effect on pest control were investigated in the laboratory and in commercial greenhouses. RESULTS: Microplitis pallidipes searched for and deposited eggs in more healthy larvae than virus‐infected larvae 3 days after viral infection. Each female parasitoid that developed in a virus‐infected host oviposited in a virus‐infected host, or emerged from a cocoon carrying virus transmitted to 4.0, 7.6 or 2.4 healthy larvae respectively. Each female parasitoid exposed to a mixture of virus and 10% honey water solution transmitted the virus to 2.2 healthy larvae. In an experiment with cabbage growing in commercial greenhouses, the pest population reduction was greater by M. pallidipes carrying SeNPV (82.3–89.7% reduction) than by parasitoids without virus (59.5–62.4% reduction). CONCLUSIONS: Control of S. exigua was greater with M. pallidipes plus SeNPV than with M. pallidipes alone. Microplitis pallidipes preferred healthy hosts to infected hosts. Parasitoids were able to complete their development in virus‐infected hosts before the hosts died from the virus infection. The parasitoid ovipositors contaminated with the virus could carry and transmit SeNPV. Copyright © 2011 Society of Chemical Industry     

利用家蚕细胞表达重组蝎毒素蛋白及其杀虫效果分析

用天然毒素蛋白杀灭害虫是农林病虫害防治的一个重要发展方向。利用家蚕核型多角体病毒(BmNPV)多角体蛋白基因的部分序列与蝎毒素蛋白基因(BmK ITa1)融合表达,SDS-PAGE分析通过融合表达策略构建的含有蝎毒素蛋白基因的重组病毒具有较高的蝎毒素蛋白表达水平,重组病毒vBmBac(polh-)PH5B/35B-BmK ITa1.6His,在家蚕BmN细胞内的BmKITa1蛋白表达水平分别为0.4、0.2 mg/mL。以家蚕和棉铃虫为供试昆虫,探讨融合蝎毒素蛋白的杀虫效果,结果表明:经口添食含融合蝎毒素蛋白的BmN细胞液对不同龄期家蚕幼虫均具有毒性,尤其对低龄幼虫的毒性较强,蚁蚕添食后的死亡率达80%;给1龄期的家蚕和棉铃虫幼虫经口添食含有不同剂量融合蝎毒素蛋白的BmN细胞破碎液,当添食剂量在2.8μg/头时,家蚕的死亡率即达100%,棉铃虫的死亡率达74%。研究结果提示:BmNPV多角体蛋白编码基因的部分序列与蝎毒素蛋白基因融合表达,可提高蝎毒素蛋白的表达水平,且重组病毒表达的蝎毒素蛋白具有较好的杀虫活性,在较低剂量下即可对低龄昆虫产生良好的杀灭效果。     

温度和寄主虫龄对单白绵绒茧蜂生长发育的影响

茶尺蠖幼虫期的重要寄生性天敌──单白绵绒茧蜂在１４和３４℃恒温下，供以寄主，无子蜂羽化。在１８～３０℃下，其幼期（卵──蛹）一般随温度上升而缩短，反之则延长；发育起点温度为９．９８±２．７８℃，有效积温２５０．００±４６．７７日度，由此推算在杭州年可发生ｌｌ～１３代。成虫寿命则随温度升高而下降，自然交温下趋势一致。寄主虫龄对该蜂的卵＋幼虫期无显著影响，而在寄主体外的预蛹＋蛹的历期则随寄主虫龄增大而缩短。     

表达短ie-1 dsRNA的转化细胞对家蚕核型多角体病毒的抑制作用

为了利用RNAi技术提高家蚕对核型多角体病毒(BmNPV)的抗性,根据BmNPV复制必需基因ie-1设计其相应的dsRNA,构建带有ie-1 dsRNA表达盒的转基因载体piggyantiIE-Neo,结果显示:表达短ie-1 dsRNA的稳定转化Bm细胞,对BmNPV的增殖表现出抑制作用,但在病毒感染后期,由于病毒恢复增殖导致RNAi的效果被掩盖。通过反向PCR分析外源DNA片段插入基因组位点,结果显示:在转化细胞中,外源DNA可通过随机整合或按照piggyBac特定的转座位点TTAA插入细胞基因组。     

蓖麻蚕核型多角体病毒解旋酶基因的克隆和序列分析

为了解柞蚕核型多角体病毒(Antheraea pernyinucleopolyhedrovirus,ApNPV)和蓖麻蚕核型多角体病毒(Philosamia cynthia riciniNPV,PcrNPV)在感染性及基因水平上的差别,克隆了PcrNPV解旋酶基因helicase,对该基因进行了测序及同源性分析。感染性试验表明,ApNPV对柞蚕和蓖麻蚕的感染率分别为78%和57%;而PcrNPV对蓖麻蚕的感染率为79%,但几乎不感染柞蚕。对PcrNPVhelicase分析的结果表明,该基因长度为3 639 bp,编码1 212个氨基酸(登录号:EU143371)。基因的同源性分析表明,PcrNPVhelicase与ApNPVhelicase的同源性最高,达98.6%,与苜蓿银纹夜蛾NPV(Autographa califorlicaNPV,AcNPV)helicase和家蚕NPV(BombyxmoriNPV,BmNPV)helicase的同源性最低,分别为58%和58.8%。PcrNPV和ApNPV的helicase基因7个保守功能域上的氨基酸序列均相同,仅在靠近DNA结合区螺旋-转角-螺旋结构处的第974位上有1个氨基酸不同,推测该位点可能与宿主域范围有关。     

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。     

松埃尺蛾空间分布型及其在实践中的应用

本文采用比较频次法、聚集度指标法研究了松埃尺蛾越冬代蛹和第一代幼虫的空间分布型。结果表明：越冬代蛹、第一代幼虫的空间分布型呈负二项分布，其分布的基本成分为个体群，个体间相互排斥。并应用Ｉｗａｏ方法分析、检验了不同虫口密度下林间调查的最适抽样数，列出序贯抽样分析表。