蓖麻蚕核型多角体病毒p6.9基因的克隆及序列分析

基于丰富蓖麻蚕核型多角体病毒(Philosamia cynthia ricininucleopolyhedrovirus,PcrNPV)分子信息的目的,对PcrNPV DNA部分片段进行了测序分析,获得1个DNA结合蛋白基因p6.9的完整序列。该基因的读码框由240个核苷酸组成,编码79个氨基酸,分子质量约为9.8 kD,转录起始位点ATG侧翼序列符合Kozak规则,其上游34bp处具有晚期基因转录的保守起始序列taag,表明该基因为晚期表达基因。将PcrNPV与12种昆虫核型多角体病毒P6.9蛋白氨基酸序列作同源性比较的结果显示:PcrNPV P6.9蛋白氨基酸序列与家蚕(Bombyx mori)NPV P6.9的同源性为59%,与柞蚕(Antheraea pernyi)NPV的同源性为100%,表明PcrNPV与ApNPV的亲缘关系很近。     

AcNPV对甜菜夜蛾二龄幼虫的生物测定

以甜菜夜蛾二龄幼虫对苜蓿银纹夜蛾核型多角体病毒（AcNPV）两种毒株采用生物测定方法进行比较。对AcNPV两毒株杀虫毒力及杀虫速率的分析表明：以甜菜夜蛾活体增殖的ACNPVA株（LD(50)=131．8PIB／头）比用草地贪夜蛾细胞株（Sf9）增殖的AcNPVB株（LD(50)=1696．8PIB／头）毒力要高10倍,而毒杀速率,也以AcNPVA株（LT(50)=4．46d）比AcNPVB株（LT(50)=6．57d）快1．5倍。     

温度对甜菜夜蛾实验种群增长的影响

为了解温度对甜菜夜蛾各的影响,测定了了甜菜夜蛾在１５．５,１８,２０,２３,２５,２７,３０,３３,３５,３８,４０℃共１１个恒温下的存活率,繁殖力和种群增长参数。     

斜纹夜蛾种群空间动态

利用频次分布法和分布型指数研究了斜纹夜蛾低中龄幼虫在菜芯Ｂｒａｓｓｉｃａ ｐａｒａｃｈｉｎｅｎｓｉｓ Ｂａｉｌｅｙ地内和种群空间分布图式,并研究了各龄幼虫在菜株上的垂直分布。结果表明：斜纹夜蛾 龄幼虫在菜株上呈聚集颁上,频次分布拟事的结果大部分样本符合负二项分布,半产本符合奈曼分布,而不符合泊松分布,频次分布拟事的结果大部样本符合负二项分布,半数样本符合奈曼分布,而不符合泊松分布,幼虫在菜株上的分     

DNA hybridization assay for detection of nucleopolyhedrovirus in whitemarked tussock moth (Orgyia leucostigma) larvae

DNA dot‐blot hybridization assays utilizing a horseradish peroxidase‐labelled whole genomic DNA probe and enhanced chemiluminescence were conducted to quantify detection thresholds of nucleopolyhedrovirus (NPV) in whitemarked tussock moth (Orgyia leucostigma) larvae. The minimum detection thresholds for an aqueous suspension of occlusion bodies (OBs), OBs added to macerates of non‐infected larvae and OBs in macerates of diseased larvae were 7.8 × 10³, 7.8 × 10³, and 1.5 × 10³ OBs, respectively. Purified viral DNA was detected at a concentration of 1.6 × 10⁻¹ ng in a 20 µl volume. The presence of pre‐occluded viral nucleocapsids and DNA, inherent to infected larvae, improved the detection threshold five‐fold compared with OBs alone. Larval tissues did not block the detection system utilized, nor did they bind non‐specifically to the probe. Detection thresholds, upon sequential hybridization of the same membrane, on average deteriorated two‐fold between the first and second hybridization and an additional six‐fold between the second and third hybridization. NPV infection was detected two days post‐inoculation (pi) in about one‐third of the larvae examined and in almost all larvae three days pi. Microscopic analysis of stained larval smears missed NPV infection in almost all larvae two days pi and about two‐thirds of the larvae three days pi. Results from the two methods of analysis were not comparable until four days pi. The detection system utilized is a reliable, efficient and simple method for the early detection of NPV infection in large numbers of larvae and may be used for further studies quantifying the role of this baculovirus in the ecology of whitemarked tussock moth populations. © 2001 Society of Chemical Industry     

DPX-MP062—a potent compound for controlling the Egyptian cotton leafworm Spodoptera littoralis (Boisd.)

The effects of DPX-MP062 [methyl 7-chloro-2,3,4a,5-tetrahydro-2-[methoxycarbonyl(4-trifluoromethoxyphenyl)carbamoyl] indeno[1,2-e][1,3,4] oxadiazine-4a-carboxylate] a broad-spectrum insecticide with a novel mode of action, on the Egyptian cotton leafworm, Spodoptera littoralis, were studied in laboratory experiments. Egg hatch was affected by high concentrations (125 mg AI litre^-1) of DPX-MP062. Larvae that hatched from treated eggs were significantly affected at concentrations of 12·5 mg AI litre^-1 and greater. Larvae were fed castor bean leaves treated with DPX-MP062; 1st-instar larvae were the most susceptible development stage. Pupation and adult formation were determined in assays with 5th-instar larvae. There was strong suppression of adult formation; 65 and 91% at 0·5 and 0·75 mg AI litre^-1, respectively. Highly affected larvae died before pupation; slightly affected ones reached pupation 2–4 days later, were smaller than larvae in the untreated control, and were sometimes unable to develop into normal adults. Comparatively high concentrations (50 and 100 mg AI litre^-1) of the test compound were necessary to affect adults by ingestion, but no effects from contact application could be determined at a concentration of 100 mg AI litre^-1. © 1998 Society of Chemical Industry     

警惕危险性害虫草地贪夜蛾入侵中国

草地贪夜蛾Spodoptera frugiperda(Smith)是一种原产于美洲的重要的毁灭性农业害虫,目前已经入侵到撒哈拉以南非洲地区及亚洲的印度,对我国构成入侵威胁。本文综述了草地贪夜蛾的生物学特征、为害、分布区域及入侵性、形态及分子鉴定方法,以及防治措施,并对其入侵中国的风险进行了预测分析,同时提出应对策略。     

Characterization of field‐evolved resistance to Bacillus thuringiensis‐derived Cry1F δ‐endotoxin in Spodoptera frugiperda populations from Argentina

BACKGROUND

Transgenic maize (Zea mays L.) event TC1507 (Herculex^® I insect protection), expressing Cry1F δ‐endotoxin derived from Bacillus thuringiensis var. aizawai, was commercialized in 2003 in the Americas. Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) susceptibility to Cry1F was monitored annually across several regions in Argentina using diagnostic concentration bioassays. Reduced performance of TC1507 maize against S. frugiperda was reported in 2013. A resistant population was established in the laboratory and the dominance of Cry1F resistance was characterized.

RESULTS

During 2012–2015, high‐survivorship of several populations was observed in the resistance monitoring program. Reciprocal crosses of a Cry1F‐resistant population with a Cry1F‐susceptible population were evaluated to calculate effective dominance (D_ML) based on mortality levels observed at 100 µg/ml Cry1F. Two additional dominance levels (D_LC and D_EC) were calculated using lethal (LC₅₀) or effective concentration (EC₅₀) derived from concentration–response bioassays. Estimates indicated that Cry1F resistance in S. frugiperda in Argentina was either highly recessive (D_ML = 0.005) or incompletely recessive (D_LC < 0.26 and D_EC < 0.19).

CONCLUSION

This study is the first documented confirmation and characterization of S. frugiperda Cry1F field‐evolved resistance in Argentina. The resistance to Cry1F in S. frugiperda populations collected in Argentina, is autosomal and incompletely recessive similar to the resistance reported in Brazil. © 2017 The Authors. Pest Management Science published by John Wiley © Sons Ltd on behalf of Society of Chemical Industry.     

斜纹夜蛾核型多角体病毒Ⅱ型(SpltMNPV Ⅱ)ORF117基因的结构和功能研究

为了研究斜纹夜蛾核型多角体病毒Ⅱ型(Splt MNPVⅡ)中ORF117基因(编码GP117蛋白)的结构和功能,根据其核苷酸序列和氨基酸序列进行了生物信息学分析,以此为基础对该基因的启动子活性和转录时相进行分析,表达纯化了GP117蛋白的部分序列以制备豚鼠来源多克隆抗体,并利用该抗体对ORF117基因的表达时相和GP117蛋白在细胞中的定位进行了分析。生物信息学分析表明该基因全长1 203 bp,编码400个氨基酸的蛋白质,预测分子量为45.5 k Da,等电点为5.06;启动子活性和转录时相分析结果表明ORF117是一个晚期表达的基因;原核表达抗原免疫制备的多克隆抗体效价可以达到1∶3 200,利用该抗体对其表达时相的分析进一步验证了ORF117为晚期表达基因,免疫荧光检测表明编码的蛋白多存在于细胞质中。