Insecticidal effect on a population of Spilarctia obliqua (Lepidoptera: Arctiidae)

Survivors of Spilarctia obliqua derived from 3^rd instars that had ingested LC₅₀ of imidacloprid (0.025%), dichlorvos (0.014%) and endosulfan (0.012%) were studied through the life table method. Survivorship was reduced in insecticide exposed populations as compared to 45‐day in the control groups. Egg hatching was significantly decreased for the insecticide treated populations. Total larval mortality was the highest for endosulfan (36.76%) as compared to the individuals that died among control groups (14.29%). Life expectancy (e_x) was decreased gradually over time and stage of development for insecticides tested and the control groups. Dichlorvos (0.014%) has caused a significant reduction in the potential fecundity i.e. 315 females/female/generation, while 415 in the unexposed cohorts. Net reproductive rate (R_o) was the lowest in endosulfan (118.47 females/female/generation) treatment followed by dichlorvos (141.97), imidacloprid (144.49) and the control groups (272.42). Similarly, the intrinsic rate of increase (r_m) was substantially decreased after exposure with endosulfan (0.124 females/female/day) as compared to 0.135 in the unexposed cohort. The finite rate of increase (λ) was not significantly different among the insecticide treatments and control groups. Mean generation time was significantly reduced after exposure to dichlorvos (37.19 days). However, 41.34 days were required to complete one generation by S. obliqua in the absence of insecticides. Based on these results, population of S. obliqua would double in 5.19 days under the influence of dichlorvos while requiring 5.13 days for unexposed cohorts.     

Losac, the first hemolin that exhibits procogulant activity through selective factor X proteolytic activation

Envenoming by the contact of human skin with Lonomia obliqua caterpillars promotes a hemorrhagic syndrome characterized by a consumptive coagulopathy. Losac (Lonomia obliqua Stuart factor activator) is a component of the bristle of L. obliqua that is probably partially responsible for the observed syndrome because it activates factor X and is recognized by an effective antilonomic serum. Here we unveil the proteolytic activity of Losac and demonstrate the feasibility of its recombinant production. On the other hand, Losac has no homology to known proteases, but it can be inhibited by PMSF, a serine protease inhibitor. Instead, it shows closer homology to members of the hemolin family of proteins, a group of cell adhesion molecules. The recombinant protein (rLosac) shortened the coagulation time of normal and deficient plasmas, whereas it was ineffective in factor X-deficient plasma unless reconstituted with this protein. rLosac was able to activate factor X in a dose- and time-dependent manner but not γ-carboxyglutamic acid domainless factor X. Moreover, phospholipids and calcium ions increased rLosac activity. Also, rLosac had no effect on fibrin or fibrinogen, indicating its specificity for blood coagulation activation. Linear double reciprocal plots indicate that rLosac follows a Michaelis-Menten kinetics. Cleavage of factor X by rLosac resulted in fragments that are compatible with those generated by RVV-X (a well known factor X activator). Together, our results validate Losac as the first protein from the hemolin family exhibiting procoagulant activity through selective proteolysis on coagulation factor X.     

Bt与EoNPV混用配比优劣性图谱分析

苏云金杆菌（Bt）和茶尺蠖核型多角体病毒（EoNPV）等,两类生物源杀虫剂常以复配混用方式应用.由于药效的迟缓,而明显表现出剂量-时间-致死作用复合特征.但目前尚缺乏与之相适应的混用配比优劣判别标准.受叶庆华等地学信息图谱分析和曹进等指纹图谱整体相似性分析理论和方法的启发,用室内人工饲养繁殖茶尺蠖2龄初—中期幼虫生物测定结果作为基础数据,分析研究了Bt与EoNPV混用的剂量-时间-致死作用复合特征,并进行了配比优劣性判别的尝试.首先将生物测定的有关结果输入SPSS统计分析软件包和Excel绘图软件,进行多因素方差分析,同时分别绘出不同药剂处理的,以剂量梯度为横轴,以累加死亡虫数为纵轴的不同观察时段害虫致死过程的曲线组图,简称时段药效信息图;再使用SPSS软件包进行单因素方差分析,提取出用于整体相似性比较的图谱单元Ⅰ和用于细节非相似性比较的图谱单元Ⅱ,且分别量化,由此得到相似值和非相似值,据此又分别算得相似系数和非相似系数;最后综合成一种混用配比优劣性总体判别指标,简称Q值.结果,Bt：EoNPV为9：1、7：3、4：1和2：3等4个混配处理的Q值依次为200、100、31.0和23.8,明显标示了其中“9：1”的Q值最大,被确认为最合理的混配处理.此结果与之前的研究相同.还对该图谱分析方法的可靠性以及应用的可行性等进行了讨论.     

Influence of host-plants on the wind dispersal and the survival of an Australian geometrid caterpillar

The wind dispersal and survival of the first instars of a native polyphagous geometrid, Ectropis excursaria (Guenée) on Citrus sinensis (L.), Cupressus macrocarpa Hartw., Eucalyptus camaldulensis Dehn., Medicago sativa (L.) and Pinus radiata D. Don., were studied. A positive correlation existed between the percentage of caterpillars remaining on the foliage in dispersal experiments and the percent survival of caterpillars on these host-plants. The results are discussed in relation to the host-selection strategy of the first instars and the advantages of dispersal are compared for adult females and young larvae.

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Résumé Les femelles d'E. excursaria ne pondent pas là où les chenilles de premier stade s'alimentent. La dispersion par le vent, a été examinée comme l'un des éléments de la sélection de la plante-hôte par les chenilles du premier stade. Cette dispersion était influencée par la nature du végétal et par l'âge du feuillage. La dispersion à partir de 3 plantes de valeurs alimentaires différentes n'a pas été modifiée par le jeûne du premier stade. Le taux de survie, sur feuilles coupées et sur plantes en pot, a été aussi influencé par la nature du végétal. Il y a eu une corrélation positive significative entre le pourcentage de chenilles de premier stade restant sur le feuillage de l'hôte lors d'expériences de dispersion et le taux de survie sur ce feuillage. On peut suggérer que le comportement alimentaire polyphage des chenilles et la fécondité élevée des femelles peuvent avoir contribué à la stratégie de dispersion larvaire, comme processus de sélection larvaire, au détriment d'une sélection du lieu de ponte par les femelles.

     

茶尺蠖水通道蛋白EoAQP1的cDNA克隆、多克隆抗体制备及亚细胞定位

【目的】水通道蛋白(aquaporin,AQP)是一种广泛存在于哺乳动物、植物和微生物中的跨膜转运蛋白,它在细胞水分运输、离子选择透过性和渗透压平衡等过程中发挥着重要的作用。本研究旨在分析茶尺蠖Ectropis obliqua Prout水通道蛋白AQP1的基因特性,在制备多克隆抗体的基础上了解其亚细胞定位分布。【方法】采用同源克隆方法并结合RACE技术克隆茶尺蠖AQP1的基因全长,通过生物信息学网站和软件分析茶尺蠖AQP1的生物学信息;通过实时荧光定量PCR技术(qRT-PCR)检测茶尺蠖AQP1在不同发育阶段和6龄幼虫不同组织中的相对表达量;通过原核表达、镍柱纯化并免疫新西兰兔制备了茶尺蠖AQP1的多克隆抗体;通过荧光显微镜观测了茶尺蠖AQP1在黑腹果蝇Drosophila melanogaster胚胎细胞S2中的定位分布,并利用多克隆抗体进行了Western blot验证。【结果】克隆并鉴定了茶尺蠖AQP1全长,将其命名为Eo AQP1(Gen Bank登录号:KT819587),Eo AQP1 c DNA全长1 826 bp,含有780 bp开放阅读框,编码259个氨基酸。系统进化树和氨基酸序列同源性比对表明,Eo AQP1在鳞翅目昆虫中高度保守。跨膜结构和水分渗透模拟表明,Eo AQP1具有经典的水分渗透模型。qRT-PCR结果表明,Eo AQP1在不同发育时期和6龄幼虫不同组织中均有表达且差异显著。亚细胞定位结果表明,Eo AQP1以圆形颗粒状成群聚集于细胞膜周边,而在细胞膜、核膜和细胞质等处均不表达。多克隆抗体Western blot检测结果表明,Eo AQP1多克隆抗体特异性较好,可用于后续相关实验。【结论】获得了Eo AQP1的c DNA序列,明确了Eo AQP1的生物学特征,阐明了Eo AQP1的时空表达特性,成功制备了Eo AQP1多克隆抗体,初步了解了Eo AQP1的亚细胞定位,为进一步研究Eo AQP1的水分渗透机理奠定了分子基础。     

茶尺蠖小RNA病毒全长cDNA克隆的构建

为了初步研究茶尺蠖小RNA病毒(Ectropis oblique picorna-like virus, EoPV)的复制机制,从被EoPV感染致死的茶尺蠖幼虫中分离并纯化病毒粒子,提取病毒RNA,根据已公布的EoPV核苷酸序列,利用基因组上单一的酶切位点,设计特异性引物,应用RT-PCR扩增出5个覆盖全长的片段。 随后采用融合PCR将5个片段拼接,最终将全长定向克隆到低拷贝质粒载体上,成功构建cDNA全长克隆p-EoPV。 双酶切及测序鉴定证明全长克隆构建成功。 与原序列比较发现,该克隆在氨基酸水平上有8个突变和1个缺失。 本研究为深入探讨EoPV病毒生物学特性、病毒复制机理等奠定了基础     

茶尺蠖病毒杀虫剂田间使用技术的研究

茶尺蠖病毒杀虫剂应在每年茶尺蠖第1、2代和第5、6代的1～2龄幼虫期使用.在7.5×109 ～ 15.0×109 PIB/hm2的使用剂量下,幼虫期的防治效果可达98%以上.采用不同的喷雾器、不同的用水量及不同的喷施方式喷施,对防治效果无影响.但挑治和丛面喷施可大幅度节约防治成本费.     

灰茶尺蛾核型多角体病毒大田防治条件的探索

为了经济有效地在大田应用EgNPV防治灰茶尺蛾 ,在室内进行EgNPV的毒力测定 ,试验表明致死中量LC50 为 1.13× 10 6PIB/mL ,90 %的致死量为 1.83× 10 7PIB/mL ,在 95%置信范围内 ,上限为 1.96× 10 7PIB/mL ,下限为 1.7× 10 7PIB/mL。用 2× 10 7PIB/mL浓度对不同龄期幼虫的感染试验表明 ,1 2龄效果最好 ,达 90 %以上。不同世代感染试验表明 :第 2、3、4、7代防治效果较好 ,高达 90 %以上 ,而第 5、6代效果不好 ,因此病毒增殖与大田防治工作应尽量避免在高温 ,干旱季节进行。另外探讨了适用大田防治的病毒增殖方法。     

不同茶园灰茶尺蠖和茶尺蠖对5种杀虫剂的抗药性监测

为了明确茶园暴食性害虫灰茶尺蠖Ectropis grisescens和茶尺蠖E. obliqua抗药性现状,制定有效的防治措施,本研究采用浸叶法分别测定了5种茶园常用杀虫剂对5个地理种群灰茶尺蠖和2个地理种群茶尺蠖的抗药性。结果表明,不同茶园灰茶尺蠖和茶尺蠖对供试药剂的抗性水平存在差异：浙江兰溪、浙江金华、河南南阳地区的灰茶尺蠖种群和浙江杭州、浙江安吉的茶尺蠖种群对联苯菊酯产生了中高等抗性;浙江杭州的茶尺蠖种群对苦参碱的抗性达到中等水平,其余供试种群对苦参碱抗性处于敏感水平;所有供试种群对虫螨腈、甲氨基阿维菌素苯甲酸盐和茚虫威这3种药剂的抗性均处于敏感水平。从化学防治的角度来看,目前茶园灰茶尺蠖和茶尺蠖对虫螨腈、甲氨基阿维菌素苯甲酸盐和茚虫威敏感性高,抗性低,推荐合理科学使用,并避免长期使用单一农药。