一种从藤粉蝶分离的微孢子虫的生物学特性和分子系统发育研究

从广州市萝岗桑园周边捕捉到的野外昆虫藤粉蝶(Delias pasithoe)成虫体内分离到1株微孢子虫,经人工接种可在家蚕体内全身性寄生。用透射电子显微镜观察到该微孢子虫为双核,极丝11～12圈,极丝同型,基本符合Nosema属微孢子虫超微结构特征。该微孢子虫在其生活史发育过程中以偶倍数核为主,以二分裂方式增殖,形成双核孢子,发育周期为108 h。基于SSU rRNA和rRNA ITS序列的相似性比较和系统进化分析进一步证实该微孢子虫与Nosema属亲缘关系最近,应被分类为Nosema属‘true’Nosema亚群。     

口蹄疫病毒非结构蛋白2C基因在昆虫细胞中的表达及应用

研究口蹄疫病毒(FMDV)非结构蛋白(NSP) 2C在区分灭活疫苗免疫动物与自然感染动物方面的意义.本研究将FMDV NSP 2C基因,克隆到穿梭载体pFast-bac- HT-B,将其转入含骨架载体Bacmid的DH10Bac,经蓝白斑筛选得到重组骨架质粒Bacmid-2C.将Bacmid-2C转染昆虫细胞Sf9,鉴定正确后,经3次增殖获得高滴度的P-3代病毒后,在High Five细胞中进行目的蛋白的表达.SDS-PAGE结果显示在High Five细胞中得到了相对分子质量约为38.93 ku的目的蛋白2C,Western blotting及Dot-ELISA结果显示,该表达产物对FMDV感染动物阳性血清有良好的反应性.以电泳纯化的2C蛋白为抗原建立间接ELISA,检测健康非免疫动物、免疫动物及FM-DV试验感染动物血清,结果表明2C-ELISA不但能区分免疫动物和感染动物的血清,而且还能检测FMDV感染早期动物血清中的2C抗体.说明昆虫细胞表达的2C蛋白可作为FMDV疫苗免疫动物与自然感染动物鉴别诊断的良好抗原.2C基因在Bac-to-Bac系统中的成功表达为建立通过检测几种NSP抗体,筛查感染及隐性带毒动物,净化畜群的方法奠定了基础.     

Avian pathogenic Escherichia coli MT78 invades chicken fibroblasts

Avian pathogenic Escherichia coli (APEC) are responsible for extraintestinal diseases, called colibacillosis, in avian species. The most severe manifestation of the disease is colisepticemia that usually starts at the respiratory tract and may result in bird death. However, it is not yet clear how APEC cross the respiratory epithelium and get into the bloodstream. In this work, we studied the interaction between 8 APEC strains (UEL31, UEL17, UEL13, UEL29, MT78, IMT5155, IMT2470, A2363) and a chicken non-phagocytic cell, the fibroblast CEC-32 cell line. We investigated the association profile, the invasion capability, the cytotoxicity effect and the induction of caspase-3/7 activation in an attempt to understand the way the pathogen gains access to the host bloodstream. Association to cells was determined after 1 h of infection, while cell invasion was determined after 4 and 24 h of infection. The cytotoxic effect of bacterial infection was measured by lactate dehydrogenase (LDH) release and the activation of the apoptotic program was verified by caspase-3/7 activation. Also, the presence of genes for adhesins, invasins and other related virulence-associated factors was verified by PCR. All bacterial strains showed similarity in relation to adhesion, LDH release and caspase-3/7 activation. However, one APEC strain, MT78, showed high invasion capability, comparable to the invasive Salmonella typhimurium strain SL1344. Since an APEC strain was capable of invading non-phagocytic cells in vitro, the same may be happening with the epithelial cells of the avian respiratory tract in vivo. CEC-32 monolayers can also provide a useful experimental model to study the molecular mechanisms used by APEC to invade non-phagocytic cells.     

上思县甘蔗主要虫害发生特点及防治对策

分析了上思甘蔗主要虫害发生原因及特点：主要是与当地气候条件关系密切,粗放的栽培管理方式与不合理的施药方法是导致虫害泛滥成灾的人为因素。提出了防治对策：通过科学的栽培管理方式与综合防治虫害措施,可达到趋利避害、高产优质的甘蔗生产目标。     

甘蔗病虫害防治技术及高效低毒农药应用

对严重影响甘蔗生产的8种甘蔗主要害虫和5种甘蔗常见流行性病害的发生危害特点进行了探讨和综述。依据甘蔗病虫害的危害特点,提出以农业防治方法为基础,物理防治灯光诱杀、生物防治为辅,减少虫源病源,统一化防为重点和抓好关键时期科学用药的综合防治措施,从而达到高效、快速、持续、有效控制甘蔗病虫害的发生危害,确保甘蔗生产持续稳定健康发展。     

海门市蔬菜病虫害测报现状及思考

为提高海门市蔬菜病虫害测报的准确率,根据海门市近几年蔬菜病虫害测报和防治实践,提出应明确测报对象．制定切实可行的测报方法,提高专业测报人员的素质,并充分利用蔬菜合作社平台开展蔬菜病虫害测报工作。     

多主棒孢菌在黄瓜、番茄和茄子寄主上致病力的分化

 对从山东、辽宁和北京蔬菜栽培地采集的黄瓜、番茄及茄子发病组织上分离的31株多主棒孢菌,和从海南橡胶上分离的10株多主棒孢菌,采用喷雾接种的方法,测定其在盆栽黄瓜、番茄和茄子上的致病力。试验结果表明,不同寄主来源菌株的致病力间存在明显的差异,寄主来源同致病力分化之间具有显著的相关性,从而证明多主棒孢菌的种内菌株存在寄主专化性的特征。相同寄主和相同地理来源的病原菌群体中致病力也存在强、中、弱的差异,说明多主棒孢菌具有明显的致病力分化现象。     

Infection processes and soft wheat response to root rot and crown rot caused by Fusarium culmorum

An isolate of the fungus Fusarium culmorum constitutively expressing green fluorescent protein was used to investigate the infection process and host response of primary seedling roots and stem base leaf sheaths of soft wheat cv. Genio. Disease progress was assessed macroscopically by visual symptoms, microscopically by confocal laser scanning microscopy (CLSM) and via gene expression analysis of fungal and wheat genes by real‐time quantitative RT‐PCR. In the roots, CLSM investigations revealed an initial intercellular and subsequent intracellular colonization by fungal hyphae. The fungus invaded the rhizodermal layer and cortex but was not seen to colonize the stele. The fungus consistently expressed TRI5 (24, 48 and 96 h post‐inoculation), indicating that trichothecenes were being synthesized throughout this phase of infection and colonization. The expression of the six host defence‐associated genes (Wheatwin 1‐2, PR1, Chitinase, PAL, WIR1 and LOX) increased early in infection and decreased during later stages. In the stem base, CLSM observations revealed the fungus sequentially penetrating though the first, second and third basal leaf sheaths. Expression of TRI5 was initiated early in the infection of each leaf sheath. The expression of the host defence‐associated genes varied over time and across leaf sheaths, and all were also expressed in leaf sheaths which had not yet been in contact with the fungus. Expression of LOX and WIR1 were particularly enhanced in the third leaf sheath.     

河北园林蚧虫名录

本文报道了河北地区为害园林植物的蚧虫8科42属73种,同时对其寄主、为害程度及分布作了介绍。     

应用白僵菌防治靖远松叶蜂的研究

利用病原菌防治害虫是森林可持续发展的重要途径。本研究采用2003年4月从山西省灵丘县下关林区油松林内采集的越冬油松毛虫DendrolimustabulaeformisTsaietLiu幼虫的僵虫尸,经实验室分离、培养和鉴定后得到的白僵菌Beauveriabassiana菌株,用2×108孢子/mL、2×107孢子/mL和2×106孢子/mL3种浓度的孢子液对靖远松叶蜂DiprionjingyuanensisXiaoetZhang幼虫分别进行室内和林间感染,试验结果显示:室内感染4龄幼虫校正致死率分别为83.37%、78.82%、62.05%,LT50分别为5.794d、6.237d、7.603d;感染5龄幼虫校正死亡率分别为68.23%、59.1%、45.79%,LT50分别为7.244d、8.414d、12.274d;林间感染幼虫校正死亡率分别为65.26%、54.21%、38.23%,LT50分别为16.788d、19.187d、30.2d。结果表明靖远松叶蜂幼虫校正死亡率与白僵菌孢子液浓度呈正相关,与LT50呈负相关,与龄期呈负相关。