斜纹夜蛾一种微孢子虫样病原的杀虫试验

1981年,我们报导了一种来自斜纹夜蛾Prodenia litura Fabrieius的微孢子虫样病原的分离提纯,室内人工致病试验,以及病征和病理学观察结果。本文报导不同虫龄斜纹夜蛾幼虫对人工饲喂孢子后的反应,以及这种微孢子虫样病原与苏芸金杆菌混合感染后的联合致病作用。     

经低温贮存的大蜡螟幼虫对线虫侵染率及侵染强度的影响

大蜡螟（Ｇａｌｅｒｉａｍｅｌｏｎｅｌａ）幼虫对昆虫病原线虫极为敏感，又易于人工标准化饲养，因此常作为昆虫病原线虫的资源调查、活体繁殖、线虫回收和线虫毒力测定等研究的试虫。大蜡螟幼虫在真菌、细菌等其它微生物研究中也有广泛用途。在实际工作中，一次试验需用...     

昆虫病原线虫的应用近况

昆虫病原线虫(Entomopathogenic nematodes)是害虫的天敌,是害虫生物防治中的一个重要因子。它的研究应用从30年代发现距今已有50多年的历史,其中携带共生细菌的斯氏线虫属Steinernema Travassos和异小杆线虫属Heterorhabditis Poinar是在害虫生物防治中最有潜力的类群。各国科学家已发表了不少线虫的生物学、生态学、大量繁殖方法和防治效果的研究报告。利用昆虫病原线虫防治害虫有以下优点:①线虫的寄主范围广,可以利用线虫防治多种害虫;②可以人工培养基大量繁殖线虫;③线虫可以主动地寻找寄主,对土栖和隐蔽的钻蛀害虫效果高;④线虫体内带有共生细菌,具高的杀虫能力,能在16至48小时内杀死寄主昆虫;⑤线虫对环境安全,有一定的抵抗力,可与一些杀虫剂混用。本文拟就日前线虫的生产、应用,影响线虫效用的因素和线虫应用的前景浅淡。     

不同施用方式下昆虫病原线虫对小地老虎的防治作用

小地老虎是重要的地下害虫,严重为害烟草、玉米、棉花等作物。为促进昆虫病原线虫在小地老虎防治中的使用,以云南当地品系与商业品系为材料,调查了3种施用方式（悬液、悬液+覆盖刨花、虫尸剂）下昆虫病原线虫防治小地老虎保护烟苗的效果,并根据后续埋入的大蜡螟死亡情况评估昆虫病原线虫在土壤中的存活情况。盆栽试验结果表明,喷施悬液7 d后,两个商业品系的昆虫病原线虫Steinernema carpocapsae All(Sc All)和Heterorhabditis indica LN2(Hi LN2)保苗效果显著。各施用方式处理保苗效果由高到低为悬液>悬液+覆盖刨花>虫尸剂。田间试验结果表明,处理7 d后,Sc All悬液处理效果优于所有品系的虫尸剂处理效果,烟苗死亡率与化学农药氯氰菊酯相当,直接施用Sc All悬液能较好地抑制小地老虎对烟苗的为害,而虫尸剂释放的昆虫病原线虫存活时间较长,可针对不同的防治窗口期结合应用。     

蚜虫上虫霉的鉴定

虫霉目Entomophthorales真菌在合适的条件下能引起广泛的流行病,是不少害虫种群自然调节的主要因素。迄今已知的160多种虫霉中,有120多种是昆虫病原。据初步统计,约40种侵染同翅目昆虫,其中20种侵染蚜虫类;约37种侵染双翘目,其中16种侵染蝇类;约19种侵染鳞翅目,13种侵染鞘翅目。由此可见,蚜虫是虫霉的重要寄主。虫霉研究在国外已有百余年历史。如今美国和法国已成功地进行了试验性生产~(13),法、美、英、苏联、澳大利亚和我国已对某些蚜虫进行了田间防治试验。在20种侵染蚜虫的虫霉中,有15种已可人工培养。在我国,蚜虫上的虫霉近年来已陆续有所采集报道,但由于虫霉目分类混乱,命名变动大,再加上资料及实验技术所限,目前在鉴定中困难不少,且有些鉴定尚有疑问。因此有必要将侵染蚜虫的20种虫霉做以介绍。     

高效新型杀松材线虫剂的筛选及其林间防效

为筛选出新型的高效松材线虫Bursaphelenchus xylophilus防治药剂,采用浸渍法室内测定了氯氟氰虫酰胺、阿维菌素、甲维盐、灭多威、特丁硫磷、甲基异柳磷和吡虫啉7种药剂对松材线虫的毒杀作用和对其繁殖能力的影响,采用高效液相色谱仪测定氯氟氰虫酰胺在马尾松体内的输导分布,并在林间开展树干注射氯氟氰虫酰胺抗松材线虫病试验。结果表明,7种药剂中氯氟氰虫酰胺对松材线虫的毒力最强,LC_(50)为0.019 mg/L,阿维菌素和甲维盐次之,LC_(50)分别为0.0213 mg/L和0.0214 mg/L。氯氟氰虫酰胺树干注射3个月内能迅速从马尾松基部传导至树梢顶部,树干边材东、南面的药剂含量显著高于西、北面,且树干不同高度的氯氟氰虫酰胺含量均高于其LC50;氯氟氰虫酰胺制剂在林间注干后,对人工接种病原松材线虫松树的防治效果为73.33%,自然感染松材线虫病松树的死亡率降至2.00%,而对照试验中人工接种松材线虫松树和自然感染松材线虫病松树的死亡率分别为93.33%和19.67%。表明氯氟氰虫酰胺制剂能有效预防松树个体的松材线虫病发生与为害,可以推广使用。     

利用大蜡螟诱集法从土壤中分离昆虫病原真菌

USINGTHE"GALLERIABAITMETHOD"FORISOLATIONOFENTOMOPATHOGENICFUNGIFROMSOILGAOSongNONGXiang-qunDENGChun-sheng(InstituteofBiologicalControl,ChineseAcademyofAgriculturalSciences,Beijing100081)利用昆虫病原真菌防治农林害虫的研究和应用，近些年来取得了很大的进展。随着公众环保意识增强，要求更多更广泛地使用真菌杀虫剂来防治害虫，使研究昆虫病原真菌的工作日益显得重要起来。通常采集菌株的方法是收集自然感病僵虫在室内保湿培养后分离。但寻找僵虫的工作量大，花费时间多且受季节影响…     

我国昆虫病原线虫的田间试验研究进展

作为害虫的自然天敌,昆虫病原线虫的寄生特性引起了人们的重视,认为是生物防治中的一支新的力量。自从八十年代引进国外大量繁殖技术以来,我国有关研究得到很大的发展,已有不少单位进行了田间应用试验,初步打开了应用研究的局面.主要情况是:1.利用线虫在土壤中比较活跃的特性,大量开展防治某个虫态生活在土中的害虫的试验,取得了防治桃小食心虫大面积试验的良好效果。桃小食心虫在我国主要为害苹果,造成重大经济损失.该虫产卵在果实表面,卵孵后幼虫钻入果心取食,幼虫老熟时随烂果落地入土化蛹。1981～1983广东省昆虫研究所和中国农科院郑州果树所根据该虫在果园土壤中化蛹、虫源地比较集中和昆虫病原线虫     

大麦虫作为替代寄主和人工饲料繁育花绒寄甲对其繁殖生物学的影响

花绒寄甲Dastarcus helophoroides是防治多种蛀干害虫天牛的重要天敌,为了解决人工大量繁育花绒寄甲的技术问题,本文选用大麦虫Zophobas atratus蛹作为繁育花绒寄甲幼虫的替代寄主,利用配置的人工饲料饲喂成虫,在实验室中连续饲养4代花绒寄甲,统计分析所繁育出的每代花绒寄甲成虫的产卵量、产卵前期时长、雌雄性比、成虫体长和体宽、体质量、卵的孵化率以及其后代幼虫的寄生率等,分析人工饲料和替代寄主对室内连续多代繁育的花绒寄甲繁殖生物学的影响。结果表明：花绒寄甲成虫的体长、体宽、体质量随着室内饲养代数的增加而减小;花绒寄甲成虫的产卵前期随着饲养代数的增加而延长,但后一种情况通过在饲料中添加维生素可有效解决。随着室内饲养花绒寄甲代数的增加和人工繁育时间的延长,平均每头花绒寄甲雌虫的产卵量增加;使用替代寄主繁殖4代后出现成虫雌雄性比下降,不符合1：1的情况。室内饲养4代的花绒寄甲卵的孵化率和初孵幼虫的寄生率几乎没有影响。通过对各个繁殖生物学指标的比较发现,饲喂改良人工饲料的花绒寄甲成虫的产卵前期在饲养多代后能明显缩短;利用大麦虫蛹繁育出的花绒寄甲后代雄虫数量明显多于雌虫;相反,利用改良人工饲料与花绒寄甲原始寄主松褐天牛幼虫配合使用,繁育出的第2、3代雌虫数量明显多于雄虫,差异显著。综上所述,利用大麦虫作为替代寄主会导致花绒寄甲后代雌雄性比下降,但可以通过改进饲料配方和利用原寄主饲养,在室内饲养多代后对花绒寄甲繁殖生物学的各项指标影响不大。因此,利用大麦虫和人工饲料等饲养技术综合应用,能够在室内人工规模化繁育花绒寄甲。     

广东水稻橙叶病病原(MLO)的越冬

研究结果表明,广东水稻橙叶病病原(MLO)的传染介体和越冬介体昆虫只有电光叶蝉(Recilia dorsalis)一种,但其卵不传递该病原到子代若虫,越冬寄主植物只有水稻的再生稻株和落粒自生稻株。在茂名病区的冬、春期间电光叶蝉的带菌虫和带菌再生稻及感病落粒自生稻始终存在,其带菌率分别为2.33％—35.11％、2.37％—17.78％和1.95％—16.67％。在茂名地区由于冬、春气温不甚低,电光叶蝉仍能继续繁殖越冬,并能不断获菌和传病,所以说该病原是动态越冬的。室内试验表明,带菌电光叶蝉的寿命可长达105—125天,而在茂名的秋收期至翌年春播期一般105天,据此推测有些带菌虫可从秋收后直接存活和传病原到春播稻苗。看麦娘(Alopecurus aequalis)用电光叶蝉人工接种并回接水稻成功。但在病区田间调查从未发现病株。以下植物人工接种不成功:野生稻(Oryzea rufipogon)、小麦(Triticum aestivum)、玉米(Zea mays)、李氏禾(Leersia hexandra)、稗草(Echinochloa crus-gallis)。     

布氏白僵菌的研究与应用

对７０年代以来布氏白僵菌的生物学、致病性、生理生化特性等基因研究以及该菌的规模征税和田间防治应用地综述。目前已发现该菌能寄生７个目的７０种昆虫,特别对鞘翅目害虫具有独特寄生效果,已在林区、农田、草地、果园中用于防治各种金龟子和天牛。在法国、瑞士等地,用布氏白僵菌连续多年防治林区的西方五月鳃金龟,取得了令人瞩目的防治效果和生态效果,日本用该菌防治杉树天牛、我国用该菌防治农田及苗圃蛴螬都取得明显成效。     

Role of the Insect Vector, Pseudotheraptus devastans, in Cassava Anthracnose Disease Development

The aim of this study was to investigate the role of Pseudotheraptus devastans in cassava anthracnose disease transmission and development. P. devastans, Dist (Het. Coriedae) insects were collected from cassava (Manihot esculenta Crantz) field plots at the International Institute of Tropical Agriculture, Ibadan, Nigeria and reared in large cages. The insects were separated at different developmental growth stages of eggs, first to fifth instar nymph, and adults. The different stages of P. devastans showed the presence of Colletotrichum gloeosporioides both externally and internally. Isolates of C. gloeosporioides derived from the insects produced cassava anthracnose disease symptoms (necrotic lesions, wilt and defoliation) 8 weeks after inoculation on two cassava clones. Re-infectivity of cassava plants by the insect-derived fungus established that P. devastans was a potential vector in anthracnose transmission. Except for the first and second instar nymphs, all nymph stages and adult insects produced significant anthracnose symptoms on cassava plants. Defoliation and lesion diameters were greatest using fifth instar nymphs and adult insects. The association between P. devastans feeding and C. gloeosporioides f.sp. manihotis, showed that feeding by P. devastans followed by fungal inoculation and vice versa resulted in more severe anthracnose symptoms than insect feeding or fungal inoculation alone. It was also observed that the influence of P. devastans damage/infection on the development of anthracnose depended on cassava cultivar resistance to both the fungus and the insect feeding.     

Insecticidal efficacy of beauveria bassiana,diatomaceous earth and fenitrothion against rhyzopertha dominica and tribolium castaneum on stored wheat

Bioassays were conducted in stored grains to evaluate the combined use of the entomopathogenic fungus Beauveria bassiana, diatomaceous earth (DE) and fenitrothion against adults of Rhyzopertha dominica and Tribolium castaneum. The insecticide agents were applied to wheat as follows: the fungus at 1 × 10⁸ conidia/kg of wheat, DE at 3 g/kg of wheat and fenitrothion at a rate of 0.15 ml/kg of wheat (25% of field dose rate). Surviving insects and progeny production were monitored at thirty-day intervals until four months. For both insect species, significantly less progeny was produced on wheat treated with fenitrothion and DE + B. bassiana formulations in comparison with controls. The effect of insecticides on the grain damage, germination power of wheat and bulk density was also evaluated. After four months, B. bassiana mixed with DE reduced the damaged insect grains by 50% in comparison with both fenitrothion and control treatments. Trials using DE caused a reduction in bulk density while there were no significant differences for germination testing between treatments.     

朱红毛斑蛾虫生真菌球孢白僵菌PfBb菌株的分离鉴定及其毒力测定

为评价从朱红毛斑蛾Phauda flammans幼虫僵虫中分离的虫生真菌在害虫生物防治中的应用潜力,通过形态学鉴定及ITS基因序列分析对分离菌株进行鉴定,并运用时间-剂量-死亡率模型分析该分离菌株对朱红毛斑蛾1~6龄幼虫的毒力。结果显示,结合形态学特征和ITS基因序列分析将该虫生真菌鉴定为球孢白僵菌Beauveria bassiana,并将该菌株命名为PfBb。球孢白僵菌PfBb菌株可侵染朱红毛斑蛾1~6龄幼虫,随着该菌株孢子液浓度的增加,其对朱红毛斑蛾幼虫的毒力逐渐增强。PfBb菌株孢子液处理朱红毛斑蛾1~5龄幼虫6 d时的LC₅₀分别为2.61×10⁴、1.04×10⁵、1.11×10⁶、1.54×10⁶和1.44×10⁷ 个/mL,处理6龄幼虫8 d时的LC₅₀为1.02×10⁷ 个/mL。随着处理时间的增加,PfBb菌株对朱红毛斑蛾1~6龄幼虫的LC₅₀和LC₉₀随之降低,表明存在剂量效应。当处理浓度相同时,朱红毛斑蛾1~6龄幼虫的僵虫率随着虫龄的增加而降低,但在较高浓度1.0×10⁷~1.0×10⁹ 个/mL处理时,1~5龄幼虫的僵虫率无显著差异,说明1.0×10⁷个/mL球孢白僵菌PfBb菌株孢子液防治朱红毛斑蛾幼虫即可达到理想的防治效果。表明球孢白僵菌PfBb菌株对朱红毛斑蛾有较大的生防应用潜力。     

球孢白僵菌定殖提高番茄对灰霉病抗性及其作用机理

为明确球孢白僵菌Beauveria bassiana定殖对番茄植株抗病性的影响及作用机理,采用灌根法将球孢白僵菌芽生孢子悬浮液接种于番茄植株内,并通过人工接种灰霉菌Botrytis cinerea评价球孢白僵菌定殖后番茄对灰霉病的抗性水平;检测灰霉菌胁迫下番茄植株不同位置叶片内球孢白僵菌的相对含量;测定番茄叶片内草酸氧化酶（oxalate oxidase,OXO）、几丁质酶（chitinase,CHI）和ATP合成酶（ATP synthase,atpA）3种抗病相关基因的表达量。结果表明,球孢白僵菌定殖能够提高番茄植株对灰霉病的抗性,接种灰霉菌第5天,番茄植株发病率、病斑直径和病情指数分别下降了61.6%、41.4%和26.4%;在灰霉菌胁迫下番茄植株内球孢白僵菌偏好于在病原菌感染位置定向聚集,并且引起植物抗病基因OXO、CHI和atpA的表达量上调。表明球孢白僵菌能通过内生定殖与植物互作提高植物抗病性,在植物病害生物防治领域有较大应用潜力。     

Cylindrocladium brown leaf spot on Howea belmoreana caused by Calonectria ilicicola (anamorph: Cylindrocladium parasiticum) in Japan

Brown leaf spot disease caused by Cylindrocladium was found on Howea belmoreana on Hachijojima Island, Tokyo, Japan, in December 2001. Typical symptoms were incited after artificial inoculation. A culture of white mycelia, isolated from leaf spot symptoms, produced reddish perithecia of a nectriaceous fungus. Based on morphological and molecular analyses, this fungus was identified as Calonectria ilicicola (anamorph: Cylindrocladium parasiticum). Pathogenicity of this fungus on five plants cultivated on Hachijojima Island was confirmed by artificial inoculation. This report is the first on Cylindrocladium brown leaf spot of H. belmoreana caused by C. ilicicola (anamorph: Cy. parasiticum).     

Long‐term survival of blast pathogen in infected rice residues as major source of primary inoculum in high altitude upland ecology

Magnaporthe oryzae is the fungal plant pathogen that causes rice blast. The sources of primary inoculum and overwintering mode of the fungus remain largely unknown. The effect of rice residues on the onset of blast epidemics and the potential for survival of M. oryzae in the residues were studied in upland conditions in Madagascar. Blast disease was observed in a 3‐year field experiment in three treatments: with either infected or uninfected rice residues on the soil surface, or without rice residues. Leaf blast incidence was significantly higher in the treatment with infected rice residues than in the two other treatments at the early stages of the epidemic. In a second set of trials, the survival of M. oryzae on rice residues was monitored. Infected rice stems were placed by lots in three places: on the mulch of rice residues, under the mulch, and buried at a depth of 10 cm in the soil. Each month, samples were taken from the field and tested for sporulation. The survival of the blast fungus decreased rapidly on the stems buried in the soil but remained high for the other conditions. Sporulation of the fungus was observed on stems left on the mulch for up to 18 months. It is concluded that under field conditions, the presence of infected rice residues could initiate an epidemic of blast. The results of this study may help in designing effective management strategies for rice residues infected by M. oryzae.     

参与烟粉虱免疫反应的clip丝氨酸蛋白酶基因分析

为了解参与烟粉虱免疫反应的clip丝氨酸蛋白酶(clip-domain serine proteases in Bemisia tabaci,Bt CLIPs)基因,对已鉴定的8个Bt CLIPs基因编码的蛋白进行序列分析,与冈比亚按蚊、家蚕、黑腹果蝇、烟草天蛾4种模式昆虫的同源蛋白进行系统进化分析,并通过q RT-PCR检测球孢白僵菌侵染烟粉虱4龄若虫后Bt CLIPs的时间表达模式。结果显示,8个Bt CLIPs基因均为含有CLIPs结构域的丝氨酸蛋白酶基因;Bt CLIP19、Bt CLIP20和Bt CLIP21独自聚在进化树的一支上;其它Bt CLIPs均与4种模式昆虫中的1种或多种同源蛋白聚在一起;与对照相比,8个Bt CLIPs基因受球孢白僵菌侵染后在不同时间的相对表达量均上调,但诱导程度不同,48 h的相对表达量达最高,其中Bt CLIP19上调表达最明显,为对照组的71倍。研究表明,8个Bt CLIPs可能参与烟粉虱对真菌侵染的免疫反应,并成为烟粉虱生物防治的新靶标。     

Biological control of the Western flower thripsFrankliniella occidentalis in cucumber using the entomopathogenic fungusMetarhizium anisopliae

Biological control of the western flower thrips (WFT)Frankliniella occidentalis, using the entomopathogenicMetarhizium anisopliae-7 (M. a-7) strain was studied in three consecutive seasons under greenhouse conditions. Cucumber plants infested with WFT were sprayed with spore suspension of the fungusM. a-7 (0.5 g m^-2), or the soil was treated with dry powder of the fungus (0.5 g m^-2); the control was without fungus application. In the 1997 spring experiment, when the cucumber plants were initially infested with only three or four insects per leaf, the spore suspension spray caused a significant reduction in growth of the thrips population compared with the other treatments and the control. However, in the 1997 summer experiment, when the plants were initially heavily infested with WFT (10–15 insects per leaf), the spray treatment caused only a modest reduction in WFT population growth, and only after 4 weeks of treatment was the reduction significant. In the 1999 experiment, with a low initial WFT population of three or four insects per leaf, the spray treatment was effective in reducing the population growth to a lower level than in the other treatments or control. TheM. a-7 strain was found to be effective in reducing the population growth of WFT under greenhouse conditions, particularly when the initial thrips population was low to moderate. http://www.phytoparasitica.org posting Nov. 4, 2001.     

棉花内生真菌CEF-373菌株对棉花黄萎病的防效及其作用机理

为明确棉花内生真菌CEF-373菌株对棉花黄萎病的防效及其作用机理,利用圆盘滤膜法和平板对扣培养法测定菌株CEF-373对大丽轮枝菌Verticillium dahliae菌丝生长的抑制作用,测定其对棉花黄萎病的温室和田间防效,并通过活性氧含量及防御基因表达情况来分析其作用机理。结果表明,菌株CEF-373的挥发性代谢产物和非挥发性代谢产物对大丽轮枝菌菌丝的生长均有显著抑制作用,抑制率最高分别可达37.75%和100.00%。用1×107CFU/mL的菌株CEF-373分生孢子悬浮液灌根后,对棉花黄萎病的温室防效可达71.12%,用质量比为3%的菌株CEF-373固体菌剂拌土栽培后,对棉花黄萎病的温室防效可达62.74%,防治作用显著。菌株CEF-373的发酵液滴灌和微生物肥料处理40 d后,对棉花黄萎病的田间防效达到最大,分别为36.23%和27.71%,而在后期有所降低。菌株CEF-373可以诱导棉花叶片中细胞活性氧的爆发;且菌株CEF-373成功激活了苯丙氨酸解氨酶、过氧化物酶、多酚氧化酶、几丁质酶和病程相关蛋白基因PR10的表达,对大丽轮枝菌的侵染具有抵御作用。表明棉花内生真菌CEF-373菌株通过抑制大丽轮枝菌生长以及诱导寄主系统抗病性来有效防治棉花黄萎病,具有较好的生物防治应用前景。