耐低水活度高毒力虫生真菌菌株选育

以球孢白僵菌Beauveria bassiana和玫烟色拟青霉Paecilomyces fumosoroseus为研究对象,利用紫外线诱变芽生孢子和低水活度条件胁迫筛选,选育出突变株Bb07240、Pf01120和Pf01160,它们在低水活度（或相对湿度）下的生长和萌发均明显优于其相应的初始菌株。生物测定表明,突变株对桃蚜的毒力明显高于其初始菌株。     

两种杀虫真菌制剂对茶小绿叶蝉的田间防效评价

于2002年盛夏在浙江遂昌一高山茶园对茶小绿叶蝉(Empoasca spp．)进行了真菌杀虫剂的田间药效试验。所用菌剂为球孢白僵菌(Beauveria bassiana)和玫烟色拟青霉(Paecilomyces fumosoroseus)的纯孢子悬乳剂及其与3％吡虫啉10％可湿剂的混配剂．各菌剂稀释500倍喷雾2次,间隔12d．结果表明。两种真菌的混配剂明显优于纯菌剂,其中球孢白僵菌混配剂的最高防效达83．4％,而玫烟色拟青霉的最高防效为71．3％．根据25d期间历次调查结果计算平均防效,球孢白僵菌混配剂达66．8％,玫烟色拟青霉混配剂为62．1％,含微量吡虫啉的矿物油为50．3％,球孢白僵菌和玫烟色拟青霉的纯菌剂分别为49．5％和19．0％,结合试验期间气候及田间种群结构特征,讨论了各处理问差异的来源及提高真菌杀虫剂控制茶小绿叶蝉效果的可能途径。     

虫生真菌玫烟色棒束孢Pf9606抗逆性的研究

采用凹玻片悬滴法测定了玫烟色棒束孢在高温、紫外照射、紫外保护剂和杀菌剂等逆境胁迫下分生孢子的存活率。结果表明:玫烟色棒束孢Pf9606分生孢子在36～45 ℃高温下连续处理24 h后,该菌孢子几乎不能存活;36～45 ℃短时高温处理后,随温度的升高和处理时间的延长,孢子存活率明显降低;随紫外照射时间的延长,玫烟色棒束孢孢子存活率明显降低,当紫外照射50 s时,孢子存活率为55.33%,当照射时间延长为180 s时,孢子不能存活;不同浓度的抗坏血酸和高浓度的荧光素钠可提高玫烟色棒束孢抗紫外照射能力,抗坏血酸的理想浓度为0.2 mg/mL,荧光素钠的理想浓度为0.9 mg/mL,而不同浓度的刚果红和氧化锌对玫烟色棒束孢没有保护作用;玫烟色棒束孢与杀菌剂72%农用硫酸链霉素、70%代森联和58%甲霜灵锰锌混用后,对该菌孢子存活影响较小,不同浓度下孢子存活率均达85%以上,而常规浓度的氟吗·锰锌对玫烟色棒束孢孢子存活有较强的抑制作用。     

耐低水活度高毒力虫生真菌菌株选育*

以球孢白僵菌Ｂｅａｕｖｅｒｉａ　 ｂａｓｓｉａｎａ和玫烟色拟青霉Ｐａｅｃｉｌｏｍｙｃｅｓ　ｆｕｍｏｓｏｒｏｓｅｕｓ为研究对象,利用紫外线诱变芽生孢子和低水活度条件胁迫筛选,选育出突变株Ｂｂ０７２４０、Ｐｆ０１１２０和Ｐｆ０１１６０,它们在低水活度（或相对湿度）下的生长和萌发均明显优于其相应的初始菌株。生物测定表明,突变株对桃蚜的毒力明显高于其初始菌株。     

琅琊山虫生真菌物种多样性和季节变化

在皖东琅琊山自然保护区设置的20个样方中共采集672份虫生真菌标本,分属于4科8属20种.优势种依次为球孢白僵菌、粉拟青霉、环链拟青霉、布氏白僵菌和玫烟色拟青霉,其中球孢白僵菌占绝对优势,相对多度高达73.8%.选用物种多度、Shannon-Wiener指数H′和Pielou均匀度E对琅琊山虫生真菌物种多样性和季节分布进行研究发现,虫生真菌数量丰富但物种多样性指数较低;夏季虫生真菌菌株最为丰富,均匀度相对较低,随着气温及降雨量的下降,菌株数量急剧下降,均匀度相对较高;不同季节白僵菌属的数量波动大,但拟青霉属波动较小.     

玫烟色棒束孢相容性农药筛选及混用对小菜蛾的防效

为了明确玫烟色棒束孢Isaria fumosoroseus与常用农药的相容性,本研究测定了7种常用农药制剂对玫烟色棒束孢菌丝生长、孢子萌发以及产孢量的影响,测试了相容性较好杀虫剂与玫烟色棒束孢混用对小菜蛾Plutella xylostella Linnaeus的防治效果。结果表明,田间使用浓度下,氯虫苯甲酰胺、双丙环虫酯和四氯虫酰胺对菌丝抑制作用相对较小,田间浓度和亚致死浓度的阿维菌素对菌丝生长具有促进作用。所有供试农药在田间使用浓度下对孢子萌发均具有明显抑制作用,抑制作用随农药浓度降低而减弱,其中噻虫啉和四氯虫酰胺对孢子萌发抑制作用相对较小。所有测试药剂对产孢量具有显著抑制作用,抑制作用随农药浓度降低而减弱,其中噻虫啉和氯虫苯甲酰胺抑制作用较小。玫烟色棒束孢3种浓度（9×107、9×106、9×105个孢子/mL）分别与次亚致死剂量四氯虫酰胺混用对小菜蛾的LT50分别为2.19 d、2.94 d、5.16 d,杀虫速度高于单用相应浓度真菌处理（2.34 d、3.29 d、5.33 d）和四氯虫酰胺单独处理（10.60 d）。综合分析结果表明,低剂量的四氯虫酰胺、噻虫啉和氯虫苯甲酰胺与玫烟色棒束孢相容性相对较好,是田间菌药混用可供选择对象;次亚致死剂量的四氯虫酰胺与真菌混用防控小菜蛾具有协同增效作用。     

比较两种昆虫病原真菌中结构相似的ATP结合匣转运蛋白的抗氧化力与多药抗性

[目的]比较分析球孢白僵菌(Beauveria bassiana)BbT1和玫烟棒束孢(Isaria fumosorosea)IfT1两种结构相似的ATP结合匣转运蛋白的生物学功能.[方法]基于Bb2860野生株构建BbT1的敲除株和回补株,并将IFT1在BbT1敲除株中异源重组表达,比较各菌株的表型变化.[结果]与野生株、回补株及异源重组株相比,敲除株对20 - 40 mmol/L过氧化氢和2-8 mmoL/L甲萘醌氧化胁迫的抵抗力下降27％ -2.1倍,对多菌灵、伊曲康唑、菌核净、放线菌酮、乙嘧酚和4-硝基喹啉N-氧化物等不同类型化学药物的抗药性下降28％ -4.7倍,对斜纹夜蛾Spodoptera litura二龄幼虫的毒力下降20％左右,而野生株、回补株及异源重组株之间无任何表型的显著差异.[结论]BbT1和IfT1是结构相似且功能一致的转运蛋白,分别是两种生防真菌多药抗性的决定因子之一,因参与抗氧化反应而对毒力有所贡献.     

两种表面活性剂对玫烟色棒束孢PF904菌株侵染小菜蛾幼虫的影响

[目的]在前期筛选试验的基础上,比较表面活性剂对玫烟色棒束孢Isaria fumosoroseus PF904菌株侵染小菜蛾Plutella xylostella 3龄幼虫的增效作用,获得可提高玫烟色棒束孢PF904防治效果的助剂.[方法]通过扫描电镜观察和室内致病力试验,测定喷施分别添加表面活性剂聚二甲基硅氧烷(OF...     

2株烟粉虱高致病力病原棒束孢菌株的毒力测定

烟粉虱(Bemisiatabaci)是世界性分布且危害严重的蔬菜花卉害虫.由于烟粉虱的抗药性及传统化学药剂施用潜在的负面影响,利用病原微生物、天敌昆虫等开展烟粉虱的生物防治一直是国内外学者们研究的重点与热点之一.本研究在室内条件下,分析了爪哇棒束孢(Isaria javanica)GZQ-1菌株和玫烟色棒束孢(Isariafumosorosea)YMS01菌株对烟粉虱(MEAM1隐种)不同虫态的致病力.结果表明,2种棒束孢昆虫病原真菌均能侵染烟粉虱各个龄期,且随着孢子浓度的升高,烟粉虱的死亡率不断上升;在1×10⁸孢子/mL时,2株棒束孢对烟粉虱2龄若虫的感染致死率分别达到92.68%和78.12%;此外,2株棒束孢菌株对2龄若虫的致病力回归方程分别为y=0.421x-1.665,y=0.33x-1.646, LC₅₀值分别为9.01×10³孢子/mL和9.74×10⁴孢子/mL, LT₅₀为3.05 d和4.56 d.综上所述,本研究所评价的2株棒束孢均对烟粉虱有较高的致病力...     

不同含水量和温度下贮存中球孢白僵菌分生孢子活力与内贮营养的衰变

将球孢白僵菌(Beauveriabassiana)BBSG8702的未干燥孢子粉(含水量58.9±1.6%)和真空冷冻干燥孢子粉(含水量7.4±0.9%)置于4℃和20℃下贮存1个月,每隔5d取样测定活孢率和孢子内贮总糖和蛋白含量,发现含水量和贮存温度交互影响孢子的活力以及内贮总糖和蛋白质的代谢水平,各组合中的活孢率一般与内贮总糖和蛋白质代谢水平均存在显著或极显著相关性.在1个月的贮存期间,4℃下冻干粉总糖含量下降13.4%,蛋白质含量下降39.2%,清水中的萌发率下降32.0%,营养液中的萌发率仅下降6.7%,而未干燥孢子粉的相同指标分别下降42.4%、66.3%、96.4%和99%;在20℃下,冻干粉的上述指标分别下降了14.1%、38.2%、55.8%和 10.4%,而未干燥孢子粉则分别下降了 43.2%、65.4%、99.4%和98.4%. 显然,含水量影响活孢率和内贮营养衰变的幅度,而温度影响衰变的速度,但内贮营养的耗尽并不立即引起孢子失活,在供给外源营养之后孢子仍能萌发.将含水量降至4.0±0.9%的冻干粉贮存1年,4℃下活孢率由初始的99.0%下降至90.2%,而20℃下贮存的前165d活孢率下降较为缓慢,但此后急剧下降,至第240d时几乎全部失活.模拟分析表明,低含水量冻干粉在4℃和20℃下贮存的半衰期(即活孢率减少一半所需的时间)分别为1006d和197d.这些结果说明,白僵菌纯孢粉的含水量     

Comparison of aerial conidia and blastospores from two entomopathogenic fungi against Diaphorina citri (Hemiptera: Liviidae) under laboratory and greenhouse conditions

This study compared the insecticidal activity of liquid culture-produced blastospores and solid substrate-produced aerial conidia of Beauveria bassiana GHA and Isaria fumosorosea ARSEF3581 strains against Diaphorina citri adults. Insects exposed to 10⁷ propagules/ml in a spray residue contact leaf bioassay died within 6 days at 25°C, with no significant differences between fungal treatments. At higher concentrations (10⁸ propagules/ml), I. fumosorosea conidia killed psyllids faster compared to its blastospore formulation, i.e. 4 versus 5 days, respectively. In greenhouse tests, the same treatments applied to infested citrus plants (2?×?10⁶ spores/ml) all significantly reduced the number of nymphs compared with the untreated controls over 3 weeks; however, only I. fumosorosea blastospores significantly reduced the number of F1 adult psyllids when compared with controls. Similar results were observed in the follow-up greenhouse test, where I. fumosorosea blastospores were the most effective treatment overall, reducing D. citri populations by about 60% after 21 days; by contrast, imidacloprid killed almost 100% of psyllids within a week in both tests. Fewer psyllids exhibited mycosis in the greenhouse (i.e. ≈20 versus?≥?87% in the laboratory). This is the first report comparing both conidial and blastospore formulations of B. bassiana and I. fumosorosea for the control of a psyllid pest. Field testing is required to determine how successful different spore formulations might be under various environmental conditions.     

Entomopathogenic fungi disturbed the larval growth and feeding performance of Ocinara varians (Lepidoptera: Bombycidae) larvae

Abstract Feeding experiments using three strains of entomopathogenic fungi, Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea were conducted with newly moulted 3rd–5th instar Ocinara varians Walker larvae in the laboratory. The mortality of larvae immersed individually in spore suspension (1 × 10⁷ spores/mL) of all the strains was ≥ 80% except 5th instar larvae treated with M. anisopliae which transformed into pupae, but did not result in adult emergence. The growth (total body mass), consumption, relative consumption rate and relative growth rate, were reduced at all three larval stages, while developmental time was extended in infected larvae with concurrent significant increase in approximate digestibility in infected larvae. Conversion of digested food (ECD) and ingested food (ECI) values declined in infected larvae as compared to the healthy larvae (control). The 5th instar larvae treated with M. anisopliae showed higher ECD and ECI values than control. Based on mortality and growth inhibition it can be suggested that all the studied fungal strains have a high potential for biocontrol and could be developed into biocontrol agents against O. varians.     

Insecticidal activity of entomopathogenic fungi (Hypocreales) for potato psyllid,Bactericera cockerelli (Hemiptera: Triozidae): Development of bioassay techniques,effect of fungal species and stage of the psyllid

The potato psyllid, Bactericera cockerelli (?ulc), is a pest of potato, tomato, and some other solanaceous vegetables and has also been incriminated in the transmission of a bacterial pathogen, Candidatus Liberibacter solanacearum, resulting in a serious disease known as ‘zebra chip’. Although there are several reports of fungal pathogens in psyllids, there are none from B. cockerelli, nor have any fungi been evaluated against it. Five isolates of fungi, one Beauveria bassiana, two Metarhizium anisopliae and two Isaria fumosorosea, were bioassayed against B. cockerelli on potato leaves under ideal conditions for the fungi. All applications were made with a Potter spray tower. With the exception of concentration-effect studies, all other applications were made using 10⁷ conidia/mL in a 2-mL aqueous suspension. All isolates except B. bassiana, produced 95–99% mortality, corrected for control mortality, in adults 2–3 days after application of conidia and 91–99% in nymphs 4 days after application. The corrected mortalities for adults and nymphs treated with B. bassiana were 53 and 78%, respectively, 4 days after application. I. fumosorosea Pfr 97 produced 95% corrected mortality in both first and late third instar nymphs. M. anisopliae (F 52) produced 96% corrected mortality in first and third instar nymphs. Pfr 97 and F 52 were evaluated for insecticidal activity against third instar B. cockerelli using 10⁵, 10⁶, and 10⁷ conidia per mL. Mortality produced by I. fumosorosea Pfr 97 ranged from 83 to 97% and that of M. anisopliae F 52 was 88 to 95% at these concentrations.     

温湿度对玫烟色棒束孢IF-1106菌株孢子萌发及对烟粉虱致病力的影响

B型烟粉虱Bemisia tabaci Gennadius是一种重要的世界性农业害虫。玫烟色棒束孢Isaria fumosorosea是防治烟粉虱的一种重要的虫生真菌。对不同温度(20、23、26、29、32℃)、不同湿度(53%、65%、75%、85%、95%)下玫烟色棒束孢IF‐1106菌株分生孢子萌发及其对烟粉虱的致病力进行了测定。结果表明:温度对孢子的萌发有显著影响,26℃时孢子萌发率最高。当相对湿度低于75%时,孢子不萌发或萌发率较低;当相对湿度为85%–95%时,孢子萌发率显著升高。26℃时,烟粉虱2龄若虫的累计死亡率最高。随着相对湿度增大,病菌的致病力增强。当相对湿度为53%–95%时,烟粉虱2龄若虫累计死亡率从54.55%增加到88.89%。玫烟色棒束孢的致病力与孢子萌发率呈正相关,但温度比相对湿度对其影响更明显。结果表明,玫烟色棒束孢IF‐1106菌株侵染烟粉虱的最佳条件是温度26℃和相对湿度大于85%。     

Medium components and culture conditions affect the thermotolerance of aerial conidia of fungal biocontrol agent Beauveria bassiana

AIMS: To produce more thermotolerable conidia of Beauveria bassiana, a well-known fungal biocontrol agent, by optimizing the medium components and culture conditions. METHODS AND RESULTS: The conidia produced on media including 0.5-6% glucose, sucrose or starch as carbon source and 50-300-microg ml(-1) Cu2+, Zn2+, Mn2+ or Fe3+ as additive to Sabouraud dextrose medium at 15-30 degrees C, pH 4-8 or KCl-adjusted water availabilities were exposed to 30-min wet heat stress at 48 degrees C. The medium components for conidial production with greatly enhanced thermotolerance included 4% glucose as optimum or 1% starch as alternative for the carbon source and < or =50-microg ml(-1) Mn2+ for the metal additive. The culture conditions were optimized as 25 degrees C and pH 5-6. Conidial thermotolerance decreased remarkably when sucrose and Fe3+ or Cu2+ were used in the cultures, but altered slightly when 50-200-microg ml(-1) Zn2+ were included. CONCLUSIONS: The tolerance of B. bassiana conidia to the thermal stress was significantly affected by the medium composition and culture conditions under which the conidia were produced. SIGNIFICANCE AND IMPACT OF THE STUDY: Proper treatment of small grains as mass production substrates for more glucose release and supplement of glucose or 50-microg ml(-1) Mn2+ are possible means to enhancing conidial thermotolerance and field persistence for improved insect control.     

白僵菌对茶小绿叶蝉田间防效的关键影响因素研究

白僵菌孢子萌发率和萌发速度是影响其田间防效的关键因素。通过对球孢白僵菌YP9(CCTCC No:M209238)孢子萌发的关键影响因素进行研究,得到以下田间应用条件:营养物质为2%麸皮浸出液(自然pH);孢子悬浮液喷洒浓度为5×10~7孢子/mL;在气温为23℃~28℃时喷洒。经昆明和无锡两试验地田间防效验证,该白僵菌在上述条件下对茶小绿叶蝉的田间防效高达65%,与茶农常用化学农药——天王星防效相当。     

Relationship between thermotolerance and hydrophobin-like proteins in aerial conidia of Beauveria bassiana and Paecilomyces fumosoroseus as fungal biocontrol agents

AIMS: This study was to illustrate the relationship between the thermotolerance and the contents of hydrophobin-like or formic-acid-extractable (FAE) proteins in aerial conidia of Beauveria bassiana and Paecilomyces fumosoroseus produced on rice-based substrate. METHODS AND RESULTS: Survival indices of 11 isolates were separately assessed as a ratio of the viability of conidia after 3-150 min thermal stress at 48 degrees C over that of unstressed conidia and fitted well to a survival model (r(2) >/= 0.97). For a given isolate, the fitted model generated an LT(50), the time for 50% viability loss under the stress. The LT(50)s of six B. bassiana isolates (10.1-61.9 min) and five P. fumosoroseus isolates (2.8-6.2 min) were correlated (r(2) = 0.81) with FAE protein contents (6.9-23.4 microg mg(-1)). The survival indices of a fixed B. bassiana isolate after 45-min thermal stress at 48 degrees C were also correlated to the FAE protein contents from conidia produced on glucose-, sucrose-, or starch-based substrate (0.79 相似文献   

Variation in virulence of Beauveria bassiana isolates and its relatedness to some morphological characteristics

Ten Beauveria bassiana isolates were characterized using the following parameters: virulence, morphological measurements, germination in solid and liquid media, in vitro growth and physiological changes in liquid media. There were significant differences in mortality of second-instar larvae of the diamondback moth (DBM), Plutella xylostella (Lepidoptera: Plutellidae) and the Colorado potato beetle (CPB), Leptinotarsa decemlineata (Coleoptera, Chrysomelidae) according to isolate used (DBM; 14.51-53.37%, CPB; 25.05-74.07%). The most virulent isolates to DBM and CPB were KCF107 (soil origin) and KCF106 (Chilo suppressalis origin), respectively. Several morphological and physiological characteristics of isolates were evaluated for a possible link to virulence against both insect species. Special mean diameter (2.45-3.58 µm) and area of conidia (5.72-10.25 µm²) were significantly different among isolates. Time for 50% of conidia to germinate (GT₅₀) varied from 11 to 21 h depending on isolate. There was no correlation to virulence of size of conidia, GT_50, red pigment production and mycelial growth. Acidity of 7-day-old broth cultures of isolates was positively correlated with virulence. Iranian isolates KCF106 and KCF107 were the most promising and virulent isolates but field testing is needed to further evaluate their potential for use in DBM and CPB management.