不同寄主及地理来源的16株绿僵菌对桃蚜的毒力比较

害虫生防真菌绿僵菌的不同种及变种被广泛应用于害虫微生物防治,但罕见以蚜虫等同翅目刺吸式害虫作为靶标。从两种绿僵菌的4个变种中精选16个不同寄主及地理来源的菌株,用喷塔接种桃蚜(Myzus persicae)无翅成蚜并在25±1℃和12L∶12D条件下饲养观察,所获生物测定数据进行时间-剂量-死亡率模型模拟分析。结果显示,高接种剂量(~1000个孢子/mm2)下7d内死亡率达67%~100%的10个菌株均为金龟子绿僵菌Metarhiziumanisopliae(Ma)及其金龟子变种M.anisopliaevar.anisopliae(Maa);而高剂量处理下仅感染致死个别或少数蚜虫的菌株包括金龟子绿僵菌及其金龟子变种、大孢变种M.anisopliaevar.majus和蝗变种M.anisopliaevar.acridum以及黄绿绿僵菌小孢变种M.flavoviridevar.minus。杀蚜活性优异的2个菌株分别为Ma 456和Maa 3332,接种后第4天的LC50分别为113和260个孢子/mm2,第5天为32和43个孢子/mm2,第6天为17和26个孢子/mm2,第7天仅11.4和19.9个孢子/mm2。这两个菌株具有用于蚜虫微生物防治的良好开发潜力。     

六株虫生真菌的生物学特性及其对油茶象甲的致病力

为了筛选防治油茶象甲的虫生真菌菌株,本研究测定了不同绿僵菌(Ma09、Ma25、Ma05、Mac985)和白僵菌(Bb01、Bb02)菌株的生长速度、产孢量以及孢子萌发率,采用孢悬液浸渍法测定了上述菌株对油茶象甲成虫和幼虫的致病力。结果表明,Ma09、Ma05、Bb01、Bb02菌株的菌落直径分别为63.13、62.55、61.99、61.90 mm(25℃、15 d),生长速度显著快于其余菌株(P0.05);Ma25、Bb01、Ma09菌株的产孢量分别为3.04×10~8、2.88×10~8、2.85×10~8孢子/cm~2,显著高于其余菌株(P0.05);Bb01和Ma09菌株的孢子萌发率较高,分别达到97.80%和96.40%。不同菌株对油茶象甲成虫和幼虫致病力差异显著,其中Bb01和Ma09在1×10~7孢子/mL时,对油茶象甲成虫的LT_(50)分别为4.64 d和4.93 d,处理后15 d累计校正死亡率均达到100.00%,僵虫率分别为76.00%和78.67%;对幼虫的LT_(50)分别为2.15 d和3.48 d,10 d累计校正死亡率均达到100.00%,僵虫率分别为81.00%和86.31%。本研究筛选的Bb01和Ma09菌株具有生长快、产孢量大、孢子萌发率高、对油茶象甲致病力强等特性,在油茶象甲的生物防治中具有潜在的应用价值。     

环境因子对努利虫疠霉产孢和孢子萌发的影响

探讨努利虫疠霉Pandoranouryi(Remaudi埁re&Hennebert)H櫣mber在田间蚜虫种群中发生与流行的规律,研究外界环境因子对感菌虫尸产孢和孢子萌发的影响。结果表明,处于水琼脂培养平板上感染努利虫疠霉的桃蚜虫尸在8~25℃的温度范围内均能产生大量的初生分生孢子,在30℃下,仅弹射极少量孢子。8℃下,孢子弹射可以持续120h,当温度高于15℃,大部分的孢子会在48h内完成弹射。相对湿度小于95%,虫尸停止产孢。20℃下,光照条件不会影响虫尸弹射孢子的总量。在8℃和30℃时,24h后处于水琼脂培养平板上的孢子萌发率分别为45.23%和61.74%,显著低于15~25℃温度范围内的孢子萌发率(95%以上)。处于叶片上的真菌孢子,当相对湿度大于74%时出现萌发,但在盖玻片的表面,当湿度低于100%时未发现孢子萌发。     

用Real-Time PCR评价花生田间金龟子绿僵菌的存活能力

为了能够对金龟子绿僵菌在花生田间的存活能力进行精确的定量研究,通过比对真菌ITS序列,设计出针对金龟子绿僵菌的特异性引物,并建立金龟子绿僵菌Real-Time PCR反应体系。结果表明,标准品在拷贝数为8.49×103–8.49×109copies/μL之间,线性关系良好,线性方程为y=-3.257x+6.969,相关系数为R=0.9980,扩增效率E=102.8%,检出痕量为20个孢子/g土壤。以Real-Time PCR方法和平板稀释法,分别对花生根部施用金龟子绿僵菌后不同时期的土壤进行定量分析,结果表明两种方法检测的金龟子绿僵菌数量变化趋势相似,施用的金龟子绿僵菌在根围与根际都呈现先快速下降后缓慢回升的趋势,90d时金龟子绿僵菌DNA量和CFU值均下降至初始的10%以下,之后出现回升;根际的金龟子绿僵菌相对地下降较慢而回升较快,在120d时可恢复到初始的52.17%和38.65%,显著高于根围的数量。试验建立的Real-Time PCR体系可用于金龟子绿僵菌土壤宿存的定量检测,而且适用于低含量的检测。     

三株金龟子绿僵菌对红火蚁的致病力测定

本研究在实验室条件下测定了金龟子绿僵菌M09、CQMa117和CQMa128三个菌株对红火蚁的侵染能力。结果表明在1.0×10~8孢子/mL浓度下M09、CQMa128和CQMa117菌株处理后10 d红火蚁的工蚁累计死亡率分别为73.3%、14.6%和55.5%;M09菌株对工蚁的LC50为3.50×106孢子/mL,8.0×107孢子/mL浓度处理LT50为4.35 d。对M09菌株侵染红火蚁幼虫和蛹的能力测定结果显示,1.0×10~8孢子/mL浓度处理后10 d该菌株对幼虫和蛹的累计侵染率为98.9%和100%,对幼虫和蛹LC50分别为6.34×10~4孢子/mL、1.01×10~4孢子/mL。以上研究证实金龟子绿僵菌M09菌株对红火蚁具较强的致病力,致死速度较快,可作为该蚁生物防治的候选菌株。     

光照、温度和pH值对小黑桫椤孢子萌发及早期配子体发育的影响

小黑桫椤(Alsophila metteniana)被列为我国国家二级保护植物。为探讨其种群数量下降原因, 作者采用无菌培养方法和显微观察技术, 在实验条件下研究了光照强度、光质、温度和pH值对小黑桫椤孢子萌发及早期配子体发育的影响。结果表明: 孢子萌发和配子体发育的最适光照强度为40–70 µmol;m^–2;s^–1, 全黑暗时孢子不萌发;白光、红光、黄光和蓝光下的萌发率分别为68.78%、65.66%、63.74%和7.51%; 白光和蓝光下配子体可以形成正常的心形原叶体, 红光和黄光下配子体发育一直处于丝状体阶段。孢子萌发和配子体发育需要在酸性土壤进行(pH值在3.7–6.7); 孢子萌发的适宜温度为20–30℃。从孢子接种到心形原叶体形成需要55 d左右。根据上述结果, 我们认为光照强度是小黑桫椤孢子萌发的必需条件, 光质是限制小黑桫椤孢子繁殖的重要原因; 光质、温度和pH值等环境因子的作用和配子体发育时间等是导致小黑桫椤种群数量日趋减少的主要原因。     

苍耳柄锈菌三裂叶豚草专化型冬孢子萌发过程及萌发条件的研究

本文采用光学显微镜和扫描电子显微镜技术及荧光染色技术,对苍耳柄锈菌三裂叶豚草专化型Puccinia xanthii sp.ambrosiae-trifidae冬孢子的萌发过程和萌发条件进行了研究。结果表明:冬孢子堆成熟时突破寄主表皮外露;在寄主上冬孢子萌发时由上细胞顶部出现皱褶和帽状物,由帽状物下伸出担子。冬孢子的上细胞和下细胞都可萌发;冬孢子在水中于25℃2h即可萌发,24h后达到萌发高峰,萌发率为12%;温度20-25℃、相对湿度97%以上、pH5-7的条件利于冬孢子萌发,光照对冬孢子萌发没有影响,木糖和乳糖对冬孢子萌发有促进作用;无机氮源营养对冬孢子萌发有抑制作用。肌醇、烟酸、核黄素及三裂叶豚草叶汁对冬孢子萌发有促进作用。     

金龟子绿僵菌菌株生长环境变量的优化

金龟子绿僵菌(Metarhizium anisopliae var.anisopliae)是一种广谱的昆虫病原真菌,提高菌株的生长速率及产孢量具有重要的意义.分别对2株金龟子绿僵菌菌株MA4与MAlml的菌丝生长及次代产孢具有影响的培养温度、培养基初始pH、装液比、光照及微量元素等环境因素进行了测定.经过优化得到2菌株最佳培养条件分别为:MA4菌株在28℃、pH 7、装液比为75ml/250ml、加入微量元素Mn全光照培养时生长最好、次代产孢量最高;MAlml菌株在28℃、pH9、装液比为75ml/250ml、加入微量元素Cu、黑暗培养时生长最好、次代产孢量最高.     

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(英文)

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%。     

金龟子绿僵菌在森林土壤中的分布及对松墨天牛致病性测定

松墨天牛是重大森林植物检疫性病害——松材线虫病的主要媒介昆虫。本研究于2005年8月～2006年8月从福建、江西两省共110个林分样区（其中松林88个样区）采集土壤样品330份,采用选择性培养基分离土壤中的金龟子绿僵菌。从21个样区的26份土样中分离出的金龟子绿僵菌占采集样区的19.1%和样品的7.9%,成菌落数（CFU）500～72500CFU/g,表明金龟子绿僵菌在森林土壤中有较为广泛的分布。对分离到的9个产孢量高的菌株,采用浸渍法（1×10⁷孢子/mL）接种3～4龄健康松墨天牛幼虫,采用跗节接种法接种2～15日龄健康成虫,测定其致病力。结果表明,MaYTTR-03、MaYTTR-04菌株对松墨天牛幼虫和成虫均有较高致病力,表现出良好的生防潜力。     

新番茄粉孢菌Oidium neolycopersici生物学特性的研究

新番茄粉孢菌Oidium neolycopersici是近几年引起番茄白粉病的主要病原菌。在国内首次对该菌的生物学特性进行了研究。结果表明：新番茄粉孢菌分生孢子萌发的适宜温度范围为20－25℃,最适相对湿度范围为80%－100%,在水滴中不能萌发;该菌对光照条件和酸碱度的要求不严格,在pH3－12时其萌发率均能达到90%以上;在碳氮源利用方面,该菌分生孢子对各种碳源均能利用,以甘露糖和半乳糖效果最好;氮源以硝态氮（硝酸钾）为佳,铵态氮、有机态氮对其萌发有抑制作用;分生孢子萌发的致死温度为44℃ 10min。     

Effects of stabilizers on shelf-life of Epicoccum nigrum formulations and their relationship with biocontrol of postharvest brown rot by Monilinia of peaches

AIM: To find a formulation of Epicoccum nigrum conidia that maintains a high viability over time and which proves efficient to biocontrol peach rot caused by Monilinia spp. METHODS AND RESULTS: We tested the effect of stabilizers and desiccants on the shelf-life of Epicoccum nigrum conidia. Conidial samples were dried for 40 min at 40 degrees C in a fluidized bed-dryer to obtain moisture contents <15%. The toxicity of additives was tested by assaying production of conidia in fermentations and germinability of the produced conidia: 50% PEG300, 10%-5% KCl (stabilizers) and 95.24% Cl(2)Ca (desiccant) significantly (P = 0.05) reduced conidial germination. To enhance shelf-life of dried conidia, nontoxic stabilizers were added at the following different stages of the production-drying process: (i) to substrate contained in bags before production, (ii) to conidial centrifuge pellets obtained after production, before filtering and drying, (iii) to conidial centrifuge pellets obtained after production, before adding talc and drying, and (iv) to conidial centrifuge pellets obtained after production, before adding silica powder and drying. Conidial germinability was tested at 0, 180 and 365 days after storage at room temperature. Shelf-life of formulations retaining the highest viability were conidia produced with 1% KCl or 50% PEG 8000, conidia dried with 2.5% methylcellulose, and conidia dried with 1% KCl + silica powder. All these formulations improved the shelf-life of E. nigrum conidia and significantly reduced brown rot on peaches. CONCLUSIONS: Our results show that additives improve the shelf-life of E. nigrum and assist controlling brown rot on peaches. SIGNIFICANCE AND IMPACT OF THE STUDY: New improved formulations of a biocontrol agent have been obtained which will improve the control of Monilinia on peach.     

The effects of some storage conditions on viability of Lecanicillium lecanii conidia to whitefly (Homoptera: Trialeurodes vaporariorum)

A study on the survival of Lecanicillium lecanii conidia in storage at room temperature was carried out. Firstly, drying methods of conidia powder were compared. Vacuum-freeze drying (VFD) was more suitable for drying conidia as compared to vacuum drying (VD) at room temperature. Vacuum-freeze drying for 24-h resulted in a water content of 5.4%, and a viability, determined as germination of conidia in 2% glucose solution after16 h, was 90.3% and the infection in greenhouse whitefly, Trialeurodes vaporariorum was about 94.7% at a dose of 1×10⁸ conidia/mL. Secondly, the factors influencing viability of conidia stored at room temperature were evaluated in the laboratory. Temperature was the most critical factor influencing conidial storage stability, among the tested factors affecting survival of conidia stored at room temperature for 6 months. Both conidial germination and infection of hosts decreased with storage temperature increasing from 15 to 35°C, and at 35°C the survival of stored conidia for 6 months was near zero. The moisture content of the conidial powder was another major factor influencing viability of stored conidia at room temperature. Conidial powder dried to about 5% moisture content showed higher viability than non-dried conidial powder. For the carriers, clay and charcoal were more suitable for storage of L. lecanii conidia at room temperature. At a room temperature of 25°C, L. lecanii conidia which were dried to 5% water content and mixed with clay or charcoal could retain about 50% survival after 6 months' storage.     

Deposition and germination of conidia of the entomopathogen Entomophaga maimaiga infecting larvae of gypsy moth,Lymantria dispar

Germination of conidia of Entomophaga maimaiga, an important fungal pathogen of gypsy moth, Lymantria dispar, was investigated on water agar and larval cuticle at varying densities. Percent germination was positively associated with conidial density on water agar but not on larval cuticle. When conidia were showered onto water agar, the rate of germination was much slower than on the cuticle of L. dispar larvae. From the same conidial showers, the resulting conidial densities on water agar were much higher than those on larval cuticle in part because many conidia adhered to setae and did not reach the cuticle. A second factor influencing conidial densities on larval cuticle was the location conidia occurred on larvae. Few conidia were found on the flexible intersegmental membranes in comparison with the areas of more rigid cuticle, presumably because conidia were physically dislodged from intersegmental membranes when larvae moved. Conidia were also exposed to heightened CO(2) to evaluate whether this might influence germination. When conidia on water agar were exposed to heightened CO(2) levels, germinating conidia primarily formed germ tubes while most conidia exposed to ambient CO(2) rapidly formed secondary conidia.     

Effect of formulation on the virulence of Metarhizium anisopliae conidia against mosquito larvae

Metarhizium anisopliae conidia were formulated with three granular carriers and nine dust diluents and stored over an 8- to 12-month period at 4° or 20°C. The virulence of formulations, with the exception of two dust preparations, was reduced significantly compared to unformulated conidia against Culex pipiens pipiens larvae. The formulation components most detrimental to conidial virulence were corn cob granules, diatomaceous earth, and two Kaolinite diluents. This was exampled by a decline in virulence from ca. 100% for unformulated conidia to 36% or below for these formulations. LT₅₀ values also increased from 2.4–2.6 days for unformulated conidia to above 6 days. In contrast, a diluent derived from dried castor oil (Thixcin R) significantly enhanced conidial virulence at several doses above that of unformulated conidia against C. pipiens larvae. Enhancement occurred whether conidia were formulated prior to storage or stored separate from the diluent and mixed prior to application. The Thixcin R formulation was more effective against Anopheles stephensi larvae, but virulence was reduced against Aedes aegypti larvae. A bentonite formulation (Bentone-38) also maintained conidial virulence effectively, but Thixcin R was a superior diluent. It was shown that conidial virulence of formulations was not correlated with differences in conidial viability. The preparations that were applied dry by a surface method were more virulent than when an aqueous suspension containing a surfactant was used. The results demonstrate the need to assess efficacy of mycoinsecticidal formulations in a virulence bioassay prior to field testing.     

Ultra-violet radiation damage to Metarhizium flavoviride conidia and the protection given by vegetable and mineral oils and chemical sunscreens

Metarhizium flavouiride conidia formulated in oil or water were exposed to simulated solar radiation. Radiation below 320 nm killed conidia and caused delays in the germination of survivors; germination was greater after 48 h of incubation than after 24 h. UV exposure of conidia formulated in oil for 2 h reduced germination from 99% to 37.5% after incubation for 48 h. Exposure of conidia in water to UV for 1 h resulted in 4.7% germination after 24 h incubation compared with 36.5% for conidia formulated in oil. The addition of 1% oxybenzone resulted in 81.9% conidial germination after 3 h exposure and 48 h incubation compared with 28.1% in oil without the sunscreen.     

Selection of surfactant compounds to enhance the dispersion of Beauveria bassiana

This study aimed to select surfactant compounds to improve the dispersion of emulsifiable conidial suspensions of Beauveria bassiana for field application while maintaining high conidia viability. Surfactant compatibility was initially evaluated by colony growth in culture media amended with increasing doses of surfactants. In a second experiment, the conidia germination rate was evaluated after temporary exposure to the compounds under laboratory conditions. The effect of the hydrophilic–lipophilic balance (HLB) of the surfactants on conidia dispersion and viability was assessed using mixtures of the most compatible products and concentration initially selected. Unitol L/20 and Ultranex NP/100 did not show deleterious effect on colony growth on culture media, while Surfom 3409 strongly inhibited conidial germination. Unitol L/20, Ultranex NP/100 and Ultratan D did not interfere in the germination of conidia during the first 10 minutes of exposure. After 120 minutes, germination was greatly reduced at a 0.5% concentration, with the exception of Ultranex NP/100. The higher conidial dispersion and germination were obtained with HLB values between 8 and 12 and lower remaining clusters at HLB 10. Moreover, there was no significant effect of HLB values on insect mortality. Unitol L/20 and Ultranex NP/100 were the most compatible surfactants in concentrations lower than 0.5%. The HLB value selected for dispersion of B. bassiana on water based formulations was 10.     

Laboratory studies on the development of a conidial formulation of Esteya vermicola

The endoparasitic nematophagous fungus, Esteya vermicola, has potential as a biocontrol agent against pinewood nematode, Bursaphelenchus xylophilus. An E. vermicola conidial formulation was developed to improve conidial resistance to ultraviolet (UV), drought and heat stress. The effective concentration of each protective additive [UV protectant [fulvic acid (FA) and skim milk (SM)]; drought protectant (sorbitol) and heat protectant (calcium chloride)] was determined based on the germination rate of E. vermicola conidia after exposure to the different stressors. A combination of 0.2% FA and 4% SM, 5% sorbitol and 0.05% calcium chloride provided the most effective protection. In addition, the concentrations of spreader–sticker and antibiotic were also decided. The final formulation could be used to improve the resistance of E. vermicola conidia to multiple stressors and to increase nematode mortality compared with unformulated conidia.     

Effect of relative humidity,temperature and fungicide on germination of conidia of Cercospora canescens caused the Cercospora leaf spot disease in mungbean

The aim of the present investigation was to determine the impact of relative humidity (RH) and temperature on conidial germination, nuclear position and effect of important fungicides on growth and conidial germination of Cercospora canescens. Germination of conidia was observed at RH range 92–100% at 5–35°C. Significant interaction between temperature and RH indicated that higher humidity and high temperature promoted quick germination both in the presence and absence of free moisture. Although in absence of free moisture at 92–95% RH higher temperatures 25–35°C promoted quick evaporation of moisture and no conidial germination. Number of germtube was increased significantly at the optimum temperature 25–30°C and higher humidity (98–100%). But higher temperature 25–35°C with lower RH did not support the conidial germination. This finding is very important for disease forecasting using meteorological data. The spray of Carbendazim as contact fungicide may not be useful since it is not effective against the conidia of C. canescens. Triadimefon did not inhibit the conidia germination but completely inhibited mycelium development at 50 μg/ml. Propriconazole inhibited both conidia germination and mycelial development. Therefore, Propiconazole may be taken as protective as well as curative spray. In non-systemic fungicide, Copper oxychloride gave anticipated result by inhibiting both conidial germination and mycelium development. Therefore, copper oxychloride can be used as protectant fungicides for Cercospora leaf spot caused by C. canescens.