日本菟丝子生防菌株的喷施条件与效果

在喷施菌制剂对日本菟丝子(Cuscuta japonica)进行生物防治时,为提高防效,孢子悬浮液喷前预浸3小时,并添加2%蔗糖,可使菌制剂孢子提早萌发,提高萌发率。即使在大气干旱的条件下,喷施后在3小时内孢子萌发率超过50%,这对菌剂孢子的侵入寄主(菟丝子)极为有利,高温高湿条件及重复多次喷洒是造成病害流行的重要因子。     

石菖蒲醇提物对番茄灰霉抑菌稳定性及防效研究

本文通过菌丝生长速率法研究石菖蒲醇提物对番茄灰霉的抑菌稳定性,采用凹玻片法和盆栽法研究其对灰霉孢子萌发的影响和温室防效,从而形成系统完整的数据基础,为石菖蒲杀菌剂的研发提供科学依据。结果表明,醇提物在不同pH和紫外照射下抑菌活性稳定,在高于78℃时抑菌活性下降,在蔗糖处理浓度高于0.08 g/mL时抑菌活性增强,在室温(22~28℃)和低温(4℃)下可长期储存;其对灰霉孢子萌发的EC50为0.0109 g/mL,EC90为0.0318 g/mL;温室防效的EC50为0.0221 g/mL,EC90为0.1533 g/mL。石菖蒲醇提物可抑制番茄灰霉菌丝生长和孢子萌发,抑菌稳定性和温室防效良好,可进一步研发为中药杀菌剂。     

球孢白僵菌两种施用方式对侵染桔小实蝇的影响及田间的防治效果

室内研究了球孢白僵菌Beauveria bassiana不同施用方式对侵染桔小实蝇Bactrocera dorsalis的影响,以及田间喷施球孢白僵菌防治桔小实蝇的效果。结果表明,两种诱集感染方式桔小实蝇成虫存活率分别为40%、41.7%,差异不显著;添加性引诱剂在白僵菌侵染桔小实蝇的过程中没有起到明显地促进感病的作用,只是缩短了病程,IIPC为0.41-0.44,说明两种诱集感染方式对桔实蝇成虫有一定的控制作用。三种施菌方式的桔小实蝇羽化率、成虫死亡率与对照差异均显著;以IIPC为评价指标,施菌方式对桔小实蝇各因子的影响大小顺序为：成虫存活率 >羽化率>化蛹率,其中以对成虫存活率的影响最大。应用化学防治和球孢白僵菌田间防治桔小实蝇的效果表明,施用2500倍40%毒死蜱效果最好,控制作用达到71.9%,其次是喷施2.0×108孢子/mL球孢白僵菌孢子悬浮液,防效达46.9%,施用2500倍4.5%高效氯氰菊酯防治效果达最差, 防效仅37.5%。     

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。     

农用硫酸链霉素防治芦笋茎枯病试验初报

通过室内孢子萌发抑制作用的测定和田间防治的试验,测定农用硫酸链霉素对芦笋茎枯病的防治作用。结果表明,农用硫酸链霉素200 ppm对芦笋茎枯病菌的孢子萌发和病斑形成与扩展均具有明显的抑制作用,田间防效达64%,略优于70%甲基托布津可湿性粉剂800倍液的防效,可作为一种防治芦笋茎枯病的生物制剂。     

不同温湿度下白僵菌对松毛虫的侵染致病效应

采用荧光显微技术和电镜扫描技术 ,对 33个温湿度组合条件下球孢白僵菌孢子在马尾松毛虫活体上的萌发、侵染进程和致病效果进行了观察与测定 ,分析了三者之间的关系 .揭示了孢子萌发速率与环境温度的关系 :V =(T - 4.2 5) /2 7.2 .其中 ,孢子萌发的起点温度为 4.2 5℃ ,有效积温为 2 7.2日度 .湿度是孢子萌发和侵染致病的主要制约因子 ,且温度和湿度具有互补与综合作用 .温度达 1 0℃以上 ,相对湿度达 95% ,侵染致病效应更为显著 .     

农用硫酸链霉素防治芦笋茎枯病试验初报

通过室内孢子萌发抑制作用的测定和田间防治的试验,测定农用硫酸链霉素对芦笋茎枯病的防治作用。结果表明,农用硫酸链霉素２００ｐｐｍ对芦笋茎枯病菌的孢子萌发和病斑形成与扩展均具有明显的抑制作用,田间防效达６４％,略优于７０％甲基托布津可湿性粉剂８００倍液的防效,可作为一种防治芦笋茎枯病的生物制剂。     

环境因子对球孢白僵菌侵染桔小实蝇致病力的影响

本文测定了温湿度、紫外线及常见杀虫剂等环境因子对球孢白僵菌B6菌株侵染桔小实蝇致病力的影响。结果表明,在19℃-31℃的温度范围内,B6菌株对桔小实蝇成虫、蛹与老熟幼虫均有致病力。在19℃-25℃条件下,桔小实蝇上述三种虫态的死亡率均随着温度的升高而增加,但在28℃和31℃温度条件下,桔小实蝇的死亡率则随着温度升高而降低。在1.8×10^8孢子/ml浓度条件下,成虫在25℃温度条件下的致死中时LT50为6.6d,在31℃的LT50为30.2d。在不同温度条件下,B6菌株对桔小实蝇成虫的致死中时差异较大。湿度越大,球孢白僵菌致病力越强,桔小实蝇的死亡速率越快。球孢白僵菌孢子萌发率随着紫外线照射时间增加而逐步降低。测定了7种常见的杀虫剂对球孢白僵菌菌落的生长影响,其中25%高效氯氟氰菊酯、2.0%阿维菌素、40%辛硫磷和10%除尽对菌落生长有促进作用。     

粉红螺旋聚孢霉67‐1厚垣孢子生物学特性的研究

粉红螺旋聚孢霉(粉红粘帚霉)是多种植物病原真菌的重寄生菌,具有巨大的生防潜力。以实验室前期分离筛选的高效粉红螺旋聚孢霉67‐1菌株为材料,研究其厚垣孢子和分生孢子的生物学特性。结果表明,以厚垣孢子为接种体,其生长速度和菌丝鲜重与以分生孢子为接种体形成的菌落没有明显差异,但产孢水平提高6倍以上。粉红螺旋聚孢霉对高温和干燥的耐受程度较差,尤其分生孢子更为敏感。当加工温度范围在40–50℃时,厚垣孢子和分生孢子的活性存在显著差异(P<0.05)。菌株67‐1的厚垣孢子耐紫外线能力明显优于分生孢子。在无保护剂条件下,紫外灯照射60s,厚垣孢子活性保持在62.6%,是分生孢子活性1倍以上。土壤抑菌作用对真菌厚垣孢子的影响相对较小,24h孢子萌发率为47.2%,而分生孢子仅为4.6%。除了百菌清,菌株67‐1厚垣孢子对多种化学农药有较强的耐受力,在药剂田间使用浓度下,厚垣孢子萌发率均大于70%,其中对15%恶霉灵、1.8%阿维菌素和3%克百威的耐受力显著高于分生孢子(P<0.05),而对井冈霉素和两种杀虫剂没有显著提高。对菌株67‐1的寄生性测定结果表明,2种孢子对核盘菌菌核的寄生能力没有差异,菌核感染达到4级的占58.9%–60.0%。106/m L浓度厚垣孢子与分生孢子对黄瓜枯萎病的防效均达到65%以上,二者没有显著差异。     

不同生物药剂对稻水象甲的毒力、拒食活性及防效分析

【目的】防治稻水象甲的现有常规化学药剂的大量施用所引发的环境问题越发突出,而其生物药剂单一,加之天敌匮乏,故开展符合绿色生产标准的低毒、低残留、无污染生物药剂的筛选已成为稻水象甲防治工作所面临的首要任务。【方法】以越冬代稻水象甲成虫为试虫,采用室内点滴、田间喷雾等方法,对5种符合绿色生产标准药剂的毒力、拒食活性、防效及药后取食量等指标进行了测定和综合评价。【结果】室内生测方面:72 h后对处理稻水象甲成虫的室内毒力显著;此外,对试验前后的取食斑面积进行比较,发现药后各药剂拒食活性差异显著,均表现出较好的拒食作用。田间药效方面:除0.6%苦参碱AS外,其他药剂对稻水象甲均表现出较好的防效,100亿个孢子·m L~(-1)白僵菌OD、1.5%除虫菊素AS、7.5%鱼藤酮EC和6%乙基多杀菌素SC这4种药剂15 d的田间防效为51.24%~82.55%;3 d后各处理组田间新增取食斑数、取食长度均显著低于对照组。【结论】在绿色水稻生产中,100亿个孢子·m L~(-1)白僵菌OD、1.5%除虫菊素AS、7.5%鱼藤酮EC和6%乙基多杀菌素SC均具有较好的推广价值,可用于稻水象甲的防治。     

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。     

The potential biocontrol agent Pseudomonas antimicrobica inhibits germination of conidia and outgrowth of Botrytis cinerea

AIMS: Antifungal metabolites of Pseudomonas antimicrobica have previously been shown to inhibit conidial germination of the grey mould pathogen Botrytis cinerea. In this study, metabolites of the bacterium have been tested at different stages of Botrytis germination to determine their effects on germ tube production and extension. METHODS AND RESULTS: Metabolites were added to conidia that had been pre-incubated for either 120 or 255 min. Pseudomonas antimicrobica inhibited B. cinerea conidial germination and caused a significant reduction in germ tube extension, irrespective of the stage of germination. Abnormal germination and a reduction in the frequency of lateral branching of the germ tubes in the presence of the metabolites were also reported, suggesting interference with normal hyphal development. CONCLUSION: The bacterium can inhibit germination of conidia and extension of germ tubes at different stages of Botrytis development. SIGNIFICANCE AND IMPACT OF THE STUDY: The antagonistic activity of the bacterium has promising implications for its use as a biocontrol agent.     

Enhanced conidial thermotolerance and field efficacy of Beauveria bassiana by saccharides

Insecticide efficacy of Beauveria bassiana conidia was improved by optimizing the concentrations of conidial heat-protective saccharides (glucose, sucrose, maltose, trehalose, α-lactose, and mannitol) using response surface methodology. Two field trials in tea gardens were carried out to control leafhopper (Empoasca vitis) by spraying B. bassiana conidia together with the optimized saccharides (0.26 g glucose, 0.28 g lactose, 0.24 g mannitol per ml). In the field studies, B. bassiana conidia were applied to control Empoasca vitis with and without saccharides and compared with bifenthrin, a pyrethroid insecticide. With the optimal concentrations of saccharides, the conidial germination rate reached 72 % and the control efficacy of the saccharides group (65.7 %) was equal to the bifenthrin group (69.4 %), which improved by about 55 %.     

Microcycle conidiation in Penicillium urticae: an ultrastructural investigation of conidial germination and outgrowth.

A cultivation system has been developed for Penicillium urticae (NRRL 2159A) which yields 'microcycle' conidiation in submerged culture. Spherical growth of conidia was initiated by incubation at 37 degrees C in a growth-favoring medium. Transfer of these enlarged conidia to a nitrogen-poor medium at 35 degrees C resulted in synchronous germination and limited outgrowth followed by roughly synchronous conidiogenesis. An ultrastructural study of the germination stage indicated nuclear migration into the emerging germ tube whose new cell wall was an extension of the parent conidium's innermost cell wall layer. Septal formation at the neck of the germ tube followed. The septal pore was filled with particulate material and the septal membranes possessed unusual linear elements in their median hydrophobic zones. The germ tube, which possessed a smooth-surfaced plasma membrane, continued to elongate with periodic septum formation. The parent conidium and later the proximal germ tube showed progressive vacuolation and the cytoplasm became largely occupied by electron-translucent material. In older cells the septal pore was blocked by Woronin bodies. Compared with normal conidial germination this microcycle' germination is far more synchronous and the resultant germling is morphologically simpler. In ultrastructural terms, however, germination appears to be identical with that obtained at 28 degrees C.     

Factors influencing pathogenicity of Fusarium tumidum on gorse (Ulex europaeus)

Factors promoting pathogenicity of Fusarium tumidum on gorse (Ulex europaeus) were determined to develop a novel strategy for delivering this potential mycoherbicide using insects as vectors of inoculum. Fusarium tumidum sprayed as a suspension of 1×10⁶ conidia mL^?1 on at least 50% of a gorse plant reduced shoot dry weight by 45% (P<0.05). A minimum of 910 viable conidia were required to cause a lesion on leaves. The leaves and flowers were more susceptible to infection than stems, spines and pods. Generally, wounding of gorse leaves and stem increased F. tumidum infection, most likely through releasing nutrients that enhanced conidial germination and hyphal growth. We showed in a separate experiment that conidial germination (93%) and germ tube length (407 µm) were greater when incubated in 0.2% gorse extract solution for 24 h than in water (62% germination, germ tube length 42 µm). Inoculation of gorse with a F. tumidum conidial suspension supplemented with 0.2% gorse extract resulted in a shoot dry weight reduction (P=0.012) equivalent to that of plants that were wounded and inoculated. It is concluded that wounding of older tissues (which mimics insect damage) is required to facilitate F. tumidum infection of mature gorse plants.     

Evaluation of Beauveria bassiana (Hyphomycetes) isolates as potential agents for control of Dendroctonus valens

Abstract The red turpentine beetle (RTB), Dendroctonus valens LeConte, as a destructive invasive pest, has become one of the most economically important forest pest in China. Effective control measures are desperately needed. Entomopathogenic fungi, such as Beauveria bassiana, have shown great potential for the management of some bark beetle species. In this study, 12 isolates of B. bassiana from bark beetle were examined for biological characteristics and virulence, to assess their potential as biocontrol agents for RTB. There were significant differences (at P= 0.05) in colony growth rate, conidial yield, conidial germination, tolerance to UV light and extracellular proteases activity among the tested B. bassiana isolates. Isolates, including Bb1801, Bb1906, Bb789 and Bb773, exhibited the best characteristics, because they have faster hyphal growth rate, higher spore production and faster spore germination, higher UV tolerance and protease (Pr1) production. The results of a pathogenicity test of B. bassiana on RTB larvae showed that most isolates of B. bassiana have demonstrated high efficacy and the highest virulent isolate was Bb1801, which killed 100% of the treated insects and had a median lethal time (LT₅₀) of 4.60 days at a concentration of 1×10⁷ conidia/mL. Therefore, isolate Bb1801 has a great potential for sustainable control of RTB in the forest. The correlation between biological characteristics and virulence of the fungal isolates is discussed and the possibility of combination of entomopathogenic fungi with semiochemicals, as one of the promising strategy for RTB control, is considered.     

白僵菌孢子悬乳剂田间控蚜效果与常温货架寿命测定

将高纯度球孢白僵菌分生孢子粉兑入一种含悬浮稳定剂及紫外保护剂的矿物油中,配成含孢量为100亿/ml的孢子悬乳剂(纯菌剂),再添加3%的吡虫啉10%可湿剂而得菌药混配剂,于2003年5月在浙江余姚进行了防治蔬菜蚜虫的田间药效试验和农户示范。在两块夏熟白菜地中,纯菌剂和菌药混配剂均用1000倍稀释液(107个孢子/ml)常规喷雾2次,并设清水对照和低用量吡虫啉对照。结果表明,混配剂在两块菜地的相对防效分别达94.5%和86.2%,纯菌剂分别达86.4%和73.1%,而低用量吡虫啉处理的最高防效仅为50%左右。在入夏豇豆、甘蓝、白菜、萝卜及梨树上用菌药混配剂1000倍稀释液对多种蚜虫进行的农户示范试验中,一次性喷雾后5~10d内对菜蚜防效为48%~68%,尤对梨二叉蚜的防效高达99%。在对含有和不含添加剂的两配方孢子悬乳剂进行的常温避光18个月贮存中,活孢率在头7个月均稳定在95%左右且未受配方影响,第10个月活孢率仍有75%左右。而后,含添加剂配方的活孢率下降加速,至16个月时基本失活;而不含添加剂配方的活孢率在第16、17及18个月时还有57%、51%和42%。     

Bioassay methods for the detection of antifungal activity by Pseudomonas antimicrobica against the grey mould pathogen Botrytis cinerea

Antagonism against the grey mould pathogen Botrytis cinerea by Pseudomonas antimicrobica was demonstrated in vitro and in vivo. Cell-free filtrates showed activity against B. cinerea growing on Potato Dextrose Agar (PDA) in a media-dependent manner with the most distinct antagonism being produced in Czapek Dox Broth (CDB). Cell-free filtrates of CDB-grown cultures also significantly reduced conidial germination of B. cinerea. An assays based on the inhibition of conidial germination was compared with two assays measuring the antagonism of mycelial growth on PDA. The conidial germination bioassay was more sensitive in the detection of this antifungal activity than the Petri dish bioassay while a bioassay using Microdetection plates did not detect antagonism due to the small loading capacity of the latter. The conidial germination bioassay was modified for detection of antibiosis on the surface of strawberry leaves. Significant reductions in percentage conidial germination were recorded on the surface of leaves of both micropropagated and glasshouse grown strawberry plants when the antifungal compounds of Ps. antimicrobica were applied to the leaf tissue with the conidia. In addition, antifungal compounds were also detectable when conidia were applied to leaf tissue which had previously been sprayed with cells of Ps. antimicrobica. These tests indicate that Ps. antimicrobica would be a suitable biocontrol agent for the control of B. cinerea.     

Laboratory and field evaluation of Metarhizium anisopliae var. anisopliae for controlling subterranean termites

The efficacy of the Metarhizium anisopliae strain ARSEF 6911 was determined in the laboratory and field against two sugarcane pests, Microtermes obesi Holmgren and Odontotermes obesus Rambur (Termitidae: Isoptera). The susceptibility of both termite species to different conidial suspensions (1 × 10(10), 1 × 10(8), 1 × 10(6) and 1 × 10(4) conidia/ml) was determined in laboratory. All conidial suspensions were able to induce mortality. Termite mortality caused by the fungal suspensions was dose dependent. There were no significant differences in the LT50 values between species. Field evaluation of M. anisopliae alone or in combination with diesel oil and thiamethoxam was carried out in two growing seasons (autumn 2005 and spring 2006) at two sites located in Punjab, Pakistan. Dipping the sugarcane setts in these suspensions was tried to determine their effects on germination and percentage of bud damage to sugarcane setts. All treatments significantly reduced termite infestation compared to the untreated control. The combined treatment of M. anisopliae and diesel oil significantly reduced insect damage by attaining higher germination > 55% and lower bud damage < 5.50% at both sites in both seasons. The results suggest that the application of M. anisopliae and diesel oil in combination might be a useful treatment option for the management of termites in sugarcane.     

Formation and Germination of Septoria tritici Secondary Conidia as Affected by Environmental Factors

The effects of medium, isolate, temperature, light and pH on the formation and germination of Septoria tritici secondary conidia were tested. Of the six media tested, the malt–yeast extract agar was the best and generated 1.82 × 10⁹ conidia/plate. The ten isolates tested showed different ability of conidia production. Darkness significantly reduced conidial formation and enhanced the transition of intermediates. The conidial germination and germ tube growth was strongly inhibited at 30°C. The suitable pH for conidial budding in malt–yeast broth (MYB) was between 5 and 9. At pH 2, 10 and 11, almost no new conidia were formed. The number of conidia reached 1.27 × 10⁸ conidia/ml after 7 days in MYB, significantly more than that in potato dextrose broth, wheat leaf extract and H₂O.