红火蚁活性真菌菌株的室内筛选研究

在实验室条件下采用浸渍法测定了15株白僵菌Beauveria spp.,15株绿僵菌Metarhizium spp.和4株淡紫紫孢菌Purpureocillium lilacinum (Thom)对红火蚁Solenopsis invicta Buren工蚁的活性,并测定分析了高活性菌株对红火蚁工蚁的毒力。结果表明,以1.0×10^8孢子mL的浓度处理红火蚁工蚁后第10天,白僵菌HHY-B、LCM1、ZYSYE-Y2、Bb034、ZG5、Bb720、Bb040、CP728和淡紫紫孢菌LYC1菌株的活性较强,红火蚁工蚁的校正死亡率为84.70%~98.79%,僵虫率为70.61%~90.38%。红火蚁工蚁在接种不同浓度的高活性菌株后,随着孢子浓度的升高,红火蚁的累积校正死亡率和僵虫率增大,致死中时(LT50)缩短,第10天白僵菌ZYSYE-Y2、CP728、HHY-B、LCM1、ZG5、Bb040、Bb720、Bb034和淡紫紫孢菌LCM1菌株对红火蚁工蚁的LC50分别为1.16×10^5、1.49×10^5、2.27×10^5、2.28×10^5、2.32×10^5、3.79×10^5、4.94×10^5、8.47×10^5和3.90×10^6孢子/mL。当孢子浓度为1.0×10^7孢子/mL时,白僵菌HHY-B、CP728、ZG5和Bb040菌株处理的最终校正死亡率分别为91.54%、94.60%、91.23%和94.65%,僵虫率为82.51%、91.74%、85.43%和80.60%,致死中时(LT50)为4.22 d、4.07 d、3.72 d和3.68 d。综合分析表明,白僵菌HHY-B、CP728、ZG5和Bb040菌株对红火蚁工蚁具有较强的活性,可作为红火蚁生物防治的候选菌株     

不同绿僵菌、白僵菌菌株对铜绿丽金龟幼虫的毒力生物测定

应用五株虫生真菌对铜绿丽金龟的幼虫的感染试验结果,将终浓度为1×107孢子/mL的孢悬液对铜绿丽金龟幼虫进行毒力测定。试验结果为:绿僵菌Ma09、Ma20,布氏白僵菌Bbr06、Bbr17,球孢白僵菌Bb202 15d的感染率分别为20.83%、14.58%、8.33%、95.83%、6.25%,其中布氏白僵菌Bbr17显示了较高的毒力,其LT50和LT95分别为10d、18d,该菌株对今后在花生蛴螬防治具有较大的应用价值。     

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。     

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。     

美国白蛾高毒力球孢白僵菌菌株筛选

【目的】美国白蛾Hyphantria cunea(Drury)是世界性检疫害虫,目前已经入侵到我国长江下游地区(从辽宁丹东至安徽芜湖)。作者在前期研究中,在芜湖地区成功获得了被球孢白僵菌Beauveria bassiana侵染的美国白蛾僵虫。本研究拟在前期已获取菌株的基础上,筛选生物学性状良好且对美国白蛾幼虫具有高致病力的球孢白僵菌菌株,以期为我国长江下游地区利用昆虫病原真菌防治美国白蛾奠定基础。【方法】对采集自美国白蛾僵虫的7株球孢白僵菌菌株(Bb01-1,Bb08-5,Bb08-6,Bb08-11,Bb22-2,Bb22-3和Bb22-5)进行生物学特性研究,以候选菌株菌落形态、生长速率、产孢量、萌发率和抗紫外能力等参数确定优良菌株;并将生物学性状良好的球孢白僵菌菌株配制成5个浓度(5×10~6,1×10~7,5×10~7,1×10~8和5×10~8孢子/m L)的孢悬液,采用浸渍法处理美国白蛾4龄幼虫,以幼虫死亡率、侵染率等指标评价菌株的致病力。【结果】供试菌株Bb22-2,Bb22-3和Bb22-5为粉状菌落,其菌落直径大,产孢早且孢子层厚;同时,这3个菌株在菌落生长速率和产孢量以及孢子萌发率、紫外照射处理后的萌发率也均显著优于其他4个菌株。毒力测定结果表明,这3个菌株对美国白蛾4龄幼虫表现出很强的致病力,各菌株处理11 d后美国白蛾幼虫校正死亡率为80%~90%。这3个菌株对美国白蛾幼虫的侵染率均高(侵染率53%~87%),并且呈现随浓度升高侵染率上升的趋势。其中,菌株Bb22-2侵染率均要高于其他2个菌株。【结论】本研究成功筛选获得了生物学性状良好且对美国白蛾高毒力的球孢白僵菌菌株。鉴于中国长江下游地区气候特点与美国白蛾生物学习性,可以预期采用昆虫病原真菌防治美国白蛾在该地区将具有很好的应用前景,但其实际应用价值尚需进一步的田间试验进行确认。     

Virulence of Metarhizium flavoviride 82 to different developmental stages of the brown planthopper, Nilaparvata lugens (Hemiptera:Delphacidae)

对水稻重要害虫褐飞虱Nilaparvata lugens目前市场上尚无一种理想的微生物杀虫剂.昆虫病原真菌具有从体壁侵入的能力因而对刺吸性害虫的防治具有优势.为此,本研究选用不同原寄主和来源地的3种昆虫病原真菌(金龟子绿僵菌Metarhizium anisopliae、黄绿绿僵菌Metarhizium flavoviride和球孢白僵菌Beauveria bassiana)的12个不同菌株,以1 100孢子/mm2孢子悬浮液进行室内毒力测定.结果表明:在参试的不同菌种12个菌株中,黄绿绿僵菌Mf82菌株对褐飞虱成虫致病力最高,10 d累计校正死亡率为83.5％,致死中时(LT50)为4.6d.其不同浓度孢子液对褐飞虱3个发育阶段有不同程度的致病力,毒力大小顺序为成虫＞高龄若虫＞低龄若虫.黄绿绿僵菌孢子液对各处理稻株褐飞虱产卵痕部位、卵粒均有侵染作用,10d侵染率分别为66.7％和51.2％,卵龄越低,侵染效果越好,卵龄为0.5d时侵染率最高.本研究表明黄绿绿僵菌Mf82菌株对褐飞虱成虫、若虫和卵均有较强的致病性,是一株极具应用潜力的生防真菌.     

白僵菌防治菜青虫的研究

通过应用6株白僵菌不同菌株对菜青虫幼虫进行室内和田间防治试验比较,结果表明,不同菌株间的毒力存在显著差异,以筛选自菜青虫僵虫尸上的Bpr3菌株毒力最强,用浓度为1×108 mL-1的孢子液对菜青虫幼虫进行室内和田间感染,室内幼虫校正死亡率达100%,LT50为2.9806 d,田间防治效果达96.6%,经大面积示范应用,取得了显著的防治效果.     

罹病棕尾别麻蝇体上白僵菌的分离、离体培养与毒力

为了有效地防治棕尾别麻蝇Boettcherisca peregrine,从罹病棕尾别麻蝇成虫上新分离出病原菌球孢白僵菌Beauveria bassiarift BBKMZW-1菌株,研究了该菌株菌落生长最适培养基为马铃薯葡萄糖琼胶培养基(PDA),最适温度范围是为20-30℃,最适pH值为7.该菌株对棕尾别麻蝇成虫的毒力测定表明:LC50值随侵染时间的延长呈下降趋势,由第4 d的1.41×1014孢子/mL降低到第9 d的4.00×105孢子/mL.不同浓度的LT50值随浓度增加而逐渐缩短,由1.07×104孢子/mL的13.70 d缩短到1.07×108孢子/mL的5.73 d.球孢白僵菌BBKMZW-1菌株是一种对棕尾别麻蝇成虫具有潜在防治作用的病原菌.     

黄绿绿僵菌Mf82悬乳剂对褐飞虱作用的时间-剂量-死亡率模型分析

为了寻找褐飞虱Nilaparvata lugens生物防治的新途径,用新分离出的黄绿绿僵菌Metarhiziumflavoviride(Mf82)菌株与实验室保存的黄绿绿僵菌、金龟子绿僵菌和球孢白僵菌3种菌种9个菌株作对比,测定了它们对褐飞虱成虫的毒力。结果表明:Mf82菌株对褐飞虱成虫的毒力最高,以1.0×108个孢子/mL的孢子液喷雾接种到褐飞虱成虫体表上,累积死亡率高达81.7%,LT50为4.6d,致病效果显著高于其他受测菌株。在此基础上研制了黄绿绿僵菌悬乳剂,并研究了其对褐飞虱的致病力。结果表明:随着黄绿绿僵菌浓度的增加,褐飞虱的累计死亡率增加,在浓度为1,048个孢子/mm2时,累计死亡率达到85.0%。利用时间-剂量-死亡率模型对数据进行处理,所建模型均顺利通过Hosmer-Lemeshow拟合异质性检验,表明模型拟合良好,并由模型估计出了该剂型对褐飞虱的致死剂量与致死时间。在接种后第7天和第9天,LC50值分别为2.1×103、9.9×102个孢子/mm2,LC90分别为7.8×104、3.7×104个孢子/mm2。黄绿绿僵菌悬乳剂对褐飞虱的致死时间与对数剂量相关,供试菌剂LT50值随着对数剂量的增加而递减,对数剂量由7.0增加到8.0时,LT50由8.9d降为5.7d。可见该黄绿绿僵菌悬乳剂对褐飞虱具有较强的毒力,在褐飞虱生物防治中具有广阔的应用前景。     

绿僵菌与3种杀虫剂混配对樟巢螟的协同作用

采用定量喷雾法测定了绿僵菌、白僵菌以及3种杀虫剂对樟巢螟Orthaga achatina低龄幼虫的室内毒力,并测定了金龟子绿僵菌Ma09与3种杀虫剂混配后的协同作用。结果显示：4种昆虫病原菌菌株均对樟巢螟2~3龄幼虫具有较好的致死效果,用绿僵菌和白僵菌孢悬液处理后,死亡高峰期在3~4 d;白僵菌GDLY9的防治效果最好,处理浓度为1×109孢子/mL下LT50和LT90分别为3.01、3.95 d,金龟子绿僵菌Ma09、3297效果相当,黄绿绿僵菌Mf985的LT50相对低一些,但是LT90优于金龟子绿僵菌Ma09和3297。金龟子绿僵菌Ma09与苯氧威、除虫脲和短稳杆菌混配后均表现出增效作用,协同指数分别为20.43、27.53和33.13。     

The Susceptibility of Boophilus annulatus (Ixodidae) Ticks to Entomopathogenic Fungi

The pathogenicity of five species of entomopathogenic fungi (Deuteromycetes, species: Beauveria bassiana, Metarhizium anisopliae, Metarhizium flavoviride, Paecilomyces fumosoroseus and Verticillium lecanii ) to the various developmental stages of Boophilus annulatus ticks was compared under laboratory conditions. M. anisopliae and B. bassiana strains were most virulent to engorged females and caused 85-100% mortality within 7-10 days post-inoculation (PI). The highest mortality of engorged females caused by other fungi reached only 25-60%. All tested fungi prevented or reduced the egg laying capability of the ticks several days before their death. Females surviving after treatment with the most virulent M. anisopliae strain (Ma-7) reached only 7-8% of their egg laying capacity as compared with the control. Other fungi caused a reduction of the weight of laid eggs by 35.4-80.8% as compared with untreated females. Only M. anisopliae and B. bassiana strains caused 70-98% mortality of the treated eggs. Unfed larvae of Boophilus annulatus were sensitive to M. anisopliae and M. flavoviride strains. The Ma-7 strain was most virulent to unfed larvae, with a mortality rate of 80.4% at a concentration of 1 ×107 spores ml -1 and 100% mortality at a concentration of 1 ×108 spores ml -1 .     

Selection of a Highly Virulent Fungal Isolate,Metarhizium anisopliae Ma TB 101, for Control of Taro Beetle,Papuana uninodis (Coleoptera: Scarabaeidae)

Fungal isolates (31 Metarhizium anisopliae var. anisopliae , five M. anisopliae var majus , three Beauveria bassiana and four B. brongniartii ) originating from a wide range of geographical locations, insect species and soil types were tested against Papuana uninodis (Coleoptera: Scarabaeidae). In a first test series, spores were applied topically to third-instar larvae and adults. The most effective strain against P. uninodis larvae and adults was Ma TB 101, a M. anisopliae isolate from adult P. woodlarkiana . For adults, strain Ma FI 384, a M. anisopliae from Popillia japonica , was almost equally effective. The 11 most effective isolates (nine M. anisopliae var. anisopliae , one B. brongniartii and one B. bassiana ) with LT values of less than 6 weeks in 50 adults and/or less than 4 weeks in larvae were tested for their efficacy against adults and larvae of P. uninodis by application of spores to soil (107 spores/g). Ma TB 101 was significantly more effective against both adults and larvae (LT ca. 10 days) than all other isolates (LT > 50 50 3 weeks). Two other M. anisopliae strains, Ma F 248 from soil and Ma FI 384 from P.japonica , were more effective than most isolates in adults. The latter three M. anisopliae isolates were tested in a concentration series against third-instar larvae and adults. Mortality was concentration related. Isolates Ma F 248 and Ma FI 384 did not achieve 50% mortality within the test period at concentrations lower than 107 spores/g of soil or feed. For concentrations of Ma TB 101 from 1 107 to 2 105 spores/g the LT ranged from 13 to 30 days in adults and 12 to 24 50 days in third-instar larvae. Accordingly, concentrations causing 50% mortality (LC ) for Ma 50 TB 101 were significantly lower than for the two other M. anisopliae isolates tested.     

Effect of Beauveria bassiana and Metarhizium anisopliae (Deuteromycetes) upon the coffee berry borer (Coleoptera: Scolytidae) under field conditions

The effect of three strains of the fungus Beauveria bassiana (Balsamo) Vuillemin and two strains of Metarhizium anisopliae (Metschnikoff) Sorokin upon the coffee berry borer, Hypothenemus hampei (Ferrari), was studied in three coffee farms at different altitudes (450-1,100 m above sea level) in Soconusco, Chiapas, Mexico. The maximum average percentage mycosis varied according to altitude. At 450 m asl (El Rincon) mycosis was 14.3% for B. bassiana and 6.3% for M. anisopliae; at 880 m asl (Santa Anita) mycosis was 40.6% for B. bassiana and 12.6% for M. anisopliae, and at 1,100 m asl (Alpujarras) 33.9% for B. bassiana and 22. 1% for M. anisopliae. The effect of fungal mycosis through time was not significant (P > 0.01) in any of the farms, but there was a significant difference between the strains of the fungus (P < 0.01); the best strains being Bb25 and Ma4 at the lower altitude, Bb26 and Ma4 for the middle altitude and Bb26 and Ma4 at the higher altitude. Environmental factors such as temperature, relative humidity and rain were not correlated with the percentage mycosis caused by B. bassiana and M. anisopliae. However, in the case of B. bassiana there was a significant, positive correlation (P < 0.01) between the infestation levels of the pest and the mycosis response of the entomopathogen.     

A new bioassay method reveals pathogenicity of Metarhizium anisopliae and Beauveria bassiana against early stages of Capnodis tenebrionis (Coleoptera; Buprestidae)

We used a newly developed bioassay method to demonstrate for the first time the potential of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae to be used for the control of neonate larvae of Capnodis tenebrionis, a major threat to stone-fruit orchards in several countries. Four B. bassiana and four M. anisopliae isolates were all pathogenic for neonate larvae of C. tenebrionis; mortality rates 10 days after inoculation by dipping in a suspension with 10(8)conidia/ml varied from 23.5% to 100%. Three of the four M. anisopliae isolates caused 100% mortality. In most cases, postmortem hyphal growth and sporulation of M. anisopliae or B. bassiana was observed covering the larvae in their galleries. The eight isolates were also evaluated for pathogenicity to C. tenebrionis eggs at the same dosage. Only two B. bassiana isolates caused significant egg hatching reduction of 84.5% and 94.5%. Our results indicate that M. anisopliae and B. bassiana may be considered as promising for a new approach to prevent larval infestations by C. tenebrionis.     

Susceptibility of different populations of ticks to entomopathogenic fungi

This study aimed to evaluate the in vitro effect of the entomopathogenic fungi Metarhizium anisopliae sensu lato (s.l.) and Beauveria bassiana sensu lato (s.l.) on two distinct populations of Rhipicephalus microplus, from two different experimental farms. Bioassays were performed with engorged females, eggs and larvae. Fungal infection was evaluated based on biological parameters of treated engorged females, percentage of hatch from treated eggs, and percentage of mortality and mean lethal time (LT90) of treated larvae. When the treatments were compared between the two populations, there were significant differences in the following parameters: pre-oviposition period, hatching period and egg production index. Moreover, the results showed that B. bassiana s.l., isolate Bb 986, was more virulent than M. anisopliae s.l., isolate 959, for engorged females, showing a control percentage of 49%. In the bioassay with eggs, the hatching percentage ranged from 3.1% to 49.5% in one population and from 3.4% to 42.7% in the other, with no significant difference between the two populations. In the bioassay with unfed larvae, the mortality percentage ranged from 91.8% to 98.7% in one population and from 71.0% to 94.0% in the other. The LT(90) varied from 19.52 to 27.51 days in one of the populations and 22.89 to 37.31 days in the other. These results suggest that populations of R. microplus show distinct variation in their susceptibility to B. bassiana s.l. and M. anisopliae s.l.     

Transfer of entomopathogenic fungi among formosan subterranean termites and subsequent mortality

Strains of the entomopathogens Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metschnikoff) Sorokin were screened for effectiveness against the Formosan subterranean termite, Coptotermes formosanus (Shiraki). Biological control methods are being considered to augment current integrated pest management schemes that involve chemical baits and nonrepellent termiticides. In this study Formosan subterranean termites selected from four colonies in the field were exposed in the laboratory to three strains of Beauveria bassiana and two strains of Metarhizium anisopliae . The exposed termites were then allowed to mingle with an equal number of unexposed termites from the same nest and mortality of the combined group was determined. One week after exposure, B. bassiana strains, ATCC 26037 and ATCC 90519, had caused significantly higher mortality than the two M. anisopliae strains, ESC 1 and II B and the remaining strain of B. bassiana , ATCC 90518. The LT₅₀ measurement for strains 26037 and 90519 was comparable at 1.8 and 2.0 days, respectively. Strains ESC 1 and 90518 caused 50% mortality at 6.5 and 11.0 days, respectively. Termites exposed to strain II B and the control group of termites did not reach 50% mortality during the 21 day experimental period. Exposure of termites to all fungal strains resulted in mortality in excess of that seen in the control groups of unexposed termites.     

Pathogenicity of entomopathogenic fungi to different developmental stages of Rhipicephalus sanguineus (Acari: Ixodidae).

The pathogenicity of 4 species of entomopathogenic fungi (Hyphomycetes species: Beauveria bassiana, Metarhizium anisopliae, Metarhizium flavoviride, and Paecilomyces fumosoroseus) to various developmental stages of Rhipicephalus sanguineus (Acari: Ixodidae) ticks was compared under laboratory conditions. The most virulent isolate, M. anisopliae-108, caused 92-96% mortality to unfed larvae and nymphs on day 7 postinfection (PI) and 100% mortality to unfed adults and engorged females on day 21 PI. The pathogenicity of M. anisopliae-108 to engorged larvae and nymphs was lower--82.6 and 60%, respectively. All tested B. bassiana, M. flavoviride, and P. fumosoroseus isolates were significantly less virulent (P < 0.05) or avirulent toward most life stages of R. sanguineus. The M. anisopliae and M. flavoviride isolates also prevented or reduced the ability of the ticks to lay eggs several days before their deaths. Female ticks infected by the fungi achieved only 11.3-60.8% of their egg-laying capacity compared with the controls.     

Attempts to control cabbage root flies Delia radicum L. and Delia floralis (Fall.) (Dipt., Anthomyiidae) with entomopathogenic fungi: laboratory and greenhouse tests

One laboratory and three greenhouse experiments were conducted to study the pathogenicity and efficacy of Finnish isolates of entomopathogenic hyphemycetous fungi against cabbage root flies. In Petri dishes, exposure to 1.5 × 10¹⁰ spores of Metarhizium anisopliae per dish caused 40–50% mortality of undifferentiated second- and third-stage larvae of Delia floralis , and 1 × 10⁷ spores per dish caused 40–50% mortality of Delia radicum larvae. In one greenhouse test, 1 × 10⁸ and 1 × 10⁹ spores of M. anisopliae and Paecilomyces fumosoroseus reduced the root damage of head cabbage by 20–70% compared with untreated controls, although this was not accompanied by significant reductions in the number of pupae. Only M. anisopliae consistently grew out of larvae and pupae of D. floralis during incubation that followed their recovery from the soil at the end of an experiment testing different formulations of M. anisopliae and Beauveria bassiana , but the frequency of the latent infections of the pest by M. anisopliae was not associated with reduced severity of damage to seedlings of head cabbage.     

Potential of some Metarhizium anisopliae isolates for control of Culex quinquefasciatus (Dipt., Culicidae)

Abstract: The potential of some isolates of Metarhizium anisopliae and Beauveria bassiana for use in the integrated management of Culex quinquefasciatus was evaluated. Metarhizium anisopliae isolate 1037 was selected in initial bioassays with a 50% lethal concentration (LC₅₀) of 1.97 × 10⁴ conidia/ml. This fungus caused higher mosquito larva mortality when applied as a conidial suspension to the surface of the water than as dry conidia, with a time to 50% lethal (LT₅₀) of 1 day compared with 3.6 days for the dry conidial application. However, results with UV- and heat-inactivated conidia did not confirm a possible role of fungal toxins in causing mortality when ingested by C. quinquefasciatus larvae. Metarhizium anisopliae did not remain active for as long as the bacterium Bacillus sphaericus isolate 2362. At 24 h after application, the fungus-induced mortality on mosquito larvae was significantly lower than the mortality caused by the bacterium. By the second day, almost no activity by the fungus was observed. Results suggest that M. anisopliae isolate 1037 has potential for use in mosquito control programmes.