Influence of plant resistance at the third trophic level: interactions between parasitoids and entomopathogenic fungi of cereal aphids

Host-plant resistance can affect herbivorous insects and their natural enemies such as parasitoids and entomopathogenic fungi. This tritrophic effect acts on interspecific interactions between the two groups of natural enemies distantly related in phylogenetic terms. The intra- and extra-host aspects of the interaction between the cereal aphid parasitoid Aphidius rhopalosiphi and the entomopathogenic fungus Erynia neoaphidis developing on the grain aphid, Sitobion avenae, on resistant and susceptible wheat (Triticum aestivum) cultivars, were studied. The competitive outcome of the intra-host interaction depended on the timing of parasitoid oviposition and fungal infection and was affected by wheat resistance. In particular, survival of the parasitoid was lower on the resistant wheat cultivar than the susceptible wheat cultivar, when the competitive outcome of the interaction was favourable for either parasitoid or fungal development. Before and after this period the influence of plant resistance was not significant. Furthermore, the extra-host interaction was not affected by the wheat cultivar, although an increase in fungal infection of S. avenae was observed when parasitoids foraged in the experimental arena with sporulating aphid cadavers compared with foraging in the absence of sporulating cadavers. Our results showed that the host plant may affect interspecific interactions between parasitoids and fungi and that these interactions depended on the timing of parasitoid oviposition and fungal infection. Received: 16 March 1998 / Accepted: 24 August 1998     

Chemical stimulation of the virulence of entomopathogenic fungi

The effect of magnesium and manganese ions on the virulence of Beauveria bassiana, Metarhizium anisopliae, Paecilomyces farinosus, and Paecilomyces fumosoroseus entomopathogenic fungi was tested. The virulence of entomopathogenic fungi to Sitona lineatus (L) weevil, pupae and larvae, was increased by tested metal ions. Mg ions increased the virulence of B. bassiana against S. linatus (L) weevils 100%. The fungi exhibited various sensitivity to these metal ions.     

Use of quantitative PCR to understand within-host competition between two entomopathogenic fungi

Interactions between the entomopathogenic fungi Zoophthora radicans and Pandora blunckii infecting larvae of Plutella xylostella were investigated. This is the first report to quantify within-host growth of one fungus in the presence of another competing for the same host resource using quantitative PCR (qPCR) at regular time points during the infection process. In larvae inoculated only with Z. radicans, there was a cumulative increase in the quantity of Z. radicans DNA throughout the time course of infection. However, in dual-inoculated larvae, there was an initial accelerated rate of growth of Z. radicans compared to when it was inoculated alone, but by the time of host death it had been effectively excluded by P. blunckii. The implications of these results for co-existence of these fungal pathogens in the field are discussed.     

Distribution,occurrence and natural invertebrate hosts of indigenous entomopathogenic fungi of Central India

We report here that, during periodical surveys of insects inhabiting diverse habitats for the collection of entomopathogenic fungi; a large number of isolates were recovered belonging to seven species, from various regions of Madhya Pradesh and Chhattisgarh forest areas and agricultural fields. The most common entomopathogenic fungi such as Beauveria bassiana, Nomuraea rileyi, Paecilomyces farinosus and Paecilomyces fumosoroseus were found to infect various insect hosts species naturally viz. Hyblaea puera, Eutectona machaeralis, Diachrysia orichalcea, Spodoptera litura, and few new insect hosts of these fungal pathogens among Indian insect population were collected for the first time from Central India, such as beetles of Agrilus species, hairy caterpillars of Lymantria species. The isolation, identification, maintenance and pathogenicity assay of these isolates was performed prior to deposition in culture collection center.     

10年肠道真菌培养结果分析

目的探讨浙江大学医学院第一附属医院住院患者肠道真菌分离状况与趋势变化。方法回顾分析2000年1月1日至2009年12月30日10年间在该院送检的住院患者粪便标本的真菌分离率、真菌种类和年度变迁变化特点。结果 10年间从送检全部疑似肠道真菌感染的2 344份粪便标本中共检到真菌1 456株,归属5属19种,每年的检出率在55.7%67.0%。分离菌株中以白色念珠菌（74.0%）占首位,其次是光滑念珠菌（11.4%）,再次是热带念珠菌（7.1%）,光滑念珠菌和热带念珠菌分离率分别从2000年的4.9%、6.6%上升到2009年的16.8%、10.6%,呈上升趋势。结论该院10年间肠道真菌的分离率较高,白色念珠菌的分离率一直位于首位,非白色念珠菌的分离率处于上升趋势。     

Occurrence of fungi in combs of fungus-growing termites (Isoptera: Termitidae, Macrotermitinae)

Fungus-growing termites cultivate their mutualistic basidiomycete Termitomyces species on a substrate called a fungal comb. Here, the Suicide Polymerase Endonuclease Restriction (SuPER) method was adapted for the first time to a fungal study to determine the entire fungal community of fungal combs and to test whether fungi other than the symbiotic cultivar interact with termite hosts. Our molecular analyses show that although active combs are dominated by Termitomyces fungi isolated with direct Polymerase Endonuclease Restriction - Denaturing Gradient Gel Electrophoresis (PCR-DGGE), they can also harbor some filamentous fungi and yeasts only revealed by SuPER PCR-DGGE. This is the first molecular evidence of the presence of non-Termitomyces species in active combs. However, because there is no evidence for a species-specific relationship between these fungi and termites, they are mere transient guests with no specialization in the symbiosis. It is however surprising to notice that termite-associated Xylaria strains were not isolated from active combs even though they are frequently retrieved when nests are abandoned by termites. This finding highlights the implication of fungus-growing termites in the regulation of fungi occurring within the combs and also suggests that they might not have any particular evolutionary-based association with Xylaria species.     

Efficiency of Nimbecidine and certain entomopathogenic fungi formulations against bean aphids,Aphis craccivora in broad bean field

This study investigates the effect of certain entomopathogenic fungi formulations (Beauveria bassiana, Verticillium lecanii, Metarhizium anisopliae and Paecilomyces fumosoroseus) compared with a botanical insecticide, Nimbecidine against Aphis craccivora in broad bean field. Bio-Catch (Verticillium lecanii) was the most effective insecticide to achieve 73.3% reduction followed by Nimbecidine (67.7%), Bio-Magic (61.6%), Priority (50.3%) and the least effective was Bio- Power (Beauveria bassiana) which caused 45.5% reduction in individual aphid populations after two sprayings at 15 days interval between the first and the second sprayings. Bio-Catch and Nimbecidine had promise compounds in controlling Aphis craccivora in faba bean fields.     

Soil management effects on entomopathogenic fungi during the transition to organic agriculture in a feed grain rotation

The growing demand for organic products creates opportunities for farmers. Information on the consequences of management practices can help farmers transition to organic and take advantage of these prospects. We examined the interaction between soil disturbance and initial cover crop on naturally occurring entomopathogenic fungi (EPF) during the 3-year transition to organic production in a feed grain rotation in central Pennsylvania. Our experiment included four systems comprised of a factorial combination of two levels of primary tillage (full vs. reduced) and two types of initial cover crop (timothy/clover vs. rye/vetch). The cropping sequence consisted of an initial cover crop, followed by soybean, and finally, maize. The entire experiment was replicated in time, with the initiation lagged by 1 year. We detected four species of EPF (Metarhizium anisopliae, Beauveria bassiana, Isaria fumosorosea, and Isaria farinosa) by bioassay of soil samples collected four times during each field season. The latter three species were detected infrequently; therefore, we focused statistical analysis on M. anisopliae. Detection of M. anisopliae varied across sampling date, year in crop sequence, and experimental start, with no consistent trend across the 3-year transition period. M. anisopliae was isolated more frequently in the systems initiated with timothy/clover cover crops and utilizing full tillage; however, we only observed a tillage effect in one temporal replicate. M. anisopliae detection was negatively associated with soil moisture, organic matter, and zinc, sulfur, and copper concentrations in the soil. This study helps to inform farmers about management effects on soil function, specifically conservation biological control.     

Occurrence of invertebrate-pathogenic fungi in a Cerrado ecosystem in Central Brazil

Occurrence of invertebrate-pathogenic fungi in a protected area of the Cerrado region of Brazil is reported. Fungi were baited with triatomines, ticks, aquatic snails or mosquito larvae from substrates or collected from infected insects. The findings underscore the importance to preserve these fungi and to investigate their potential for vector control.     

Dissemination of entomopathogenic fungi using Busseola fusca male as vector

The stem borer, Busseola fusca (Fuller), is an important pest of maize Zea mays L. and sorghum Sorghum bicolor (L.) in eastern and southern Africa. To control this pest, biological control methods including the use of entomopathogenic fungi are being considered. The pathogenicity of one isolate of Metarhizium anisopliae (Metsch.) Sorok. and one isolate of Beauveria bassiana Bals. (Vuill.) were first tested on different developmental stages of B. fusca including eggs, neonate, 2nd and 3rd-instar larvae. Both fungal isolates were pathogenic to all the stages tested. However, differences in mortality were observed among larvae that hatched from treated egg masses. Experiments were conducted thereafter to test whether B. fusca males could serve as a vector for fungal conidia to contaminate B. fusca females and subsequently eggs and larvae. Results demonstrated that B. fusca males successfully transferred inoculum to females during copulation, which in turn transmitted it to the eggs they laid on maize plants, resulting in the decrease of leaf damages.     

A PCR-based tool for the cultivation-independent monitoring of Pandora neoaphidis

Pandora neoaphidis is one of the most important fungal pathogens of aphids and has a great potential for use in biocontrol. Little is known on how this fungus persists in an area and in particular on its overwintering strategies. It is hypothesized that natural areas play an important role for survival and that soil may serve as a source of inoculum for new aphid populations in spring. To test these hypotheses, a cultivation-independent PCR-based diagnostic tool was developed, that allows the detection of P. neoaphidis in the environment. Two P. neoaphidis specific PCR primer pairs were designed, targeting sequences in the ribosomal RNA gene cluster. Specificity of both primer pairs was demonstrated with P. neoaphidis and non-target close entomophthoralean relatives. Moreover, single amplicons of expected sizes were obtained with both primer pairs from various environmental sample types, including aphid cadavers, plant material, and soil. The PCR-based diagnostic tool was applied to investigate the persistence of P. neoaphidis in soil samples obtained in 2004/2005 from a nettle field harboring infected aphids in fall 2004. P. neoaphidis was detected in every sample collected in November 2004 and March 2005, suggesting an overwintering stage of P. neoaphidis in top soil layers. The developed cultivation-independent PCR-based tool will be valuable for further investigation of the ecology of P. neoaphidis and for the development and future implementation of management strategies against aphids involving conservation biocontrol.     

Characterization in biological traits of entomopathogenic nematodes isolated from North China

In a series of bioassays, thirty-one isolates that were collected from diverse locations in northern China and the laboratory kept isolate Steinernema carpocapsae All, were compared in order to select superior isolates for biological control of Bradysia odoriphaga. Virulence of the isolates against B. odoriphaga was significantly different among nematode isolates. Tolerance of infective juveniles (IJs) to heat, cold, and desiccation differed significantly among and within species. Strains from S. carpocapsae, S. ceratophorum, S. longicaudum, Heterorhabditis indica, and H. bacteriophora were more heat tolerant than strains from S. feltiae, S. hebeiense, S. monticolum, and H. megidis. Heterorhabditis megidis, H. bacteriophora, and S. carpocapsae showed better cold tolerance than the other species. High desiccation tolerance was recorded for S. carpocapsae, S. hebeiense, and S. ceratophorum. The infectivity of IJ of these species against Galleria mellonella larvae was not significantly different between the treated and non-treated IJ after the nematodes had been exposed to 40 °C for 2 h, −5 °C for 8 h or 25% glycerin for 72 h. Nematode survival was significantly affected by exposure time and IJ concentration when exposed to 40 °C or −5 °C. All nematode isolates lost their infectivity against G. mellonella after exposure to −5 °C for 16 h, except for H. megidis LFS10, which had a low infectivity of 3.3%. A hierarchical classification analysis classified the isolates in four main clusters. The fourth cluster, composed of 13 isolates, grouped the isolates that scored well for most traits.     

Laboratory investigations on the potential of entomopathogenic fungi for biocontrol of Helicoverpa armigera (Lepidoptera: Noctuidae) larvae and pupae

The susceptibility of third instar Helicoverpa armigera to seven strains of three entomopathogenic fungal species, i.e. Metarhizium anisopliae, Beauveria bassiana and Paecilomyces fumosoroseus, was tested under laboratory conditions using the larval immersion method. High efficacies ranging from 68 to 100% corrected mortality were recorded with more profound effects in treatments with B. bassiana and P. fumosoroseus strains. The median lethal concentration (LC₅₀) for L3 was 6.0×10⁵ in M. anisopliae 79, 1.5×10⁵ in B. bassiana 124 and 4.2×10⁴ in P. fumosoroseus 14. These three strains were further used to characterize the age-dependent mortality of different larval stages (L2-L5) and the effect against pupae of H. armigera. Larval stages did not differ in their mortality but differed i in median lethal time, with shorter values recorded in the second instar. Tested fungi also caused a high reduction between 74.4 and 100% in the emergence of pupae using the soil inoculation method and the pupal immersion technique. All three fungal species, especially P. fumosoroseus, have a high potential for biocontrol of H. armigera larvae and also as a soil treatment targeting the pupae.     

中国虫霉目真菌及其寄主昆虫名录更新

报道了中国虫霉目Entomophthorales真菌共计79种,隶属2科11属,并对其所涉及的寄主昆虫名称进行了规范与更新,且给出了各寄主的分类地位和中英文名称对照。     

Mycorrhizal and dark septate endophytic fungi of <Emphasis Type="Italic">Pedicularis</Emphasis> species from northwest of Yunnan Province,China

Colonization of mycorrhizal and root endophytic fungi in 14 Pedicularis species from northwest of Yunnan Province, southwest China, was examined. These species included: Pedicularis gracilis Wall., Pedicularis longipes Maxim., Pedicularis axillaris Franch., Pedicularis cephalantha Franch., Pedicularis tenuisecta Franch., Pedicularis tapaoensis Tsoong, Pedicularis likiangensis Franch., Pedicularis dichotoma Bonati, Pedicularis yui Li, Pedicularis rhinanthoides Schrenk, Pedicularis rex C.B. Clarke, Pedicularis longiflora Rudolph., Pedicularis siphonantha Don, and Pedicularis oxycarpa Franch., among which nine are endemic to China (one to Yunnan). Three types of potentially beneficial fungi associated with roots of these species were observed, namely, arbuscular mycorrhizal fungi, ectomycorrhizal fungi, and dark septate endophytic fungi (DSEF), with DSEF as the most common colonizers. An unexpected high colonization level was detected in this hemiparasitic genus. Of the 19 sampling sites examined, 10 gave colonization frequency of above 50% and 6 showed a colonization index of above 50. Heavy colonization suggested a significant ecological role of these fungi and their potential to be applied to successful cultivation of these intractable plants.     

Defense mechanism of the termite, Coptotermes formosanus Shiraki, to entomopathogenic fungi

Termites, Coptotermes formosanus Shiraki, reared individually, were highly susceptible to entomopathogenic fungi, Paecilomyces fumosoroseus and Beauveria brongniartii and Metarhizium anisopliae, while termites reared in groups were highly resistant. Quantitative assays with an epifluoresent microscope revealed a significant difference in the number of conidia attachments among three entomopathogenic fungi. The conidia of B. brongniartii and P. fumosoroseus bound to termite cuticles more effectively than M. anisopliae conidia. Our results also suggested that self-grooming behavior is less effective, but mutual grooming is very effective in the removal of conidia from cuticles of their nestmates. Statistical analysis of removal rates indicated that conidia of P. fumosoroseus and B. brongniartii were removed more rapidly than M. anisopliae conidia from termite cuticles.     

Occurrence of keratinophilic fungi anddermatophytes on domestic birds in Nigeria

Feathers, nails and beaks of one hundred and twenty common birds in Nigeria, Chicken [50], Ducks [20], Turkeys [15] and Pigeons[35], were examined using the soil plate technique for their mycoflora.15 species of fungi were recovered and they belong to the genera Chrysosporium, Trichophyton, Microsporum, Aspergillus, Fusarium, Mucor, Rhizopus, Penicillium and Trichoderma. Microsporum gypseum was the species most frequently isolated (35% of the samples). The most common genus was Chrysosporium and C. keratinophilum was the species with the highest frequency in the genus (28.3%). The species isolated included potential pathogens and mycotoxin producing fungi (Aspergillus flavus, A. niger, Fusarium oxysporum). This revised version was published online in June 2006 with corrections to the Cover Date.     

转Cry1Ac抗虫棉对土壤虫生真菌群落的影响

利用大蜡螟Galleria mellonella L.诱菌法对转Cry1Ac抗虫棉GK-12及其亲本棉花SM-3的根系区域土壤的虫生真菌种类和数量进行监测,评价转Cry1Ac基因对棉田土壤虫生真菌的影响。实验结果显示:检测到的虫生真菌种类,在转基因棉GK-12及其亲本棉SM-3之间无显著差异,均为球孢白僵菌(Beauveria bassiana)和金龟子绿僵菌(Metarhizium anisopliae);在棉花开花初期、盛花期、结铃期或吐絮期,土壤虫生真菌数量在抗虫棉GK-12和亲本SM-3之间存在显著的差异,其他生育期无显著差异;在距棉花主根不同距离(5 cm\15 cm\25 cm)的区域,土壤虫生真菌数量和种类在抗虫棉GK-12和亲本SM-3之间无显著差异。     

Effects in vitro of nine fungicides on growth of entomopathogenic fungi

The effects of nine fungicides were evaluated in vitro on the entomopathogenic fungi Beauveria bassiana, Conidiobolus coronatus, C. thromboides, Metarhizium anisopliae, Paecilomyces farinosus, P. fumosoroseus, Scopulariopsis brevicaulis and Verticillium lecanii. The susceptibility of the fungi to the fungicides varied. The dithiocarbamate derivations zineb + copper oxychloride, and mancozeb completely inhibited germination of C. coronatus, C. thromboides, B. bassiana, P. farinosus, M. anisopliae and V. lecanii. The fungicides triadimefon, copper oxychloride, metalaxyl, sulfur, sulfur + nitrothal‐isopropyl and hymexazol exhibited various effects on the fungi. Usually, fungistasis was more pronounced at 15°C than at 25°C and the inhibitory effects were in direct proportion to the dosage of active ingredient (recommended field rate, 10‐fold higher and 10‐fold lower). In a few combinations, fungi partially overcame or even recovered from the initial inhibition of growth which might have resulted from delayed germination. In other cases, inhibition of growth occurred only after a delay and its intensity increased with time. Stimulation of fungal growth when it occurred was rarely permanent. Generally, adverse effects were much greater against the entomophthoraleans C. coronatus and C. thromboides than against the Hyphomycetes. Extrapolation of the results to practice and to the field situation is difficult. Nevertheless, the data presented give an idea of possible side‐effects in nature.     

基于吸虫塔（Suction Trap）的蚜虫测报预警网络的构建

吸虫塔（suction trap）是用来监测麦类蚜虫和大豆蚜Aphis glycines迁飞种群动态的大型植保测报设备,在欧洲和北美洲已经呈网络安装分布,为蚜虫的预警和防控提供重要依据。作者借鉴欧美吸虫塔的工作原理和设计,设计并生产了符合我国地域特征的吸虫塔设备,在东北、华北、华中、华东、西北等地布点安装了21台,用于监控我国麦类蚜虫和大豆蚜的迁飞动态,初步形成覆盖我国小麦主产区和大豆主产区的吸虫塔网络系统。此吸虫塔总高8.8m,由底部轴流风机运转产生的负压在塔顶部形成吸力,将迁飞经附近的小型昆虫吸入塔管,最后落入下部的样品收集瓶中,以此获得其迁飞的动态数据。吸虫塔网络的构建和完善,不仅为麦类蚜虫和大豆蚜的监控提供支撑,同时也为其它小型迁飞性昆虫监测、种群动态、生物多样性、生物信息学等研究提供数据。