蚧虫的病原真菌及其在生物防治中的潜力

蚧虫（半翅目：蚧总科）是农林果树和花卉的一类重要害虫。作者综述了寄生蚧虫的虫生真菌及其在生物防治中的潜力。总结了昆虫病原真菌作为生物杀虫剂的研究历史,并将其划分为3个发展阶段,即开创阶段、缓慢发展阶段和快速发展阶段。讨论了该领域在中国的研究现状。列出了世界上目前已记录的蚧虫病原真菌,包括55属140种,及其寄主蚧虫的名录。对虫生真菌未来的研究和开发提出了4点建议。     

Entomopathogenic fungi-mediated biological control of the red palm weevil Rhynchophorus ferrugineus

The red palm weevil (RPW), Rhynchophorus ferrugineus, is an important pest of palms, and difficult to control by conventional methods. Therefore, microbial control is an alternative strategy for controlling RPW. Herein, a total of 15 entomopathogenic fungi (EPFs) were subjected to primary pathogenicity screening against last stage of RPW larvae. The preliminary data showed that four Beauveria bassiana isolates (JEF-484, 158, 462 and 507) and one Isaria fumosorosea isolate (JEF-014) resulted in 100 % mortality within 5–10 days post inoculation (d.p.i.), respectively. According to the time required for RPW mortality, JEF-484, 158, 462 and 014 were further subjected to bioassays using 10⁷ conidia/ml suspensions by spraying method. Based on the results, JEF-484 showed the highest mortality and shortest LT₅₀ on the last stage of RPW larvae, followed by JEF-158. The two isolates also showed good conidial production and high thermal stability compared to the other isolates. Therefore, JEF-484 and JEF-158 were selected for bioassays against RPW egg and the last larval stage with different concentrations of 10⁵, 10⁶ and 10⁷ conidia/ml conidial suspensions by spraying method. For the bioassay at the egg stage, JEF-158 showed a significantly higher ovicidal effect than JEF-484. In the larval bioassay, both EPF isolates showed a dosage-dependent effect on the RPW larvae. JEF-484 caused higher mortality in RPW larvae than JEF-158. In summary, the combination of the 2 promising EPF isolates might provide an opportunity for the practical microbial control of RPW at different life stages in palm tree fields.     

Dual biocontrol potential of the entomopathogenic fungus Akanthomyces muscarius against Thaumetopoea pityocampa and plant pathogenic fungi

Akanthomyces spp. species are known for their capacity to biocontrol of certain insects and plant pathogens; however, their ability to biocontrol the pine processionary (Thaumetopoea pityocampa) and certain phytopathogenic fungi belonging to the genera Fusarium and Curvularia have not been studied before. In this study, a strain from Akanthomyces muscarius was isolated from wheat grains and then identified by morphological and molecular tests. The strain was further studied for its capacity to control Thaumetopoea pityocampa larvae through dose-mortality tests, and its ability to control some phytopathogenic fungi strains of the genera Fusarium and Curvularia was studied through direct confrontation tests. Dose-mortality tests at three concentrations of Akanthomyces muscarius against the first instar larvae revealed a mortality of 92.15% after 11 days for the concentration of 2.3 × 10⁶ conidia.ml⁻¹, with a median lethal concentration of 7.6 x10³ conidia.ml^1. Our isolate also showed antifungal activity against these phytopathogenic fungi with inhibition rates ranging from 39.61% to 52.94%. Akanthomyces muscarius proved to be a promising biocontrol agent for plant pests and diseases.     

昆虫病原微生物及其在蝗灾治理中的应用

本文重点介绍了蝗虫痘病毒、蝗虫微孢子虫、蝗噬虫霉、绿僵菌、白僵菌、米曲霉等的生物学特性、侵染机制、流行病学和应用潜力。蝗虫痘病毒和蝗噬虫霉分别由于生产成本过高和分生孢子在离体条件下存活时间过短而难以开发应用;蝗虫微孢子虫和绿僵菌已广泛应用到蝗灾治理中,有关绿僵菌致病机理的研究最深入全面;白僵菌也已成功应用到蝗灾治理中;介绍了一种新病原——米曲霉对飞蝗的毒力以及产孢量和耐热性等生物学特性。最后,提出蝗虫新病原微生物资源的开发、提高生产工艺水平、研发新的制剂和延长储存时间等是今后蝗虫微生物农药的发展方向。     

Adenylate cyclase orthologues in two filamentous entomopathogens contribute differentially to growth,conidiation, pathogenicity,and multistress responses

Adenylate cyclase (AC) is a core element of cAMP signalling network. Here we show functional diversity and differentiation of Beauveria bassiana AC (BbAC) and Metarhizium robertsii AC (MrAC). Severe growth defects occurred in ΔBbAC and ΔMrAC grown on nutrition-rich SDAY and several minimal media but were largely alleviated by adding cAMP to SDAY. Conidial yield increased greatly in ΔBbAC but decreased in ΔMrAC. During colony growth, ΔBbAC was highly sensitive to oxidation, high osmolarity, cell wall perturbation, carbendazim fungicide, Mn²⁺, Zn²⁺, Fe³⁺, and EDTA but more tolerant to Cu²⁺ while ΔMrAC showed higher osmotolerance, decreased sensitivity to Fe³⁺, and null response to carbendazim or cell wall stress despite similar responses to oxidation and other metal ions. Conidial UV-B resistance decreased by 32 % in ΔBbAC and 22 % in ΔMrAC despite little change in their theromotolerance. Median lethal time (LT₅₀) estimates of ΔBbAC and ΔMrAC against susceptible insects were 10.9 and 1.4 d longer than those from wild-type strains respectively. All the phenotypic changes were restored to wild-type levels by each gene complementation. Taken together, BbAC and MrAC regulated differentially conidiation, pathogenicity, and multistress responses in B. bassiana and M. robertsii, thereby making different contributions to their biocontrol potential.     

虫生真菌分子致病机理及基因工程改造研究进展

虫生真菌侵染寄主昆虫的复杂过程可分为体表附着、体壁穿透及体内定殖和致死等不同阶段.近年来,以金龟子绿僵茵(Metarhizium anisopliae)和球孢白僵菌(Beauveria bassiana)为代表的基因功能研究取得了长足的进展,从不同角度阐明了虫生真菌的分子致病机理;同时,基因工程技术的应用为昆虫病原真菌的遗传改良和选育高毒力杀虫菌株开辟了新的途径.对近年来昆虫病原真菌侵染寄主的分子对策及基因工程改造的研究进展进行了综述,并就进一步研究虫生真菌的毒力基因及功能进行了探讨.     

虫生真菌分子致病机理及基因工程改造研究进展

虫生真菌侵染寄主昆虫的复杂过程可分为体表附着、体壁穿透及体内定殖和致死等不同阶段。近年来, 以金龟子绿僵菌(Metarhizium anisopliae)和球孢白僵菌(Beauveria bassiana)为代表的基因功能研究取得了长足的进展, 从不同角度阐明了虫生真菌的分子致病机理; 同时, 基因工程技术的应用为昆虫病原真菌的遗传改良和选育高毒力杀虫菌株开辟了新的途径。对近年来昆虫病原真菌侵染寄主的分子对策及基因工程改造的研究进展进行了综述, 并就进一步研究虫生真菌的毒力基因及功能进行了探讨。     

Potential of a combination of entomopathogenic fungal strains and a non-ionic surfactant to control the fall armyworm (Spodoptera frugiperda)

The fall armyworm, Spodoptera frugiperda, is an emerging invasive pest in Taiwan that feeds on a wide range of crops and causes serious damage. Herein, an entomopathogenic fungal library (EFLib) was constructed to identify potential microbes to control FAW. Twenty-eight indigenous entomopathogenic fungi (EPF) were isolated and investigated for their potential pathogenicity, with Metarhizium pinghaense (Mp-NCHU-124) and Beauveria bassiana (Bb-NCHU-157) exerting dose-dependent effects on the 4th instar FAW larvae. The non-ionic surfactant Silwet L-77 rapidly killed FAW larvae after spraying at a concentration of 300 mg/kg and the toxic effect of Silwet L-77 on FAW larvae was dose-dependent. When the EPF isolates (10⁶ conidia/mL) were applied to FAW larvae in combination with the non-ionic surfactant Silwet L-77 (30–90 mg/kg), the mortality rate dramatically increased and the LT₅₀ reduced, with increased fungal mycosis (Mp-NCHU-124: 38% to 72% and Bb-NCHU-157: 20 to 62%), indicating the high compatibility of EPF with the non-ionic surfactant. Thus, the Silwet L-77+EPF combined formulation has potential for practical field application for FAW pest control and sustainable agriculture in the future.     

Assessing the Potential of Entomopathogenic Nematodes to Control the Grape Root Borer Vitacea polistiformis (Lepidoptera: Sesiidae) Through Laboratory and Greenhouse Bioassays

Seventeen entomopathogenic nematode species and strains were evaluated for virulence to the grape root borer, Vitacea polistiformis (Harris) in laboratory and greenhouse bioassays. Heterohabditis bacteriophora strain GPS11 and H. zealandica strain X1 produced a larval mortality rate of over 85% of larvae embedded in the root cambium in laboratory bioassays. The nematode species H. marelata and H. bacteriophora strain Oswego produced mortality rates of over 75%. Of the Steinernema species tested, S. carpocapsae strain 'All' performed the best with a mortality rate of 69%. All other nematode species and strains tested, with the exception of S. bicornutum , produced some degree of larval mortality. In the greenhouse bioassays, 93% control was achieved with H. zealandica strain X1 applied at 4 ×109 infective juveniles (IJs) acre1 -1 (9.88 ×10 9 IJs ha -1 ). H. bacteriophora strain GPS11 successfully reproduced in grape root borer larvae. The numbers of IJs produced within infected larvae were related to larval size. The survival rate of neonate larvae on grape root sections was 61%, which thus provides a means to rear the neonate larvae for bioassays.     

Enhanced biological control potential of the entomopathogenic nematode,Steinernema carpocapsae,applied with a protective gel formulation

Entomopathogenic nematodes (EPNs) are adapted to subterranean environments, and are prone to damage by UV light and desiccation. EPNs, Steinernema carpocapsae, combined with a protective gel, and anti-UV ingredients, have potential for above-ground pest management. We (1) ascertained whether the gel could provide protection to EPNs at low concentrations when applied in direct sun, (2) determined if other ingredients added to the gel increased efficacy, and (3) quantified retention and survival of EPNs applied to foliage with the gel. EPNs in 1% protective gel caused higher host mortality (60%) than other treatments (2–37%). UV protection provided by titanium dioxide (TD) and octyl methoxycinnamate with 1% protective gel solution was tested outdoors; these formulations resulted in higher host mortality (43% and 25%) than other treatments (2–7%). After 8?h in the greenhouse, 0.25% protective gel solution had the highest percentage of live EPNs on leaves. The gel at low concentration protects EPNs, and addition of TD enhanced the protective properties of the formulation. It is important to continually improve options for different growing systems and insect pest behaviour. TD added to a low-concentration formulation of the protective gel makes this application technique more viable for growers to use.     

First steps towards biological control of the pear gall midge (Contarinia pyrivora) with the insect pathogenic fungus Metarhizium brunneum

Gall midges are important pests in many crops. In fruit, they are difficult to control due to their life cycle, which takes place partially within the fruit. Here, we provide the first successful laboratory experiment to infect pear gall midge (Contarinia pyrivora) with the insect pathogenic fungus Metarhizium brunneum. We developed a procedure for sampling larvae, maintaining them in the laboratory and subjecting them to the fungus. We demonstrated that dipping larvae in a fungus suspension or adding a fungus suspension to the soil result in significant fungus induced mortality of the pear gall midge. An immune response in treated larvae was recorded proving that there was a real pathogenesis. Finally, we discuss next steps and a strategy for field experiments.     

厌氧真菌纤维降解酶及其应用潜力的研究进展

反刍动物的瘤胃是已知的纤维降解能力最强的天然发酵罐,其发酵粗纤维的能力很大程度上依赖于其中栖息的各类微生物的功能。厌氧真菌作为瘤胃内的一类低丰度菌群,最先定殖到宿主动物摄入的纤维质饲料上,并通过分泌大量高效的碳水化合物活性酶降解粗纤维。然而,由于缺乏足够的基因组信息和有效的厌氧真菌遗传操作系统,目前国内外对厌氧真菌分泌的纤维降解酶及其降解机制的研究进展有限。本文对厌氧真菌的分类及已发表的基因组信息进行概述,介绍了各类纤维降解酶及纤维小体的组成结构和催化机制特点,并对纤维降解酶在生物质能、饲料处理、纺织造纸及食品加工等方面的应用进行总结。研究厌氧真菌纤维降解酶的作用特性,将有助于完善其在瘤胃环境中有效竞争资源并降解粗纤维的知识体系,也将进一步了解其生物技术应用潜力,为工业生产中应用酶制剂提供新思路。     

The potential of Beauveria bassiana inoculum formulated into a polymeric matrix for a microbial control of spruce bark beetle

Two local strains of Beauveria bassiana originally isolated from naturally infected spruce bark beetles in Slovakia were tested for their virulence to Ips typographus (IT) and for their compatibility with a polymeric matrix composed of low-molecular polyethylene. Conidia could be homogenously immobilized in the low-molecular polyethylene matrix with no adverse effect on their viability and infectivity. At constant temperature (25°C), viability of immobilized conidial decreased only by 1–2% after 7 or 14 days when compared with non-formulated conidia. In field conditions, viability of conidia formulated in the matrix was even significantly higher than non-formulated conidia 35 days after their application in traps. Conidia incorporated into the polymeric matrix were infective to IT adults in laboratory bioassays. Mean values of LC₅₀ for native conidia (0.72–2.05?×?10⁶ conidia?ml^?1) and conidia immobilized in the polymeric matrix (0.64–1.03?×?10⁵ conidia?mm^?2) demonstrated high virulence. The efficacy of the local strains was significantly higher than that of B. bassiana strains from mycoinsecticides (Boverol^®, Botanigard^® ES and Naturalis-L^®). Results showed potential of this polymeric material for its use in microbial control of IT when mixed with conidia of B. bassiana.     

In vitro studies on the potential for biological control of Aspergillus flavus and Fusarium moniliforme by Trichoderma species

Culture filtrates of Trichoderma viride and Trichoderma harzianum were inhibitory of Fusarium moniliforme and, to a lesser extent, Aspergillus flavus. The degree of inhibition was, however, dependent on the carbon or nitrogen source incorporated into the medium. Scanning electron microscopy revealed the development of abnormal fruiting structures on exposure to some Trichoderma culture filtrate, while macroscopically, growth restriction and, in the case of A. flavus, altered colony colouration were observed. Based on the results of inverted colony culture, it would appear that some isolates of Trichoderma produce inhibitory volatile compounds. The production of possible antibiotics was also demonstrated. The aggressive behaviour (towards A. flavus and F. moniliforme) demonstrated by Trichoderma spp. may be partly explained by the liberation of extracellular enzymes by these fungi. An isolate of T. viride exhibited amylolytic, pectinolytic, proteolytic and cellulolytic activity. Based on the results of the present investigation, Trichoderma spp. are potential candidates for biocontrol of some mycotoxin-producing fungi, but there exists some doubt as to their osmotolerance within the air-dry seed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Entomopathogenic fungi as a potential control agent against the lesser mealworm, <Emphasis Type="Italic">Alphitobius diaperinus</Emphasis> in broiler houses

We tested the pathogenicity of 18 Metarhizium anisopliae (Metschn.) Sorokin isolates and 22 Beauveria bassiana (Balsamo) Vuillemin isolates against Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) larvae and adults. The efficacy of the most virulent isolate—M. anisopliae K—was evaluated in containers with a concrete bottom covered with wood shavings, under simulated poultry house conditions. Application of conidia of this isolate to the shavings or directly to the concrete bottom reduced the yield of larvae in 8–15 time compared with the control. In another test, the mortality of mature larvae placed on previously inoculated shavings or bottom reached 80–90% within 14 days, compared with 14% in the control. The residual activity of conidia kept at 28°C retained its initial level during 14 days post-inoculation, but declined after three weeks. Based on our data M. anisopliae has considerable potential for the control of A. diaperinus.

根结线虫放线菌及其生物防治活性研究

从感染植物根部的根结线虫卵和雌虫中,分离得到放线菌20株。形态、细胞壁氨基酸组分和16S rRNA基因序列分析表明:分离菌株分属于链霉菌属(Streptomyces)、诺卡氏菌属(Nocardia)和假诺卡氏菌属(Pseudonocardia),其中链霉菌占80.0%。分离菌株对根结线虫Meloidogynespp.卵的平均寄生率、卵的孵化率、幼虫死亡率分别为54.1%、40.4%和26.2%。根据体外测试的结果,选择具有高致病力的3株链霉菌(1-17,2-6,9-47)和1株诺卡氏菌(5-1)进行温室番茄防效测试,其生防效率分别为31.4%、37.7%、56.4%和42.4%。     

Occurrence of phytoalexins and phenolic compounds in endomycorrhizal interactions,and their potential role in biological control

This paper will review work mainly done during the last twenty years on the involvement of phytoalexin and phenolic compounds in mycorrhizal interactions. It has been observed that phytoalexins and associated molecules accumulate in roots after mycorrhizal infection, but less intensively and more slowly than in pathogenic interactions. Following mycorrhizal infection, enzymes of phenylpropanoid metabolism have been shown to be activated differentially. Some flavonoids and isoflavonoids have been reported to stimulate in vitro germination of mycorrhizal fungi or in vitro mycorrhizal infection, but their biological significance in signalling between the two symbiotic partners, and in biocontrol of plant disease by arbuscular mycorrhizal fungi, have not yet been elucidated.     

Weed biological control in California,USA: review of the past and prospects for the future

Weed biological control in California, USA began in 1940 with the release of a native scale insect on native Opuntia spp. on Santa Cruz Island, just offshore from mainland California. Since then, a total 39 weed species have been targets of biological control releases in California. Releases on 11 weed targets were transfer experiments where agents from related weed hosts were released on a new host. Most of the transfer experiment introductions failed but one weed was successfully controlled. Of the other 28 weeds, release sites for three species were destroyed and for six species releases are too recent to score, but for 19 weeds, their level of control was rated as: complete control (three species), substantial control (five species), and partial (six species), and negligible control (five species). Overall, 42% of the projects provide successful control, a result lower than observed in other countries worldwide. Since 1940, 77 species of agents have been released: 54 species established, 12 species failed to establish, six species had their release sites destroyed, and five species are too early to determine. Establishment rate was 82% but the rate differed among taxonomic orders. Individual agents were scored according to level of impact on their host and Coleoptera obtained the highest average impact score and Diptera the lowest. Mean impact scores over time showed a substantial drop in the 1980s but later increased. Future research efforts that emphasize introduction of high impact agents will further support development of this critical weed control method for California.     

Exotic ladybirds for biological control of herbivorous insects – a review

Since the late 19th century, exotic ladybirds (Coleoptera: Coccinellidae) have been used extensively for suppressing herbivorous insects of economic importance. In recent decades, the introduction of non‐native biological control (BC) agents has been greatly limited due to the awareness of the potential non‐target effects of introductions. Nonetheless, recent episodes of biological invasions of economically important pests have raised the need to carefully consider whether the expected benefits of pest control go beyond the possible environmental risks of introduction. To better understand the factors that contributed to successful BC programs, here we review the literature behind classical and augmentative BC using exotic ladybirds. Additionally, by means of case studies, we discuss the BC efficacy of selected exotic species, e.g., Coccinella septempunctata L., Harmonia axyridis (Pallas), and Hippodamia variegata (Goeze), and their position within the communities of predators in the introduced areas of USA, Canada, and Chile. In Europe, much of the research on exotic ladybirds has been conducted on the undesired impact of H. axyridis. Therefore, we summarize the risk assessment data for this species and review the field research investigating the ecological impact on European aphidophagous predators. According to the BIOCAT database of classical BC programs, 212 ladybird species belonging to 68 genera have been released in about 130 years of BC activity, with 14.6% of introductions having resulted in partial, substantial, or complete control of the target pest. However, because post‐release evaluation of establishment and BC success has not always been conducted, this rate could underestimate the successful cases. Among other factors, ladybird establishment and pest suppression mostly depend on (1) intrinsic factors, i.e., high voracity, synchronized predator‐prey life cycle, and high dispersal ability, and (2) extrinsic factors, i.e., adaptability to the new environment and landscape composition. This review contributes to improved understanding of ladybirds as exotic BC agents.