医学昆虫的病原真菌*

昆虫病原真菌在自然环境中对害虫数量起着抑制作用,在害虫的微生物防治中也占有独特的地位。长期以来,在应用昆虫病原真菌防治人畜害虫方面研究进展较为缓慢,六十年代以来才有较大的发展。本文介绍了近十年的发展概况,并讨论了影响这个研究领域迅速发展的原因及病原真菌在生物防治中应用的潜力。认为尽管至今对这类真菌了解还较少,但通过今后加强有关方面的研究工作,其应用前景是广阔的。     

利用黄粉虫分离土壤昆虫病原真菌

利用黄粉虫Tenebrio molitorL.作为寄主从土壤中诱感并分离昆虫病原真菌。结果显示,从吉林市的18个土样中分离出球孢白僵菌(Beauveria bassiana)、金龟子绿僵菌(Metarhiziumanisopliae)和玫烟色拟青霉(Paecilomyces fumosoroseus)。土样中的昆虫病原真菌检出率为77.78%。这表明利用黄粉虫分离土壤中的昆虫病原真菌是一种简单、有效的方法。     

昆虫病原真菌紫外线诱变育种研究进展

昆虫病原真菌是控制害虫发生与危害的重要生物防治因子之一,优良的菌种是生物防治的关键。紫外线诱变育种是昆虫病原真菌菌种选育中最常用的方法之一,具有易操作、效果好、作用时间长等优点。本文以白僵菌、绿僵菌、玫烟色棒束孢等三种常用的昆虫病原真菌为例,从诱变育种的材料、方法和应用等方面综述了昆虫病原真菌紫外线诱变育种的研究现状,以期为昆虫病原真菌的生物防治研究提供参考。     

PFGE技术及其在昆虫病原真菌研究中的应用

染色体是真核生物遗传物质的载体,控制着生物体的遗传、变异和发育,研究染色体的组型和行为对于更好地改造、利用真核生物是至关重要的。昆虫病原真菌在自然界广为分布,作为害虫生物防除剂、中药材、经济昆虫病原真菌等被利用和研究［１～３］。然而,极大部分昆虫病原真菌属于半知菌类,缺乏有性生殖世代,核分裂时观察不到染色体凝缩,同时由于其染色体很小,又不具有足够数目的基因标记,所以难以用核染色和杂交等方法进行核型分析和遗传育种的研究,也难以用遗传学方法和细胞学方法测定其染色体数目和大小。另一方面,昆虫病原真菌Ｄ…     

峨眉山自然保护区的虫草及其它昆虫病原真菌I.

采自四川省峨眉山自然保护区的虫草及其相关昆虫病原真菌,其中除有常见的蛹虫草Ｃｏｒｄｙｃｅｐｓｍｉｌｉｔａｒｉｓ（Ｖｕｉｌｌ）,Ｆｒ粉被虫草（ＣｏｒｄｙｃｅｐｓｐｒｕｉｎｏｓａＰｅｔｃｈ）蝉拟青霉Ｐａｅｃｉｌｏｍｙｃｅｓｃｉｃａｄａｅ（Ｍｉｑｕｅｌ）Ｓａｍｓｏｎ,球孢白僵菌Ｂｅａｕｖｅｒｉａｂａｓｓｉａｎａ（Ｂａｌｓ．）Ｖｕｉｌｌ等昆虫病原真菌外,将主要描述峨眉虫草新种（Ｃｏｒｄｙｃｅｐｓｅｍｅｉｅ     

基因工程改良在昆虫病原真菌中的应用

昆虫病原真菌是自然界控制害虫种群的主要生物因子,其研究开发越来越受到重视。但由于存在致死速度慢、防效不稳定、对环境条件要求高等缺点,限制了昆虫病原真菌的应用。近年来,通过基因工程技术对昆虫病原真菌进行遗传改造,提高菌株的致病性和毒力,创造高效、稳定的工程菌株取得了较大进展。对昆虫病原真菌选择标记、遗传转化方法、基因工程改良及应用等方面最新研究进展进行了综述。     

昆虫病原线虫研究概况

昆虫病原线虫 (ｅｎｔｏｍｏｐａｔｈｏｇｅｎｉｃｎｅｍａｔｏｄｅｓ)是本世纪发展起来的一种有潜能的生物防治因子[1] 。它具有寄主范围广、能主动寻找寄主、对人畜及环境安全无毒 ,并能人工大量培养等优点。因此 ,在农药污染日益严重、害虫抗药性发展迅速的今天 ,昆虫病原线虫已成为可持续发展农业的迫切需要 ,受到国际生防领域的重视。特别是斯氏科线虫和小杆科线虫 ,已成为当前国际生防领域研究热点之一。本文简要概述了线虫的生物学特性、分类及致病机理等方面的研究 ,并结合目前存在的问题探讨了昆虫病原线虫的研究方向和发展前景。1…     

我国大陆寄生蚜虫的病原真菌

蚜虫的微生物防治日益成为新的研究热点。病原真菌作为一类重要的生防资源,在蚜虫的微生物治理中占有举足轻重的地位。我国的微生物生防工作开始较晚,但近几十年发展迅速。文章综合已发表的文献,介绍了我国大陆寄生蚜虫的病原真菌的种类及分布,并对其流行概况作了简要的描述与分析。     

Host range of Neozygites floridana isolates (Zygomycetes: Entomophthorales) to spider mites

Neozygites floridana (Weiser & Muma) (Zygomycetes: Entomophthorales) has been reported infecting naturally at least 18 species of tetranychids worldwide. However, the host range of N. floridana is unknown. Epizootics caused by this pathogen to tetranychid populations indicate that N. floridana has the potential to be used as a biological control agent. However, the virulence and specificity of species and strains of Neozygites need to be assessed in the laboratory to reveal its potential as a biological control agent. N. floridana isolates are currently been investigated in Brazil as biological control agents against the tomato red mite, Tetranychus evansi Baker & Pritchard, and the two-spotted spider mite, Tetranychus urticae Koch. The pathogenicity of five strains of N. floridana obtained from T. urticae, T. evansi and T. ludeni Zacher was assessed against populations of Mononychellus tanajoa (Bondar), Schizotetranychus sacharum Flechtmann & Baker, Tetranychus abacae Baker & Pritchard and Tetranychus armipenis Flechtmann & Baker, in addition to the species from which the fungus was obtained. Mummified mites were placed on leaf discs of the host plant of each tetranychid to promote fungal sporulation, and after 24 h the mites were transferred to the leaf discs. Contamination, infection and mummification were evaluated daily for seven days after confinement. Each isolate was pathogenic to three or four out of the six spider mite species tested. However, except for isolate ESALQ1421, all isolates caused higher levels of infection and significant mummification only to the tetranychid species from which they were collected. None of the isolates was pathogenic to S. sacharum and only one isolate infected T. abacae. Alternative hosts may be important for N. floridana survival in tropical regions where resting spores are rarely found.     

中国昆虫病原线虫一新纪录

采用分子生物学和形态学相结合的方法,对采自广东省翁源县的编号为GDa71的昆虫病原线虫进行分类鉴定.该线虫各虫期的主要形态特征、测量值以及基于ITS1和部分28S rDNA两段序列,分别构建系统进化树均显示它与印度异小杆线虫Heterorhabditis indica相似,是后者的一个品系,为中国新纪录.     

Beauveria Bassiana Pathogenicity to the Citrus Rust Mite Phyllocoptruta Oleivora

The pathogenicity of Beauveria bassiana to one of the major pests of citrus crops, Phyllocoptruta oleivora, was assessed by inoculating mites with different concentrations of conidia (1×10⁶, 5×10⁶, 1×10⁷, 5×10⁷ and 1×10⁸). Treated mites were kept at controlled conditions (25 ± 0.5°C, 12 h photoperiod and 98% relative humidity) and mite survivorship was evaluated daily. Mortality was found to increase in time and was dependent on the conidia concentration, with values ranging from 24 to 91% for the lowest and highest conidia concentration, respectively. The calculated LC₅₀ on the fifth day was 4.23×10⁶ conidia/ml. Mean lethal time was 3.98, 9.79, 3.09 and 2.74 days for 5×10⁶, 1×10⁷, 5×10⁷ and 1×10⁸ conidia/ml, respectively. Conidia were found to adhere all over the mite body surface, especially at the anal region, where vegetative mycelium was found entering the mite body. We noticed the formation of small crystals inside the mite’s body that were produced during colonization of the body cavity by the fungus. This is the first report of B. bassiana pathogenicity for this species.     

Variability in azygospore production among Entomophaga maimaiga isolates

This study describes in vitro and in vivo azygospore production by nine isolates of Entomophaga maimaiga, a fungal pathogen of the gypsy moth, Lymantria dispar. The three E. maimaiga isolates that consistently produced azygospores in vitro were also strong producers of azygospores in vivo. However, two additional isolates that were strong azygospore producers in vivo did not produce azygospores in vitro. Isolates that produced azygospores in vitro produced both conidia and azygospores more frequently in vivo than isolates not producing azygospores in vitro. In vitro azygospore production varied over time as well as by isolate. After >2 years of cold storage, while three isolates continued in vitro azygospore production, three isolates no longer produced azygospores in vitro.     

Ocurrence of the antibiotic producing bacterium Burkholderia sp. in colonies of the leaf-cutting ant Atta sexdens rubropilosa

Fungus garden material from recently established Atta sexdens rubropilosa colonies (6-12 months old) was sampled to detect antibiotic producing microorganisms that inhibited the growth of pathogens of insects and of the fungus gardens but did not affect their mutualistic fungus. A bacterium with activity against the entomopathogenic fungus Beauveria bassiana was isolated from 56% of the gardens tested (n=57) and identified from its biochemical profile and from 16S and 23S ribosomal DNA sequences as a member of the genus Burkholderia. The ant-associated Burkholderia isolates secreted a potent, anti-fungal agent that inhibited germination of conidia of the entomopathogenic fungi B. bassiana, Metarhizium anisopliae, of the saprophytic Verticillium lecanii, and also of a specialist fungus garden Escovopsis weberi. Growth of the ant's mutualist fungus was unaffected.     

Susceptibility of the filbertworm (Cydia latiferreana, Lepidoptera: Tortricidae) and filbert weevil (Curculio occidentalis, Coleoptera: Curculionidae) to entomopathogenic nematodes

The objective of this study was to determine the susceptibility of the two primary direct insect pests of hazelnuts in Oregon to three species of entomopathogenic nematodes. The entomopathogenic nematodes (Heterorhabditis marelatus Pt. Reyes, Steinernema carpocapsae All and Steinernema kraussei L137) were used in laboratory soil bioassays to determine their virulence against filbertworm, Cydia latiferreana (Walsingham) (Lepidoptera: Tortricidae) and filbert weevil, Curculio occidentalis (Casey) (Coleoptera: Curculionidae). All three nematode species were infective in laboratory bioassays. Infectivity ranged from 73-100% and 23-85% for filbertworm and filbert weevil, respectively. Field results were similar to those found in the laboratory with filbertworm larvae being more susceptible to nematode infection.     

Efficacy of entomopathogenic bacteria for control of Musca domestica

The aim of this study was to evaluate the larvicidal activity, and sub lethal effects of entomopathogenic bacteria Brevibacillus laterosporus, Bacillus thuringiensis var. israelensis, B. thuringiensis var. kurstaki, and a commercial formulation of Bacillus sphaericus on Musca domestica. Bacterial suspensions were prepared in different concentrations and added to the diet of newly-hatched larvae which were monitored until the adult stage. The larvae were susceptible to the B. laterosporus, B. thuringiensis var. israelensis, and B. thuringiensis var. kurstaki bacteria in varied concentration levels. These bacteria have larvicidal and sub lethal effects on the development of flies, reducing both adult size, and impairing the reproductive performance of the species.     

Isolation and molecular characterization of Bacillus thuringiensis found in soils of the Cerrado region of Brazil,and their toxicity to Aedes aegypti larvae

This study investigated the potential of Bacillus thuringiensis isolates obtained in the Cerrado region of the Brazilian state of Maranhão for the biological control of Aedes aegypti larvae. The isolates were obtained from soil samples and the identification of the B. thuringiensis colonies was based on morphological characteristics. Bioassays were run to assess the pathogenicity and toxicity of the different strains of the B. thuringiensis against third-instar larvae of A. aegypti. Protein profiles were obtained by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Polymerase chain reaction assays were used to detect the toxin genes found in the bacterial isolates. Overall, 12 (4.0%) of the 300 isolates obtained from 45 soil samples were found to present larvicidal activity, with the BtMA-104, BtMA-401 and BtMA-560 isolates causing 100% of mortality. The BtMA-401 isolate was the most virulent, with the lowest median lethal concentration (LC₅₀) (0.004 × 10⁷ spores/mL), followed by the Bacillus thuringiensis var. israelensis standard (0.32 × 10⁷ spores/mL). The protein profiles of BtMA-25 and BtMA-401 isolates indicated the presence of molecular mass consistent with the presence of the proteins Cry4Aa, Cry11Aa and Cyt1, similar to the profile of Bacillus thuringiensis var. israelensis IPS-82. Surprisingly, however, none of the cry and cyt genes analyzed were amplified in the isolate BtMA-401. The results of the present study revealed the larvicidal potential of B. thuringiensis isolates found in the soils of the Cerrado region from Maranhão, although further research will be necessary to better elucidate and describe other genes associated with the production of insecticidal toxins in these isolates.