虫生真菌分子生物学研究进展*

虫生真菌分子生物学的在研究自开展以来,有20多种与杀虫毒力相关的蛋白酶得到纯化,而与之相对应的基因也分别得到克隆、测序。陆续的基因克隆表明,虫生具菌中的重要毒力基因以基因族的形式存在,在精确的表达调控下以适应不同的寄主种类和不同的环境条件。基因工程研究表明,以增加毒力基因拷贝的方式可显提高虫生真菌的杀虫速率,带有外源基因标记的菌株也是环境释放示综研究的良好材料及方法。     

虫生真菌深层培养分生孢子的喷雾干燥技术

虫生真菌制剂田间应用的一个关键问题是液体深层培养中产生的分生孢子（下简称液生孢子）或芽孢子的稳定性。最近,Ｓｔｅｐｈｅｎ等（见Ｂｉｏｃｏｎｔｒｏｌ　Ｓｃｉｅｎｃｅ　ａｎｄＴｅｃｈｎｏｌｅｇｙ,１９９８,８：３－１１）发展了一种喷雾干燥技术。采用这一技术可使芽抱子和／或液生抱子干燥,又不显著降低孢子的活力。这项研究是在实验室用小型喷雾干燥器进行的。１保护剂的筛选测定了６种保护剂对金龟子绿僵菌Ｍｅｔａｒ－ｈｉｚｉｕｍ　ａｎｉｓｏｐｌｉａｅ　（Ｍｅｔｓｃｈｎｉｋｏｖ）ＳｏｒｏｋｉｎＭａ９７株的存活力的…     

虫生真菌育种工作研究进展

虫生真菌在害虫的生物防治中有着极其重要的作用,在植物病害和杂草的生物防治、保健品、医药和化妆品等领域的开发应用上也具有很大的潜力.目前虫生真菌的育种工作主要采用自然选育、人工诱变、准性生殖、原生质体技术、基因工程技术等方法获得优良菌株.对虫生真菌的育种方法及其应用进展进行了综述,同时评述了各种育种方法的优缺点,分析了今后虫生真菌的育种方向.     

虫生真菌研究与应用的发展与机遇——2012年虫生真菌专刊序言

我国从事虫生真菌基础及应用研究的人员众多,各方面均取得了重要的研究成果.2012年“虫生真菌专刊”收集了国内关于虫生真菌资源、基因克隆及活性成分代谢等方面的最新研究进展.进一步的基础研究,应加强致病基因功能和真菌-昆虫分子相互作用等研究,以提高真菌杀虫剂或药用虫生真菌的应用效率.     

应用虫生真菌防治烟粉虱的现状

烟粉虱Bemisia tabaci属世界性分布的大害虫.在一定条件下虫生真菌是控制烟粉虱种群的最有效天敌.本文就几种常见虫生真菌:蜡蚧轮枝菌Verticilliumlecanii、座壳孢Aschersoniaspp.、玫烟色拟青霉Paecilomycesfumosoroseus、球孢白僵菌Beauveria bassiana等防治烟粉虱的应用现状以及它们与其它天敌的关系进行了综述,以期对烟粉虱虫生真菌的合理利用提供参考.     

山西省虫生真菌生态多样性研究

为探讨山西省主要植被区的虫生真菌资源区系分布特点 ,于 1996～ 2 0 0 1年连续对山西省境内芦芽山、庞泉沟、蟒河 3个国家级保护区的虫生真菌进行了生态多样性研究。采集的 4 6 9份昆虫标本中有 2 76份分离出真菌 ,总检出率为 5 8.8%。根据菌落形态、营养体、孢子等特征将 12 4株虫生真菌初步鉴定到属 ,其中 110株已鉴定到种和专化型 ,共计 10属 2 5种。对各地区和各生境中真菌类群进行统计比较的结果表明 ,各保护区内虫生真菌物种资源都很丰富 ,广泛分布在不同的生境中 ,但具有明显的地域性分布特点。从菌株数量来看 ,从大到小依次为蟒河(45株 ) >庞泉沟 (40株 ) >芦芽山 (39株 ) ;从属的种类来看 ,蟒河地区 (10属 )标本中的类群略多于其他 2个自然保护区 ;但从属内种类多样性来看 ,蟒河 (18种 )明显比其他 2个自然保护区丰富。白僵菌 (Beauveriaspp .)、绿僵菌 (Metarhiziumspp .)等优势类群具有明显的垂直分布带谱和小生境分布特点 ,中等海拔区域 (10 0 0～ 2 0 0 0m)分离菌株数最多 ,占到 6 6 %以上 ,包含种类最丰富 ,几乎囊括了所有种类 ,混交林、阔叶林和溪边灌林中 ,虫生真菌种类丰富 ,分布均匀 ,尤以混交林中菌株最多 ,共 2 3株 ,占到 37% ,种类也最丰富 ,包含 8种     

真菌的分子生物学

随着对真菌及真菌病诊断与治疗研究的深入,近年来在真菌分子生物学研究方面也有不少进展,如在真菌超微结构、细胞壁的特殊构型与致病的关系、真菌的染色体倍性与数目,以及其遗传信息等方面。     

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.     

Occurrence of entomopathogenic fungi in migratory alate aphids in Yunnan Province of China

Occurrence of entomopathogenic fungi in migratory alate aphids was investigated by trapping air-borne aphids on cabbage and wheat plants and observing the insects for fungal infection. Potted cabbage and wheat plants put 30 m apart on yellow cloth, were placed on the top of a tall building (six-story building) in Yunnan Province, China away from cultivated fields or other vegetation. Seven species of aphids trapped on the plants collected and monitored for fungal infection in the laboratory at a daily basis from 19 February to 31 December, 2003. Aphids were trapped every day during the experimental period. A total of 3,681 aphids were trapped on cabbage and 3,028 on wheat. Myzus persicae (Sulzer), Brevicoryne brassicae (L.), and Sitobion avenae (F.) were the major aphid species accounting for 94.7 and 93.1% of the insect specimens landing on cabbage and wheat, respectively. Other aphis species encountered included, Acyrthosiphon pisum (Harris), Lipaphis erysimi (Kaltenbach), Pemphigus sinobursaris Zhang, and Rhopalosiphum padi (L.). Preference for host plant played significant role in attraction of aphids, especially of the three major species. Accumulated over the whole experimental period nearly 33% of the aphids landing on cabbage and 37% landing on wheat were infected with fungi. Erynia neoaphidis Remaudiere and Hennebert, Entomophthora planchoniana Cornu, and Beauveria bassiana (Balsamo) Vuilemin were the major fungal pathogens encountered and accounted for nearly 95% of the infested aphids on cabbage and 97% on wheat. Other entomopathogenic fungi encountered included Zoophthora radicans (Brefeld) Batko, Conidiobolus obscurus (Hall and Dunn) Remaudiere, and Neozygites fresenii (Nowakowski), Paecilomyces fumosoroseus (Wize) Brown and Smith, and Aspergillus flavus Link. Although the number of aphids trapped throughout the experimental period did not vary consistently with temperature or humidity, the fungal was directly related to relative humidity. Higher insect mortality, especially due to Entomophthorales, positively related relative humidity.     

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.     

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.     

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.     

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。     

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.     

Levels of diversity in endomycorrhizal fungi (Glomales,Zygomycetes) and their role in defining taxonomic and non-taxonomic groups

Diversity in glomalean fungi is manifested at the molecular, morphological, and ecological levels. Characters at any of these levels can be ordered into hierarchical patterns defining taxonomic groups if they are conserved enough to be heritable through geologic time in all descendants of a common ancestor. At present, only morphological characters associated with mode of spore formation and in subcellular structure of spores are sufficiently stable and diverse to recognize at least 150 species. Ontogenetic comparisons indicate that species integrity, despite asexual reproduction, is the result of rigid internal constraints imposed on variation during the process of spore subcellular differentiation. Epigenetic factors dominate because the differentiation sequence is linear and each new stage is causally linked to preceding stages. Some morphological characters of the fungal mycelium also exist, but they define more inclusive groups at the family level and above. Most diversity in the mycorrhizae consists of life-history traits associated with abundance and architecture of fungal components, their rate of formation and longevity, and their cost in the symbiosis. These characters participate in processes at the molecular and ecological levels, so they are autonomous from morphological determinants. They often are labile or affected by external environmental conditions, so fewer stable taxonomic characters are likely to be discovered. Instead, molecular and ecological diversity has greater potential to define; (a) niche specificity of organisms/populations and (b) causal processes linked to host-fungus compatibility and mycorrhizal efficiency. Any taxonomic characters that relate to mycorrhizal functions will come only from comparative studies involving organisms from shared habitats rather than those having shared spore morphologies.