In vivo development and pathogenicity of Ascosphaera proliperda (Ascosphaeraceae) to the alfalfa leafcutting bee, Megachile rotundata

First instar larvae of the leafcutting bee, Megachile rotundata, were fed on either artificial or natural provisions containing spores of Ascosphaera proliperda. Two isolates were used as a source of inocula: one originated from in vitro isolates obtained while culturing what was thought to be pure spores of A. aggregata, the second originated from in vitro cultures from Denmark. Histological and scanning electron microscopy studies revealed that the spores germinated in the gut lumen and the developing hyphae invaded all tissues, after which they penetrated through larval integument and began the sexual phase of the life cycle aerially. Virtually all fungus-exposed larvae developed symptoms of disease regardless of source of inoculum, type of provision, and spore dose (1.5 × 10³ to 3 × 10⁶) per insect. It was concluded that the fungus was pathogenic to the alfalfa leafcutting bee under laboratory conditions and future studies should be conducted to determine its etiology, cross infectivity, and natural distribution in other bee taxa.     

ϵ-N-Trimethyllysine desaminase de Neurospora crassa

?-N-Trimethyl-l-lysine is transformed into 2-keto-6-N-trimethylhexanoic acid by a soluble deaminase present in the mycelium of Neurospora crassa. The activity of the enzyme increases with the age of the mycelium with an optimum after 10 days of culture. ?-N-Trimethyllysine deaminase has been purified 200-fold by differential centrifugation and successive column chromatography on DEAE Sephadex and Ultrogel AcA-34. Some of its properties have been studied. This is the first enzyme reported to metabolise ?-N-trimethyllysine.     

漳江口红树林沉积物可培养真菌多样性

红树林真菌由于其独特的生态特征、多样性特点和丰富的新型生物活性代谢产物而引起了广泛的关注。本文以漳江口红树林保护区4个采样点的沉积物为研究对象，采用8种筛选培养基（RBM、PDA、CDA、Martin、YM、SDA、ISP2和R2A）分离可培养真菌，根据ITS序列分析对其进行物种鉴定，并利用多样性指数分析评价不同采样点真菌群落的差异。结果共分离到274株真菌，隶属于2门12纲23目39科52属，其中优势属为枝顶孢属（Acremonium）（20.8%）和青霉属（Penicillium）（11.3%），其次是枝孢属（Cladosporium）（7.3%）和帚枝霉属（Sarocladium）（7.3%）。8种筛选培养基中，分离出真菌种属类型最丰富的培养基是RBM，其次是PDA。根据Shannon-Wiener多样性指数（H′）、Simpson优势度指数（D）、Magalef丰富度指数（R）和Pielou均匀度指数（J）的分析结果，在属的水平上，距海洋的位置越近，真菌的群落多样性越高，真菌分布越均匀。     

Isolation and functional characteristics of the fungus Hypoxylon spp. Sj18 with biocontrol potential

A fungus with biocontrol potential was isolated from the roots of hickory trees. The strain named sj18 was classified as a member of the genus Hypoxylon (Hypoxylaceae) after multigene phylogenetic analysis (beta-tubulin gene, internal transcribed spacer, 28S large subunit ribosomal RNA gene, and RNA polymerase II subunit gene). The strain grew well on a PDA with an optimum temperature range between 32 and 34 °C. The fungus had obvious inhibitory effects on Botryosphaeria dothidea, Colletotrichum gloeosporioides, and Gibberella moniliformis in fumigation experiments on solid agar plates. In an inoculation experiment of Chinese cabbage, the fungus was also found to have an obvious repellent effect on cabbage caterpillars. In vitro experiments on Petri dishes showed that the fermentation broth of the sj18 strain could kill 100% of Bursaphelenchus xylophilus within 8 h even if the fermentation broth was diluted 8 times. The inoculation test of Arabidopsis thaliana showed that the fungus could promote the lateral root formation of plants and significantly increase their aboveground biomass. Through the analysis of solid phase microextraction (SPME), it was found that the main volatile components of the fermentation products were azulene 65.39% (61.77% + 3.62%), caryophyllene 7.41%, and eucalyptol 6.83% according to the peak area ratio. Therefore, sj18 can be used as a candidate for the further research and development of biocontrol agents.     

Development of a PCR-based Assay for the Detection of Plasmodiophora brassicae in Soil

A single-tube nested polymerase chain reaction (STN PCR) method was developed for detecting the causal agent of clubroot disease, Plasmodiophora brassicae. Outer primer PBTZS-2 (5′-CCGAATTCGCGTCAGCGTGA-3′) to amplify a 1457 bp-fragment from P. brassicae DNA and nested primers, PBTZS-3 (5′-CCACGTCGATCACGTTGCAAT-3′) and PBTZS-4 (5′-GCTGGCGTTGATGTACTGGAA-TT-3′), to amplify a 398 bp-fragment internal of the 1457 bp-fragment were used for the STN PCR. The 398 bp-fragment was amplified from as little as 1 fg of P. brassicae DNA with the STN PCR. A protocol for extracting P. brassicae DNA directly from soil was developed. By using the protocol, DNA was extracted from artificially infested soil containing various numbers of P. brassicae resting spores and the resulting DNA was used as template for the STN PCR. As little as one resting spore of P. brassicae per g of soil was detectable with the STN PCR. The STN PCR was applied to naturally infested soil from 3 fields and one canal bed. The 398 bp-fragment was amplified from soil of 2 fields and the canal bed. To improve the detection of P. brassicae, the STN PCR products were subjected to second PCR amplification (double PCR) using the nested primers PBTZS-3 and PBTZS-4. The double PCR amplification generated a single 398 bp-DNA band which was visualized clearly on the agarose gel for all the 4 soil samples tested. A combination of the STN PCR and the double PCR appears a useful assay method for detecting P. brassicae resting spores in field soil.     

白腐菌菌体对染料的生物吸附脱色及机理研究

目的：研究了白腐菌菌体吸附染料特性、影响因素及吸附机理。方法：采用分光光度法、吸附热特性、傅立叶变换红外光谱（FTIR）分析等系统地对菌体吸附特性及机理进行研究。结果：白腐菌BP对不同类型的染料有不同的吸附效果,240min内染料RBBR脱色率能达82.35%。菌体对RBBR的合适吸附条件为：温度28℃、转速100r/min、菌体粒径小于60目。吸附符合Freun-dlich模式,为多分子层吸附。菌体吸附染料主要通过菌体表面的羟基、羧基、胺基及磷酸基团与染料分子以共价键、离子交换或氢键结合来进行。结论：利用白腐菌菌体能有效的对部分染料进行吸附脱色。     

Identification of spores in the polycentric anaerobic gut fungi which enhance their ability to survive

Two new isolates of the gut fungi were obtained from the rumen digesta and faeces of a cow. These isolates, designated Anaeromyces following rDNA typing, displayed a polycentric growth habit but differed from all other gut fungi in that they were able to survive in the laboratory for considerable periods without the need for sub-culture. Light microscopy of preparations from old liquid-grown cultures revealed the presence of DNA-containing spores with two or four chambers. A comparative evaluation of the growth produced when fresh media were inoculated with a sample originating from young or old cultures revealed that active growth was delayed with the inoculum from the older culture. We propose that the chambered spores observed in these cultures provide an alternative path in the life cycle of these fungi and may function as a resting stage within the anaerobic environment of the herbivore gut.     

Sur la presence de derives ε-N-methyles de la lysine a l'etat libre dans Neurospora crassa

By using two different isotopically labelled precursors and ion exchange aminoacid analysis, it is shown that the methyl group of methionine is incorporated into ε-N-mono-, di- and trimethylated derivatives of lysine found in the free state in the cells of Neurospora crassa. The possibility that lysine methylation can take place not only via protein bound-lysine but also directly on the free aminoacid, or a derivative is considered.     

Effect of repeated subculturing on agar and passaging through an insect host on pathogenicity,morphology, and growth rate of Verticillium lecanii

Repeated subculturing on different agars of single- and multispore isolates of a strain of Verticillium lecanii resulted in physiological and gross morphological changes but no significant attenuation of virulence. Furthermore, passaging through an aphid host, Macrosiphoniella sanborni, did not enhance virulence. Precautions are suggested to avoid any changes in characteristics of fungal strains in axenic culture.     

Study on the potential of cold-active lipases from psychrotrophic fungi for detergent formulation

Lipases from psychrotrophic fungal isolates BPF4 and BPF6 identified as Penicilium canesense and Pseudogymnoascus roseus respectively were characterized for their compatibility towards laundry detergent. BPF4 and BPF6 lipases showed maximum activity at pH 11 and 9 respectively and at 40?°C. The residual activities at 20?°C and 4?°C of BPF4 lipase were 35% and 20% and of BPF6 lipase were 70% and 20?°C respectively. Both the enzymes were stable at 4?°C, 20?°C and 40?°C for 2?h losing at the most 20% of activities. Both the enzymes were metalloenzymes with activity enhancement by nearly threefold by Ca²⁺. Contrary to BPF6 lipase, BPF4 enzyme was not stimulated by EDTA nor inhibited, rather stimulated by SDS and Triton X-100 by 125% and 330% respectively. Both the lipases showed minor to moderate inhibition by NaClO₃ and H₂O₂, and exhibited nearly 90% residual activity after 1?h of incubation in selected detergent brands thus indicating potential for their inclusion in detergent formulation thereby facilitating cold-washing as a step towards mitigation of climate change.     

西洋参真菌病害的生物防治

从北京市怀柔区西洋参种植基地中共采集西洋参根围土样20份,分离到一株对西洋参真菌病害有较强拮抗作用的菌株,经PDA平板拮抗试验表明,该菌对西洋参真菌病害的抑制力均在90％以上;田间试验结果表明,该菌对西洋参病害具有一定的防治效果,使四年生西洋参的平均发病率降低了54．2％。     

黄栌根内生真菌分离鉴定及拮抗真菌筛选

采用常规分离法从不同地区健康黄栌根系中共分离得到33株内生真菌,其优势类群主要包括无孢菌群、腐质霉属、青霉属等,而且不同地区黄栌的根系内生真菌种类和数量有一定差异。内生真菌对黄栌枯萎病病原——大丽轮枝孢的拮抗试验结果表明:团炭角菌、黑乌霉属、假丝酵母属的菌株抑制作用较强,其它内生真菌菌株对大丽轮枝孢均有不同程度的抑制作用。     

基于DNA复合条形码技术的蝗虫肠道共生真菌多样性研究

利用DNA复合条形码技术,研究了11个样本的蝗虫肠道共生真菌的多样性。结果显示:ITS在所研究的物种中鉴定了5门16纲29目40属2786 OTU真菌。肠道真菌群落组成分析结果显示:所有物种肠道真菌类群中含量最高的是木耳菌目和银耳目,其中斑翅蝗科的真菌类群多样性相对最高,斑腿蝗科的真菌类群多样性相对最低,表明各蝗虫肠道之间存在着明显的菌群多样性变化。α多样性分析结果显示:斑翅蝗科的共生真菌群落丰富度和多样性最高,斑腿蝗科的则最低。β多样性分析结果显示:(1)同科的各个种的肠道真菌群落结构差异性较小,不同科的种的肠道真菌群落结构差异性较大;(2)剑角蝗科的肠道真菌群落结构与其他物种的相似性均相对较低,而且在两个不同取样地得到的中华剑角蝗的真菌群落结构相似性也相对较低。聚类分析结果显示:(1)同科的蝗虫肠道真菌首先聚到一起,且群落相似性也相对较高;(2)布勒掷孢酵母属、内疣衣属和外瓶霉属3个属在蝗虫肠道真菌中是优势菌属。     

Anaeromyces mucronatus nov. gen., nov. sp. A new strictly anaerobic rumen fungus with polycentric thallus

A new species of strictly anaerobic fungus was isolated from the cow rumen. It is characterized by a polycentric thallus, a polynuclear rhizomycelium, mucronate zoosporangia and uniflagellated zoospores. It is also singular in that the sporocysts do not react to the specific lectins of L-fucose, N-acetyl-D-galactosamine and diacetyl chitobiose. These characteristics justify the creation of a new genus.     

Paeonol produced by Chaetomium sp., an endophytic fungus isolated from Paeonia suffruticosa

In order to research the relationship between endophytic fungus and active ingredients in medicinal Paeonia suffruticosa, a total of 57 fungal isolates were isolated from the roots, stems, leaves and buds of medicinal plant Paeonia ostii; mycelium was collected after these fungal isolates were fermented on PDA medium for a few days; then the mycelium products were extracted; their extracts were analyzed by gas chromatography–mass spectrometry. With this method, a strain endophytic fungi named J1-2 which can produce paeonol was screened. Paeonol produced by J1-2 was analyzed by using a high resolution mass spectrometer (HRMS) and nuclear magnetic resonance (NMR). The potential paeonol-procucing named J1-2 was identified Chaetomium based on morphological characteristics and ITS sequence analysis. The current research initially indicates that endophytic fungi can affect the potency of peony. At the same time it also indicates that the numerous endophytic fungi inside the medicinal Paeonia suffruticosa are precious resource for the pharmaceutical natural products that are originally from the Paeonia suffruticosa.     

Haliphthoros milfordensis isolated from black tiger prawn larvae (Penaeus monodon) in Vietnam

A marine fungus was isolated from the black tiger prawn Penaeus monodon at Nha Trang, Vietnam, on March 20, 2001 and named isolate NJM 0131. The fungus was identified as Haliphthoros milfordensis from the characteristics of asexual reproduction, and its physiological characteristics were investigated. Although the optimum temperature for growth of the isolate was 25°–30°C, the fungus grew at a wide range of temperatures (15°–40°C). H. milfordensis grew well in 50%–100% seawater, but poorly in PYG agar containing 1.0%–5.0% NaCl and KCl. The fungus grew at a wide range of pH (4.0–11.0) with the optimum pH value of 7.0–9.0. The isolate also showed pathogenicity to swimming crab larvae (Portunus trituberculatus) by artificial infection, but mortality was not high. This is the first report of disease in the black tiger prawn P. monodon in Vietnam caused by H. milfordensis.     

霉菌固定化综述

分别对霉菌的固定化方法、固定化生物反应器及固定化霉菌发酵的应用等方面的研究进展进行了综述