球壳孢目真菌个体发育研究Ⅰ:壳二胞等四属

人工培养条件下,用光学显微镜和透射电子显微镜对球壳孢目（Sphaerosidales）的Ascochyta,Phyllosticta,Phomopsis,Septoria四属进行个体发育研究,明确了其产孢方式：Ascochyta和Phyllosticta为内壁芽生瓶梗式;Phomopsis为全壁芽生瓶梗式;Septoria为全壁芽生合轴式,这一结果为属级分类提供了可靠的依据。产孢方式是属内共同具有的稳定特征,分生孢子的形成方式类型、产孢细胞及分生孢子的形态是划分属的首要标准。以寄主植物属为基础确定的同属大多数种在个体发育上无明显差别,分生孢子的形态及大小是分种的一个重要依据。     

Isolation and identification of an anticancer drug,taxol from <Emphasis Type="Italic">Phyllosticta tabernaemontanae</Emphasis>, a leaf spot fungus of an angiosperm, <Emphasis Type="Italic">Wrightia tinctoria</Emphasis>

Phyllosticta tabernaemontanae, a leaf spot fungus isolated from the diseased leaves of Wrightia tinctoria, showed the production of taxol, an anticancer drug, on modified liquid medium (MID) and potato dextrose broth (PDB) medium in culture for the first time. The presence of taxol was confirmed by spectroscopic and chromatographic methods of analysis. The amount of taxol produced by this fungus was quantified using high performance liquid chromatography (HPLC). The maximum amount of taxol production was recorded in the fungus grown on MID medium (461 μg/L) followed by PDB medium (150 μg/L). The production rate was increased to 9.2 × 10³ fold than that found in the culture broth of earlier reported fungus, Taxomyces andreanae. The results designate that P. tabernaemontanae is an excellent candidate for taxol production. The fungal taxol extracted also showed a strong cytotoxic activity in the in vitro culture of tested human cancer cells by apoptotic assay.     

Microbial transformation of zaluzanin-D

Microbial transformation of zaluzanin-D using different fungi gave 11,13-dihydrozaluzanin-C, zaluzanin-C, 4,16,11,13 - tetrahydro zaluzanin-C, estafiatone, dihydroestafiatol and dihydroestafiatone.     

鸭跖草叶点霉粗毒素的提取方法和产毒条件的研究

以石油醚、乙酸乙酯、四氯化碳三种极性不同的溶剂对鸭跖草叶点霉病原菌的菌丝和培养液进行萃取,获取粗毒素。结果表明,叶点霉菌产生的毒素既有胞外毒素,又有胞内毒素,且用极性中等的乙酸乙酯萃取效果最好。大豆培养基和PSK分别是适合菌株生长和毒素分泌的固体培养基和液体培养基。确定有利于叶点霉产毒的温度为32℃、培养时间为14d、培养方式为150r/min振荡培养。     

鸭跖草生防菌叶点霉F-3菌株生物学特性的研究

主要研究影响鸭跖草生防菌叶点霉菌株菌丝生长和孢子萌发的条件。研究表明,PDA培养基和鸭跖草培养基是该菌生长的适宜培养基,该菌生长的温度范围为20~35℃,最适温度为25℃,分生孢子萌发的温度范围为25~35℃,最适温度为30℃;pH为6时生长最快,偏酸偏碱有利于孢子的萌发,在pH为10时萌发最好。F-3菌株在供试的6种碳源中生长速度相差不大,在不同氮源中差异显著,以硝酸铵为氮源生长最好。     

Expression of Recombinant Aequorin as an Intracellular Calcium Reporter in the Phytopathogenic Fungus Phyllosticta ampelicida

Conidia of Phyllosticta ampelicida germinate only after they have made contact with a substratum. Previous work has shown that external free calcium must be available to the spore for germination to be initiated. Transgenic strains of P. ampelicida expressing apo-aequorin, a calcium-sensitive luminescent protein, were developed to monitor cytoplasmic free Ca(2+) ([Ca(2+)]c). Transformants were verified by PCR and Southern hybridization. Apo-aequorin production was quantified for each of 21 transformants. The transformant that emitted the most light per unit of protein was found to contain 0.59 mg apo-aequorin/g total protein. To ascertain the feasibility of aequorin-based [Ca(2+)]c quantification, [Ca(2+)]c changes were measured in mycelia during various physiologically perturbing treatments: exposure to high concentrations of external Ca(2+), hypoosmotic shock, and mechanical perturbation. This is the first report of a plant pathogenic fungus for which aequorin-based Ca(2+) measurement protocols have been developed.     

Epidemiology,identification and disease management of grape black rot and potentially useful metabolites of black rot pathogens for industrial applications – a review

Phyllosticta ampelicida (teleomorph: Guignardia bidwellii) is the fungal plant pathogen that causes black rot on grapevine. It is able to infect all green, expanding grape tissues. Black rot poses a threat to both yield and wine quality; a severe attack can virtually cause a complete crop loss. The fungus is native to North America, and was spread to Europe at the end of the 19th century. With the beginning of the 21st century, an increasing importance of the disease was observed in several European winegrowing regions. Successful black rot control strategies combine sanitary measures, cultural techniques, growing cultivars with reduced susceptibility and the use of effective fungicides. Berries are most susceptible to infections between flowering and bunch closure and consequently, fungicide applications against black rot need to focus on this period. In this paper, forecast models and decision support systems that help to achieve satisfactory control with a minimum of fungicide input are presented. Black rot in grapevine was reported to be well controlled in field experiments by a broad range of fungicide classes including quinone outside inhibitors (98 ± 3% efficacy), demethylation inhibitors (98 ± 3% efficacy) and dithiocarbamates (92 ± 7% efficacy). Average efficacies of other fungicide classes tested so far ranged from 32 to 69%; meta data on the efficacy of fungicides in numerous field trials are given. At present, black rot causes major problems primarily if vineyard management and fungicide use were reduced or abandoned. Organically managed vineyards are highly affected. Hence, site and cultivar selection as well as cultural measures are of the highest importance especially in organic viticulture. Recent studies showed that some strains formerly classified as G. bidwellii in fact belong to a distinct species (P. parthenocissi). Black rot pathogens produce some phytotoxic secondary metabolites, such as phenguignardic acid, guignardic acid, alaguignardic acid and the guignardianones A, E and F, which could stimulate the development of new herbicides of natural origin.     

Taxonomical re-evaluation of Phoma-like soybean pathogenic fungi

Coelomycetous fungi classified in Ascochyta, Phoma, and Phyllosticta have been recorded from spots on leaves and pods of soybeans. Based on the Genealogical Concordance Phylogenetic Species Concept, the authors suggest the re-evaluation of the taxonomic status of Phoma sojicola (syn. = Ascochyta sojicola) and Phyllosticta sojicola. In spite of the former delimitation of Phoma sojicola based on small differences in morphological features, it has proved to be identical to Phoma pinodella. Similarly, it was also confirmed that Phyllosticta sojicola was identical to Phoma exigua var. exigua. The authors supply tools for identification of Phoma-like fungi by combined conventional and molecular methods. Protein-encoding genes (tef1 and β-tubulin) were successfully applied within the Phoma genus to infer phylogenetic relationships.     

Detection of a Bacterium Associated with a Leaf Spot Disease of Maize in Brazil

A leaf spot disease of maize occurring in Brazil in the 1980s was described as being caused by the ascomycete Phaeosphaeria maydis (P. Henn) Rane. Payak and Renfro (imperfect form Phyllosticta sp.). Disease symptoms were dark-green water-soaked spots that later became necrotic lesions. There are no reports at present in the literature of re-infection by the fungus under controlled conditions, casting doubt on the true identity of the pathogen. In this study, cytological analyses of lesions at the initial stages did not detect the presence of fungal structures. Bacterial colonies with yellow pigmentation were isolated from the lesions, which reacted positively in hypersensitivity tests in tobacco plants. Maize plants were inoculated with the isolated bacteria. After 72 h incubation in a dew chamber, plants were transferred to a greenhouse, where they remained until evaluation. Typical symptoms of the disease were observed 5–7 days after inoculation of plants, only on treatments inoculated with the bacteria. The bacterium was re-isolated, which suggests its involvement in the initial phases of disease. The bacterium was identified as Pantoea ananas (synonym Erwinia ananas ).     

Extensive host range of an endophytic fungus, <Emphasis Type="Italic">Guignardia endophyllicola</Emphasis> (anamorph: <Emphasis Type="Italic">Phyllosticta capitalensis</Emphasis>)

Isolation of endophytic species of Guignardia (anamorph: Phyllosticta) from healthy leaves of 94 plants (91 species and 3 varieties) in 69 genera, 42 families, was carried out in a test site (Kyoto Herbal Garden) to investigate the host range of Guignardia endophyllicola (anamorph: Phyllosticta capitalensis). Species of Guignardia and Phyllosticta were isolated from the leaves of 67 plants (66 species and 1 variety) belonging to 54 genera, 38 families. Among them, 53 isolates from different plants belonging to 43 genera in 36 families were similar in morphology, and sequence analysis of internal transcribed spacer (ITS) regions of ribosome DNA revealed these isolates to be conspecific with G. endophyllicola. In addition, this fungus was isolated from leaves of various plants collected in different places in Japan and Thailand. Thus, this endophytic fungus has been revealed to live within various vascular plants, angiosperms, gymnosperms, and pteridophytes.