中国拟盘多毛孢属真菌的七个新组合

作者按照Steyaert(1949)和Sutton(1980)的分类观点,对1987—1988年从我国广西、江西、浙江等地采集的Pestalotiopsis属真菌进行了研究,确认了拟盘多毛孢属真菌的7个新组合种,即Pestalotiopsis carveri(Guba)comb nov.,Pestalotiopsis elasticae(Koord.)comb.nov.,Pestalotiopsis langloisii(Guba)comb.nov.,Pestalotiopsis oleandri(Guba)comb.nov.,Pestalotiopsis sinensis(shen)comb.nov.,Pestalotiopsis sydowiana(Bres.)comb.nov.和Pestalotiopsis zahlbruckneriana(Bres.)comb.nov.     

健康与凋萎病杨梅树体及根围菌群的差异

凋萎病是制约杨梅产业发展的严重病害。为了有效防控凋萎病,本研究分析了杨梅健康和感染凋萎病树体各部位及根表土和根围土中细菌和真菌群落的丰富度与多样性的差异。结果表明: 与健康树相比,病树根围土、根表土、根、枝干、枝皮和叶片的细菌和真菌丰富度均发生了显著变化,其中,根表土细菌和枝皮内真菌的丰富度和多样性均显著降低,而枝皮内细菌和根表土的真菌丰富度和多样性均显著升高。病树各部位及根表、根围土细菌和真菌的优势菌相对丰度在门、纲和属水平上发生了明显的变化,在病树枝干、根和根表土中的假单胞菌属及根表土、根围土中的镰刀菌属的相对丰度明显降低,病树根表土及根围土中青霉菌属的相对丰度明显增加。与凋萎病菌同属的拟盘多毛孢菌在病树根内显著减少,而在其他位置均大量增殖,其相对丰度与多数相对丰度较高的真菌呈正相关。本研究结果将为开发杨梅凋萎病的生态改良、培育健康树体和生物防治技术提供有效的理论依据。     

Genomic and transcriptomic analysis of the endophytic fungus Pestalotiopsis fici reveals its lifestyle and high potential for synthesis of natural products

Background

In recent years, the genus Pestalotiopsis is receiving increasing attention, not only because of its economic impact as a plant pathogen but also as a commonly isolated endophyte which is an important source of bioactive natural products. Pestalotiopsis fici Steyaert W106-1/CGMCC3.15140 as an endophyte of tea produces numerous novel secondary metabolites, including chloropupukeananin, a derivative of chlorinated pupukeanane that is first discovered in fungi. Some of them might be important as the drug leads for future pharmaceutics.

Results

Here, we report the genome sequence of the endophytic fungus of tea Pestalotiopsis fici W106-1/CGMCC3.15140. The abundant carbohydrate-active enzymes especially significantly expanding pectinases allow the fungus to utilize the limited intercellular nutrients within the host plants, suggesting adaptation of the fungus to endophytic lifestyle. The P. fici genome encodes a rich set of secondary metabolite synthesis genes, including 27 polyketide synthases (PKSs), 12 non-ribosomal peptide synthases (NRPSs), five dimethylallyl tryptophan synthases, four putative PKS-like enzymes, 15 putative NRPS-like enzymes, 15 terpenoid synthases, seven terpenoid cyclases, seven fatty-acid synthases, and five hybrids of PKS-NRPS. The majority of these core enzymes distributed into 74 secondary metabolite clusters. The putative Diels-Alderase genes have undergone expansion.

Conclusion

The significant expansion of pectinase encoding genes provides essential insight in the life strategy of endophytes, and richness of gene clusters for secondary metabolites reveals high potential of natural products of endophytic fungi.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-014-1190-9) contains supplementary material, which is available to authorized users.     

一株新的拟盘多毛孢菌液体培养特性研究

对影响红豆杉内生真菌拟盘多毛孢属菌株H619生长的11个发酵因素进行试验。结果表明该菌适宜生长的C源、N源、C/N、pH值、Ca~(2 )和Mg~(2 )浓度分别约为蔗糖、玉米粉、25:1～35:1、7～8、1.5‰、0.07‰;最优培养条件约为装料比50 mL/250 mL、接种量10%、转速176 r/min,温度25℃,发酵周期7 d。探讨了各因素影响菌丝生长的显著性,为后续研究提供了实验依据。     

Description and identification of two new diseases of guariroba palm (Syagrus oleraceae) in Brazil

Guariroba is a palm species native to central Brazil. Seedlings of guariroba with leaf spots of unknown aetiology were found in Patos de Minas, Minas Gerais, Brazil. The leaf spots were manifest as two different symptom types: the first lesion type consisted of necrotic spots with a rounded to elongate shape, with a light colour and dark edges; the second lesion type had a rounded shape, was dark brown in colour with a light brown edge. The objective of this study was to elucidate the aetiology of both diseases. The likely causal agents were isolated and Koch's postulate fulfilled. Subsequently, the ITS region of rDNA from both micro‐organisms were amplified and sequenced. According to the morphological characteristics and molecular analyses, the fungal species were identified as Pestalotiopsis adusta (causing necrotic spots with a rounded to elongate shape, with a light colour and dark edges), and Alternaria tenuissima (causing lesions with a rounded shape, dark brown in colour with a light brown edge). Identification of the causal organisms of these two diseases will help guide management approaches that might be tested to reduce impact of the disease on Guariroba, including the use of fungicides and cultural approaches.     

Genomic Analysis of the Mycoparasite Pestalotiopsis sp. PG52

Pestalotiopsis sp. is a mycoparasite of the plant pathogen Aecidium wenshanense. To further understand the mycoparasitism mechanism of Pestalotiopsis sp., we assembled and analyzed its genome. The genome of Pestalotiopsis sp. strain PG52 was assembled into 335 scaffolds and had a size of 58.01 Mb. A total of 20,023 predicted genes and proteins were annotated. This study compared PG52 with the mycoparasites Trichoderma harzianum, Trichoderma atroviride, and Trichoderma virens. This study reveals the entirely different mycoparasitism mechanism of Pestalotiopsis compared to Trichoderma and reveals this mycoparasite’s strong ability to produce secondary metabolites.     

黄精褐斑病的病原生物学特性

为明确发生在贵州省施秉县的黄精叶斑类病害的病原菌,通过形态学和分子生物学方法对病原菌进行了鉴定,并对其生物学特性进行了初步研究。形态特征及rDNA-ITS、β-tubulin和tef1多基因序列分析表明,该病原菌为棕榈拟盘多毛孢Pestalotiopsis trachicarpicola。生物学特性研究结果表明,该菌菌丝体适宜生长温度为15-30℃,最适温度为28℃;最适pH值为5;以葡萄糖为碳源、酵母浸膏为氮源比较适合菌丝体的生长;菌丝体生长的最佳培养基为PDA;光照对菌丝体生长无明显影响;菌丝体致死温度为45℃。     

Sinopestalotiollides A–D,cytotoxic diphenyl ether derivatives from plant endophytic fungus Pestalotiopsis palmarum

Four new diphenyl ether derivatives, sinopestalotiollides A–D (1–4), one new natural α-pyrone product (11), as well as twelve known compounds (5–1?7), were obtained from the ethyl acetate extract of the endophytic fungus Pestalotiopsis palmarum isolated from the leaves of medicinal plant Sinomenium acutum (Thunb.) Rehd et Wils. The structures were elucidated by HR-ESI-MS and NMR spectrometry data. Bioassay experiments revealed that compounds 1–4 and 11 exhibited strong to weak cytotoxicities against three human tumor cell lines Hela, HCT116 and A549.     

Detection of cross reactive antigens between Pestalotiopsis theae and tea leaves and their cellular location

Among the 12 varieties of tea tested against three isolates of Pestalotiopsis theae, causal agent of grey blight disease, Teen Ali-17/1/54 and TV-23 were found to be highly susceptible while CP-1 and TV-26 were resistant under identical conditions. Leaf antigens were prepared from all the tea varieties, three isolates of P. theae and a non-pathogen of tea (Bipolaris tetramera). Polyclonal antisera were raised against mycelial suspensions of P. theae (isolate Pt-2) and leaf antigens of Teen Ali-17/1/54 and CP-1. These were compared an immunodiffusion test and enzyme-linked immunosorbent assay to detect cross reactive antigens (CRA) shared the host and the parasite. CRA were found among the susceptible varieties and isolates of P. theae (Pt-1, 2 and 3). Such antigens were not detected between isolates of P. theae and resistant varieties, B. tetramera and tea varieties or isolates of P. theae. Indirect staining of antibodies using fluorescein isothiocyanate (FITC) indicated that in cross sections of tea leaves, the CRA was concentrated in the epidermal cells and mesophyll tissues. CRA was present in the young hyphal tips of the mycelia and on the setulae and appendages of the conidia of P. theae.     

Analysis of beta-tubulin cDNAs from taxol-resistant Pestalotiopsis microspora and taxol-sensitive Pythium ultimum and comparison of the taxol-binding properties of their products

The anti-cancer drug taxol binds to β-tubulin in assembled microtubules and causes cell cycle arrest in animal cells; in contrast, in fungi, the effect of taxol varies. For instance, the taxol-producer Pestalotiopsis microspora Ne32, an ascomycete, is resistant to taxol (IC₅₀ greater than 11.7 μM), whereas Pythium ultimum, an oomycete, is sensitive to taxol (IC₅₀ 0.1 μM). In order to understand the differential fungal response to taxol, we isolated cDNAs encoding β-tubulin from both P. microspora and P. ultimum. The deduced amino acid sequence of β-tubulin from P. microspora is very similar to those from other Ascomycetes, many of which are resistant to taxol. The sequence of β-tubulin from P. ultimum is very similar to those from Oomycetes and non-fungal organisms, many of which are sensitive to taxol. To examine the interaction between taxol and fungal microtubules, binding studies were performed with fungal cells, using [³H]taxol. The labeled taxol was found to bind specifically to P. ultimum, but not to P. microspora. In addition, the amount of [³H]taxol specifically bound to P. ultimum was reduced by the microtubule-depolymerizing drug thiabendazole, in a dose-dependent manner. These results suggest efficient binding of taxol to microtubules in P. ultimum, but not in P. microspora, and are consistent with the differential taxol sensitivity of these two organisms. Finally a comparison of previously characterized taxol binding sites in various β-tubulin sequences showed that β-tubulins of taxol-sensitive organisms, including P. ultimum, contain Thr219, but β-tubulins of resistant organisms, including P. microspora, contain Asn or Gln at this position, suggesting an important role for residue 219 in the interaction between taxol and β-tubulin. Received: 16 March 1999 / Accepted: 21 August 1999