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Purification and characterization of an exo-polygalacturonase from Pycnoporus sanguineus 血螺外聚半乳糖醛酸酶的纯化及特性研究
Pub Date : 2009-12-01 DOI: 10.1016/j.mycres.2009.09.007
Emma N. Quiroga , Melina A. Sgariglia , César F. Molina , Diego A. Sampietro , José R. Soberón , Marta A. Vattuone

The present work describes the purification and characterization of a novel extracellular polygalacturonase, PGase I, produced by Pycnoporus sanguineus when grown on citrus fruit pectin. This substrate gave enhanced enzyme production as compared to sucrose and lactose. PGase I is an exocellular enzyme releasing galacturonic acid as its principal hydrolysis product as determined by TLC and orcinol-sulphuric acid staining. Its capacity to hydrolyze digalacturonate identified PGase I as an exo-polygalacturonase. SDS-PAGE showed that PGase I is an N-glycosidated monomer. The enzyme has a molecular mass of 42 kDa, optimum pH 4.8 and stability between pH 3.8 and 8.0. A temperature optimum was observed at 50–60 °C, with some enzyme activity retained up to 80 °C. Its activation energy was 5.352 cal mol−1. PGase I showed a higher affinity towards PGA than citric pectin (Km = 0.55 ± 0.02 and 0.72 ± 0.02 mg ml−1, respectively). Consequently, PGase I is an exo-PGase, EC 3.2.1.82.

本工作描述了一种新的细胞外多半乳糖醛酸酶,PGase I的纯化和特性,该酶是由生长在柑橘果实果胶上的血蝎螺(Pycnoporus sanguineus)产生的。与蔗糖和乳糖相比,这种底物能提高酶的产量。通过薄层色谱法和orcinol-硫酸染色测定,PGase I是一种细胞外酶,其主要水解产物为半乳糖醛酸。它水解二半乳糖酸的能力鉴定了PGase I是一种外聚半乳糖酸酶。SDS-PAGE显示PGase I是一个n -糖苷化单体。酶的分子量为42 kDa,最适pH为4.8,稳定性在pH 3.8 ~ 8.0之间。在50-60°C的温度下,酶的活性保持在80°C。其活化能为5.352 cal mol−1。PGase I对PGA的亲和力高于柠檬酸果胶(Km分别为0.55±0.02和0.72±0.02 mg ml−1)。因此,PGase I是一个外显子PGase, EC 3.2.1.82。
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引用次数: 12
Characterization of the cell wall of the ubiquitous plant pathogen Botrytis cinerea 普遍存在的植物病原菌灰霉病细胞壁的特性
Pub Date : 2009-12-01 DOI: 10.1016/j.mycres.2009.09.006
Dario Cantu, L. Carl Greve, John M. Labavitch, Ann L.T. Powell

The ascomycete Botrytis cinerea is a destructive and ubiquitous plant pathogen and represents a model organism for the study of necrotrophic fungal pathogens. Higher fungi possess a complex and dynamic multilayer cell wall involved in crucial aspects of fungal development, growth and pathogenicity. Plant resistance to microbial pathogens is determined often by the capacity of the plant to recognize molecular patterns associated with the surface of an interacting microbe. Here we report the chemical characterization of cell walls from B. cinerea during axenic growth. Neutral sugars and proteins constituted most of the mass of the B. cinerea cell walls, although chitin and uronic acids were detected. Glucose was the most abundant neutral sugar, but arabinose, galactose, xylose and mannose also were present. Changes in cell wall composition during culture were observed. As the culture developed, protein levels declined, while chitin and neutral sugars increased. Growth of B. cinerea was associated with a remarkable decline in the fraction of its cell wall material that was soluble in hot alkali. These results suggest that the cell wall of B. cinerea undergoes significant modifications during growth, possibly becoming more extensively covalently cross-linked, as a result of aging of mycelia or in response to decreasing nutrient supply or as a consequence of increasing culture density.

子囊菌灰霉病菌(Botrytis cinerea)是一种普遍存在的破坏性植物病原体,是研究坏死性真菌病原体的模式生物。高等真菌具有复杂和动态的多层细胞壁,参与真菌发育、生长和致病性的关键方面。植物对微生物病原体的抗性通常是由植物识别与相互作用微生物表面相关的分子模式的能力决定的。在这里,我们报道了灰绿杆菌在无菌生长过程中细胞壁的化学特性。中性糖和蛋白质构成了葡萄球菌细胞壁的大部分质量,尽管检测到几丁质和糖醛酸。葡萄糖是最丰富的中性糖,但也存在阿拉伯糖、半乳糖、木糖和甘露糖。观察培养过程中细胞壁组成的变化。随着培养的发展,蛋白质水平下降,而几丁质和中性糖增加。灰霉病菌生长时,其细胞壁可溶于热碱的部分显著下降。这些结果表明,B. cinerea细胞壁在生长过程中经历了显著的修饰,可能由于菌丝老化或营养供应减少或培养密度增加而变得更广泛的共价交联。
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引用次数: 42
Fungal biomass associated with the phyllosphere of bryophytes and vascular plants 苔藓植物和维管植物叶层相关的真菌生物量
Pub Date : 2009-11-01 DOI: 10.1016/j.mycres.2009.08.001
M.L. Davey , L. Nybakken , H. Kauserud , M. Ohlson

Little is known about the amount of fungal biomass in the phyllosphere of bryophytes compared to higher plants. In this study, fungal biomass associated with the phyllosphere of three bryophytes (Hylocomium splendens, Pleurozium schreberi, Polytrichum commune) and three vascular plants (Avenella flexuosa, Gymnocarpium dryopteris, Vaccinium myrtillus) was investigated using ergosterol content as a proxy for fungal biomass. Phyllosphere fungi accounted for 0.2–4.0 % of the dry mass of moss gametophytes, representing the first estimation of fungal biomass associated with bryophytes. Significantly more fungal biomass was associated with the phyllosphere of bryophytes than co-occurring vascular plants. The ergosterol present in moss gametophytic tissues differed significantly between species, while the ergosterol present in vascular plant leaf tissues did not. The photosynthetic tissues of mosses had less associated fungal biomass than their senescent tissues, and the magnitude of this difference varied in a species-specific manner. The fungal biomass associated with the vascular plants studied varied significantly between localities, while that of mosses did not. The observed differences in phyllosphere community biomass suggest their size could be affected by host anatomical and physiological attributes, including micro-niche availability and chemical host defenses, in addition to abiotic factors like moisture and nutrient availability.

与高等植物相比,苔藓植物层层中真菌生物量的数量知之甚少。以麦角甾醇含量为指标,研究了3种苔藓植物(Hylocomium splendens, Pleurozium schreberi, Polytrichum commune)和3种维管植物(Avenella flexuosa, Gymnocarpium dryopteris, Vaccinium myrtillus)层球真菌生物量。层球真菌占苔藓配子体干质量的0.2 - 4.0%,是苔藓植物真菌生物量的首次估算。与共生维管植物相比,苔藓植物层球的真菌生物量显著增加。苔藓配子体组织中麦角甾醇含量在种间差异显著,而维管植物叶片组织中麦角甾醇含量无显著差异。与衰老组织相比,苔藓光合组织的相关真菌生物量更少,而且这种差异的大小因物种而异。与维管植物相关的真菌生物量在不同地区差异显著,而与苔藓相关的真菌生物量则无显著差异。观察到的层圈群落生物量差异表明,除了水分和养分有效性等非生物因素外,它们的大小还可能受到宿主解剖和生理属性(包括微生态位有效性和宿主化学防御)的影响。
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引用次数: 59
Species delimitation in the European species of Clavulina (Cantharellales, Basidiomycota) inferred from phylogenetic analyses of ITS region and morphological data 基于ITS区系统发育分析和形态学资料的欧洲Clavulina (cantharelllales,担子菌科)种划分
Pub Date : 2009-11-01 DOI: 10.1016/j.mycres.2009.08.008
Ibai Olariaga , Begoña M. Jugo , Koldo García-Etxebarria , Isabel Salcedo

The identification of the conventionally accepted species of Clavulina (Cantharellales, Basidiomycota) in Europe (Clavulina amethystina, Clavulina cinerea, Clavulina cristata, and Clavulina rugosa) is often difficult and many specimens are not straightforwardly assignable to any of those four species, which is why some authors have questioned their identity. In order to assess the status of those species, a morphological examination was combined with the molecular analysis of the ITS region. The same six major clades were obtained in the Bayesian and parsimony phylogenetic analyses, and all six clades were well-supported at least by one of the analyses. Morphological characters, such as the overall branching pattern, the presence and intensity of grey colour, the cristation of the apices, and basidiospore size and shape were to various extents correlated with the phylogenetic signal obtained from the ITS region. The congruence between the molecular analyses and morphology, rather than geographical origin, suggests the existence of several species that can be delimited using a combined phylogenetic and morphological species recognition. The analyses revealed that C. cristata and C. rugosa are well-delimited species. In contrast, more than one taxa could be subsumed under the names C. amethystina and C. cinerea, the taxonomical complexity of which is discussed. The ITS region is proved to be adequate to separate phylogenetic species of Clavulina.

欧洲的Clavulina (Cantharellales, Basidiomycota) (Clavulina amethystina, Clavulina cinerea, Clavulina cristata和Clavulina rugosa)的传统物种鉴定通常是困难的,许多标本不能直接归属于这四个物种中的任何一个,这就是为什么一些作者质疑它们的身份。为了评估这些物种的地位,形态学检查与ITS区域的分子分析相结合。在贝叶斯和简约系统发育分析中得到了相同的6个主要分支,并且所有6个分支至少有一个分析得到了很好的支持。形态学特征,如总体分枝模式、灰色的存在和强度、顶端的嵴、担子孢子的大小和形状等,在不同程度上与ITS区域获得的系统发育信号相关。分子分析和形态学之间的一致性,而不是地理起源之间的一致性,表明存在几个物种,可以使用系统发育和形态物种识别相结合来划分。分析结果表明,冠蝉和冠蝉属属的分界较好。相比之下,C. amethystina和C. cinerea可归入一个以上的分类群,并讨论了它们的分类复杂性。ITS区被证明足以区分棒柳属的系统发育种。
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引用次数: 27
Mycological Research News 真菌学研究新闻
Pub Date : 2009-11-01 DOI: 10.1016/j.mycres.2009.10.004
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引用次数: 0
Putative source of the invasive Sirex noctilio fungal symbiont, Amylostereum areolatum, in the eastern United States and its association with native siricid woodwasps 美国东部侵入性夜蛾真菌共生体Amylostereum areolatum的推测来源及其与本地毒蜂木蜂的关系
Pub Date : 2009-11-01 DOI: 10.1016/j.mycres.2009.08.012
Charlotte Nielsen , David W. Williams , Ann E. Hajek

Two genotypes of the fungal symbiont Amylostereum areolatum are associated with the invasive woodwasp Sirex noctilio first found in North America in 2004. S. noctilio is native to Europe but has been introduced to Australasia, South America and Africa where it has caused enormous losses in pine plantations. Based on nucleotide sequence data from the intergenic spacer region (IGS) of the nuclear ribosomal DNA, the A. areolatum genotypes found in North America are most similar to genotypes found in Europe, and not to genotypes from the southern hemisphere. Although two IGS strains of A. areolatum were found in North America it cannot be stated whether A. areolatum was introduced to North America from Europe once or twice based on our study. Genetic groupings formed by sequencing data were in most cases supported by vegetative compatibility groups (VCGs). Other siricid woodwasp species in the genus Sirex are native to North America. The North American native Sirex edwardsii emerging from the same tree as S. noctilio carried the same strain of A. areolatum as S. noctilio. The North American native Sirex sp. ‘nitidus’ collected outside the geographical range of S. noctilio carried a unique strain within A. areolatum. Our findings of A. areolatum in the native North American species, S. sp. ‘nitidus’, contrast with the previous view that A. areolatum was not present in North America before the accidental invasion of S. noctilio.

真菌共生体areolatum Amylostereum的两个基因型与2004年首次在北美发现的入侵木黄蜂Sirex noctilio有关。山松原产于欧洲,但已被引入大洋洲、南美洲和非洲,在那里它造成了巨大的损失。根据核糖体DNA基因间间隔区(IGS)的核苷酸序列数据,发现北美的沙纹松果基因型与欧洲的基因型最相似,而与南半球的基因型不相似。虽然在北美地区发现了两株沙棘,但根据我们的研究,还不能确定沙棘是从欧洲传入北美的一次还是两次。在大多数情况下,由测序数据形成的遗传分组由营养相容性组(vcg)支持。其他Sirex属的木蜂种类原产于北美。北美本土的edward Sirex与S. noctilio来自同一棵树,携带与S. noctilio相同的A. areolatum品系。在S. noctilio地理范围之外采集的北美本地Sirex sp. ' nitidus '在a . areolatum中携带一种独特的菌株。我们在北美本土物种S. sp. ' nitidus '中发现了沙纹线虫,这与之前关于沙纹线虫在S. noctilio意外入侵之前在北美不存在的观点形成了对比。
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引用次数: 54
Halotolerant and halophilic fungi 耐盐和嗜盐真菌
Pub Date : 2009-11-01 DOI: 10.1016/j.mycres.2009.09.002
Nina Gunde-Cimerman , Jose Ramos , Ana Plemenitaš

Extreme environments have for long been considered to be populated almost exclusively by prokaryotic organisms and therefore monopolized by bacteriologists. Solar salterns are natural hypersaline environments characterized by extreme concentrations of NaCl, often high concentrations of other ions, high uv irradiation and in some cases extremes in pH. In 2000 fungi were first reported to be active inhabitants of solar salterns. Since then many new species and species previously known only as food contaminants have been discovered in hypersaline environments around the globe. The eukaryotic microorganism most studied for its salt tolerance is Saccharomyces cerevisiae. However, S. cerevisiae is rather salt sensitive and not able to adapt to hypersaline conditions. In contrast, some species like Debaryomyces hansenii, Hortaea werneckii, and Wallemia ichthyophaga have been isolated globally from natural hypersaline environments. We believe that all three are more suitable model organisms to study halotolerance in eukaryotes than S. cerevisiae. Furthermore, they belong to different and distant taxonomic groups and have developed different strategies to cope with the same problems of ion toxicity and loss of water.

长期以来,人们一直认为极端环境几乎完全由原核生物居住,因此被细菌学家所垄断。太阳盐碱地是一种天然的高盐环境,其特征是NaCl浓度极高,其他离子浓度通常也很高,紫外线照射强度高,在某些情况下ph值极高。2000年,真菌首次被报道为太阳盐碱地的活跃居民。从那时起,在全球高盐环境中发现了许多新物种和以前只被称为食物污染物的物种。研究最多的耐盐真核微生物是酿酒酵母菌。然而,酿酒酵母对盐非常敏感,不能适应高盐环境。相比之下,一些物种如Debaryomyces hansenii, Hortaea werneckii和Wallemia ichthyophaga已经在全球范围内从自然高盐环境中分离出来。我们认为这三种生物都比酿酒酵母更适合研究真核生物的耐盐性。此外,它们属于不同和遥远的分类群,并发展出不同的策略来应对相同的离子毒性和水分流失问题。
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引用次数: 271
The genus Laetiporus (Basidiomycota, Polyporales) in East Asia 东亚的担子菌属(担子菌科,多孢子目)
Pub Date : 2009-11-01 DOI: 10.1016/j.mycres.2009.08.014
Yuko Ota , Tsutomu Hattori , Mark T. Banik , Gregor Hagedorn , Kozue Sotome , Sawako Tokuda , Yasuhisa Abe

Relationships among East Asian, North American and European Laetiporus sulphureus s. lat., a cosmopolitan brown rot species complex, were assessed with phylogenetic analyses and incompatibility tests. Three East Asian taxa, Laetiporus cremeiporus sp. nov., Laetiporus montanus and Laetiporus versisporus, are described and illustrated as well as compared with related taxa from Southeast Asia, North America and Europe. Phylogenetic analyses showed that L. cremeiporus and L. versisporus are clearly distinct species among Laetiporus taxa. The three conifer inhabiting species, Laetiporus conifericola, Laetiporus huroniensis and L. montanus, are closely related to each other. The European population of L. montanus exhibits two sequence variants of the EF1α: one is the same as observed in L. sulphureus in Europe and the other is that observed in East Asian population of L. montanus. A key to the known species of Laetiporus in the northern hemisphere is provided.

东亚、北美和欧洲硫斑蝶的亲缘关系。采用系统发育分析和不亲和性试验对一世界性褐腐病种复合体进行了评价。本文描述了东亚的三个分类群,即cremeiporus sp. nov.、Laetiporus montanus和Laetiporus versisporus,并与东南亚、北美和欧洲的相关分类群进行了比较。系统发育分析表明,cremeiporus和versisporus在李氏菌类群中是明显不同的种。三种针叶树栖种松柏Laetiporus conifericola、胡氏Laetiporus huroniensis和montanus之间亲缘关系密切。L. montanus欧洲种群的EF1α序列有两个变异,一个与欧洲的L. sulphureus相同,另一个与东亚的L. montanus相同。提供了北半球已知的雷氏菌种类的钥匙。
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引用次数: 34
Description of Cryptosporiopsis kienholzii and species profiles of Neofabraea in major pome fruit growing districts in the Pacific Northwest USA 美国西北太平洋地区主要梨果种植区kienholzii隐孢子虫的描述和neofabraa的物种概况
Pub Date : 2009-11-01 DOI: 10.1016/j.mycres.2009.08.013
Robert A. Spotts , Keith A. Seifert , Kelly M. Wallis , David Sugar , Chang Lin Xiao , Maryna Serdani , Jose L. Henriquez

The objectives of this study were i) to give a taxonomic description of a fungus phylogenetically related to Neofabraea and assign the name Cryptosporiopsis kienholzii to this fungus, ii) to expand previous Neofabraea species profiles from infected apple and pear fruit collected from major pome fruit production districts in Oregon and Washington, and iii) to determine the sensitivity of Neofabraea alba, Neofabraea malicorticis, Neofabraea perennans, and C. kienholzii to a range of fungicides. A name is given herein to the anamorph of the fungus previously called ‘Neofabraea sp. nov.’ in the literature, and the fungus is designated as C. kienholzii. The teleomorph of this fungus has never been observed in vivo or in vitro. N. alba, N. perennans, and C. kienholzii accounted for 61.3, 35.6, and 3.1 %, respectively, of 608 isolates obtained from pear fruit and 6.0, 81.3, and 12.7 % of 150 isolates from apple fruit. N. alba was the most common species in Oregon and N. perennans was most common in Washington. N. malicorticis was not found in any of the districts and may be limited to the wet climates west of the Cascade mountain range. C. kienholzii occurs in pome fruit orchards from southern Oregon to North Central Washington, and this is the first report of C. kienholzii in the state of Washington. The four most effective fungicides for control of bull's-eye rot caused by all species were thiabendazole, thiophanate-methyl, pyrimethanil, and pyraclostrobin + boscalid. Other fungicides gave acceptable control of some species of Neofabraea but not others. There was good agreement of the microbioassay with fungicide evaluations using wound-inoculated pear fruit for five of seven fungicides, but the microbioassay was not consistent enough for determination of the sensitivity of Neofabraea species to new fungicides.

本研究的目的是:(1)对一种与Neofabraea系统发育相关的真菌进行分类描述,并将其命名为隐孢子虫(Cryptosporiopsis kienholzii);(2)从俄勒冈州和华盛顿州主要的梨果产区收集的受感染的苹果和梨果实中扩展以前的Neofabraea种谱;(3)确定Neofabraea alba、Neofabraea malicorticis、Neofabraea perennans和C. kienholzii对一系列杀菌剂的敏感性。本文给以前在文献中称为“Neofabraea sp. nov.”的真菌的变形命名,并且该真菌被指定为C. kienholzii。这种真菌的远变形从未在体内或体外观察到过。在梨果实608株分离株中,白僵菌占61.3%、35.6%和3.1%;在苹果果实150株分离株中,白僵菌占6.0、81.3%和12.7%。在俄勒冈州最常见的是白杨,在华盛顿州最常见的是多年生白杨。在任何地区都没有发现N. maliccortis,可能仅限于喀斯喀特山脉以西的潮湿气候。C. kienholzii发生在俄勒冈州南部到华盛顿州中北部的梨树果园,这是华盛顿州首次报道的C. kienholzii。对所有病原菌引起的牛眼腐病防治效果最好的4种杀菌剂分别是噻苯达唑、甲基硫代菌酯、嘧菌酯和嘧菌酯+ boscalid。其他杀菌剂对某些种类的新真菌有可接受的控制作用,但对另一些则没有。在7种杀菌剂中,有5种杀菌剂的微生物测定结果与梨伤口接种的杀菌剂评价结果吻合较好,但在测定新真菌对新杀菌剂的敏感性方面,微生物测定结果不够一致。
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引用次数: 38
Are the microcyclic rust species Puccinia melampodii and Puccinia xanthii conspecific? 微环锈菌是同种锈菌吗?
Pub Date : 2009-11-01 DOI: 10.1016/j.mycres.2009.08.009
Marion K. Seier , Louise Morin , Marlien van der merwe , Harry C. Evans , Ángel Romero

The taxonomic relationship between two microcyclic rust species, Puccinia melampodii and Puccinia xanthii, recorded from a number of Asteraceae hosts, was explored by comparing teliospore morphology, host specificity and sequence data. Teliospore morphology varied between and within individual rust accessions but, in general, teliospores of P. xanthii were larger than those of P. melampodii. Field observations and cross-inoculation greenhouse studies showed that P. melampodii accessions were highly specific to their original host species and unable to establish compatible interactions with related Asteraceae species. Though small amounts of genetic variation were detected between P. melampodii and P. xanthii accessions, the ITS and TEF phylogenetic analyses comprising several other rust species grouped these accessions within a well supported clade. Our data indicate that despite differences between P. xanthii and P. melampodii accessions these rusts cannot readily be separated at the species level, supporting earlier taxonomic conclusions and the recent proposal to group them into a morphospecies. Based on host-specificity data, the name Puccinia xanthii var. parthenii-hysterophorae is proposed to accommodate all Mesoamerican records of P. melampodii associated with the host Parthenium hysterophorus.

通过对菊科不同寄主的端孢子形态、寄主特异性和序列数据的比较,探讨了两种微环锈病(Puccinia melampodii和Puccinia xanthii)的分类学关系。端孢子形态在不同的锈病株和不同的锈病株之间存在差异,但一般而言,黄氏霉的端孢子大于黑足霉。田间观察和交叉接种温室研究表明,黑足假单胞虫对其原寄主种具有高度特异性,无法与菊科近缘种建立亲和互作关系。虽然在黑足假单锈菌和黄腐假锈菌之间检测到少量遗传变异,但包括其他几种锈菌的ITS和TEF系统发育分析将这些菌株归为一个支持良好的分支。我们的数据表明,尽管黄腐斑孢锈菌和黑腐斑孢锈菌在物种水平上存在差异,但这些锈菌不能轻易地在物种水平上分开,这支持了早期的分类结论和最近将它们归类为形态种的建议。基于宿主特异性数据,我们提出将所有与宿主Parthenium hysterophorus相关的中美洲melampodii命名为Puccinia xanthii var. Parthenium -hysterophorae。
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引用次数: 23
期刊
Mycological research
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