拟盘多毛孢属真菌的新种*

报导拟盘多毛孢属真菌的8个新种。这8个新种是:近似状拟盘多毛孢Pestalotiopsis affinis、山姜拟盘多毛孢P. alpiniae、见血封喉拟盘多毛孢P. antiaris、五桠果拟盘多毛孢P. dilleniae、广西拟盘多毛孢P. kuwangsiensis、莲拟盘多毛孢P. nelumbinis、木荷拟盘多毛孢P. schimae、和神秘果拟盘多毛孢P. synsepali。新种的模式标本保存在广西大学植保系标本室。     

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.     

四川主要松树内生拟盘多毛孢

从5种松树健康的枝、叶、干中共分离了2,400个组织块,得到199株内生拟盘多毛孢Pestalotiopsis,形态学和分子生物学鉴定分属于11个种,平均分离率为8.29%。所得11种内生拟盘多毛孢都是首次在四川的松属植物中分离得到,其中葡萄生拟盘多毛孢P.uvicola和山桃草拟盘多毛孢P.gaurae为首次在松树上分离得到。基于ITS序列的分子系统发育分析显示松树内生拟盘多毛孢种类之间存在一定的遗传差异,部分内生拟盘多毛孢与病原拟盘多毛孢的ITS序列具有很高的同源性。     

内生拟盘多毛孢菌化学成分研究进展

内生真菌已经成为学者寻找具有显著活性药物的重要来源。本文对近年来从内生拟盘多毛孢菌(Pestalotiopsis spp.)中发现的活性代谢产物进行总结,按其结构特点主要包括了萜类,生物碱类,异香豆素和香豆素类,醌类和半醌类,色酮类和脂类等,并简要阐述了一些代谢产物的活性。     

蓝莓拟盘多毛孢枝枯病的病原菌

近期在山东省蓝莓种植区发现1种枝枯病害。为明确该致病菌,通过病原菌组织分离和接种试验获得2个致病菌株。通过形态学观察和rDNA‐ITS序列分析,确定该病原菌为棒状拟盘多毛孢Pestalotiopsis clavispora。这是棒状拟盘多毛孢所致蓝莓枝枯病在国内的首次报道。     

大熊猫体表一株海南拟盘多毛孢菌株

中国保护大熊猫研究中心雅安碧峰峡基地的一只大熊猫患皮肤病,从皮屑中分离到一株拟盘多毛孢菌GYMXP201108。进行了形态学观察和ITS序列分析,并构建系统进化树。结果表明,该菌株顶端附属丝短,1–3根,多数不具柄,与海南拟盘多毛孢Pestalotiopsis hainanensis模式菌株形态一致,ITS区序列分析也表明与海南拟盘多毛孢的序列相似性最高,为99.6%,因此鉴定为海南拟盘多毛孢,GenBank ID为KF551573。首次报道了海南拟盘多毛孢能在动物体表生存。     

一株具有高除草活性的真菌菌株

以从湖南张家界国家森林公园的发病毛竹上分离的一株病原真菌为研究材料,经生物测定发现该菌产生的粗毒素具有很强的除草活性,具有开发为生物除草剂的潜力。根据形态学特征和rDNA ITS测序分析,该菌为小孢拟盘多毛孢Pestalotiopsis microspora。     

一株产紫杉醇内生真菌YN6的分离及鉴定

从云南红豆杉（Taxus yunnanensis）树皮内表皮中分离得到75株内生真菌,采用基于紫杉醇合成关键酶10-去乙酰巴卡亭Ⅲ-10-O-乙酰基转移酶（10-deacetylbaccatin Ⅲ-10-O-acetyl transferase,DBAT）和C-13苯丙氨基侧链CoA乙酰基转移酶（C-13-phenylpropanoid side chain-CoA acyltransferase,BAPT）基因为标志分子的快速筛选方法获得一株可产紫杉醇的内生真菌YN6,并通过高效液相色谱法和质谱法对其紫杉醇进行分析.同时,通过对内生真菌YN6的形态特征分析以及18S rDNA序列分析将其初步鉴定为拟盘多毛孢属（Pestalotiopsis sp.）真菌. 拟盘多毛孢YN6的紫杉醇产量约为120~140 μg/L,是目前已报道的紫杉醇产量较高的野生菌株之一. 拟盘多毛孢YN6的发现为微生物发酵生产紫杉醇提供了潜在的优良种质资源.     

麻风树叶枯病菌的鉴定

作者于2009年在贵州麻风树栽培基地栽培的麻风树上发现一种叶部新病害,定名为麻风树叶枯病。该病主要危害植株叶片,叶片受害后初期产生椭圆形或不规则病斑,后期病斑连成片,常引起叶片过早脱落。从贵州罗甸麻风树栽培基地采集了16个叶枯病标样,经分离培养获得13个真菌分离物。通过致病性测定,证明菌株PE06为麻风树叶枯病的病原菌。通过形态学观察及其rDNA-ITS序列分析,将贵州麻风树叶枯病的病原菌鉴定为小孢拟盘多毛孢菌Pestalotiopsis microspora,这也是首次在麻风树叶片上发现由该病菌引起的病害。     

广西柳树桑寄生内生真菌的分离鉴定与抗肿瘤活性菌株筛选

该研究从广西钦州市采集健康的柳树桑寄生的根、茎和叶中分离并纯化内生真菌,对真菌进行形态学鉴定,提取内生真菌的DNA,采用真菌ITS序列对内生真菌进行分子鉴定。利用A549和H460细胞作为抗肿瘤活性指示细胞,采用MTT法测定真菌乙酸乙酯提取物的抗肿瘤活性。经过初步分离分析,从柳树桑寄生中纯化出27株内生真菌,经鉴定它们分别属于7个目9个属15个种。拟盘多毛孢属和间座壳属为优势属,其中拟盘多毛孢属全部定植于寄生根;其次为新壳梭孢属、拟茎点霉属和球座菌属,各分离到3株;其他包括青霉属、镰刀菌属、炭疽菌属和派伦霉属,各分离到1株。抗肿瘤活性研究表明,有一株与Pestalotiopsis protearum的ITS序列相似性达100%的拟盘多毛孢属菌株Gen24表现有抑制肿瘤细胞A549和H460生长的特性,在真菌乙酸乙酯浸提物浓度为800μg·mL~(-1)时,对A549细胞的抑制率达到了56.92%,对H460细胞的抑制率达到了70.11%。该研究结果表明广西柳树桑寄生内生真菌较丰富,在寄主中的分布表现了一定的组织特异性,而且还存在一些具有抗肿瘤活性的菌株及其活性物质可供进一步深入研究。     

Wasalexins A and B, new phytoalexins from wasabi: isolation, synthesis, and antifungal activity

The chemical structure determination of two phytoalexins from wasabi (Wasabia japonica, syn. Eutrema wasabi), a plant resistant to virulent isolates of the blackleg fungus [Leptosphaeria maculans (Desm.) Ces. et de Not., asexual stage Phoma lingam (Tode ex Fr.) Desm.], as well as their synthesis and antifungal activity towards isolates of P. lingam and P. wasabiae is reported.     

New sesquiterpenic phytotoxins establish unprecedented relationship between different groups of blackleg fungal isolates

A comprehensive search for sesquiterpenic metabolites produced by isolates of the blackleg fungus [Leptosphaeria maculans (Desm.)] Ces. et de Not. [asexual stage Phoma lingam (Tode ex Fr.) Desm.] revealed that an isolate pathogenic on both canola and brown mustard (IBCN 18) and two isolates pathogenic on brown mustard (Laird 2 and Mayfair 2) produced similar sesquiterpenes. The isolation, chemical structure elucidation, and phytotoxicity of these new sesquiterpenes with silphinene and selinene type skeletons is reported. This is the first time that an isolate virulent on canola and brown mustard is found to produce metabolites characteristic of both virulent (sirodesmins) and avirulent (phomalairdenones) L. maculans/P. lingam. In the context of grouping the various isolates of L. maculans/P. lingam, this work suggests an additional pathogenicity group comprising isolates that produce both sirodesmins and phomalairdenones and are virulent on both canola and brown mustard.     

Detoxification of the phytoalexin brassinin by isolates of Leptosphaeria maculans pathogenic on brown mustard involves an inducible hydrolase

Brassinin is a phytoalexin produced by plants from the family Brassicaceae that displays antifungal activity against a number of pathogens of Brassica species, including Leptosphaeria maculans (Desm.) Ces. et de Not. [asexual stage Phoma lingam (Tode ex Fr.) Desm.] and L. biglobosa. The interaction of a group of isolates of L. maculans virulent on brown mustard (Brassica juncea) with brassinin was investigated. The metabolic pathway for degradation of brassinin, the substrate selectivity of the putative detoxifying hydrolase, as well as the antifungal activity of metabolites and analogs of brassinin are reported. Brassinin hydrolase activity was detectable only in cell-free homogenates resulting from cultures induced with brassinin, N'-methylbrassinin, or camalexin. The phytoalexin camalexin was a substantially stronger inhibitor of these isolates than brassinin, causing complete growth inhibition at 0.5mM.     

The phytopathogenic fungi Leptosphaeria maculans and Leptosphaeria biglobosa: chemotaxonomical characterization of isolates and metabolite production in different culture media

Previous molecular chemotaxonomic analyses of isolates of the plant pathogenic fungus Leptosphaeria maculans (Desm.) Ces. et de Not. (asexual stage Phoma lingam (Tode ex Fr.) Desm.) in a chemically defined medium suggested that this species complex was composed of at least three distinct groups. Subsequently, a group within L. maculans was classified as Leptosphaeria biglobosa, on the basis of morphologic characteristics and the lack of sexual crossing. To obtain clarification regarding the metabolite profiles of the various groups or species of blackleg fungi, the objectives of this work were (i) to determine the chemical structures of metabolites produced by Canadian V isolates and Polish-type isolates in potato dextrose broth (PDB) and (ii) to determine the chemotaxonomic relationship among French isolates of L. biglobosa and among Canadian W isolates and Thlaspi isolates of L. maculans. Here, we report for the first time that Canadian V isolates grown in PDB produced 2,4-dihydroxy-3,6-dimethylbenzaldehyde, a metabolite never reported from L. maculans, but none of the usual phytotoxins (sirodesmins). In addition, we report a new metabolite, 2-[2-(5-hydroxybenzofuranyl)]-3-(4-hydroxyphenyl)propanenitrile, from Polish-type isolates of L. maculans grown in PDB and the metabolite profiles of 16 Thlaspi isolates. The metabolite profiles of Thlaspi isolates indicate that these are part of two distinct groups, the Polish W group and the Canadian W group, i.e., L. biglobosa. Finally, we demonstrate that the metabolite profiles of the French isolates classified as L. biglobosa are similar to those of Canadian W isolates.     

Phytoalexins from Thlaspi arvense, a wild crucifer resistant to virulent Leptosphaeria maculans: structures, syntheses and antifungal activity

Phytoalexins are inducible chemical defenses produced by plants in response to diverse forms of stress, including microbial attack. Our search for phytoalexins from cruciferous plants resistant to economically important fungal diseases led us to examine stinkweed or pennycress (Thlaspi arvense), a potential source of disease resistance to blackleg. We have investigated phytoalexin production in leaves of T. arvense under abiotic (copper chloride) and biotic elicitation by Leptosphaeria maculans (Desm.) Ces. et de Not. [asexual stage Phoma lingam (Tode ex Fr.) Desm.], and report here two phytoalexins, wasalexin A and arvelexin (4-methoxyindolyl-3-acetonitrile), their syntheses and antifungal activity against isolates of P. lingam/L. maculans, as well as the isolation of isovitexin, a constitutive glycosyl flavonoid of stinkweed, having antioxidant properties but devoid of antifungal activity.     

Phomapyrones from blackleg causing phytopathogenic fungi: isolation, structure determination, biosyntheses and biological activity

The isolation and structure determination of phomapyrones D-G, three 2-pyrones and a coumarin, from a group of isolates of the fungal pathogen Leptosphaeria maculans (Desm.) Ces. et de Not., asexual stage Phoma lingam (Tode ex Fr.) Desm, is reported. As well, phomenin B, infectopyrone, and polanrazines B and C were also obtained for the first time from these isolates. In addition, based on results of incorporations of 13C-labeled acetate and malonate, and deuterated methionine, a polyketide pathway is proposed for the biosyntheses of phomapyrones.     

Camalexin induces detoxification of the phytoalexin brassinin in the plant pathogen Leptosphaeria maculans

The impact of the phytoalexins camalexin and spirobrassinin on brassinin detoxification by Leptosphaeria maculans (Desm.) Ces. et de Not. [asexual stage Phoma lingam (Tode ex Fr.) Desm.], a pathogenic fungus prevalent on crucifers, was investigated. Brassinin is a plant metabolite of great significance due to its dual role both as an effective phytoalexin and as an early biosynthetic precursor of the majority of the phytoalexins produced by plants of the family Brassicaceae (Cruciferae). The rate of detoxification of brassinin in cultures of L. maculans increased substantially in the presence of camalexin, whereas spirobrassinin did not appear to have a detectable effect. In addition, the brassinin detoxifying activity of cell-free extracts obtained from cultures incubated with camalexin was substantially higher than that of control cell-free extracts or cultures incubated with spirobrassinin, and correlated positively with brassinin oxidase activity. The discovery of a potent synthetic modulator of brassinin oxidase activity, 3-phenylindole, and comparison with the commercial fungicide thiabendazole is also reported. The overall results indicate that brassinin oxidase production is induced by camalexin and 3-phenylindole but not by spirobrassinin or thiabendazole. Importantly, our work suggests that introduction of the camalexin pathway into plants that produce brassinin might make these plants more susceptible to L. maculans.     

Phomalairdenone: a new host-selective phytotoxin from a virulent type of the blackleg fungus Phoma lingam

The chemical structure and bioactivity of phomalairdenone (7), a new sesquiterpenic host-selective phytotoxin produced by an unusual virulent type isolate of the blackleg fungus [Phoma lingam, perfect stage Leptosphaeria maculans (Desm.) Ces. et de Not.] are reported.     

Isolation, structure determination, and phytotoxicity of unusual dioxopiperazines from the phytopathogenic fungus Phoma lingam.

The isolation, chemical structure elucidation, and bioactivity of polanrazines B-F, five new dioxopiperazines produced by isolates of the blackleg fungus [Phoma lingam, perfect stage Leptosphaeria maculans (Desm.) Ces. et de Not.] originating from Poland, are reported. Polanrazines C and E showed moderate but selective toxicity, causing necrotic and chlorotic lesions (1-3 mm diameter) on brown mustard leaves.     

HPLC analyses of cultures of Phoma spp.: differentiation among groups and species through secondary metabolite profiles

The metabolite profiles of 26 isolates of the blackleg fungus (Leptosphaeria maculans (Desm.) Ces. et de Not., asexual stage Phoma lingam (Tode ex Fr.) Desm.), obtained from diverse parts of the world (part of the International Blackleg Crucifer Network collection), were studied utilizing specific culture conditions, HPLC analysis, and a set of chemical markers. This fungus is the causative agent of blackleg disease of brassica oilseeds; a virulent strain of the pathogen has caused significant rapeseed (Brassica napus L., and B. rapa L.) and canola (B. napus L., and B. rapa L.) losses in Canada, and is also considered a serious agricultural problem worldwide. Effective surveys of blackleg epidemics require simple and reliable analytical methodology to differentiate among the diverse groups of isolates. The chemical analysis of phytotoxins and related secondary metabolites is perhaps one of the most discriminating and the least ambiguous methods for differentiation of Phoma blackleg isolates. Following HPLC analyses, the 26 isolates could be placed in three main groups, irrespective of country of origin: isolates producing phomamide and sirodesmins, isolates producing indolyl dioxopiperazines, and isolates producing polyketides. Discussion of the implications of our findings and suggestions for species reclassification are provided.