BACKGROUND & AIMS: ETHNOPHARMACOLOGICAL RELEVANCE:Tinospora cordifolia (Willd. Hook. f. & Thomson; family: Menispermaceae), has a long history of use in various traditional medicinal systems including "Ayurveda". It is reported to possess anticancer, antidiabetic, antimicrobial, antispasmodic, and antiinflammatory activities. T. cordifolia has also been well documented for production of various bioactive metabolites and their antioxidant activity, but the microorganisms associated with it have been least explored for the same properties. AIM OF THE STUDY:Aim of the present study was to evaluate antioxidant and in vivo genoprotective potential of phenolic compounds produced by an endophytic fungus Cladosporium velox TN-9S isolated from T. cordifolia. MATERIALS AND METHODS:The isolate of C. velox TN-9S was cultivated in malt extract medium and extracted with ethyl acetate. Total phenol content was determined by Folin Ciocalteu reagent. The antioxidant activity was measured in terms of DPPH and FRAP assay. The in vivo genoprotective activity was assessed using fish Channa punctatus as model. Identification of phenolic compounds was carried out using RP-HPLC. The fungal extract was evaluated for biosafety using Salmonella typhimurium His- strain and CHO cell lines for mutagenicity and cytotoxicity, respectively. RESULTS:The total phenolic content in the ethyl acetate extract of the fungus was determined to be 730μg gallic acid equivalent/mL. The extract evinced significant antioxidant activity with IC50 value of 22.5µg/mL in DPPH scavenging assay. The phenolic extract showed good in vivo genoprotective activity against the genetic damage induced in fish C. punctatus after treatment with a non-ionic surfactant 4-nonylphenol. RP-HPLC analysis revealed the presence of peaks corresponding to various phenolic compounds in the extract. Mutagenicity and cytotoxicity results revealed the extract to be nonmutagenic and non cytotoxic in nature. CONCLUSION:The results indicate the potential of an endophytic C. velox isolated from T. cordifolia as a producer of phenolic compounds with antioxidant and genoprotective activities which could be exploited in pharmaceutical industry. The ability of endophytes to produce similar compounds as the host, is also revealed in the present study.

背景与目标：

BACKGROUND & AIMS: :A 26-year-old woman currently treated for systemic lupus erythematosus with steroid therapy presented with sudden onset of right hemiplegia. Computed tomography of the brain showed a large frontoparietal ring-enhanced lesion with perifocal edema. Stereotactic aspiration of the lesion revealed Cladosporium bantianum. The size of the abscess did not reduce in spite of optimum antifungal treatment. The abscess was subsequently excised through a frontoparietal craniotomy. At follow up after 24 months, there was no recurrence of the abscess. Cerebral Cladosporium bantianum infection is usually refractory to antifungal agents and the prognosis is very poor. This patient had the longest survival period in a case of Cladosporium brain abscess so far reported.

背景与目标： : 一名26岁的妇女目前接受类固醇治疗的系统性红斑狼疮，表现为右偏瘫突然发作。大脑的计算机断层扫描显示大额顶环形增强病变并伴有病灶周围水肿。病变的立体定向抽吸显示为bantianum枝孢菌。尽管进行了最佳的抗真菌治疗，但脓肿的大小并未减少。随后通过额顶开颅手术切除脓肿。24个月后随访，无脓肿复发。bantianum脑枝孢菌感染通常对抗真菌药物难治，预后很差。该患者在迄今为止报道的枝孢菌性脑脓肿病例中生存期最长。

BACKGROUND & AIMS: :Effectors are microbial-derived secreted proteins with an essential function in modulating host immunity during infections. CfAvr4, an effector protein from the tomato pathogen Cladosporium fulvum and the founding member of a fungal effector family, promotes parasitism through binding fungal chitin and protecting it from chitinases. Binding of Avr4 to chitin is mediated by a carbohydrate-binding module of family 14 (CBM14), an abundant CBM across all domains of life. To date, the structural basis of chitin-binding by Avr4 effector proteins and of recognition by the cognate Cf-4 plant immune receptor are still poorly understood. Using X-ray crystallography, we solved the crystal structure of CfAvr4 in complex with chitohexaose [(GlcNAc)6] at 1.95Å resolution. This is the first co-crystal structure of a CBM14 protein together with its ligand that further reveals the molecular mechanism of (GlcNAc)6 binding by Avr4 effector proteins and CBM14 family members in general. The structure showed that two molecules of CfAvr4 interact through the ligand and form a three-dimensional molecular sandwich that encapsulates two (GlcNAc)6 molecules within the dimeric assembly. Contrary to previous assumptions made with other CBM14 members, the chitohexaose-binding domain (ChBD) extends to the entire length of CfAvr4 with the reducing end of (GlcNAc)6 positioned near the N-terminus and the non-reducing end at the C-terminus. Site-directed mutagenesis of residues interacting with (GlcNAc)6 enabled the elucidation of the precise topography and amino acid composition of Avr4's ChBD and further showed that these residues do not individually mediate the recognition of CfAvr4 by the Cf-4 immune receptor. Instead, the studies highlighted the dependency of Cf-4-mediated recognition on CfAvr4's stability and resistance against proteolysis in the leaf apoplast, and provided the evidence for structurally separating intrinsic function from immune receptor recognition in this effector family.

背景与目标： : 效应物是微生物来源的分泌蛋白，在感染过程中具有调节宿主免疫力的重要功能。CfAvr4是番茄病原体fulvum Cladosporium的效应蛋白，也是真菌效应家族的创始成员，通过结合真菌几丁质并保护其免受几丁质酶的侵害来促进寄生。Avr4与几丁质的结合是由家族14 (CBM14) 的碳水化合物结合模块介导的，该模块是生命所有领域的丰富CBM。迄今为止，对Avr4效应蛋白结合几丁质和同源Cf-4植物免疫受体识别的结构基础仍知之甚少。使用x射线晶体学，我们以1.95的分辨率解决了CfAvr4与壳聚糖六糖 [(GlcNAc)6] 配合物的晶体结构。这是CBM14蛋白及其配体的第一个共晶体结构，进一步揭示了Avr4效应蛋白和CBM14家族成员结合 (GlcNAc)6的分子机制。结构表明，两个CfAvr4分子通过配体相互作用并形成三维分子三明治，该三维分子三明治将两个 (GlcNAc)6分子封装在二聚组件中。与先前对其他CBM14成员所做的假设相反，壳聚糖六糖结合结构域 (ChBD) 延伸到CfAvr4的整个长度，(GlcNAc)6的还原端位于N端附近，而非还原端位于C端。与 (GlcNAc)6相互作用的残基的定点诱变能够阐明Avr4的ChBD的精确形貌和氨基酸组成，并进一步表明这些残基不能单独介导Cf-4免疫受体对CfAvr4的识别。相反，研究强调了Cf-4-mediated识别对CfAvr4的稳定性和对叶片质外体中蛋白水解的抵抗力的依赖性，并提供了在该效应子家族中将内在功能与免疫受体识别在结构上分离的证据。

BACKGROUND & AIMS: :Hydrophobins are amphipathic molecules which form part of fungal cell walls and extracellular matrices and perform a variety of roles in fungal growth and development. The tomato pathogen Cladosporium fulvum has six hydrophobin genes, HCf-1 to -6. We have devised an epitope tagging approach for establishing hydrophobin localization during growth in culture and in plants. In this paper we localize HCf-2, -3, -4 and -5 and compare the data to our previous observations for HCf-1 and -6. In culture, HCf-1, -2, -3 and 4 localize to conidia and also appear on aerial hyphae. HCf-4 is unique in that it appears on submerged hyphae. HCf-5 expression is tightly regulated and appears on aerial hyphae early on during growth. Only HCf-1, -3 and -6 were observed during infection; HCf-3 appears on both conidia and emerging germ tubes. We also show that HCf-6 is secreted and coats surfaces under and around growing hyphae and demonstrate the effect of deleting HCf-6 on the adhesion of germinating C. fulvum conidia to glass slides.

背景与目标： 疏水蛋白是两亲性分子，形成真菌细胞壁和细胞外基质的一部分，并在真菌的生长和发育中起多种作用。番茄病原菌叶枝孢菌有6个疏水蛋白基因，HCf-1为-6。我们设计了一种表位标记方法，用于在培养物和植物的生长过程中建立疏水蛋白定位。在本文中，我们定位了HCf-2，-3，-4和-5，并将数据与我们先前对HCf-1和-6的观察结果进行了比较。在培养物中，HCf-1，-2，-3和4定位于分生孢子，也出现在空中菌丝上。HCf-4的独特之处在于它出现在淹没的菌丝上。HCf-5表达受到严格调节，并在生长早期出现在气生菌丝上。在感染期间仅观察到HCf-1，-3和-6; HCf-3出现在分生孢子和新出现的芽管上。我们还表明，HCf-6是分泌的，并在生长的菌丝下面和周围覆盖表面，并证明了删除HCf-6对发芽的C. fulvum分生孢子与载玻片的粘附性的影响。

BACKGROUND & AIMS: :Tomato (Lycopersicon esculentum Mill.) is an important vegetable crop grown in Liaoning Province, China. In May 2012, dark brown, angular lesions were observed on lower, older leaves of 5-month-old tomato plants of cv. Bi Jiao in commercial greenhouses in Dawa County, which is administered by Panjin City in Liaoning Province. The irregularly shaped lesions varied in size from 1 to 7 mm in diameter. The necrotic lesions were usually surrounded by a yellow halo. Sporulation was rarely seen on the leaf surfaces. This contrasts with tomato leaf mold caused by Passalora fulva, in which the conidia develop as a velvety brown patch in lesions. By July 2012, the same disease was found in research greenhouses of Shenyang Agriculture University and Liaoning Academy of Agricultural Sciences. The incidence of symptoms was 30 to 40%. To identify the pathogen, leaf pieces (3 to 5 mm) with both infected and healthy portions were taken at the edge of lesions and surface-disinfected by placing them in 75% ethanol for 5 s, then transferred to a 0.1% aqueous mercuric chloride solution for 30 s and rinsed with sterilized water three times. The sections were placed on potato dextrose agar (PDA) at 25°C in the dark. Ten pure fungal cultures were obtained from single spores. For studies of microscopic morphology, isolates were grown on synthetic nutrient agar (SNA) in slide cultures. Conidia ranged in shape from subglobose or ovoid (2 to 4 × 2 to 3 μm) to subcylindrical (4.5 to 17.8 × 2.4 to 4.5 μm). Conidiophores were straight to slightly flexuous with typical nodes. The internal transcribed spacer (ITS) region from isolate PJ12-36 was amplified using primers ITS1 and ITS4 and sequenced (GenBank Accession No. KC137278). The 560-bp amplicons had 99% identity to C. oxysporum (JQ775499). On the basis of morphological characteristics (1) and nucleotide homology, the isolate was identified as C. oxysporum. Koch's postulates were fulfilled in the laboratory on fully expanded leaves of 5-week-old tomato plants 'Moneymaker' inoculated with C. oxysporum conidial suspensions (107 conidia ml-1). Eight seedlings were incubated in an illuminated incubator at 25°C for 8 to 10 days with 85% relative humidity. Characteristic lesions that developed on inoculated leaves were similar in appearance to those observed on diseased tomato plants in the greenhouse. C. oxysporum was reisolated from lesions and its identity was confirmed by morphological characteristics. C. oxysporum was previously reported as the causal agent of a leaf spot disease of pepper (2) and greenhouse tomato (4). To our knowledge, this is the first report of C. oxysporum causing disease on tomato foliage in China. It is noteworthy that C. oxysporum has led to a decline in pepper yield in northern regions of China (3). In addition, pathogenicity tests showed that the isolate W10-02 from pepper and the isolate PJ12-36 from tomato could each cause damage to the opposite host, producing symptoms similar to those observed on the host of origin. Studies are needed on strategies to manage C. oxysporum in crops, because its prevalence may cause yield loss both on tomato and pepper in northern regions of China. References: (1) K. Bensch et al. Stud. Mycol. 67:1, 2010. (2) A. M. Hammouda. Plant Dis. 76:536, 1992. (3) X. Y. Huang et al. Plant Dis. 96:1072, 2012. (4) J. S. Lamboy and H. R. Dillard. Plant Dis. 81:228, 1997.

背景与目标： : 番茄 (Lycopersicon esculentum Mill。) 是中国辽宁省种植的重要蔬菜作物。在2012年5月中，在5个月大的cv番茄植株的下部，较老的叶子上观察到深棕色的角状病变。辽宁省盘锦市管辖的大洼县商业大棚中的毕娇。不规则形状的病变的直径从1到7毫米不等。坏死性病变通常被黄色光环包围。在叶片表面很少见到孢子形成。这与Passalora fulva引起的番茄叶霉菌形成对比，其中分生孢子在病变中发展为天鹅绒般的棕色斑块。经2012年7月，在沈阳农业大学和辽宁省农业科学院的研究温室中发现了相同的疾病。症状发生率为30 ~ 40%。为了鉴定病原体，将具有感染和健康部分的叶块 (3至5毫米) 在病变边缘处取出，并通过将其置于75% 乙醇中5 s进行表面消毒，然后转移到0.1% 的氯化汞水溶液中30 s，并用无菌水冲洗三次。将切片在黑暗中于25 °C放置在马铃薯葡萄糖琼脂 (PDA) 上。从单个孢子中获得十种纯真菌培养物。为了研究微观形态，在玻片培养物中的合成营养琼脂 (SNA) 上生长了分离株。分生孢子的形状范围从近球形或卵形 (2至4 × 2至3μm) 到近圆柱形 (4.5至17.8 × 2.4至4.5 μ m)。分生孢子笔直至略带弯曲，具有典型的节。使用引物ITS1和ITS4扩增来自分离PJ12-36的内部转录间隔区 (ITS) 区域并测序 (GenBank登录号KC137278)。560 bp扩增子与尖孢菌 (JQ775499) 具有99% 的同一性。根据形态学特征 (1) 和核苷酸同源性，分离株被鉴定为尖孢杆菌。Koch的假设是在实验室中对5周龄番茄植物 “moneymaker” 的完全膨胀的叶子进行的，该叶子接种了C. oxysporum分生孢子悬浮液 (107 conidi 1毫升-1)。将八个幼苗在25 °C的照明培养箱中于85% 相对湿度下孵育8至10天。在接种的叶子上形成的特征性病变在外观上与在温室中患病的番茄植物上观察到的相似。从病变中重新分离出C. oxysporum，并通过形态学特征确认其身份。C. oxysporum以前被报道为辣椒 (2) 和温室番茄 (4) 的叶斑病的病原体。据我们所知，这是中国首次报道的C. oxysporum引起番茄叶片疾病。值得注意的是，尖孢菌导致中国北方地区辣椒产量下降 (3)。此外，致病性测试表明，从胡椒中W10-02的分离物和从番茄中PJ12-36的分离物都可能对相反的宿主造成损害，产生与在来源宿主上观察到的症状相似的症状。需要对农作物中处理C. oxysporum的策略进行研究，因为它的流行可能会导致中国北部地区番茄和辣椒的单产损失。参考文献 :( 1) K。Bensch等人。Mycol。67:1，2010。(2) A.M。哈姆穆达。植物Dis。76:536，1992。(3) X。Y。黄等人。植物Dis。96:1072，2012。(4) J。S。Lamboy和h.r.迪拉德。植物Dis。81:228，1997。

BACKGROUND & AIMS: :The purpose of this study was to examine the usefulness of electron paramagnetic resonance spectroscopy (EPR) to estimate zinc and copper ions biosorption from the environment by pigmented soil fungi Cladosporium cladosporioides. The existence of a low amount of pheomelanin, besides eumelanin, in C. cladosporioides samples was proved by the analysis of shape of their EPR spectra. Concentration of o-semiquinone free radicals in crude mycelium was 2.4x10(17) spin/g. Changes in free radicals system of C. cladosporioides cultured in the presence of Zn2+ and Cu2+ were analysed. Both magnetic and chemical interactions of zinc and copper ions with free radicals in C. cladosporioides melanin were found. Magnetically interacting diamagnetic Zn2+ ions increased the concentration of o-semiquinone free radicals in melanin existing in C. cladosporioides mycelium, whereas paramagnetic Cu2+ ions decreased this concentration. Chemical interactions of Zn2+ and Cu2+ ions decreased the free radical concentrations in C. cladosporioides melanin. Homogeneously distributed free radicals in C. cladosporioides melanin rise its activity in biosorption processes.

背景与目标： : 这项研究的目的是检查电子顺磁共振波谱 (EPR) 估算有色土壤真菌Cladosporium cladosporioides对环境中锌和铜离子生物吸附的有用性。通过分析其EPR光谱的形状，证明了除真黑色素外，在枝孢菌素样品中还存在少量的pheomelanin。粗菌丝体中o-半醌自由基的浓度为2.4x10(17) spin/g。分析了在Zn2和Cu2存在下培养的枝孢杆菌自由基系统的变化。发现了枝孢杆菌黑色素中锌和铜离子与自由基的磁性和化学相互作用。磁性相互作用的抗磁性Zn2离子增加了C. cladosporioides菌丝体中存在的黑色素中o-半醌自由基的浓度，而顺磁性Cu2离子则降低了该浓度。Zn2和Cu2离子的化学相互作用降低了C. cladosporioides黑色素中的自由基浓度。黑色素C. cladosporioides中均匀分布的自由基增加了其在生物吸附过程中的活性。

BACKGROUND & AIMS: :We sequenced and compared the genomes of the Dothideomycete fungal plant pathogens Cladosporium fulvum (Cfu) (syn. Passalora fulva) and Dothistroma septosporum (Dse) that are closely related phylogenetically, but have different lifestyles and hosts. Although both fungi grow extracellularly in close contact with host mesophyll cells, Cfu is a biotroph infecting tomato, while Dse is a hemibiotroph infecting pine. The genomes of these fungi have a similar set of genes (70% of gene content in both genomes are homologs), but differ significantly in size (Cfu >61.1-Mb; Dse 31.2-Mb), which is mainly due to the difference in repeat content (47.2% in Cfu versus 3.2% in Dse). Recent adaptation to different lifestyles and hosts is suggested by diverged sets of genes. Cfu contains an α-tomatinase gene that we predict might be required for detoxification of tomatine, while this gene is absent in Dse. Many genes encoding secreted proteins are unique to each species and the repeat-rich areas in Cfu are enriched for these species-specific genes. In contrast, conserved genes suggest common host ancestry. Homologs of Cfu effector genes, including Ecp2 and Avr4, are present in Dse and induce a Cf-Ecp2- and Cf-4-mediated hypersensitive response, respectively. Strikingly, genes involved in production of the toxin dothistromin, a likely virulence factor for Dse, are conserved in Cfu, but their expression differs markedly with essentially no expression by Cfu in planta. Likewise, Cfu has a carbohydrate-degrading enzyme catalog that is more similar to that of necrotrophs or hemibiotrophs and a larger pectinolytic gene arsenal than Dse, but many of these genes are not expressed in planta or are pseudogenized. Overall, comparison of their genomes suggests that these closely related plant pathogens had a common ancestral host but since adapted to different hosts and lifestyles by a combination of differentiated gene content, pseudogenization, and gene regulation.

背景与目标： : 我们对真菌真菌植物病原体的基因组进行了测序和比较，它们在系统发育上密切相关，但具有不同的生活方式和宿主。尽管两种真菌都与宿主叶肉细胞密切接触者在细胞外生长，但Cfu是一种生物滋养植物，可感染番茄，而Dse是一种半生物滋养植物，可感染松树。这些真菌的基因组具有相似的一组基因 (两个基因组中基因含量的70% 是同源物)，但大小差异显著 (Cfu >61.1-Mb; Dse 31.2-Mb)，这主要是由于重复内容的差异 (Cfu中的47.2% 与Dse中的3.2%)。不同的基因组建议最近适应不同的生活方式和宿主。Cfu包含一个 α-番茄酶基因，我们预测该基因可能是番茄碱解毒所必需的，而Dse中不存在该基因。每个物种都有许多编码分泌蛋白的基因，Cfu中重复序列丰富的区域富含这些物种特异性基因。相反，保守基因提示共同的宿主血统。Cfu效应基因 (包括Ecp2和Avr4) 的同源物存在于Dse中，分别诱导Cf-Ecp2和Cf-4-mediated超敏反应。令人惊讶的是，与毒素dothistromin (可能是Dse的毒力因子) 产生有关的基因在Cfu中是保守的，但它们的表达明显不同，而Cfu在植物中基本上没有表达。同样，Cfu的碳水化合物降解酶目录与坏死性或半生物营养物更相似，并且比Dse具有更大的果胶分解基因库，但是其中许多基因在植物中不表达或被假基因化。总体而言，它们的基因组比较表明，这些密切相关的植物病原体具有共同的祖先宿主，但由于通过区分基因含量，假基因化和基因调控的组合适应了不同的宿主和生活方式。

BACKGROUND & AIMS: :As part of a worldwide survey of the indoor mycobiota about 520 new Cladosporium isolates from indoor environments mainly collected in China, Europe, New Zealand, North America and South Africa were investigated by using a polyphasic approach to determine their species identity. All Cladosporium species occurring in indoor environments are fully described and illustrated. Fourty-six Cladosporium species are treated of which 16 species are introduced as new. A key for the most common Cladosporium species isolated from indoor environments is provided. Cladosporium halotolerans proved to be the most frequently isolated Cladosporium species indoors.

背景与目标： : 作为对室内真菌生物群进行的全球调查的一部分，通过使用多相方法确定其物种身份，对主要在中国，欧洲，新西兰，北美和南非收集的来自室内环境的约520种新的枝孢菌进行了调查。对室内环境中发生的所有枝孢属物种进行了全面描述和说明。处理了46种枝孢属物种，其中16种被引入新物种。提供了从室内环境中分离出来的最常见的枝孢菌物种的关键。被证明是室内最常分离的枝孢菌属物种。

BACKGROUND & AIMS: :Cladosporium cladosporioides is a dematiaceous fungus with coloured mycelia and conidia due to the presence of dark pigments. The purpose of this study was to characterize the dark pigments synthetized by Cladosporium sp. LPSC no. 1088 and also to identify the putative polyketide synthase (pks) gene that might be involved in the pigment biosynthesis. Morphological as well as molecular features like the ITS sequence confirmed that LPSC 1088 is Cladosporium cladosporioides. UV-visible, Fourier Transform Infrared (FTIR) and Electron Spin Resonance (ESR) spectroscopy analysis as well as melanin inhibitors suggest that the main dark pigment of the isolate was 1,8 dihydroxynaphthalene (DHN)-melanin-type compound. Two commercial fungicides, Difenoconazole and Chlorothalonil, inhibited fungal growth as well as increased pigmentation of the colonies suggesting that melanin might protect the fungus against chemical stress. The pigment is most probably synthetized by means of a pentaketide pathway since the sequence of a 651 bp fragment, coding for a putative polyketide synthase, is highly homologous to pks sequences from other fungi.

背景与目标： : Cladosporium cladosporioides是一种灰质真菌，由于存在深色色素，具有有色菌丝体和分生孢子。这项研究的目的是表征由Cladosporium sp. LPSC no.1088合成的深色色素，并鉴定可能参与色素生物合成的推定聚酮化合物合酶 (pks) 基因。形态以及类似ITS序列的分子特征证实了LPSC 1088是枝孢菌属。紫外可见、傅里叶变换红外 (FTIR) 和电子自旋共振 (ESR) 光谱分析以及黑色素抑制剂表明，分离物的主要深色色素是1,8二羟基萘 (DHN)-黑色素型化合物。两种商业杀菌剂，苯醚甲康唑和百菌清，抑制了真菌的生长，并增加了菌落的色素沉着，这表明黑色素可以保护真菌免受化学胁迫。由于编码推定的聚酮化合物合酶的651 bp片段的序列与来自其他真菌的pks序列高度同源，因此最有可能通过喷酮肽途径合成色素。

BACKGROUND & AIMS: :Microorganisms based biosynthesis of nanomaterials has triggered significant attention, due to their great potential as vast source of the production of biocompatible nanoparticles (NPs). Such biosynthesized functional nanomaterials can be used for various biomedical applications. The present study investigates the green synthesis of silver nanoparticles (Ag NPs) using the fungus Curvularia pallescens (C. pallescens) which is isolated from cereals. The C. pallescens cell filtrate was used for the reduction of AgNO3 to Ag NPs. To the best of our knowledge C. pallescens is utilized first time for the preparation of Ag NPs. Several alkaloids and proteins present in the phytopathogenic fungus C. pallescens were mainly responsible for the formation of highly crystalline Ag NPs. The as-synthesized Ag NPs were characterized by using UV-Visible spectroscopy, X-ray diffraction and transmission electron microscopy (TEM). The TEM micrographs have revealed that spherical shaped Ag NPs with polydisperse in size were obtained. These results have clearly suggested that the biomolecules secreted by C. pallescens are mainly responsible for the formation and stabilization of nanoparticles. Furthermore, the antifungal activity of the as-prepared Ag NPs was tested against Cladosporium fulvum, which is the major cause of a serious plant disease, known as tomato leaf mold. The synthesized Ag NPs displayed excellent fungicidal activity against the tested fungal pathogen. The extreme zone of reduction occurred at 50 μL, whereas, an increase in the reduction activity is observed with increasing the concentration of Ag NPs. These encouraging results can be further exploited by employing the as synthesized Ag NPs against various pathogenic fungi in order to ascertain their spectrum of fungicidal activity.

背景与目标： : 基于微生物的纳米材料的生物合成已引起广泛关注，因为它们具有作为生物相容性纳米颗粒 (NPs) 生产的巨大潜力。这种生物合成的功能性纳米材料可用于各种生物医学应用。本研究使用从谷物中分离出的真菌弯孢菌 (C. pallescens) 研究了银纳米颗粒 (Ag NPs) 的绿色合成。使用C. pallescens细胞滤液将AgNO3还原为Ag np。据我们所知，C. pallescens首次用于制备Ag NPs。植物病原真菌C. pallescens中存在的几种生物碱和蛋白质主要负责形成高度结晶的Ag np。通过紫外可见光谱，x射线衍射和透射电子显微镜 (TEM) 对合成的Ag np进行了表征。TEM显微照片显示，获得了尺寸多分散的球形Ag np。这些结果清楚地表明，由C. pallescens分泌的生物分子主要负责纳米颗粒的形成和稳定。此外，还测试了所制备的Ag NPs对fulvum的抗真菌活性，这是严重的植物病害 (称为番茄叶霉病) 的主要原因。合成的Ag np对测试的真菌病原体具有出色的杀菌活性。还原的极端区域发生在50μl，而随着Ag NPs浓度的增加，还原活性增加。通过使用as合成的Ag np对抗各种病原真菌，可以进一步利用这些令人鼓舞的结果，以确定其杀菌活性谱。

BACKGROUND & AIMS: :The presence of Cladosporium fulvum (syn. Passalora fulva), causal agent of tomato leaf mold, was confirmed in the two main greenhouse-production areas for tomato in Argentina. Using both morphological characters and internal transcribed spacer sequencing, we confirmed the presence of physiological races of this pathogen. A diagnostic multiplex polymerase chain reaction (PCR) was also developed, using primers derived from C. fulvum avirulence (Avr) genes. In all, 20 isolates of Cladosporium spp. were obtained as monospore cultures and 12 were identified as C. fulvum. By this method, we showed that, of these 12 isolates, 5 were race 0 (carrying functional Avr2, Avr4, Avr4E, and Avr9 genes) and 7 were race 2 (lacking the Avr2 gene). Race identity was confirmed by testing their virulence on a set of tomato differentials carrying different Cf resistance genes. All Avr genes could be amplified in single or multiplex PCR using DNA isolated from in vitro grown monospore cultures but only three Avr could be amplified when genomic DNA was isolated from C. fulvum-infected necrotic leaf tissue.

背景与目标： : 在阿根廷的两个主要的番茄温室生产区中，证实了番茄叶霉病的成因叶枝孢 (syn. Passalora fulva) 的存在。使用形态学特征和内部转录间隔区测序，我们证实了该病原体的生理小种的存在。还使用源自黄连无毒性 (Avr) 基因的引物开发了诊断性多重聚合酶链反应 (PCR)。共分离出20株枝孢菌。获得了单孢子培养物，并鉴定了12种为C。fulvum。通过这种方法，我们发现，在这12个分离株中，有5个是0种族 (携带功能性Avr2，Avr4，Avr4E和Avr9基因)，而7个是2种族 (缺少Avr2基因)。通过在一组携带不同Cf抗性基因的番茄差异上测试其毒力，确认了种族身份。使用从体外生长的单孢子培养物中分离的DNA，可以在单次或多重PCR中扩增所有Avr基因，但是当从感染了黄连的坏死叶组织中分离基因组DNA时，只能扩增三个Avr。

BACKGROUND & AIMS: :Disease resistance in plants is commonly activated by the product of an avirulence (Avr) gene of a pathogen after interaction with the product of a matching resistance (R) gene in the host. In susceptible plants, Avr products might function as virulence or pathogenicity factors. The AVR9 elicitor from the fungus Cladosporium fulvum induces defense responses in tomato plants carrying the Cf-9 resistance gene. This 28-residue beta-sheet AVR9 peptide contains three disulfide bridges, which were identified in this study as Cys2-Cys16, Cys6-Cys19, and Cys12-Cys26. For this purpose, AVR9 was partially reduced, and the thiol groups of newly formed cysteines were modified to prevent reactions with disulfides. After HPLC purification, the partially reduced peptides were sequenced to determine the positions of the modified cysteines, which originated from the reduced disulfide bridge(s). All steps involving molecules with free thiol groups were performed at low pH to suppress disulfide scrambling. For that reason, cysteine modification by N-ethylmaleimide was preferred over modification by iodoacetamide. Upon (partial) reduction of native AVR9, the Cys2-Cys16 bridge opened selectively. The resulting molecule was further reduced to two one-bridge intermediates, which were subsequently completely reduced. The (partially) reduced cysteine-modified AVR9 species showed little or no necrosis-inducing activity, demonstrating the importance of the disulfide bridges for biological activity. Based on peptide length and cysteine spacing, it was previously suggested that AVR9 isa cystine-knotted peptide. Now, we have proven that the bridging pattern of AVR9 is indeed identical to that of cystine-knotted peptides. Moreover, NMR data obtained for AVR9 show that it is structurally closely related to the cystine-knotted carboxypeptidase inhibitor. However, AVR9 does not show any carboxypeptidase inhibiting activity, indicating that the cystine-knot fold is a commonly occurring motif with varying biological functions.

背景与目标： : 植物中的抗病性通常在与宿主中匹配抗性 (R) 基因的产物相互作用后，由病原体的无毒 (Avr) 基因的产物激活。在易感植物中，Avr产物可能起着毒力或致病性因子的作用。来自真菌枯枝菌的AVR9激发子在携带Cf-9抗性基因的番茄植物中诱导防御反应。该28个残基 β-折叠AVR9肽包含三个二硫键，在本研究中被鉴定为Cys2-Cys16，Cys6-Cys19和Cys12-Cys26。为此，AVR9被部分还原，并且新形成的半胱氨酸的硫醇基团被修饰以防止与二硫化物反应。HPLC纯化后，对部分还原的肽进行测序以确定修饰的半胱氨酸的位置，该半胱氨酸源自还原的二硫键。所有涉及具有游离硫醇基团的分子的步骤均在低pH下进行，以抑制二硫键加扰。因此，与碘乙酰胺修饰不同，N-乙基马来酰亚胺修饰半胱氨酸是优选的。在 (部分) 减少天然AVR9时，Cys2-Cys16桥选择性地打开。所得分子进一步还原为两个一桥中间体，随后完全还原。(部分) 还原的半胱氨酸修饰的AVR9物种几乎没有或没有坏死诱导活性，表明二硫键对生物活性的重要性。根据肽长度和半胱氨酸间距，以前曾建议AVR9是胱氨酸结肽。现在，我们已经证明AVR9的桥接模式确实与胱氨酸结肽的桥接模式相同。此外，为AVR9获得的NMR数据表明，它在结构上与胱氨酸结羧肽酶抑制剂密切相关。然而，AVR9没有显示出任何羧肽酶抑制活性，表明胱氨酸结折叠是具有不同生物学功能的常见基序。

BACKGROUND & AIMS: :The metalworking industry is responsible for one of the most complex and difficult to handle oily effluents. These effluents consist of cutting fluids, which provide refrigeration and purification of metallic pieces in the machining system. When these effluents are biologically treated, is important to do this with autochthonous microorganisms; the use of these microorganisms (bioaugmentation) tends to be more efficient because they are already adapted to the existing pollutants. For this purpose, this study aimed to use two indigenous microorganisms, Epicoccum nigrum and Cladosporium sp. for metalworking effluent treatment using an air-lift reactor; the fungus Aspergillus niger (laboratory strain) was used as a reference microorganism. The original effluent characterization presented considerable pollutant potential. The color of the effluent was 1495 mg Pt/L, and it contained 59 mg/L H2O2, 53 mg/L total phenols, 2.5 mgO2/L dissolved oxygen (DO), and 887 mg/L oil and grease. The COD was 9147 mgO2/L and the chronic toxicity factor was 1667. Following biotreatment, the fungus Epicoccum nigrum was found to be the most efficient in reducing (effective reduction) the majority of the parameters (26% COD, 12% H2O2, 59% total phenols, and 40% oil and grease), while Cladosporium sp. was more efficient in color reduction (77%).

背景与目标： : 金属加工行业是最复杂，最难处理的油性废水之一。这些废水由切削液组成，可在加工系统中提供金属零件的制冷和净化。当对这些废水进行生物处理时，对本地微生物进行此处理很重要; 这些微生物的使用 (生物增强) 往往更有效，因为它们已经适应了现有的污染物。为此，本研究旨在使用两种本地微生物，即epicoccupm nigrum和Cladosporium sp。使用气举反应器进行金属加工废水处理; 将黑曲霉 (实验室菌株) 用作参考微生物。原始的废水特性具有相当大的污染物潜力。出水颜色为1495 mg Pt/L，含59 mg/L H2O2、53 mg/L总酚、2.5 mgO2/L溶解氧 (DO) 和887 mg/L油脂。COD为9147 mgO2/L，慢性毒性因子为1667。在生物处理之后，发现真菌epicoccupm nigrum在减少 (有效减少) 大多数参数 (26% COD，12% H2O2，59% 总酚以及40% 油和油脂) 方面最有效，而Cladosporium sp。在减少颜色方面更有效 (77%)。

BACKGROUND & AIMS: :In December 2011, symptoms typical of Cladosporium leaf spot caused by Cladosporium variabile (4) were observed in organic "baby leaf" spinach (Spinacia oleracea) crops of the cultivars Amazon, Missouri, Tasman, and Tonga in the Imperial Valley (Imperial County, CA and Yuma County, AZ). Leaves had small, circular lesions (1 to 3 mm in diameter), some of which had progressed to necrotic, bleached lesions surrounded by a thin dark margin. The incidence of symptoms in affected crops was ≤20%. Fungal isolates resembling C. variabile were recovered by surfacesterilizing sections (5 mm2) of symptomatic leaf tissue in 0.6% NaOCl, triple-rinsing the sections in sterile water, and plating the sections onto water agar and potato dextrose agar amended with 100 ppm chloramphenicol (cPDA). Single-spore transfers made onto cPDA were maintained at 24 ± 2°C with a natural day/night cycle. Each isolate produced slow growing cultures consisting of dense masses of dark conidiophores (≤350 μm long) with chains of up to three dematiaceous (olive) conidia, and almost no mycelium. Torulose (coiled) aerial hyphae developed from the apices of conidiophores after 5 to 7 days, and distinguished the isolates as C. variabile, not C. macrocarpum (2,4). Pathogenicity was tested for each of six single-spore isolates using 36-dayold plants of the spinach cultivar Carmel. The plants were enclosed in clear plastic bags overnight and inoculated the next day with the isolates of C. variabile by atomizing approximately 30 ml of a spore suspension (1.0 × 106 conidia/ml in sterile water amended with 0.01% Tween 20) of the appropriate isolate onto the upper and lower leaf surfaces of each of five plants/isolate. Five control plants were inoculated similarly with sterile water + 0.01% Tween 20. The plants were resealed in plastic bags for 72 h and then placed on a greenhouse bench. Pinpoint, sunken lesions developed within 4 to 7 days on the leaves of plants inoculated with each of the six test isolates. Lesions developed into dry, circular spots typical of Cladosporium leaf spot. Symptoms were not observed on control plants. After 20 days, C. variabile was reisolated from lesions caused by all six isolates, but not from control plants. Although Cladosporium leaf spot has been reported in the Salinas Valley of California (4), to our knowledge, this is the first report of the disease on spinach crops in the Imperial Valley of California and Arizona, the primary winter, fresh market spinach production region of the United States. Inoculum of C. variabile may have been introduced to this region on spinach seed lots (3), because even seed infestation levels <0.1% could lead to seed transmission (1) under the dense planting populations (≤9 million seeds/ha) and overhead irrigation typical of "baby leaf" spinach crops in this region. Fungicides can be used to manage Cladosporium leaf spot in conventional spinach crops (1), but management in certified organic crops may be more challenging. References: (1) L. J. du Toit et al. Fung. Nemat. Tests 59:V115, 2004. (2) M. B. Ellis. Page 315 in: Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Surrey, England, 1971. (3) P. Hernandez-Perez. Page 79 in: Management of Seedborne Stemphylium botryosum and Cladosporium variabile Causing Leaf Spot of Spinach Seed Crops in Western Washington, MS thesis, Pullman, WA, 2005. (4) P. Hernandez-Perez and L. J. du Toit. Plant Dis. 90:137, 2006.

背景与目标： : 在2011年12月中，在帝国谷 (加利福尼亚州帝国县和尤马县) 的亚马逊，密苏里州，塔斯曼和汤加品种的有机 “小叶” 菠菜 (Spinacia oleracea) 作物中观察到由variabile (4) 引起的枝孢菌叶斑病的典型症状。叶片具有小的圆形病变 (直径为1至3毫米)，其中一些已发展为坏死的漂白病变，周围有薄的深色边缘。受影响作物的症状发生率 ≤ 20%。通过在0.6% NaOCl中对有症状的叶组织的切片 (5平方毫米) 进行表面酯化处理，在无菌水中三次冲洗切片，并将切片铺到用100 ppm氯霉素 (cPDA) 修正的水琼脂和马铃薯葡萄糖琼脂上，回收类似变种梭菌的真菌分离物。在cPDA上进行的单孢子转移保持在24 ± 2 °C的自然昼夜周期。每个分离株产生缓慢生长的培养物，由密集的深色分生孢子团 (≤ 350 μ m长) 组成，链最多为三个脱毛 (橄榄) 分生孢子，几乎没有菌丝体。5至7天后，从分生孢子的顶端发育出Torulose (盘绕的) 气生菌丝，并将分离物区分为变种C. variabile，而不是C. macrocarpum (2,4)。使用菠菜品种卡梅尔 (Carmel) 的36天龄植物对六个单孢子分离株中的每一个进行了致病性测试。将植物封闭在透明的塑料袋中过夜，并在第二天通过将大约30毫升的孢子悬浮液 (1.0 × 106分生孢子/ml在无菌水中，用0.01% 吐温20修正) 雾化到适当的分离株上，接种变种梭菌。五个中的每一个的上下叶表面植物/分离物。用无菌水 + 0.01% 吐温20类似地接种五个对照植物。将植物重新密封在塑料袋中72小时，然后放在温室长凳上。精确定位，在接种了六个测试分离株的植物的叶子上，在4至7天内形成了凹陷的病变。病变发展为典型的枝孢叶斑病的干燥圆形斑点。在对照植物上未观察到症状。20天后，从所有六个分离株引起的病变中重新分离出变种梭菌，但未从对照植物中分离。尽管已经在加利福尼亚的萨利纳斯山谷 (4) 报道了枝孢菌叶斑病，但据我们所知，这是加利福尼亚帝国谷和亚利桑那州菠菜作物的首次报道，这是该病的主要冬季，新鲜市场菠菜生产地区美国。在菠菜种子批次 (3) 上，可能已将变种梭菌的接种物引入该区域，因为即使种子侵染水平 <0.1% 也可能导致该地区 “小叶” 菠菜作物在密集种植种群 (≤ 900万种子/公顷) 和塔顶灌溉下的种子传播 (1)。杀菌剂可用于管理常规菠菜作物中的枝孢菌叶斑病 (1)，但在认证有机作物中的管理可能更具挑战性。参考文献 :( 1) L。J、杜托特等人。冯。内马特。测试59:V115，2004。(2) M。B.埃利斯。第315页: 脱色菌丝菌。英联邦真菌学研究所，萨里，英格兰，1971。(3) P。埃尔南德斯-佩雷斯。第79页: 华盛顿州西部菠菜种子作物叶斑病的管理，MS论文，华盛顿州普尔曼，2005。(4) P。埃尔南德斯-佩雷斯和L。J、杜托特。植物Dis。90:137，2006。

BACKGROUND & AIMS: :The binaphthyl derivative cladosporol 3 was supplied from 60 to 200 mg l(-1) to shaken cultures of Cladosporium cf. cladosporioides. Compared to blank, fungal biomass was not affected by adding cladosporol till 100 mg l(-1): it rather increased at higher ratios between 150 and 200 mg l(-1). The production of the major pentacyclic metabolite 1, a cytokine production and tyrosine kinase inhibitor, was enhanced tenfold when cladosporol was supplied at the highest ratio (200 mg l(-1)) to shaken growing cultures of the fungus. The bioconversion of cladosporol to cladosporol D through reductive cleavage of the epoxide group was also observed. Interest in this kind of metabolites lies in their potential activity vs DNA topoisomerase I.

背景与目标： : 将联萘基衍生物cladosporol 3从60至200 mg l(-1) 提供给摇匀的枝孢菌属 (cladosporioides) 的培养物。与空白相比，添加cladosporol直至100 mg l(-1) 不会影响真菌生物量: 它在150和200 mg l(-1) 之间的较高比率下增加。当以最高比率 (200 mg l(-1)) 向真菌的振荡生长培养物供应cladosporol时，主要的五环代谢物1 (细胞因子产生和酪氨酸激酶抑制剂) 的产生提高了十倍。还观察到通过环氧化物基团的还原裂解将cladosporol生物转化为cladosporol D。对这种代谢物的兴趣在于它们与DNA拓扑异构酶I的潜在活性。