New species in Aspergillus section Terrei

Section Terrei of Aspergillus was studied using a polyphasic approach including sequence analysis of parts of the β-tubulin and calmodulin genes and the ITS region, macro- and micromorphological analyses and examination of extrolite profiles to describe three new species in this section. Based on phylogenetic analysis of calmodulin and β-tubulin sequences seven lineages were observed among isolates that have previously been treated as A. terreus and its subspecies by Raper & Fennell (1965) and others. Aspergillus alabamensis, A. terreus var. floccosus, A. terreus var. africanus, A. terreus var. aureus, A. hortai and A. terreus NRRL 4017 all represent distinct lineages from the A. terreus clade. Among them, A. terreus var. floccosus, A. terreus NRRL 4017 and A. terreus var. aureus could also be distinguished from A. terreus by using ITS sequence data. New names are proposed for A. terreus var. floccosus, A. terreus var. africanus, A. terreus var. aureus, while Aspergillus hortai is recognised at species level. Aspergillus terreus NRRL 4017 is described as the new species A. pseudoterreus. Also included in section Terrei are some species formerly placed in sections Flavipedes and Versicolores. A. clade including the type isolate of A. niveus (CBS 115.27) constitutes a lineage closely related to A. carneus. Fennellia nivea, the hypothesized teleomorph is not related to this clade. Aspergillus allahabadii, A. niveus var. indicus, and two species originally placed in section Versicolores, A. ambiguus and A. microcysticus, also form well-defined lineages on all trees. Species in Aspergillus section Terrei are producers of a diverse array of secondary metabolites. However, many of the species in the section produce different combinations of the following metabolites: acetylaranotin, asperphenamate, aspochalamins, aspulvinones, asteltoxin, asterric acid, asterriquinones, aszonalenins, atrovenetins, butyrolactones, citreoisocoumarins, citreoviridins, citrinins, decaturins, fulvic acid, geodins, gregatins, mevinolins, serantrypinone, terreic acid (only the precursor 3,6-dihydroxytoluquinone found), terreins, terrequinones, terretonins and territrems. The cholesterol-lowering agent mevinolin was found in A. terreus and A. neoafricanus only. The hepatotoxic extrolite citrinin was found in eight species: A. alabamensis, A. allahabadii, A. carneus, A. floccosus, A. hortai, A. neoindicus, A. niveus and A. pseudoterreus. The neurotoxic extrolite citreoviridin was found in five species: A. neoafricanus, A. aureoterreus, A. pseudoterreus, A. terreus and A. neoniveus. Territrems, tremorgenic extrolites, were found in some strains of A. alabamensis and A. terreus.     

New taxa in Aspergillus section Usti

Based on phylogenetic analysis of sequence data, Aspergillus section Usti includes 21 species, inclucing two teleomorphic species Aspergillus heterothallicus (= Emericella heterothallica) and Fennellia monodii. Aspergillus germanicus sp. nov. was isolated from indoor air in Germany. This species has identical ITS sequences with A. insuetusCBS 119.27, but is clearly distinct from that species based on β-tubulin and calmodulin sequence data. This species is unable to grow at 37 °C, similarly to A. keveii and A. insuetus. Aspergillus carlsbadensis sp. nov. was isolated from the Carlsbad Caverns National Park in New Mexico. This taxon is related to, but distinct from a clade including A. calidoustus, A. pseudodeflectus, A. insuetus and A. keveii on all trees. This species is also unable to grow at 37 °C, and acid production was not observed on CREA. Aspergillus californicus sp. nov. is proposed for an isolate from chamise chaparral (Adenostoma fasciculatum) in California. It is related to a clade including A. subsessilis and A. kassunensis on all trees. This species grew well at 37 °C, and acid production was not observed on CREA. The strain CBS 504.65 from soil in Turkey showed to be clearly distinct from the A. deflectus ex-type strain, indicating that this isolate represents a distinct species in this section. We propose the name A. turkensis sp. nov. for this taxon. This species grew, although rather restrictedly at 37 °C, and acid production was not observed on CREA. Isolates from stored maize, South Africa, as a culture contaminant of Bipolaris sorokiniana from indoor air in Finland proved to be related to, but different from A. ustus and A. puniceus. The taxon is proposed as the new species A. pseudoustus. Although supported only by low bootstrap values, F. monodii was found to belong to section Usti based on phylogenetic analysis of either loci BLAST searches to the GenBank database also resulted in closest hits from section Usti. This species obviously does not belong to the Fennellia genus, instead it is a member of the Emericella genus. However, in accordance with the guidelines of the Amsterdam Declaration on fungal nomenclature (Hawksworth et al. 2011), and based on phylogenetic and physiological evidence, we propose the new combination Aspergillus monodii comb. nov. for this taxon. Species assigned to section Usti can be assigned to three chemical groups based on the extrolites. Aspergillus ustus, A. granulosus and A. puniceus produced ustic acid, while A. ustus and A. puniceus also produced austocystins and versicolorins. In the second chemical group, A. pseudodeflectus produced drimans in common with the other species in this group, and also several unique unknown compounds. Aspergillus calidoustus isolates produced drimans and ophiobolins in common with A. insuetus and A. keveii, but also produced austins. Aspergillus insuetus isolates also produced pergillin while A. keveii isolates produced nidulol. In the third chemical group, E. heterothallica has been reported to produce emethallicins, 5'-hydroxyaveranthin, emeheterone, emesterones, 5'-hydroxyaveranthin.     

黑曲霉菌渣资源化再利用途径分析

黑曲霉菌渣是黑曲霉发酵法生产葡萄糖酸钠后产生的废弃物,其资源化利用已成为企业扩大生产亟需解决的问题。对该菌渣有效成分检测得知,该菌渣有害金属元素均小于城镇垃圾农用控制标准、有机-无机复混肥料控制标准和鱼粉、猪、家禽添加剂预混合饲料卫生指标,有机质含量符合有机肥料中的有机质含量指标,总糖、粗纤维、粗蛋白、脂肪等营养成分含量较多,同时也含有丰富的N、P、K、Ca、Mg、Fe等元素,可从有机肥料和禽畜饲料方向进行资源化利用,有利于减轻环境污染及实现变废为宝,具有积极的社会、经济和生态效益。     

New and revisited species in Aspergillus section Nigri

Four new species, Aspergillus eucalypticola, A. neoniger, A. fijiensis and A. indologenus are described and illustrated. Aspergillus eucalypticola was isolated from Eucalyptus leaf from Australia, and is related to A. tubingensis and A. costaricaensis, but could clearly be distinguished from them based on either β-tubulin or calmodulin sequence data. Aspergillus eucalypticola produced pyranonigrin A, funalenone, aurasperone B and other naphtho-γ-pyrones. Aspergillus neoniger is also a biseriate species isolated from desert sand in Namibia, and mangrove water in Venezuela, which produces aurasperone B and pyranonigrin A. Aspergillus fijiensis is a uniseriate species related to A. aculeatinus, and was isolated from soil in Fiji, and from Lactuca sativa in Indonesia. This species is able to grow at 37 °C, and produces asperparalines and okaramins. Aspergillus indologenus was isolated from soil, India. This species also belongs to the uniseriate group of black aspergilli, and was found to be related to, but clearly distinguishable from A. uvarum based on β-tubulin, calmodulin and ITS sequence data. Aspergillus indologenus produced the insecticidal compounds okaramins A, B, H, and two types of indol-alkaloids which have not been structure elucidated. Two other species, A. violaceofuscus and A. acidus, are revalidated based on molecular and extrolite data. Aspergillus violaceofuscus was found to be related to A. japonicus, and produced some of the same interesting indol-alkaloids as A. indologenus, and also produced several families of partially characterised extrolites that were also found in A. heteromorphus. Aspergillus acidus (previously known as A. foetidus var. pallidus and A. foetidus var. acidus) is also a valid species, while A. foetidus is a synonym of A. niger based on molecular and physiological data. Two other species described previously, A. coreanus and A. lacticoffeatus, were found to be colour mutants of A. acidus and A. niger, respectively. Methods which could be used to distinguish the two closely related and economically important species A. niger and A. awamori are also detailed. Although these species differ in their occurrence and several physiological means (elastase activities, abilities to utilise 2-deoxy-D-glucose as sole carbon source), our data indicate that only molecular approaches including sequence analysis of calmodulin or β-tubulin genes, AFLP analysis, UP-PCR analysis or mtDNA RFLP analysis can be used reliably to distinguish these sibling species. Aspergillus section Nigri now includes 26 taxa.     

日本曲霉和黑曲霉发酵甘蔗糖蜜和酵母粉生产低聚果糖

在培养日本曲霉(Aspergillus japonicus-FCL)和黑曲霉(Aspergillus niger ATCC 20611)的过程中,研究了培养基中不同浓度的蔗糖(3～25%w/v)和蛋白胨(2～5%w/v)对曲霉的影响以探索较优配方。此外,甘蔗糖蜜(3.5～17.5%w/v总糖量)和酵母粉(1.5～5%w/v)被当作两种菌株的营养素替代品用于研究。探索这些培养基的目的是用来分析细胞生长趋、果糖基转移酶产酶和低聚果糖(FOS)产物。果糖基转移酶活性和低聚果糖产量利用高效液相色谱进行检测分析。两种菌株在3%(w/v)蔗糖和3%(w/v)蛋白胨的培养基中酶反应产物含量最高,或者在含总糖3.5%(w/v)的甘蔗糖蜜和1.5%(w/v)酵母粉的培养基中可以达到同等效果。研究表明,甘蔗糖蜜和酵母粉在酶和低聚果糖(约60%w/w)的生产中与蔗糖和蛋白胨的效果一样好。     

普洱茶发酵过程中外源接种黄曲霉产毒研究

以云南大叶种晒青茶为原料,接种产毒黄曲霉进行模拟普洱茶发酵试验,并在发酵结束时通过LC-MS/MS法对茶样进行黄曲霉毒素检测,研究普洱茶发酵过程中黄曲霉的生长及产毒情况。结果表明,在普洱茶发酵过程中,接种的黄曲霉能在茶叶中生长繁殖,初期生长较快,但随发酵时间的延长,黄曲霉在茶叶中的生长明显受到抑制,其数量逐渐下降。发酵终止时,未在茶样中检测出黄曲霉毒素。     

黄曲霉对花生和玉米的侵染研究

黄曲霉(Aspergillus flavus)极易侵染花生、玉米等多种农作物,并且在侵染后产生具有致癌、致畸、致突变作用的有毒代谢产物——黄曲霉毒素.黄曲霉的侵染和黄曲霉毒素污染不仅发生在作物生长环节,而且在收获、干燥、储运等过程也会发生,严重威胁农产品消费安全和人畜生命健康.因此,解析黄曲霉对粮油作物的侵染过程及侵染...     

Geographical distribution of Aspergillus flavus in peanut harvest period in China

In order to grasp the distribution of Aspergillus fiavus in the soil of peanut production areas in China, A. fiavus biomarkers were tested on 555 soil samples from 37 sampling points in 17 provinces, peanut fields in four agroecological zones(Southern area, Yangtze River Basin, Northern area, Northeast area). The results showed that(1) the cultivation amount of A. fiavus per gram of soil in the Yangtze River Basin is 1.30 times that of the southern area, 1.56 times that of the northern area, and...     

广东花生产区土壤黄曲霉产毒菌落分布初步研究

采用稀释倒平板法对广东10个花生主产地土壤中黄曲霉的数量以及分离菌株的产毒能力进行了分析。结果表明：全省各地花生种植土壤中黄曲霉的数量存在较大的差异,其中清远市土壤中的黄曲霉数量最多,达160 cfu/g干土壤,惠州和广州地区次之,均为100 cfu/g干土壤,汕头地区最少,在该地区抽取的土壤样品中未分离出黄曲霉菌株。在分离出来的黄曲霉菌株中,广州地区有10株产毒菌株、清远地区17株、惠州地区16株、云浮地区8株、河源地区8株。     

Identification of resistance to Aspergillus flavus infection in cotton germplasm

Natural resistance in cottonseed to Aspergillus flavus infection has not been explored to date. A green fluorescent protein (GFP) expressing A. flavus strain was used to assess the resistance of seed from 35 cotton varieties from Gossypium arboreum, G. barbadense, and G. hirsutum. Mature cotyledons devoid of seed coat were wounded, inoculated, and assessed for innate resistance to A. flavus infection. Of the initial 35 varieties tested, we observed a range of resistance to infection in representatives of all three species. A subset of 15 representatives was further analyzed. Within this group, G. arboreum cultivar A2 186 and G. hirsutum cultivar SA 1582 were most resistant to fungal infection. The most susceptible cultivar in this group was G. hirsutum SA 1595. The remaining 12 representatives tested in the secondary screen (3 G. arboreum, 3 G. barbadense, and 6 G. hirsutum lines) exhibited intermediate resistance. We did not observe any relationship between species and resistance. Within each species, there was a range of responses to fungal infection. Future studies using this methodology to screen additional diploid and tetraploid cotton lines may enable us to identify naturally resistant germplasm that can be used to develop cotton with enhanced resistance to fungal infection.     

抗氯嘧磺隆黑曲霉乙酰乳酸合成酶(ALS)酶学特性研究

本文比较了不同黑曲霉(Aspergillus niger)菌株对大豆田除草剂氯嘧磺隆的耐药性,并且对抗性菌株中靶标酶乙酰乳酸合成酶(ALS)的酶学特性进行了研究。结果发现TR-H为抗性菌株,其ALS酶活性显著高于其它菌株。酶学特性分析显示,TR-H菌株的最佳培养时间为84h,最佳反应温度为35℃,最适反应pH为7.3,酶反应的初速度范围为0～60min。以丙酮酸钠为反应底物时,其米氏常数(Km)为39.41mmol/L,最大反应速度(Vmax)为232.2μg/mL·h。     

Sesquiterpenoids from the Mangrove-Derived Aspergillus ustus 094102

Four new drimane sesquiterpenoids (1–4) and three known ones (5–7) were isolated from the fermentation broth of the mangrove-derived Aspergillus ustus 094102. Compound 5 was further resolved as four purified compounds 5a–5d. By means of extensive spectroscopic and ECD analysis as well as the chemical transformation, their structures were identified as (2R,3R,5S,9R,10S)-2,3,9,11-tetrahydroxydrim-7-en-6-one (ustusol F, 1), (2R,3R,5R,9S,10R)-2,3,11-trihydroxydrim-7-en-6-one (9-deoxyustusol F, 2), (3S,5R,9R,10R)-3,11,12-trihydroxydrim-7-en-6-one (ustusol G, 3), (5S,6R,9S,10S, 11R,2′E,4′E)-(11-dideoxy-11-hydroxystrobilactone A-6-yl)-5-carboxypenta-2,4-dienoate (ustusolate H, 4), ((5S,6R,9S,10S)-strobilactone A-6-yl) (2E,4E)-6,7-dihydroxyocta-2,4-dienoate (ustusolate I, 5), (2′E,4′E;6′,7′-erythro)-ustusolate I (5a) and (2′E,4′E;ent-6′,7′-erythro)-ustusolate I (5b), (2′E,4′E,6′R,7′R)-ustusolate I (5c) and (2′E,4′E,6′S,7′S)-ustusolate I (5d), (5S,6R,9S,10S,2′E,4′E)-(strobilactone A-6-yl)-5-carboxypenta-2,4-dienoate (ustusolate J, 6), and (2S,5S,9R,10S)-2,9,11-trihydroxydrim-7-en-6-one (ustusol B, 7), respectively. Compound 5 showed antiproliferation against the human tumor cells CAL-62 and MG-63 with the IC₅₀ values of 16.3 and 10.1 µM, respectively.     

Cytosolic streaming in vegetative mycelium and aerial structures of Aspergillus niger

Aspergillus niger forms aerial hyphae and conidiophores after a period of vegetative growth. The hyphae within the mycelium of A. niger are divided by septa. The central pore in these septa allows for cytoplasmic streaming. Here, we studied inter- and intra-compartmental streaming of the reporter protein GFP in A. niger. Expression of the gene encoding nuclear targeted GFP from the gpdA or glaA promoter resulted in strong fluorescence of nuclei within the vegetative hyphae and weak fluorescence in nuclei within the aerial structures. These data and nuclear run on experiments showed that gpdA and glaA are higher expressed in the vegetative mycelium when compared to aerial hyphae, conidiophores and conidia. Notably, gpdA or glaA driven expression of the gene encoding cytosolic GFP resulted in strongly fluorescent vegetative hyphae and aerial structures. Apparently, GFP streams from vegetative hyphae into aerial structures. This was confirmed by monitoring fluorescence of photo-activatable GFP (PA-GFP). In contrast, PA-GFP did not stream from aerial structures to vegetative hyphae. Streaming of PA-GFP within vegetative hyphae or within aerial structures of A. niger occurred at a rate of 10–15 μm s^-1. Taken together, these results not only show that GFP streams from the vegetative mycelium to aerial structures but it also indicates that its encoding RNA is not streaming. Absence of RNA streaming would explain why distinct RNA profiles were found in aerial structures and the vegetative mycelium by nuclear run on analysis and micro-array analysis.Key words: aerial hypha, Aspergillus, conidia, conidiophore, cytoplasmic streaming, development, fungus, vegetative mycelium     

Antiviral Cyclopropane Acids from Deep-Sea-Derived Fungus Aspergillus sydowii

Four novel monocyclic cyclopropane acids, namely, sydocyclopropanes A–D (1–4), along with one known congener hamavellone B (5), were isolated from the Aspergillus sydowii MCCC 3A00324 fungus, which was isolated from the deep-sea sediment. The gross structures of novel compounds were established by detailed analyses of the spectroscopic data (HRESIMS and NMR spectra), and their absolute configurations were resolved on the basis of the quantum chemical calculations of ECD and NMR data, in association with DP4+ probability analyses. Sydocyclopropanes A–D, featuring the 1,1,2,3-tetrasubstituted cyclopropane nucleus with different lengthy alkyl side chains, were discovered in nature for the first time. All compounds exhibited antiviral activities against A/WSN/33 (H1N1), with IC₅₀ values ranging from 26.7 to 77.2 μM, of which compound 1 exhibited a moderate inhibitory effect (IC₅₀ = 26.7 μM).     

Neuronal Modulators from the Coral-Associated Fungi Aspergillus candidus

Three new p-terphenyl derivatives, named 4″-O-methyl-prenylterphenyllin B (1) and phenylcandilide A and B (17 and 18), and three new indole-diterpene alkaloids, asperindoles E–G (22-24), were isolated together with eighteen known analogues from the fungi Aspergillus candidus associated with the South China Sea gorgonian Junceela fragillis. The structures and absolute configurations of the new compounds were elucidated on the basis of spectroscopic analysis, and DFT/NMR and TDDFT/ECD calculations. In a primary cultured cortical neuronal network, the compounds 6, 9, 14, 17, 18 and 24 modulated spontaneous Ca²⁺ oscillations and 4-aminopyridine hyperexcited neuronal activity. A preliminary structure–activity relationship was discussed.     

花生种质资源对黄曲霉菌侵染的反应

对来自我国不同生态区的700份花生种质进行抗黄曲霉菌侵染的评价。鉴定结果表明,700份花生种质中,大部分对黄曲霉菌侵染表现高感,低侵染率（50％以下）种质仅12份,其中EF7284的侵染率最低,为28．57％。低侵染率花生种质均来自南方生态区的珍珠豆型材料,本文对低侵染率花生种质的主要植物学,农艺性状及品质性状作了分析评价。     

Secondary metabolites from an algicolous Aspergillus versicolor strain

Two new compounds, asperversin A (1) and 9ξ-O-2(2,3-dimethylbut-3-enyl)brevianamide Q (2), and nine known compounds, brevianamide K (3), brevianamide M (4), aversin (5), 6,8-di-O-methylnidurufin (6), 6,8-di-O-methylaverufin (7), 6-O-methylaverufin (8), 5α,8α-epidioxyergosta-6,22-dien-3β-ol (9), ergosta-7,22-diene-3β,5α,6β-triol (10), and 6β-methoxyergosta-7,22-diene-3β,5α-diol (11), were obtained from the culture of Aspergillus versicolor, an endophytic fungus isolated from the marine brown alga Sargassum thunbergii. The structures of these compounds were established by spectroscopic techniques. Compounds 4, 7 and 8 exhibited antibacterial activities against Escherichia coli and Staphyloccocus aureus, and 7 also showed lethality against brine shrimp (Artemia salina) with an LC₅₀ value of 0.5 μg/mL.     

Methylthio-Aspochalasins from a Marine-Derived Fungus Aspergillus sp.

Two novel aspochalasins, 20-β-methylthio-aspochalsin Q (named as aspochalasin V), (1) and aspochalasin W (2), were isolated from culture broth of Aspergillus sp., which was found in the gut of a marine isopod Ligia oceanica. The structures were determined on the basis of NMR and mass spectral data analysis. This is the first report about methylthio-substituted aspochalasin derivatives. Cytotoxicity against the prostate cancer PC3 cell line and HCT116 cell line was assayed using the MTT method. Apochalasin V showed moderate activity at IC₅₀ values of 30.4 and 39.2 μM, respectively.     

花生品质性状与抗黄曲霉侵染关系的研究

以对黄曲霉具有不同抗性的花生品种为材料,通过人工黄曲霉接种测定和品质性状的分析,探讨花生品质性状与抗黄曲霉的关系。结果表明,花生种子胰蛋白酶抑制剂活性的高低(PTI值)与品种黄曲霉侵染率呈显著负相关;而种子可溶性糖、蔗糖和蛋白质含量与品种抗黄曲霉侵染能力无关,花生种子胰蛋白酶抑制剂活性的高低可以作为花生抗黄曲霉育种选育的品质性状标记。