黄曲霉群中三个种的分生孢子扫描电镜观察

利用扫描电子显徽镜,观察了米曲霉(Aspergillus ot yzae)、黄曲霉(A. flavus)和寄生曲霉(A. paraslttcus)的分生孢子的表面结构。米曲霉的分生孢子表面呈瘤状突起,瘤块大,边缘呈波纹状,有明显的层叠结构,酷似大脑。黄曲霉的分生孢子表面虽也有不规则的突起,但小而疏,不像米曲霉那样呈瘤脑状;而寄生曲霉的分生孢子表面纹饰与前者均不相同,其瘤状突起大,而末端尖,整个形状像一朵大丽花。三个种的分生孢子表面纹饰有明显差异。扫描电镜观察对于鉴定曲霉是一种行之有效的方法。     

甘草药材上的污染真菌类群及其产毒素特性

对药材市场上霉变甘草样品的污染真菌进行分析,共得到4属7种真菌,包括Penicillium、Aspergillus、Fusarium、Mucor属,其中Penicillium polonicum、Aspergillus parasiticus以及P.crustosum是优势真菌。采用高效液相色谱-质谱联用技术对优势菌菌株产黄曲霉毒素及赭曲霉毒素A的特性进行检测。结果表明A.parasiticus主要产生黄曲霉毒素(AFG2、AFG1、AFB2、AFB1)和赭曲霉毒素A(OTA);而Penicillium polonicum主要产生赭曲霉毒素A(OTA)。     

黄曲霉毒素生物防控菌的筛选及鉴定

筛选黄曲霉毒素生物防控菌,为黄曲霉毒素的生物防控提供支持。以花生原产地土壤为材料,采用牛津杯法筛选所需菌株。对筛选出的拮抗菌株进行抑制产毒曲霉菌株的生长、产孢、降解黄曲霉毒素实验。筛选出2株黄曲霉毒素生防细菌,编号21-1-2、17-3,经鉴定,拮抗菌21-1-2为枯草芽胞杆菌,拮抗菌17-3为地衣芽胞杆菌。分别对拮抗菌对曲霉孢子萌发的抑制、抑制黄曲霉的生长和菌丝延长以及减少黄曲霉毒素的产生、对黄曲霉毒素的分解作用等几个方面进行研究,结果表明,拮抗菌可以明显抑制产毒曲霉孢子的萌发、生长、菌丝的延长,减少黄曲霉毒素的产生以及分解黄曲霉毒素。     

米蛋白分解菌A.0—6的菌种鉴定及其与米曲霉As.3.951菌株的比较

为了进一步认识经多年选育得到的Ａ·０—６米蛋白分解菌,对其进行了形态及培养特性和生理特性的研究试验,认为Ａ·０—６菌株为半知菌类,丛梗孢目,丛梗孢科,曲霉属,黄曲霉群的米曲霉。与在酿造工业生产上应用多年的米曲霉Ａｓ．３．９５１相比,Ａ·０—６菌株具有蛋白酶活力高,米蛋白分解能力强,生长快,易培养等特点,是值得推广应用的优良菌株。     

酱油曲霉蛋白酶活力的提高

蛋白酶在工业上被广泛地应用于皮革、毛皮、丝绸、医药、食品和酿造等行业。霉菌生产的蛋白酶在豆酱和酱油生产上的应用已经具有相当悠悠的历史。曲霉菌,例如米曲霉、黄曲霉、栖土曲霉等,一般能分泌2～3种蛋白酶(酸性、中性及碱性),其中以中性和碱性酶为主。黑曲酶则以产酸性蛋白酶为主。酱油曲霉所产的蛋白酶也以中性     

曲霉高效基因编辑技术研究进展

曲霉Aspergillus spp.是自然界中分布极为广泛的一大类真菌,在工业上如黑曲霉等已经被人们广泛利用于食品添加剂等生产。因此,曲霉在工业、农业、医药领域均发挥着重要的作用。然而,有些曲霉如黄曲霉能够分泌致癌物黄曲霉毒素从而污染农产品;在临床上还有一类以烟曲霉为主的引起侵袭性真菌感染的条件致病曲霉菌。因此,曲霉就像一把双刃剑影响着人们的生活。曲霉菌丝具有发达的隔膜形成多细胞菌丝体并能在分生孢子梗上产生大量的分生孢子进行无性繁殖。研究曲霉的基因编辑技术对于控制医学和农业上有害曲霉的增殖和促进工业上有益曲霉的生长和代谢都具有非常重要的意义。长期以来,同源重组和随机整合一直是被用于研究曲霉基因功能的传统基因编辑方法,然而其操作费时费力且效率低、难以达到预期的要求。随着第三代基因编辑技术CRISPR/Cas9即成簇的规律间隔的短回文重复序列及其相关系统CRISPR/Cas9（Clustered regulatory interspaced short palindromic repeats-associated protein 9）建立以来,CRISPR/Cas9以编辑高效、操作简便等优势被广泛应用到不同物种中。目前,世界上多个研究团队已经建立了有效地用于不同曲霉物种的CRISPR-Cas9 基因编辑体系。本文概述了有关曲霉基因编辑的历史和进展,以期为尚未建立完整遗传编辑体系的其他曲霉物种或者丝状真菌引入高效CRISPR-Cas9体系提供帮助。     

黄曲霉和米曲霉的多相鉴定方法

【背景】黄曲霉(Aspergillus flavus)和米曲霉(Aspergillus oryzae)形态特征相近,基因组高度相似,较难区分。【目的】旨在总结一套准确鉴别二者的分类方法。【方法】利用22株标准菌株对传统形态学、产毒培养基、酶联免疫毒素检测、系统发育分析、产毒基因检测等5种鉴别方法分别进行验证。【结果】各鉴定方法的结果存在异同,单一的鉴定方法容易出现假阴性或假阳性结果。【结论】利用单一方法区分黄曲霉和米曲霉具有潜在风险,多相鉴定方法可以准确鉴别二者。     

云南省可培养丝孢真菌资源的调查研究Ⅱ

本文继续报道曲霉属的16个分类群：杂色曲霉原变种,杂色曲霉疣梗变种,聚多曲霉,焦曲霉,土曲霉原变种,土曲霉金色变种,黄柄曲霉,温特曲霉,黄曲霉原变种,黄曲霉柱头变种,溜曲霉,炭黑曲霉,黑曲霉,泡盛曲霉,塔宾曲霉和日本曲霉。     

黄曲霉群菌种产生黄曲霉毒素B1的调查研究

本文对来自我国20个省、市、自治区的不同基物上分离的和中国科学院微生物研究所菌种保藏室以及其他单位提供的黄曲霉群菌种,经随机选取82株进行了黄曲霉毒素B_1的测定,证明在测试的9个已知分类群中产生黄曲霉毒素B_1的菌种只限于寄生曲霉和黄曲霉,另外4株种名未定者也能产生此种毒素。在黄曲霉中产毒菌株约占30％(28.3％),其在GAN(葡萄糖硝酸铵)和大米培养基中的黄曲霉毒素B_1的最高产量分别为133,333.3和160,000.0ppb。总的来说,大体上可以反映在我国一般基物上黄曲霉产毒菌株存在的现状。在实验过程中,还对黄曲霉群菌种在GAN和大米培养基中黄曲霉毒素B_1的产量和产毒菌株数作了比较。发现在大米培养基中黄曲霉毒素B_1的产量高于GAN,而且测试的黄曲霉产毒菌株在这两种培养基中均各有不能产毒的菌株,因此,在测定产毒菌株时,若仅采用其中一种产毒培养基,往往会有漏掉产毒菌株的可能性。     

黄曲霉群菌种产生黄曲霉毒素B_1的调查研究

本文对来自我国20个省、市、自治区的不同基物上分离的和中国科学院微生物研究所菌种保藏室以及其他单位提供的黄曲霉群菌种,经随机选取82株进行了黄曲霉毒素B_1的测定,证明在测试的9个已知分类群中产生黄曲霉毒素B_1的菌种只限于寄生曲霉和黄曲霉,另外4株种名未定者也能产生此种毒素。在黄曲霉中产毒菌株约占30％(28.3％),其在GAN(葡萄糖硝酸铵)和大米培养基中的黄曲霉毒素B_1的最高产量分别为133,333.3和160,000.0ppb。总的来说,大体上可以反映在我国一般基物上黄曲霉产毒菌株存在的现状。在实验过程中,还对黄曲霉群菌种在GAN和大米培养基中黄曲霉毒素B_1的产量和产毒菌株数作了比较。发现在大米培养基中黄曲霉毒素B_1的产量高于GAN,而且测试的黄曲霉产毒菌株在这两种培养基中均各有不能产毒的菌株,因此,在测定产毒菌株时,若仅采用其中一种产毒培养基,往往会有漏掉产毒菌株的可能性。     

散囊菌属的两个新种

本文报道两个新种,肋状散囊菌(Eurotium costiforme)和少疣散囊菌(E.parviver-ruculosum)。前者的主要特征是子囊孢子的凸面有许多网结的肋状突起;在查氏琼脂上产生大量的闭囊壳,但分生孢子结构极少。后者在查氏琼脂上生长很局限;子囊孢子大,凸面近于平滑或有少量小疣等主要特点。     

曲霉及其有性型散囊菌的新分类群和新记录种

在收集和分离我国曲霉工作中,发现一个国内新纪录和三个新分类群:1.埃及曲霉,国内新记录。2.合阳曲霉新种,与爪甲曲霉相近,但颜色和具刺不育菌丝有所不同。3.温特曲霉烟色变种,与原变种的主要区别在于颜色为蓝灰色并具异常膨大的梗基。4.瘤突散囊菌新种,其子囊孢子的纹饰独特,凸面具粗疏瘤状突起。     

分离自西藏的两个散囊菌新种

本文报道两个新种:粪生散囊菌(Eurotium fimicola)和旱生散囊菌(Eurotium aridicola),它们不同于已报道的该属诸种。     

Taxonomic comparison of three different groups of aflatoxin producers and a new efficient producer of aflatoxin B1, sterigmatocystin and 3-O-methylsterigmatocystin, Aspergillus rambellii sp. nov

Accumulation of the carcinogenic mycotoxin aflatoxin B, has been reported from members of three different groups of Aspergilli (4) Aspergillus flavus, A. flavus var. parvisclerotigenus, A. parasiticus, A. toxicarius, A. nomius, A. pseudotamarii, A. zhaoqingensis, A. bombycis and from the ascomycete genus Petromyces (Aspergillus section Flavi), (2) Emericella astellata and E. venezuelensis from the ascomycete genus Emericella (Aspergillus section Nidulantes) and (3) Aspergillus ochraceoroseus from a new section proposed here: Aspergillus section Ochraceorosei. We here describe a new species, A. rambellii referable to Ochraceorosei, that accumulates very large amounts of sterigmatocystin, 3-O-methylsterigmatocystin and aflatoxin B1, but not any of the other known extrolites produced by members of Aspergillus section Flavi or Nidulantes. G type aflatoxins were only found in some of the species in Aspergillus section Flavi, while the B type aflatoxins are common in all three groups. Based on the cladistic analysis of nucleotide sequences of ITS1 and 2 and 5.8S, it appears that type G aflatoxin producers are paraphyletic and that section Ochraceorosei is a sister group to the sections Flavi, Circumdati and Cervini, with Emericella species being an outgroup to these sister groups. All aflatoxin producing members of section Flavi produce kojic acid and most species, except A. bombycis and A. pseudotamarii, produce aspergillic acid. Species in Flavi, that produce B type aflatoxins, but not G type aflatoxins, often produced cyclopiazonic acid. No strain was found which produce both G type aflatoxins and cyclopiazonic acid. It was confirmed that some strains of A. flavus var. columnaris produce aflatoxin B2, but this extrolite was not detected in the ex type strain of that variety. A. flavus var. parvisclerotigenus is raised to species level based on the specific combination of small sclerotia, profile of extrolites and rDNA sequence differences. A. zhaoqingensis is regarded as a synonym of A. nomius, while A. toxicarius resembles A. parasiticus but differs with at least three base pair differences. At least 10 Aspergillus species can be recognized which are able to biosynthesize aflatoxins, and they are placed in three very different clades.     

分离自赤豆的青霉新种

本文报道青霉属一个新种,分离自赤豆,特点是分生孢子无色,命名为无色青霉(Penicillium incoloratum sp.nov.).模式和来自模式的活培养物都保存在中国科学院微生物研究所。     

弯孢虫疫霉的分离和鉴定

1984—1987年,在福建的南平、福州、南安等地持续发生了由弯孢虫疫霉(Eryniacurvlspora)感染而引起库蚊(culex sp.)成虫死亡的流行病,病原菌分生孢子梗掌状分枝;分生孢子浅绿色,窄而长,略弯曲,单核,双囊壁,含有多个小脂肪粒,大小29.7—39.6×13.2—19.8(平均32.5×16.1)μm,长宽比1.5—3.0(平均2.0);次生分生孢子梨形,基部为钝的乳突,中间有一大的脂肪粒,孢子大小16.6—19.9×11.6—14.9(平均18.7×13.7)μm;休眠孢子球形,棕褐色,直径23.2—33.2(平均29.5)μm;有假根及囊状体。     

Aspergillus salviicola,a new species from imported spice

A new species ofAspergillus, A. salviicola, has been isolated from Turkish sage, an imported spice in Japan. The species, characterized by white to rosy buff conidial heads, pinkish smooth-walled conidiophores, large biseriate aspergilla, globose smooth-walled conidia, absence of sclerotia and thermotolerant growth, is considered to represent an interface species in the subgenusCircumdati.     

古尼虫草分生孢子阶段的分离和鉴定

用古尼虫草[Cordyceps gunnii(Berk.)Berk.]的子座和内菌核作组织分离,用成熟子座自然发射的子囊孢子作孢子分离,经多批次重复操作结果皆获得同一种真菌培养物。鉴定表明这种真菌是拟青霉属的一个新种——古尼拟青霉(Paecilomyces gunnii Liang sp.nov.)。它的主要特征是,在察氏琼脂上菌落白色至灰色,背面棕色;分生孢子梗短,多从气生菌丝上长出,一般长60μm;瓶梗7—12(-19)×2—3(-4)μm,有再育现象;分生孢子多数拟椭圆形或梭形,表面具细刺,(1.6-)2.6—4.0(-4.8)×(1.2-)1.6—2.5(-3.5)μm,平均大小为4.0×2.6μm;厚垣孢子近球形,光滑;5.5—7.2×3.2—5.5μm。32℃以上不生长。     

Molasses supplementation promotes conidiation but suppresses aflatoxin production by small sclerotial Aspergillus flavus

AIMS: To find a supplemental ingredient that can be added to routinely used growth media to increase conidial production and decrease aflatoxin biosynthesis in small sclerotial (S strain) isolates of Aspergillus flavus. METHODS AND RESULTS: Molasses was added to three commonly used culture media: coconut agar (CAM), potato dextrose agar (PDA), and vegetable juice agar (V8) and production of conidia, sclerotia, and aflatoxins by A. flavus isolate CA43 was determined. The effect of nitrogen sources in molasses medium (MM) on production of conidia, sclerotia and aflatoxins was examined. Water activity and medium pH were also measured. Conidia harvested from agar plates were counted using a haemocytometer. Sclerotia were weighed after drying at 45 degrees C for 5 days. Aflatoxins B(1) and B(2) were quantified by high-performance liquid chromatography. Addition of molasses to the media did not change water activity or the pH significantly. Supplementing CAM and PDA with molasses increased conidial production and decreased aflatoxins. Two-fold increased yield of conidia was found on MM, which, like V8, did not support aflatoxin production. Adding ammonium to MM significantly increased the production of sclerotia and aflatoxins, but slightly decreased conidial production. Adding urea to MM significantly increased the production of conidia, sclerotia and aflatoxins. CONCLUSIONS: Molasses stimulated conidial production and inhibited aflatoxin production. Its effect on sclerotial production was medium-dependent. Water activity and medium pH were not related to changes in conidial, sclerotial or aflatoxin production. Medium containing molasses alone or molasses plus V8 juice were ideal for conidial production by S strain A. flavus. SIGNIFICANCE AND IMPACT OF THE STUDY: Insight into molecular events associated with the utilization of molasses may help to elucidate the mechanism(s) that decreases aflatoxin biosynthesis. Targeting genetic parameters in S strain A. flavus isolates may reduce aflatoxin contamination of crops by reducing the survival and toxigenicity of these strains.     

Isolation and characterisation of Aspergillus awamori BS05, a root-knot-nematode-trapping fungus

Nematode-trapping fungi are important biocontrol agents against parasitic nematodes through adhesive or mechanical hyphal traps. Aspergillus awamori, a root-knot-nematode-trapping fungus from tomato rhizosphere soil, was identified based on morphology and molecular characteristics of internal transcribed spacer DNA sequence. Conidial heads were white to black brown, loosely globose, and 72–127 μm in diameter. Conidiophores usually arose from the foot cell of basal mycelium, straight, and 960–1730 × 10.2–13.4 μm, hyaline to pale brown, not constricted below the vesicles; vesicles hemispherical to elongate, 43–56 μm in diameter, black brown, fertile over the upper half to two-thirds. Aspergilla were biseriate, and metulae were variable, 12–26 × 3.8–4.7 μm; phialides were 8.2–9.4 × 2.5–3 μm. Conidia were globose or subglobose, 3.6–4.8 μm in diameter, rough, grey brown and parallel in chains. A. awamori BS05 showed 44.9% control efficacy against Meloidogyne incogtina in pot experiments which suggests it as a potential biocontrol agent against Meloidogyne. This is the first report on A. awamori as nematode-trapping fungus.