江西三种危害食用菌的绿色霉菌生物学特性研究

本文报道了江西三种危害食用菌的绿色霉菌的生物学特性。它们是:绿色木霉(Trichoderma Viride pers exFn)。产黄青霉(Penicillium Chrysoenum Thom)和构巢曲霉(Aspergillus nidulcms(Eidzm)Winter)。     

1株黄曲霉颉颃菌的鉴定及其抑菌活性成分分析

【目的】筛选黄曲霉颉颃菌,探究其最佳培养条件及有效抗菌成分,为实际生产中应用生物防治方法抑制黄曲霉污染提供新的理论依据。【方法】通过平板对峙法及分子生物学鉴定方法对待测菌株进行筛选鉴定。以抑菌率作为衡量标准,判断所得黄曲霉颉颃菌的最佳培养温度、培养时间、接种量、转速、pH及装液量。通过硫酸铵沉淀法从颉颃菌无菌发酵上清液中提取蛋白,观测粗提物对黄曲霉菌丝生长的影响。通过离子交换色谱和凝胶过滤色谱进一步纯化粗提蛋白,并进行质谱测序及序列对比分析,筛选鉴定颉颃菌产生的有效抑菌成分。【结果】筛选到1株解淀粉芽孢杆菌B10(菌种保藏号CCTCCNO:M2018353)。最佳培养条件为温度40℃,培养时间36 h,接种量3%,转速120 r/min, pH 6.0,装液量30 mL,最大抑菌率可达46.56%。其粗提蛋白能有效抑制黄曲霉生长,破坏菌丝正常形态,并从中筛选出几丁质结合蛋白、壳聚糖酶、葡聚糖酶等8种可能的有效抑菌成分。【结论】本试验分离鉴定出1株抑制黄曲霉正常生长发育的解淀粉芽孢杆菌B10,其产生的多种蛋白成分对黄曲霉有显著抑菌活性。     

Impedimetric evaluation of compounds for antifungal activity versus Aspergillus niger

The procedure described utilized a microbial monitoring system (bactometer) to measure impedimetrically the inhibitory activity of known antifungal compounds (2-(thiocyanomethylthio)benzothiazole [TCMTB], 3-iodo-2-propynylbutyl carbamate [IPBC], chlorothalonil, tebuconazole, and copper-8-quinolinolate [Cu-8]) on growth of Aspergillus niger Teigh. in a liquid medium. The MIC values determined were based on the activity of the compounds to inhibit a change in the capacitance of the medium during a 96-h period after inoculation. The MIC values obtained with the bactometer were compared with MIC values derived from the inhibition of radial growth of A. niger on the same liquid medium solidified with agar. The MIC values for all the compounds tested were substantially lower when determined with the bactometer than on agar. The impedimetric evaluation of compounds for antifungal activity offers advantages of speed, sensitivity, and data collection.     

高通量测序技术结合传统方法筛选及鉴定奶酒中曲霉菌

奶酒含有丰富的营养,且具有驱寒活血、开胃消食等保健功效。但由于传统的可培养方法对其微生物全貌的认识具有一定的局限性。因此,本文采用高通量测序技术,对奶酒的真菌生物多样性进行检测。结果表明,酵母菌属(Saccharomyces)和曲霉属(Aspergillus)是奶酒最主要的真菌群落,其中,酵母菌属为第一丰度的真菌,丰度约为62%。第二丰度的真菌为曲霉属,丰度为33%。通过进一步分离并采用16S鉴定出了一株曲霉菌,这可为奶酒发酵剂的开发提供依据。     

黑曲霉发酵酱油渣的研究

试验研究了黑曲霉的耐盐能力,试验结果表明:NaCl浓度在0-120 g/l以内,黑曲霉生长速率随NaCl浓度的增加先增加后减小,当NaCl浓度为40 g/l时黑曲霉生长速率达到最大值,为2.08 cm/d;并且以酱油渣为试验材料,研究了黑曲霉对酱油渣中粗纤维成分的降解能力,试验结果表明,黑曲霉能高效降解酱渣中半纤维素,30 d降解率达到了84.28%;同时对纤维素和木质素也有一定程度降解,30 d时降解率分别为23.86%和9.43%。本研究结果为酱油渣的饲料化利用提供了必要的数据支持。     

花生黄曲霉生长预测模型的研究

通过对分离自花生的黄曲霉LYYSS03-3在不同温度(15-35℃)、不同相对湿度(水活度分别为0.5,0.7,0.9)下的菌落生长特性进行检测,采用Boltzmann和Logistic 模型对黄曲霉的生长曲线进行拟合,结果表明,在适宜温湿度阶段,花生黄曲霉的生长曲线更适合用Boltzmann模型拟合.     

茶叶微生物固态发酵中咖啡碱降解途径初探

为探究微生物作用下咖啡碱的降解产物与途径,将普洱茶发酵中筛选鉴定的Aspergillus sydowii NRRL250（聚多曲霉）、Aspergillus pallidofulvus NRRL4789、Aspergillus sesamicola CBS137324和Penicillium mangini CBS253.31等优势菌株分别接种至晒青毛茶进行单菌种固态发酵,并采用高效液相色谱（HPLC）测定咖啡碱、可可碱、茶碱的含量,探究微生物对咖啡碱代谢的影响;另外,基于UHPLC-QTOF-MS代谢组学技术,以灭菌处理组（ST组）和原料组（RM组）为对照,对聚多曲霉接种发酵样进行代谢组学分析。结果表明,A. pallidofulvus NRRL4789、A. sesamicola CBS137324和Penicillium mangini CBS253.31等优势菌株对咖啡碱等嘌呤类碱代谢均无显著影响,而在聚多曲霉接种发酵中,咖啡碱含量显著下降（P<0.05）,降幅达83.89%;茶碱含量显著增加（P<0.05）,发酵末期含量为(25.03±1.17) mg·g^-1;而可可碱保持基本稳定。由此可知,聚多曲霉对咖啡碱降解代谢有显著影响。采用UHPLC-QTOF-MS方法检出茶碱、3-甲基黄嘌呤、1,7-二甲基黄嘌呤等9种与咖啡碱降解相关的代谢物。在聚多曲霉作用下,茶碱、3-甲基黄嘌呤、1,7-二甲基黄嘌呤、7-甲基黄嘌呤含量显著提高（P<0.05）。茶碱、3-甲基黄嘌呤、1,7-二甲基黄嘌呤和1-甲基黄嘌呤与咖啡碱及其相关代谢物的N-脱甲基化途径相关。1,7-二甲基尿酸、1-甲基尿酸与咖啡碱相关代谢物的氧化途径相关。由此可知,聚多曲霉为降解普洱茶咖啡碱的优势菌株,且具有将咖啡碱转化为茶碱的潜在能力;在咖啡碱降解代谢过程中,存在聚多曲霉作用下的N-脱甲基化和氧化,并以N-脱甲基化为主。     

黑曲霉NRRL3135菌株植酸酶基因在毕赤酵母GS-115系统中的表达

将黑曲S（Aspergillus riger）NRRL3135中的植酸酶基因转入毕赤酵母GS-115中,并整合到染色体DNA上,然后进行诱导表达。通过SDS-PAGE对表达产物进行鉴定,发现重组毕赤酵母GS-115分泌了1个约100kD的蛋白,分泌的蛋白较大。用脱糖基化酶endoglycosidase H降解后,形成了约50kD的蛋白,与植酸酶基因产物的理论值相符。通过对表达产物的酶学性质研究,发现该表达产物在pH2．5和pH5．5时具有较高植酸酶活性,同时该表达产物的最适温度约为55℃。这说明植酸酶得到正确的分泌表达。     

Distribution and genetic diversity among Aspergillus flavus isolates across three agro-ecologies essential for maize cultivation in Ghana

Fungal mycotoxins are important contaminants of agricultural commodities that pose serious concerns to producers, consumers and exporters. Aflatoxin is a carcinogenic metabolite produced by the fungi Aspergillus flavus and A. parasiticus. These fungi and other mould species living in the soil contaminate several crops including maize. This study was undertaken with the following objectives: (i) to assess the presence of A. flavus and other moulds in Ghanaian soil; (ii) to determine the distribution and density of A. flavus under three agro-ecologies noted for major maize production; and (iii) to assess the effect of percentage of carbon (C) and nitrogen (N) and soil pH on A. flavus population densities and diversity in the isolates found across the agro-ecologies. The data showed seven moulds that were common across the agro-ecologies. Significant differences (P < 0.05) were observed in the A. flavus density and distribution within and across these agro-ecologies. Fumesua soils recorded the highest levels of A. flavus (1.185 × 10³ cfu g⁻¹) while Akomadan recorded the least (9.76 × 10² cfu g⁻¹). Percentage available C, N and soil pH did not significantly influence A. flavus density. The A. flavus isolates identified in this study varied in genetic sequence within the aflatoxin gene cluster, but these differences were not distinguishable by origin.     

糖化酶工业株的鉴定及糖化酶基因的克隆

对4种糖化酶工业株的rDNA片段进行克隆,在对比其rDNA-ITS1序列与GenBank数据库的基础上,结合形态学观察对菌株进行了鉴定。结果显示,4种菌株分别为黑曲霉、无花果曲霉、米曲霉和灰色小克银汉霉。通过Me-gAlign得到的进化树表明,前3种菌株亲缘关系较近,三者与灰色小克银汉霉的亲缘关系较远。进一步克隆了黑曲霉、无花果曲霉和米曲霉的糖化酶基因,并在毕赤酵母中实现了分泌表达。