Kizis Dimosthenis, Natskoulis Pantelis, Nychas George-John E, Panagou Efstathios Z
Laboratory of Microbiology and Biotechnology of Foods, Department of Food Science and Human Nutrition, Agricultural University of Athens, Athens, Greece.
PLoS One. 2014 Apr 7;9(4):e93923. doi: 10.1371/journal.pone.0093923. eCollection 2014.
A study on the occurrence of Aspergillus section Nigri species on grapes from four traditional grape-producing areas in Greece during the 2011/2012 vintage, and their capability to produce OTA was conducted. One hundred and twenty-eight black aspergilli isolates were characterised at the species level initially by the use of morphological criteria in accordance with appropriate keys, followed by molecular characterisation performed with Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) of the 5.8 ribosomal RNA gene Internal Transcribed Spacer region (5.8 rRNA ITS). Restriction enzyme digestion of the ITS amplicons using the HhaI, HinfI and RsaI, endonucleases distinguished eleven different patterns of restriction fragment length polymorphism (RFLP), four for each of the HhaI and RsaI digests and three for HinfI. From a total number of 128 individual isolates, 124 were classified into four Aspergillus species corresponding to A. carbonarius, A. tubingensis, A. japonicus and A. ibericus, and the remaining 4 were classified as members of the A. niger aggregate. A. carbonarius and A. tubingensis being the main representative species were equally counted, with higher geographical representation of the former in southern and the latter in northern regions, respectively. All isolates were tested for their ochratoxigenic potential by use of High Performance Liquid Chromatography (HPLC) and Enzyme Linked Immuno Sorbent Assay (ELISA), resulting in significant interspecies differences in OTA production.
针对2011/2012葡萄收获季希腊四个传统葡萄种植区葡萄上黑曲霉属物种的发生情况及其产生赭曲霉毒素A（OTA）的能力开展了一项研究。最初依据合适的检索表采用形态学标准对128株黑色曲霉分离株进行种水平鉴定，随后利用5.8核糖体RNA基因内转录间隔区（5.8 rRNA ITS）的聚合酶链反应-限制性片段长度多态性分析（PCR-RFLP）进行分子鉴定。使用限制性内切酶HhaI、HinfI和RsaI对ITS扩增子进行酶切，区分出11种不同的限制性片段长度多态性（RFLP）模式，其中HhaI和RsaI酶切各有4种，HinfI酶切有3种。在总共128株分离株中，124株被归类为四种曲霉属物种，分别对应于黑曲霉、管囊曲霉、日本曲霉和伊比利亚曲霉，其余4株被归类为黑曲霉复合体成员。作为主要代表物种的黑曲霉和管囊曲霉数量相当，前者在南部地区地理代表性更高，后者在北部地区地理代表性更高。使用高效液相色谱法（HPLC）和酶联免疫吸附测定法（ELISA）对所有分离株的产赭曲霉毒素潜力进行检测，结果显示不同物种在OTA产生方面存在显著差异。
