腐烂葡萄中微生物多样性及真菌毒素测定分析

以天津、北京、杭州等10个地区的葡萄园以及市场中的75份腐烂葡萄为研究对象，应用分子生物学手段进行葡萄表面微生物（细菌和真菌）多样性的核酸测序（包括16S区域扩增子和ITS区域扩增子）分析；同时基于次级代谢产物，使用高效液相色谱（HPLC）法对葡萄样本进行了21种真菌毒素的测定分析，以初步摸清主要风险危害，锁定风险菌株。结果表明，对表面微生物的核酸测序共获得细菌16S有效OTU信息732个和真菌ITS有效OTU信息4 336个，构建了葡萄表面菌相分子数据库，其中包括367种细菌和256种真菌，其主要病原菌为青霉、黑曲、链格孢菌、杂色曲霉、灰葡萄孢、炭疽和盾壳霉；通过真菌毒素测定分析，共检测出4种真菌毒素，分别为交链孢毒素AOH、交链孢毒素AME、交链孢毒素TeA和交链孢毒素TEN。

Analysis of Microbial Diversity and Mycotoxins in Rotting Grapes

With 75 rotten grapes from vineyards and markets in Tianjin, Beijing, Hangzhou and other regions as research objects, sequencing(including 16S-rDNA and ITS regional amplicons)was conducted to investigate the microbial(bacterial and fungal)diversity on the surface of grapes. At the same time, based on secondary metabolites, high-performance liquid chromatography(HPLC)method was used to analyze 21 mycotoxins in grape samples to figure out the main risk hazards and determine the risk strains. The results showed that comparing with the bacteria 16S database and fungi ITS database, 732 bacterial and 4 336 fungal effective OTUs were obtained respectively by sequencing, and a molecular database of grape surface microbes, including 367 kinds of bacteria and 256 kinds of fungi was constructed. The main pathogens were Penicillium, Aspergillus niger, Alternaria, Aspergillus versicolor, Botrytis cinerea, Bacillus anthraci and Coniothyrium minitans. Mycotoxin assay showed that a total of four mycotoxins were detected, including alternariol(AOH), alternariol methyl ether (AME), tenuazonic acid (TeA), and tentoxin (TEN).