基于生物质谱技术的寒区黄曲霉产毒基因检测及表达分析研究

Aspergillus flavus and aflatoxins can contaminate many important crops, resulting in serious agricultural product safety problems. The distribution of A. flavus isolates significantly varied among the districts and agroecological zones of China. The incidence of toxigenic A. flavus strains may be associated with the geographical location and climate. The project is carried out on the detection and analysis of the relationship between differently expressed genes and aflatoxin production in cold regions by the reaction of RT-PCR.The MALDI-TOF MS is applied to simultaneously detection of multiplex gene expression in aflatoxin-producing strains, and drawing the specific mass spectrum of strain with produces aflatoxin in cold regions. Focusing on the expression of genes involved in aflatoxin biosynthesis, a novel biomass-based method to rapid and accurate detection aflatoxin-producing gene is developed. The project ascertains the difference of aflatoxin-producing strains at the level of gene expression, provides the molecular biology method of discriminating toxigenic and non-toxigenic strains, and explores the inner workings of aflatoxin biosynthesis in cold regions. The goal of the project is expected to provide the theory basis for deeply researching the molecular mechanism of toxin production in fungi, and the technlogical basis for the application of mass spectrometry to the detection of toxigenic fungi.

黄曲霉及其产生的黄曲霉毒素能污染多种农作物并引起严重的农产品安全问题。我国黄曲霉菌的分布呈现明显的地域生态特征。黄曲霉菌的产毒概率跟地理位置和气候密切相关。本研究拟利用RT-PCR方法检测寒区黄曲霉产毒相关基因的表达情况并进行分析，明确基因表达差异与产毒的相关性；利用MALDI-TOF MS方法同时检测产毒菌株多重基因表达情况，绘制寒区黄曲霉产毒菌株的特异性质谱检测峰图。旨在探明黄曲霉毒素生物合成过程中差异基因的表达关系，并建立一种基于生物质谱技术的寒区黄曲霉产毒基因快速、准确检测方法。本研究在基因表达水平探索不同菌株产生黄曲霉毒素的差异，提供鉴别产毒菌株的分子生物学方法，探索寒区黄曲霉毒素生物合成的特殊规律，为研究产毒真菌的产毒分子机制提供理论依据，也为质谱技术在产毒真菌基因检测上的应用提供技术基础。