Abstract:
The Penicillium genus exhibits a broad global distribution and holds substantial economic value in sectors including agriculture, industry, and medicine. Particularly in agriculture, Penicillium species significantly impact plants, causing diseases and contamination that adversely affect crop yields and quality. Timely detection of Penicillium species is crucial for controlling disease and preventing mycotoxins from entering the food chain. To tackle this issue, we implement a novel species identification approach called Analysis of whole GEnome (AGE). Here, we initially applied bioinformatics analysis to construct specific target sequence libraries from the whole genomes of seven Penicillium species with significant economic impact: P. canescens, P. citrinum, P. oxalicum, P. polonicum, P. paneum, P. rubens, and P. roqueforti. We successfully identified seven Penicillium species using the target we screened combined with Sanger sequencing and CRISPR-Cas12a technologies. Notably, based on CRISPR-Cas12a technology, AGE can achieve rapid and accurate identification of genomic DNA samples at a concentration as low as 0.01 ng/µL within 30 min. This method features high sensitivity and portability, making it suitable for on-site detection. This robust molecular approach provides precise fungal species identification with broad implications for agricultural control, industrial production, clinical diagnostics, and food safety.
摘要:
青霉属在全球范围内分布广泛,在包括农业在内的行业中具有重要的经济价值。工业,和医学。尤其是在农业方面,青霉物种显著影响植物,造成疾病和污染,对作物产量和质量产生不利影响。及时检测青霉属物种对于控制疾病和防止霉菌毒素进入食物链至关重要。为了解决这个问题,我们实施了一种新的物种鉴定方法,称为整个GEnome分析(AGE)。这里,我们最初应用生物信息学分析从七个具有重大经济影响的青霉物种的全基因组中构建特定的靶序列库:P.citrinum,草酸磷,P.polonicum,P.paneum,P.rubens,还有P.roqueforti.我们使用筛选的靶标结合Sanger测序和CRISPR-Cas12a技术成功鉴定了7种青霉菌。值得注意的是,基于CRISPR-Cas12a技术,AGE可以在30分钟内以低至0.01ng/µL的浓度快速准确地鉴定基因组DNA样品。该方法具有高灵敏度和便携性,使其适合现场检测。这种强大的分子方法提供了精确的真菌物种鉴定,对农业控制具有广泛的意义。工业生产,临床诊断,和食品安全。