Three new species-specific primer sets were designed to detect C. truncatum, C. gloeosporioides species complex, and C. scovillei, which were reported as the cause of chili anthracnose disease. These new primers were designed based on the sequence data of the nuclear ribosomal DNA (rDNA) region to amplify and identify short size target DNA (<200 bp). The short size target DNA is more efficient for DNA hybridization, and will improve the sensitivity and specificity of DNA assay. The amplicon size obtained for C. gloeosporioides species complex, C. truncatum, and C. scovillei, was 66, 112, and 168 bp, respectively. Each primer set could identify its specific species of Colletotrichum as confirmed by the absence of PCR product with DNA of two other Colletotrichum species and DNA from different fungal genera, indicating high specificity. The sensitivity test was employed, and the results showed that these new primer sets could strongly amplify DNA at the concentration in femto- to picogram levels. Furthermore, these primer sets can be used to amplify and identify Colletotrichum species DNA directly from infected seeds and fruit tissue. The short PCR products obtained by the new primers have the potential to improve the sensitivity in DNA biosensors applications.

设计三个新的种属特异性引物检测C.元宝枫，炭疽菌种复杂，和C. scovillei，其报道为辣椒炭疽病疾病的原因。根据核糖体DNA（rDNA）区的序列数据设计这些新引物，以扩增和鉴定短尺寸的目标DNA（<200 bp）。短尺寸的目标DNA对于DNA杂交更有效，并且将提高DNA测定的灵敏度和特异性。C. gloeosporioides物种复合体，C. truncatum和C. scovillei获得的扩增子大小分别为66、112和168 bp。每个引物组都可以识别其特定种类的炭疽菌通过与其他两种炭疽菌物种的DNA和来自不同真菌属的DNA的PCR产物不存在而得到证实，表明具有高特异性。进行了敏感性测试，结果表明这些新的引物组可以在毫微微至皮克级的浓度下强烈扩增DNA。此外，这些引物组可用于直接从受感染的种子和果实组织中扩增和鉴定炭疽菌物种的DNA。通过新引物获得的短PCR产物具有提高DNA生物传感器应用灵敏度的潜力。