Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresource, College of Plant Protection, Hainan University, Haikou, Hainan Province, 570228, P.R. China.
Chinese Academy of Sciences, South China Botanical Garden, Tianhi District, Guangzhou, 510000, Guangdong Province, P.R. China.
Plant Dis. 2018 Jul;102(7):1426-1433. doi: 10.1094/PDIS-10-17-1672-RE. Epub 2018 May 14.
Colletotrichum gossypii is a causal agent of anthracnose in upland cotton (Gossypii hirsutum). C. gossypii belongs to the C. gloeosporioides species complex. However, the high level of genetic similarity among the species within the C. gloeosporioides species complex makes the development of a diagnostic assay for C. gossypii challenging. Furthermore, the spore size and cultural characteristics of C. gossypii from different geographical areas can vary. In this study, we examined the morphological variance, growth pattern, and pathogenicity of C. gossypii and developed a molecular diagnostic assay to detect C. gossypii in cotton plants from different regions of China. To overcome any ambiguity in morphological and pathogenic characteristics, a set of primers targeting the β-tubulin (TUB) gene of C. gossypii was designed. Single-nucleotide polymorphisms (SNP) were used to identify C. gossypii at the species level using DNA sequence alignments of the TUB gene. The SPSCG/F and SPSCG/R primer pair only amplified C. gossypii, and was able to amplify C. gossypii in mixtures of other Colletotrichum spp., even when DNA concentrations were up to 10 times lower than that of the other species. This is the first report of the development of SNP-based markers for the specific identification of C. gossypii.
炭疽菌（Colletotrichum gossypii）是陆地棉（Gossypii hirsutum）炭疽病的病原体。炭疽菌属于炭疽菌复合种。然而，炭疽菌复合种内种间的遗传相似性水平较高，使得炭疽菌的诊断检测方法的开发具有挑战性。此外，不同地理区域的炭疽菌的孢子大小和培养特征可能会有所不同。在这项研究中，我们研究了炭疽菌的形态变异、生长模式和致病性，并开发了一种分子诊断检测方法，以检测来自中国不同地区的棉花植株中的炭疽菌。为了克服形态和致病性特征的任何歧义，我们设计了一组针对炭疽菌β-微管蛋白（TUB）基因的引物。使用 TUB 基因的 DNA 序列比对来鉴定炭疽菌复合种内的单核苷酸多态性（SNP）。SPSCG/F 和 SPSCG/R 引物对仅扩增炭疽菌，并且即使其他 Colletotrichum spp.的 DNA 浓度比炭疽菌低 10 倍，也能够在其他 Colletotrichum spp.的混合物中扩增炭疽菌。这是首次报道基于 SNP 的标记物用于炭疽菌的特异性鉴定。
