He Jiao, Sun Mei-Ling, Li De-Wei, Zhu Li-Hua, Ye Jian-Ren, Huang Lin
Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, China.
The Connecticut Agricultural Experiment Station Valley Laboratory, Windsor, CT, USA.
Pest Manag Sci. 2023 Mar;79(3):980-988. doi: 10.1002/ps.7260. Epub 2022 Nov 14.
Anthracnose is one of the most widespread and destructive diseases on Chinese fir. Colletotrichum cangyuanense, Colletotrichum fructicola, Colletotrichum gloeosporioides, and Colletotrichum siamense are the causal agents of anthracnose on Chinese fir. A rapid and accurate diagnosis of different pathogens is critical for the disease management.
Phylogenetic tree and sequence similarity analysis showed that the single-locus ApMat provides superior phylogenetic information and is an appropriate target to distinguish C. cangyuanense, C. fructicola, C. gloeosporioides, and C. siamense. The real-time PCR assays with the primer sets of MQ-F/R, 1#C-F/R, YK-F/R, and WZ-F/R, and corresponding TaqMan probes of MQ-P, 1#C-P, YK-P, and WZ-P were specific for C. cangyuanense, C. fructicola, C. gloeosporioides, and C. siamense, respectively. The sensitivity tests showed that the lowest amount of gDNA that the PCRs can detect was 1 ng of genomic DNA. The validity of the assays was confirmed by directly detecting the pathogens from both the fungal culture and infected Chinese fir.
These results demonstrated the potential of the TaqMan real-time PCR targeting the ApMat gene to provide rapid, specific, and reliable molecular detection of C. fructicola, C. gloeosporioides, C. siamense, and C. cangyuanense, respectively. The data also provided a reference solution for the identification of species within Colletotrichum gloeosporioides species complex (CGSC), which share similar morphological characteristics. © 2022 Society of Chemical Industry.
炭疽病是杉木上分布最广、危害最大的病害之一。沧源炭疽菌、果生炭疽菌、胶孢炭疽菌和暹罗炭疽菌是杉木炭疽病的病原菌。快速准确地诊断不同病原菌对于病害防治至关重要。
系统发育树和序列相似性分析表明，单基因座ApMat提供了优越的系统发育信息，是区分沧源炭疽菌、果生炭疽菌、胶孢炭疽菌和暹罗炭疽菌的合适靶点。使用引物对MQ-F/R、1#C-F/R、YK-F/R和WZ-F/R以及相应的TaqMan探针MQ-P、1#C-P、YK-P和WZ-P进行的实时荧光定量PCR检测分别对沧源炭疽菌、果生炭疽菌、胶孢炭疽菌和暹罗炭疽菌具有特异性。敏感性测试表明，PCR能够检测到的最低基因组DNA量为1 ng。通过直接从真菌培养物和感染的杉木中检测病原菌，证实了该检测方法的有效性。
这些结果证明了靶向ApMat基因的TaqMan实时荧光定量PCR分别对果生炭疽菌、胶孢炭疽菌、暹罗炭疽菌和沧源炭疽菌进行快速、特异性和可靠分子检测的潜力。这些数据也为形态特征相似的胶孢炭疽菌种复合体（CGSC）内的物种鉴定提供了参考解决方案。© 2022化学工业协会。
