基于全基因组开发小麦矮腥黑粉菌特异性分子标记OA

Wheat dwarf smut,caused by Tilletia controversa(TCK),is a dangerous fungal wheat disease that is highly destructive and difficult to eradication,possessing a significant impact.The disease is known as"No.1 disease of wheat",and is listed as a quarantine pest by European Plant Protection Organisation(EPPO),Australia,China and other countries/regions and organizations.Due to difficulties for distinguishing TCK from other closely related species such as Tilletia laevis in levels of morphological characteristics and molecular biology,misdiagnosis easily happen in quarantine inspections.In this study,TCK-specific molecular markers were developed at the genomic level,and four candidate genes,namely OG0008430,OG0008601,OG0008603 and OG0008610,were screened.Eight pairs of primers and four probes based on these four candidate genes were designed.Validation of combinations among the primers and probes were verified,and two sets of"triple+single"real-time fluorescence PCR methods were established.This combined method can effectively distinguish target species(TCK)from closely related species,achieving accurate identification of TCK,and it can also be used to distinguish different species within the genus Tilletia.