Abies georgei var. smithii (Viguie & Gaussen) is a dominant conifer along the southeastern margin of the Qinghai–Tibet Plateau, where heart rot often develops covertly, complicating forest health monitoring and disease management. Fomitopsis subpinicola B.K. Cui, M.L. Han & Shun Liu is an important causal agent of heart rot affecting A. georgei var. smithii in this region, yet rapid, field-deployable molecular diagnostics of this pathogen remain limited. Here, we developed and evaluated two TEF1α-based isothermal platforms for specific detection of F. subpinicola: RAA and LAMP. To reduce potential cross-reactivity, TEF1α sequences from representative taxa within the F. pinicola species complex and closely related non-complex species were aligned for primer/probe design. Candidate RAA primers were screened by gel electrophoresis to select an optimal pair, and two LAMP primer sets were compared by specificity testing to identify the best-performing set. Both assays specifically detected F. subpinicola with no cross-amplification in the tested non-target fungi. Limits of detection were 9.97 copies/μL for fluorescent RAA (25 min), 9.97 × 102 copies/μL for RAA-LFD (15 min), and 9.97 × 103 copies/μL for LAMP (35 min). In 30 increment core samples from A. georgei var. smithii, all methods consistently detected samples with obvious decay, while fluorescent RAA additionally yielded positives in some apparently asymptomatic samples, indicating promise for early or low-abundance screening. Together, these assays constitute a tiered and application-oriented detection system, enabling flexible selection of diagnostic approaches according to sensitivity requirements, operational conditions, and field surveillance needs for heart rot of A. georgei var. smithii.
Kong et al. (Wed,) studied this question.