Rapid diagnosis of Fusarium root rot in soybean caused by Fusarium equiseti or Fusarium graminearum using loop-mediated isothermal amplification (LAMP) assays

Abstract

Rapid diagnostic assays for Fusarium root rot in soybean, caused by Fusarium equiseti or Fusarium graminearum were developed using the target gene CYP51C. Both assays amplified the target gene in 60 min at 62°C, after which they were assessed for specificity and sensitivity. Specificity was evaluated against other Fusarium spp., other fungal species, and oomycetes. A positive yellow-green color (by the naked eye) or intense green fluorescence (under ultraviolet light) was observed only in the presence of F. equiseti or F. graminearum after adding SYBR Green I, whereas other strains showed either no color change or weak fluorescence. The detection limit of the CYP51C-Fe-LAMP assay was 10 pg. μL −1 genomic DNA per reaction, and as few as four conidia per gram of soil could be detected; and the detection limit of the CYP51C-Fg-LAMP assay was 100 pg. μL −1 genomic DNA per reaction, and 40 conidia per gram of soil could be identified. The assays also detected F. equiseti or F. graminearum from inoculated soybean tissues and diseased plants in the field. These results suggest that CYP51C-Fe-LAMP assay and CYP51C-Fg-LAMP assay are effective in rapidly diagnosing soybean root rot caused by F. equiseti and F. graminearum.

Rapid and sensitive detection of Acidovorax citrulli in cucurbit seeds by visual loop‐mediated isothermal amplification assay

“…Data from this study indicated that the detection limit of the LAMP assay was 1 pg genomic DNA, in accordance with other reports of LAMP methods (Lu, Zhang, Wang, & Zheng, ), and the sensitivity was greater than the previous report for LAMP assay used to detect A. citrulli (Oya et al, ). Compared to the conventional PCR, LAMP provides reliable detection as specific as conventional PCR but increases the sensitivity by 100‐fold.…”

Peer Review #1 of "Endophytic fungal communities associated with field-grown soybean roots and seeds in the Huang-Huai region of China (v0.1)"

“…We found many fungal genera/species in healthy soybean roots that are well-known pathogen species of soybean and/or other plants such as wheat. For example, Fusarium, Macrophomina, Rhizoctonia, and Alternaria cause root rot or foliar and stem blight in soybeans (Lu et al 2015a;Lu et al 2015b;Ramos et al 2016;Yamamoto et al 2017). Our preliminary pathogenicity assays for some of these isolates showed that many of them may be truly asymptomatic in soybean seedlings, while some were able to cause disease symptoms, indicating that they may be latent pathogens that cause disease under stress or at a later time during plant growth.…”

Rapid Diagnosis of Soya Bean Root Rot Caused by Fusarium culmorum Using a Loop‐Mediated Isothermal Amplification Assay

“…2015a), F. equiseti and F. graminearum (Lu et al. 2015b). However, a report of LAMP assay for detecting F. culmorum is lacking.…”

“…lycopersici (Almasi et al 2013), F. oxysporum f. sp. ciceris (Ghosh et al 2015), F. oxysporum (Lu et al 2015a), F. equiseti and F. graminearum (Lu et al 2015b). However, a report of LAMP assay for detecting F. culmorum is lacking.…”