Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Phoma macdonaldii, the Causal Agent of Sunflower Black Stem

Phoma macdonaldii, the causal agent of sunflower black stem, severely affects sunflower yield and quality. A rapid and sensitive detection method is necessary for diagnosis of this disease. In this study, a loopmediated isothermal amplification (LAMP) assay was developed for rapid detection of the pathogen from diseased sunflower tissues. The LAMP primers were designed to target the rDNA region of the fungus. The reaction condition was optimized to 60 degrees C water baths for 45 min. The detection limit of the LAMP assay was 100 fg DNA or 10 conidia/g seeds. The LAMP assay was validated by detecting P. macdonaldii from infected sunflower tissue samples, including leaves, stems, and seeds, and applying to seed samples randomly collected from sunflower fields. This LAMP assay will be useful for estimating disease prevalence and implementing sustainable management of sunflower black stem.

Automated summary

A rapid and sensitive detection method based on LAMP technology was successfully developed in this study for accurate detection of the causal agent of sunflower black stem. This method not only has the characteristics of rapid and sensitive detection, but also can be applied to different types of infected tissues and seed samples, which is of great significance for assessing the prevalence and implementing sustainable management of the disease.