Pathogenic analyses of fungus strains isolated from medicinal Fritillaria przewalskii Maxim. bulb rot

Abstract

Bulb rot causes a great loss of rare perennial medicinal Fritillaria przewalskii Maxim., which is exacerbated with growth year and seriously constrains the plant productivity, but the pathogens responsible for the disease were still unknown. In this paper, we attempted to explore the potential pathogens leading to bulb rotting and their occurrence patterns in artificial domesticated F. przewalskii. Fungus strains were isolated from the bulb rot spot of the 3‐ to 5‐year‐old diseased F. przewalskii plants by tissue separation, further assayed for pathogenicity according to Koch's law and finally identified by both morphological and molecular methods. Seven pathogenic strains (F1‐F7) were detected in 5‐year‐old rot bulbs, six (F1‐F6) in 4‐year‐old rot bulbs and four (F1, F2, F5, F6) in 3‐year‐old rot bulbs. All of the strains were able to infect bulbs by stabbing and some exhibited varying levels of aggressiveness. Relative to the non‐stabbing controls, the bulbs stab‐inoculated with F5, F2, F7, F4, F1, F6, and F3 showed 76.65%, 75.15%, 71.44%, 40.37%, 39.09%, 36.87%, and 34.93% rot after 8 days, respectively. Phylogenetic analysis revealed that these seven strains were clustered into Bionectria ochroleuca (F1, F3, F4), Fusarium oxysporum (F2, F7), Fusarium tricinctum (F5), and Clonostachys rosea (F6). The two species of Fusarium had the strongest pathogenicity, followed by Bionectria ochroleuca and Clonostachys rosea. Although leading to low bulb rot incidence by stab inoculation, F1 showed the highest isolation rate (48.9%) among all strains. Thus, the edible and medicinal bulbs of F. przewalskii are susceptible to synergetic contamination by these seven pathogens at some point after their third year of growth, which has contributed to the species endangered status, with the two strains of Fusarium being the predominant pathogens. To our knowledge, this is the first report on the seven strains of four fungal species causing F. przewalskii bulb rot in China.