Isolation, Identification and Characterization of Leptosphaerulina trifolii, the Causative Agent of Alfalfa Leptosphaerulina Leaf Spot in Inner Mongolia, China

• Published in: Agronomy (Basel) 2024-06, Vol.14 (6), p.1156
• Main Authors: Huo, Hongli, Huangfu, Jiuru, Song, Peiling, Zhang, Dongmei, Shi, Zhidan, Zhao, Lili, Li, Ziqin, Zhou, Hongyou
• Format: Article
• Language: English
• Subjects: agar; agronomy; Alfalfa; Carbon; Carbon sources; China; Disease; fungi; Growth conditions; Identification; leaf spot; Leafspot; Leaves; Leptosphaerulina leaf spot; Leptosphaerulina trifolii; Medicago sativa; Morphology; mycelium; Nitrogen; oats; Pathogenicity; Pathogens; Phylogenetics; Phylogeny; RNA polymerase; Signs and symptoms; starch; statistical analysis; Trees; yeast extract; Yeasts
• ISSN: 2073-4395; 2073-4395
• DOI: 10.3390/agronomy14061156

Leptosphaerulina leaf spot, caused by Leptosphaerulina trifolii, is a major disease of alfalfa (Medicago sativa), leading to noticeable losses. From 2022 to 2023, we collected samples of alfalfa with symptoms of the disease from different locations in Inner Mongolia, China. Nine fungal isolates recovered from these samples were identified through morphological traits and a maximum likelihood phylogeny based on concatenated partial sequences of ITS, 28S, and rpb2. A pathogenicity test on alfalfa confirmed the pathogenicity of the isolates on alfalfa. Analysis of physiological traits of L. trifolii revealed optimal mycelium growth at 20 °C and a pH range of 5 to 7, with soluble starch as the preferred carbon source and yeast extract as the optimal nitrogen source. The pathogen thrived in V8-juice agar and oat agar media. This study confirms L. trifolii as the causative agent of Leptosphaerulina leaf spot of alfalfa in Inner Mongolia and provides valuable insights into its optimal growth conditions. These findings enhance the understanding and management of this disease in alfalfa fields.