Rhizopus oryzae Causes the Leaf Rot Disease of Epimedium sagittatum in Guizhou, China

HomePlant DiseaseAhead of PrintRhizopus oryzae Causes the Leaf Rot Disease of Epimedium sagittatum in Guizhou, China PreviousNext DISEASE NOTE OPENOpen Access licenseRhizopus oryzae Causes the Leaf Rot Disease of Epimedium sagittatum in Guizhou, ChinaShihua Zhou, Yanping Gao, Jiuchun An, Lang Qin, Xiaohong Ou, Lanping Guo, Weike Jiang, Tao Zhou, and Qing-Song YuanShihua ZhouResource Institute for Chinese and Ethnic Materia Medica, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China, Yanping GaoResource Institute for Chinese and Ethnic Materia Medica, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China, Jiuchun AnResource Institute for Chinese and Ethnic Materia Medica, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China, Lang QinResource Institute for Chinese and Ethnic Materia Medica, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China, Xiaohong OuResource Institute for Chinese and Ethnic Materia Medica, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China, Lanping GuoNational Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, Beijing 100700, China, Weike JiangResource Institute for Chinese and Ethnic Materia Medica, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China, Tao Zhou†Corresponding authors: Q.-S. Yuan; E-mail Address: [email protected] and E-mail Address: [email protected], and T. Zhou; E-mail Address: [email protected]https://orcid.org/0000-0001-8508-9355Resource Institute for Chinese and Ethnic Materia Medica, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China, and Qing-Song Yuan†Corresponding authors: Q.-S. Yuan; E-mail Address: [email protected] and E-mail Address: [email protected], and T. Zhou; E-mail Address: [email protected]https://orcid.org/0000-0002-9713-099XResource Institute for Chinese and Ethnic Materia Medica, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, ChinaNational Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, Beijing 100700, ChinaAffiliationsAuthors and Affiliations Shihua Zhou1 Yanping Gao1 Jiuchun An1 Lang Qin1 Xiaohong Ou1 Lanping Guo2 Weike Jiang1 Tao Zhou1 † Qing-Song Yuan1 2 † 1Resource Institute for Chinese and Ethnic Materia Medica, Guizhou University of Traditional Chinese Medicine, Guiyang 550025, China 2National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, Beijing 100700, China Published Online:1 Apr 2024https://doi.org/10.1094/PDIS-10-23-2241-PDNAboutSectionsView articlePDFSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat View articleEpimedium sagittatum is a collective term for herbaceous plants belonging to the family Berberidaceae. Their dried leaves and stems have significant therapeutic properties, including tumor inhibition, hypertension control, and coronary heart disease control (Ke et al. 2023; Zhao et al. 2019). In 2021 and 2022, plants with similar leaf rot symptoms ranging from 30 to 55% were observed in Congjiang County, Guizhou Province. The initial symptoms of the disease manifested locally on the leaf, with yellowing on the surface edge of the affected tissue, browning in the middle part, and brownish white discoloration in the innermost part. As the disease progressed, the entire infected leaf gradually softened, while the veins remained intact. Ultimately, the leaf withered and dehisced. Nine samples with typical symptoms were collected from Congjiang County (26.598°N, 106.707°E), Guizhou Province. Twenty-seven fungi were isolated, including 10 isolates of Rhizopus and 17 isolates of seven other genera. The isolate YYH-CJ-17 had many sporangia that turned brownish gray to black on potato dextrose agar after culturing for 5 days in the dark at 25°C. The branches of mycelia were finger or root shaped. The sporangium was spherical or nearly spherical and 60 to 250 μm in diameter, and sporangiospores were elliptical or spherical and 4 to 8 μm in diameter. The obtained 547-bp internal transcribed spacer (ITS) fragment (accession no. OR225970) and 1,231-bp elongation factor 1-alpha (EF-1α) region (accession no. OR242258) from the isolate YYH-CJ-17 were compared with the sequences in the NR database using the BLAST tool provided by NCBI, which revealed more than 99.5% identity (query cover more than 98%) with the sequences of ITS (accession no. MF522822.1) and EF-1α (accession no. AB281541.1) of Rhizopus oryzae Went & H.C. Prinsen Geerlings (Gao et al. 2023; Zhang et al. 2023). The phylogenetic tree constructed with the ITS and EF-1α gene sequences demonstrated that the strain YYH-CJ-17 clustered with R. oryzae in the same branch, and the bootstrap value was greater than 99%. Based on the morphological characteristics and ITS and EF-1a sequences, the isolate YYH-CJ-17 was identified as R. oryzae. Pathogenicity tests were performed on detached healthy leaves and living plants of E. sagittatum. Healthy leaves of E. sagittatum were subjected to inoculation with the isolate YYH-CJ-17 with 5 × 105 CFU/ml concentration in sterile culture dishes. The progression of the disease was marked by gradual softening of the infected leaves and expansion of lesions, which ultimately produced blackish brown sporangium. Furthermore, the E. sagittatum living plants were sprayed with 5 × 105 CFU/ml conidial suspension of the isolate YYH-CJ-17, with ddH2O as a negative control, and then were cultivated at 25°C and 90% relative humidity for 21 days in a greenhouse. This assay found that the E. sagittatum leaves treated with the isolate YYH-CJ-17 exhibited the same symptoms observed on plants in fields. The fungus reisolated from the inoculated leaves was identified as R. oryzae by ITS sequencing, and its sequence blasted with the NR database showed the highest match with the ITS sequence (accession no. MF522822.1), thus fulfilling Koch's postulates. R. oryzae has been identified as a causative agent of a diverse array of host diseases, including leaf mildew of tobacco, fruit rot of yellow oleander and pears, and soft rot of bananas (Arif et al. 2017; Khokhar et al. 2019; Kwon et al. 2012; Pan et al. 2021). To the best of our knowledge, this is the first report of leaf rot on E. sagittatum caused by R. oryzae in China, which will provide a clear prevention and management target for the leaf rot disease of E. sagittatum.The author(s) declare no conflict of interest.References:Arif, S., et al. 2017. Can. J. 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Gao contributed equally to this work.Funding: This project was supported by the Ability Establishment of Sustainable Use for Valuable Chinese Medicine Resources (grant number 2060302), the National Technical System of Traditional Chinese Medicine Industry (grant number CARS-21), High-Level Innovative Talents of Guizhou Province of China (Qian Ke He Platform and Talent [2018]5638-2), Innovation Group Major Research Projects (Qian Jiao He KY Zi [2018]022), Guizhou Provincial Key Laboratory of Molecular Pharmacognosy (Qian Jiao Ji [2022]021), and Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences (CI2021B013).The author(s) declare no conflict of interest.DetailsFiguresLiterature CitedRelated Just PublishedSubscribeISSN:0191-2917e-ISSN:1943-7692 Metrics Article History Published: 1 Apr 2024First Look: 6 Feb 2024Accepted: 2 Feb 2024 Information© 2024 The American Phytopathological SocietyFundingAbility Establishment of Sustainable Use for Valuable Chinese Medicine ResourcesGrant/Award Number: 2060302National Technical System of Traditional Chinese Medicine IndustryGrant/Award Number: CARS-21High-Level Innovative Talents of Guizhou Province of ChinaGrant/Award Number: [2018]5638-2Innovation Group Major Research ProjectsGrant/Award Number: [2018]022Guizhou Provincial Key Laboratory of Molecular PharmacognosyGrant/Award Number: [2022]021China Academy of Chinese Medical SciencesGrant/Award Number: CI2021B013KeywordsEpimedium sagittatumleaf rot diseaseRhizopus oryzaeThe author(s) declare no conflict of interest.PDF download