First Report of Sclerotinia sclerotiorum Causing Hibiscus rosa-sinensis Stem Rot in Guangxi, China.

Abstract

Hibiscus rosa-sinensis, native to the south of China, is currently planted as an important landscaping tree species in more than 100 countries around the world. Since 2012, an unknown stem rot disease of H. rosa-sinensis has occurred sporadically in a few green belts of Nanning, Guangxi, China. In February 2023, the incidence rate of the disease in the southern part of the city (108°38'E, 22°77'N) reached 5-8%. The pathogen mainly infected the stems near the soil line and aboveground stems. Initially, brown spots appeared and developed into long strips of large spots around the stem, slightly sunken. Later, the diseased tissue cortex presented longitudinal cracks and the vascular bundle tissue was exposed like silk hemp. White mycelium appeared on the diseased stem surfaces under high humidity conditions, eventually maturing into hard black sclerotia (1.5 to 11.0 mm in length). The leaves turned yellow and the whole plant finally died. For fungal isolation, seven diseased plants distributed within 800 square meters were collected, and 35 symptomatic stem sections were surface disinfect with alcohol for 30s, 0.08% NaClO for 1 min, triple rinsed with sterile distilled water, and cultured in potato dextrose agar (PDA) medium at 28℃. Sclerotinia-like colonies were consistently isolated from all diseased tissues and four isolates (Z1-Z4) were purified (Bolton et al. 2006). Irregular white immature sclerotia were produced after 5 to 7 days on the edges of the plates and turned black after 7 to 14 days, with a size of 1.8 to 4.6 × 1.2 to 3.4 mm (avg. 3.3 × 2.4 mm, n = 20). For molecular characterization, three gene regions (ITS, CaM and Mcm7) were amplified (White et al. 1990; Carbone et al. 1999; Schmitt et al. 2009) and sequenced (GenBank accession nos.: ITS: OR016764 to OR016767; CaM: OR257811 to OR257814; Mcm7: OR345318 to OR345321). The sequences of three analyzed DNA fragments shared 100% identity with sequences of Sclerotinia sclerotiorum strains (accession nos. JN013184, AF341304, KF545468). To fullfill Koch's postulates, healthy H. rosa-sinensis nursery stocks at the six months stage were individually planted in plastic pots at 25±3℃. The base of the stem and upper three branches of each plant were wounded with a sterile needle and inoculated with 5-mm discs of mycelium grown on PDA, then the inoculation sites of stem bases were covered with one layer nursery substrate and those of branches were wrapped with transparent tape to maintain the humidity. Three plants were inoculated with each isolate. As a control, three plants were inoculated with PDA discs. All the inoculated plants with mycelial discs developed characteristic symptoms 5 to 8 days after inoculation. The inoculation sites appeared white mycelium and the leaves sagged and wilted. Later, black sclerotia appeared on the diseased stem and the whole plant withered, while the control plants remained symptomless. Fungal cultures reisolated from symptomatic plants were morphologically identical with the cultures used as inoculum. Sclerotinia sclerotiorum has only been reported from H. rosa-sinensis in Taiwan (Tai 1979). The pathogen is a widely distributed fungus, causing many economically important diseases on various plants (Hossain et al. 2023). To our knowledge, this is the first report of S. sclerotiorum causing H. rosa-sinensis stem rot in Chinese Mainland, laying the foundation for monitoring its occurrence and spread.