First report of rootstalk rot of Hibiscus mutabilis caused by Fusarium oxysporum in China.

Abstract

H. Mutabilis (Cotton rose or confederate rose) is a deciduous shrub in the Malvaceae family, with ornamental, medicinal and edible values (Fan et al. 2015). In May to August 2020, 40.4% of potted plants of H. mutabilis were found to have root and stalk rot in Chengdu Botanical Garden (E104°7'28″, N30°45'57″). At first the leaves of affected H. mutabilis turned yellow and wilted, followed by the stem and root cortex became dark brown and rotten. Finally, the whole plant died within two months. Root and stem produced white mycelium when the humidity exceeded 90%. Samples taken from the lesions were surface disinfested for 3 min in 4% sodium hypochlorite, rinsed in sterile water and plated on potato dextrose agar (PDA), 35 single-spore cultures with similar morphology isolated from symptomatic tissues were obtained and subcultured. After seven days at 25°C in the dark, the mycelium of a representative culture MFR1 covered the entire plate surface (9 cm diameter). The aerial mycelium of cultures were white and fluffy at first and produced lavender pigment on the back of the cultures in the later stage. After seven days, the cultures produced abundant sickle-shaped macroconidia which have 3 to 5 septations and some oval or oblong microconidia which have 0 to 1 septation. Macroconidia 22.35~46.67 μm (mean 32.11 μm) in length and 4.32~7.72 μm (mean 5.21 μm) in width (n = 100). Microconidia 7.10~21.85 μm (mean 11.62 μm) in length and 2.76~6.84 μm (mean 4.20 μm) in width (n = 100). Based on these characteristics, isolates were tentatively identified as Fusarium sp. (Crous et al. 2021). Pathogenicity was tested on 1-year-old potted seedlings of H. mutabilis by root-zone irrigation inoculation in Sichuan Agricultural University (Jia et al.2019). Conidia suspension (1×107conidia/mL,collected from MFR1 )was poured into the soil along the plant roots. The same amount of distilled water was poured around the roots of the control plants. All inoculated and control plants were incubated in the greenhouse (about 25 ± 2°C). The experiment was performed three times. Within 25 days after inoculation, all plants inoculated with pathogens showed symptoms similar to those in the field, whereas the controls remained symptomless. The pathogen was reisolated from all inoculated plants, and the cultural and morphological characteristics were the same as those of the original isolate. After DNA extraction and PCR amplification, the translation elongation factor 1-alpha (TEF) and RNA polymerase II second largest subunit (RPB2) genes of a representative culture MFR1, were sequenced (O'Donnell et al. 2010) and deposited in GenBank (accession numbers OK334295 and ON316728, respectively). The TEF and RPB2 sequences were 99.7% and 99.39% identical to those of F. oxysporum (MN892354 and MZ198892). The result was confirmed by multilocus phylogenetic analysis. Through morphological identification and molecular analyses, the pathogen was identified as F. oxysporum. F. oxysporum is known to infect cotton (Dowd et al.2004), soybean (Ellis et al.2016) and banana (Fourie et al.2011) among other hosts, but it is the first report of F. oxysporum infecting H. mutabilis in China or worldwide. This disease seriously reduced the survival rate of H. mutabilis and may become an important reason to hinder the increase of H. mutabilis in potted seedlings stage. Moreover, the findings will provide theoretical basis to solve the bottleneck problem affecting the popularization and propagation of H. mutabilis.