First Report of Powdery Mildew Caused by on in China.

Symphyotrichum subulatum (Michx.) G. L. Nesom (syn. Aster subulatus), an annual herb in the Asteraceae family, is native to North America. Nowadays, it has become an invasive weed in several provinces of China, including Jiangsu, Zhejiang, and Sichuan (Li and Xie, 2002). Despite being invasive, this species holds significance in Chinese medicine, where it is used for the external treatment of eczema and swollen sore poison (Hu, 2020). In June 2023, symptoms of powdery mildew were observed in S. subulatum populations in Deyang and Nanchong, Sichuan Province, China. About 32.73% among 55 surveyed S. subulatum plants showed signs of infection. Symptoms initially appeared as small, scattered white powdery patches on the leaves, which enlarged and coalesced over time. Subsequently, hyphal growth forming extensive conidia covered up to 90% of the leaf area on both surfaces (Fig. S1A, B), and the infected leaves withered and fell off (Fig. S1A). A specimen was archived at China West Normal University (SsPM-ZL). Conidiophores were cylindrical and erect, 66.4 to 183.2 µm (avg. 108.2±40.8 μm) in length (n=30) (Fig. S1C). Conidia, produced singly, were ellipsoid-ovoid to nearly cylindrical, measuring 29.5 to 36.7 μm in length (avg. 32.9±2.6 μm) and 16.0 to 19.9 μm in width (avg. 17.4±1.3 μm), lacking distinct fibrosin bodies (n=30) (Fig. S1D). Under a scanning electron microscope, turgid conidia displayed reticulate wrinkles on the surface, with gentle contractions or bulges at both poles (Fig. S1E, F). Based on these characteristics, the powdery mildew fungus was consistent with the genus Golovinomyces (Bradshaw et al. 2022a). To confirm the identity of the causal fungus of specimen (SsPM-ZL), the calmodulin (CAM), RNA polymerase II subunit (RPB2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), glutamine synthetase (GS), and the internal transcribed spacer (ITS) were amplified using PMCAMF/R, PMRPB2F/R, PMGAPDH1/3R, PMGSF/R, and PM5/ITS4 primers (Bradshaw et al. 2022b), and deposited in GenBank (CAM: OR761878; RPB2: OR761881; GAPDH: OR761879; GS: OR761880; ITS: OR758452). BLAST analysis showed 99 to 100% identity with the sequences of Golovinomyces ambrosiae (FH00941234) for CAM (ON101658, 99.65%), RPB2 (ON119165, 100%), GS (ON075690, 99.78%), and ITS (ON073876, 99.47%). Phylogenetic analysis was performed in MEGAX with maximum likelihood method (Kumar et al. 2016) and clustered SsPM-ZL into the G. ambrosiae clade with a 100% bootstrap support value based on the concatenated sequences of CAM, RPB2, GAPDH, GS and ITS (Fig. S2). Combining morphological and phylogenetic analyses, SsPM-ZL was identified as Golovinomyces ambrosiae. To evaluate pathogenicity, leaves of 3 healthy potted S. subulatum plants (3 leaves per plant) were inoculated by gently pressing them with diseased leaves, while 3 non-contact plants were used as control. Plants in two groups were incubated in separate greenhouses maintained at 27±1°C, with a photoperiod of 14 hours and a relative humidity of 80%. After 7 days, the inoculated plants exhibited symptoms of powdery mildew (Fig. S1H, J), while the control plants remained asymptomatic (Fig. S1G, I). Morphological characteristics of the artificially induced powdery mildew were consistent with those on naturally infected plants. Powdery mildew caused by G. ambrosiae has been reported affecting Helianthus tuberosus (Huang et al. 2017) and Bidens pilosa (Mukhtar et al. 2022) in China. To our knowledge, this is the first report of powdery mildew caused by G. ambrosiae on S. subulatum in China. Our finding will provide the fundamental knowledge for future powdery mildew diagnosis and the development of potential control strategies.