First Report of Sawadaea polyfida Causing Powdery Mildew of Acer palmatum in China
Acer palmatum Thunb. is an important colorful leaf ornamental tree species widely distributed in Japan, Korea and China (Carlos et al. 2016). In October 2019, powdery mildew was observed on leaves of A. palmatum planted at Qixia Mountain Park and the campus of Nanjing Forestry University, Nanjing, Jiangsu, China. The powdery mildew infected and colonized leaves, covering both leaf surfaces with white mycelia, giving affected plants an unsightly appearance. Nearly 17.4% of the plants (87/501) exhibited these signs and symptoms. Fresh specimens were collected and examined for the identification of the pathogen. Photos were taken with a ZEISS Axio Imager A2m microscope and a scanning electronic microscope. Chasmothecia were scattered or aggregated on the upper and lower surfaces of the leaves, blackish brown, oblate, 157.5 to 238.1 × 152.3 to 217.8 μm (n=30), with numerous appendages (100 to 200). Appendages were often (1−) 2 to 3 times branched from the middle of the stalk, uncinate to circinate at the apex, hyaline, aseptate, 30.0 to 70.8 × 4.1 to 8.2 μm (n=30). Asci were 11 to 21 per chasmothecium (n=30), long oval, oval, oblong, with short stalk or sessile, 80.6 ± 8.6 × 40.3 ± 4.0 um (n=30) in length, 6 to 8 spored (n=30). Ascospores were ovoid, 18.2 ± 1.6 × 11.1 ± 1.2 μm (n=30). Microconidiophores were 25 to 50 × 4.0 to 5.5 μm, producing microconidia in chains. Microconidia were ellipsoidal, subglobose, 8.7 ± 0.6 × 7.2 ± 0.6 μm (n=30). Macroconidia were not observed. Based on the morphological characteristics, the fungus was identified as Sawadaea polyfida (C.T. Wei) R.Y. Zheng & G. Q. Chen (Zheng and Yu 1987). To confirm the causative species identity, a representative voucher specimen collected and deposited at Nanjing Forestry University was used for a molecular analysis. Mycelia and conidia were collected from diseased leaves and genomic DNA of the pathogen was extracted and the internal transcribed spacer region (ITS) was amplified with primers ITS1/ITS4 (White et al. 1990). The resulting sequence of 461 bp was deposited in GenBank (accession no. MW255383). BLAST result showed that this sequence fully agreed with a sequence of S. polyfida [AB193381.1 (ITS), identities = 461/461 (100%)]. A maximum likelihood phylogenetic analyses using IQtree v. 1.6.8 with the ITS sequence placed this fungus in the S. polyfida clade. Based on the morphology and phylogeny, the fungus was identified as S. polyfida (Hirose et al. 2005; Zheng and Yu 1987). Pathogenicity was tested through inoculation by gently pressing the naturally infected leaves onto healthy ones of three potted A. palmatum seedlings wih five leaves. Healthy leaves from three other seedlings served as control. Inoculated and control seedlings were placed in separate growth chambers maintained at 20 ± 2°C, 70% humidity, with a 16 h/8 h light/dark period. Symptoms developed 8 days after inoculation. The powdery mildew developing on the inoculated seedlings was sequenced and confirmed as S. polyfida. The control leaves did not develop powdery mildew. S. polyfida has been reported on Acer catalpifolium in China (Zheng and Chen 1980), A. amoenum, A. australe, A. japonicum, A. palmatum, A. shirasawanum, and A. sieboldianum in Japan (Hirose et al. 2005; Meeboonet al. 2015), as well as A. takesimense in Korea (Lee et al. 2011). To the best of our knowledge, this is the first report of powdery mildew caused by S. polyfida on A. palmatum in China. These results form the basis for developing effective strategies for monitoring and managing this disease.