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Mahonia fortunei belongs to Berberidaceae, and is often used as a potted plant in Chinese tradition medicine (Li et al. 2015). In March 2019, leaf blight was observed on this species in the campus of Anhui Agricultural University (31°51'51″N; 117°15'31″E) in Hefei City, Anhui Province, China. The average disease incidence was 35% over 20 m2 planting area. Symptoms of the disease are easily apparent, with irregular, dark brown necrosis at the edge of the leaves. To identify the pathogen, symptomatic leaves were collected from three symptomatic leaves from three plants and cut into small pieces. Leaf pieces from the margin of the diseased tissues were surface sterilized for 1 min in 75% ethanol, rinsed three times with sterile distilled water, and subsequently, plated onto potato dextrose agar (PDA) and incubated in dark at 28°C for 4 days. The colonies of three isolates were obtained and appeared morphologically similar in agar media. The cultures were initially white, gradually becoming dark gray, and could grow to the edges of plates (90mm diameter) four days after subculturing. After 7 days on media, the colony was induced for sporulation by UV for 15min, and conidia were observed after 2 weeks. Conidia were nonseptate, hyaline, and oval, measuring 16.9 to 25.3 μm (mean 21.4 μm) × 5.0 to 8.4 μm (mean 6.8 μm) (n = 50). The isolate was identified as Botryosphaeria dothidea preliminarily according to the morphological characteristics (Zhai et al. 2014). For accurate identification, DNA was extracted from the mycelia. The internal transcribed spacer (ITS), β-tubulin (TUB) and Ef1-a were amplified and sequenced using primers ITS1/ITS4, Bt2a/Bt2b and 983f /2228r, respectively (Zhai et al. 2014). The 543-bp ITS (GenBank accession no. MK850215), 673-bp TUB (no. MN370930) and 1069-bp Ef1-a (no. MN598070) showed >99% identity with those of B. dothidea KP183180 (Xu et al. 2015), KU306116 (Lisboa et al. 2016) and DQ767637.1 (Schoch et al. 2006), respectively. Based on cultural characteristics and phylogenetic analysis, the current isolate associated with leaf blight of M. fortunei was identified as B. dothidea. The representative strain was deposited at the China Center for Type Culture Collection (CCTCC) as CCTCC AF 2019004 SX03. Pathogenicity test was performed on three healthy M. fortunei on campus. Three branches of each plant were selected and all leaves on branches were wounded with dissecting needle by puncturing, then inoculated with a spore suspension (105 conidia/mL) by a hand-held spray bottle, using sterile water as control (Huang et al. 2019). In order to maintain high relative humidity, the inoculated leaves were sealed with transparent plastic bags for 3 days. Fourteen days after inoculation, symptoms (leaf blight) were similar to those on infected leaves on campus, while the controls remained healthy. The experiments were conducted three times and the pathogen was reisolated from inoculated leaves and was confirmed as B. dothidea based on morphological and molecular analyses. To our knowledge, this is the first report of B. dothidea causing marginal leaf necrosis on M. fortunei in China. This study provides a preliminary basis for identifying the causal agent of this plant disease.

Ligustrum japonicum is a small evergreen tree belonging to the Oleaceae and widely grown in China as a landscape ornamental and medicinal plant (Oh et al. 2021). In April 2021, a leaf spot disease was observed on Ligustrum japonicum in the campus of Anhui Agricultural University (31°51'4″N; 117°14'54″E), in Hefei City, Anhui Province, China. Approximately 50% of L. japonicum infections showed symptoms of round to oval, brown to dark brown lesions surrounded by a yellow halo. Diseased leaves were collected to determine the caused pathogen. Nine tissue pieces from three symptomatic leaves were surface sterilized with 2% NaClO for 1-2 minutes, followed by 75% ethanol for 1 minute, and then rinsed with sterile water for three times. The tissues samples were plated on potato dextrose agar（PDA）medium and incubated at 28 °C for 3 days. Seven fungal isolates were obtained from the plated tissues; the fungal hyphae were initially white and finally gray brown with flocculent aerial mycelia. Conidia were solitary or in chains, with various shapes, mostly subglobose. The size was (12.0-30.0) μm long and (6.0～12.0) μm wide (n=20). The cultural and morphological characteristics of these isolates were similar to those of Alternaria alternata (Simmons et al. 2007; Garibaldi et al. 2020). For accurate identification, genomic DNA was extracted from the mycelia of representative isolate (JSNZ). The internal transcribed spacer (ITS), 18S nrDNA (SSU), translation elongation factor 1-alpha (Tef1-α) and endopolygalacturonase (endoPG) sequences were amplified with the primer pairs of ITS1/ITS4, NS1/NS4, EFI-728F/EFI-986R and EPG-specific/EPG-3b, respectively (Woudenberg et al, 2013). The sequences were deposited in GeneBank under accessions MZ360963 (ITS), MZ677478 (SSU), OK274117(Tef1-α) and OK513186 (endoPG). BLAST analysis of the sequences of ITS, SSU, Tef1-α and endoPG showed >99% identity with those of A. alternata MK108918(561/601bp), KX609765 (1030/1035bp), LC132712 (281/281bp) and MT185591(459/483bp), respectively. A neighbor-joining phylogenetic tree was generated based on the concatenated data from sequences of ITS, SSU and Tef1-α using MEGA5.1, which clustered the present isolate with A. alternata strain CBS916.96 with high bootstrap support (100%). Based on cultural characteristics and phylogenetic analysis, the current isolate associated with leaf spot of L. japonicum was identified as A. alternata. Pathogenicity test was performed on three healthy L. japonicum on campus. Three healthy leaves of each plant were wounded with one sterile needle and inoculated with a 5-mm-diameter mycelial plug using sterile PDA plugs as control. The inoculated plants were covered with plastic bags and sprayed with water every 24 hours to maintain a high temperature and humidity environment. The experiment was repeated three times. After 12-days of incubation, symptoms were apparent on pathogen-inoculated plants, while the control plants remained asymptomatic. A. alternata was reisolated from inoculated leaves and matched the morphological and molecular characteristics of the original isolates, thus fulfilling Koch's postulates. To the best of our knowledge, this is the first report of a leaf spot disease caused by A. alternata in L. japonicum in China. Its identification will establish a foundation for managing the disease in China.

Berberis fortune (Lindl.) is commonly used in Chinese traditional medicine (Liu et al. 2020). In April 2020, white powdery colonies covering up to 100% of both upper leaf surfaces and calyces were observed on this species growing on Anhui Agricultural University campus (31°51'51″N; 117°15'31″E) in Hefei City, Anhui Province, China. Sporulating mycelia were white and effuse. Conidiophores were erect, with straight, cylindrical foot cells, 20 to 26 × 9 to 12 μm (average: 24 × 11 µm) (n = 30), followed by one to three shorter cells, and producing conidia in chains. Conidia were ellipsoid-ovoid, subcylindrical, and measured 27 to 36 × 12 to 16.5 µm (average: 32.4 × 14.1 µm) (n = 50). For accurate identification, DNA was extracted from the mycelia, which were collected by scraping symptomatic leaves. The internal transcribed spacer (ITS) was amplified and sequenced using primers ITS1/ITS4. The 623-bp ITS (GenBank accession no. MT449013) showed 99% identity with those of Erysiphe berberidis LC010057 (Takamatsu et al. 2015), KY661153 and KY660920. Based on morphological characteristics and phylogenetic analysis, the powdery mildew fungus on B. fortunei was identified as E. berberidis (Glawe, D. A. 2003). Ten leaves on an asymptomatic B. fortunei were inoculated by gently pressing diseased leaves against the surface of healthy leaves. Ten non-inoculated plants served as controls. All plants were maintained in a greenhouse at 22 to 25°C and >80% relative humidity. Inoculated plants developed powdery mildew colonies after 14 days, whereas uninoculated plants remained healthy. Morphological and molecular characters of the powdery mildew fungus on artificially inoculated plants were identical to those on naturally infected B. fortune. Previously in Siberia, Russia, powdery mildew on woody plants has been reported to be caused by E. berberidis (Tomoshevich M. A. 2019). However, this is the first report of powdery mildew caused by E. berberidis on B. fortunei in China. Its identification will establish a foundation for controlling the disease in China.