This study led to the discovery of three entomopathogenic fungi associated with Kuwanaspis howardi, a scale insect on Phyllostachys heteroclada (fishscale bamboo) and Pleioblastus amarus (bitter bamboo) in China. Two of these species belong to Podonectria: P. kuwanaspidis X.L. Xu & C.L. Yang sp. nov. and P. novae-zelandiae Dingley. The new species P. kuwanaspidis has wider and thicker setae, longer and wider asci, longer ascospores, and more septa as compared with similar Podonectria species. The morphs of extant species P. novae-zelandiae is confirmed based on sexual and asexual morphologies. Maximum likelihood and Bayesian inference analyses of ITS, LSU, SSU, tef1-α, and rpb2 sequence data provide further evidence for the validity of the two species and their placement in Podonectriaceae (Pleosporales). The second new species, Microcera kuwanaspidis X.L. Xu & C.L. Yang sp. nov., is established based on DNA sequence data from ITS, LSU, SSU, tef1-α, rpb1, rpb2, acl1, act, cmdA, and his3 gene regions, and it is characterized by morphological differences in septum numbers and single conidial mass.

Photinia × fraseri is a well-known green plant mainly distributed in the Yangtze River and Yellow River Basin, east and southwest of China (Guan et al. 2013). In October 2020, typical leaf blight s...

Magnolia coco (Lour.) DC. is an ornamental shrub and widely cultivated in southern China (Nana et al. 2017). In April 2020, leaf blight symptoms were observed on the leaves of M. coco in the Chengdu campus of Sichuan Agricultural University (30°42′19.92″N, 103°51′30.61″E, 493 m) where didn’t have great protection, with roughly 70% leaves per plant were diseased. The initial symptoms presented on the leaf apex, which was manifested as dark brown spots surrounded with obvious yellowish halo (Fig. 1). As the disease progressed, spots gradually enlarged and coalesced covering the leaf, and severe infection finally caused leaf necrosis and plant decline. Four specimens from different diseased plants were used for pathogen isolation and morphological observation. Four fungal isolates were obtained from four specimens, following Chomnunti et al. (2014). Colonies on potato dextrose agar (PDA) medium were initially white and then light brown to dark brown. Pycnidia measured 284-427 × 326-554 μm (x=372.8 μm × 476.1 μm, n=20), and were brownish-black to black, broadly globose to irregular. The pycnidial wall measured 16-27 μm wide (n=20) and was composed of hyaline to brown cells of textura angularis. Conidiophores were absent, and the conidiogenous cells are pear-shaped, colorless, and smooth. Conidia measured 5-8 × 4-6 μm (x=6.5 μm × 4.6 μm, n=50), and were elliptical or subglobose, thick-walled, aseptate, hyaline, smooth, brown. These asexual structures were similar to Nothophoma quercina (Syd. & P. Syd.) Qian Chen & L. Cai described by Chen et al. (2017). The genomic DNA of representative isolate SICAUCC 21-0011 was extracted, and the internal transcribed spacers (ITS), 28S large subunit rDNA (LSU), RNA polymerase II large subunit 2 (RPB2), and beta-tubulin (TUB2) regions were amplified using the primer pairs ITS5/ITS4, LR0R/LR5, FRPB2-5F/FRPB2-7cR, and T1/BT4R, respectively. The accession numbers deposited in GenBank were MW541930 (ITS), MW541934 (LSU), MW883395 (RPB2), and MW883394 (TUB2). Nucleotide BLAST showed high homology with the sequences of N. quercina, viz. GU237900 (ITS, 485/486, 99.79%), EU754127 (LSU, 862/862, 100%), KT389657 (RPB2, 593/596, 99.49%), and GU237609 (TUB2, 333/335, 99.40%). Phylogenetic analyses based on a combined dataset showed 100% bootstrap support values in a clade with N. quercina complexes (Fig. 2). Four healthy potted plants (2-years-old) with 15 to 20 leaves per plant were sprayed with conidial suspension (105 conidia/mL) prepared from 4-week-old cultures of SICAUCC 21-0011, which incubated on PDA at 25℃, onto the wounded sites via pin-prick inoculation described by Desai et al. (2019). Another four plants were sprayed with sterilely distilled water as controls. Inoculated plants were cultured in a growth chamber (25℃, 95% relative humidity, and 12-h photoperiod). About 30 days later, brown spots were found on the inoculated leaves, which were similar to those observed in the field. There were no symptoms on the control plants, and the pathogen was re-isolated from the diseased leaves and characterized morphologically. N. quercina has been reported on Photinia × fraseri Dress, Aucuba japonica, Malus micromalus, and Chaenomeles sinensis (Mohamed et al. 2019, Lv et al. 2020, Zou et al. 2021). To our knowledge, this is the first report of leaf blight on M. coco caused by N. quercina. M. coco is one of the important ornaments in the courtyard, street, and park in China, and the risk of this pathogen needs further exploration and effective control measures should be made. Qian Zeng, Yicong Lv, and Xinyue Li contributed equally to this work.

Aucuba japonica Thunb. is an ornamental shrub and widely cultivated in China (Zheng et al. 2019). In October 2019, leaf blight symptoms were observed in nearly 120 A. japonica plants in Chengdu cam...

Phyllostachys aureosulcata McClure 'Spectabilis' C.D. Chu. et C.S. Chao is predominantly native to subtropical to warm temperate areas and is widely cultivated for landscaping in China (Neményi et al. 2015). In November 2020 (10 – 16 ℃), culm blight symptoms were observed on P. aureosulcata 'Spectabilis' in Wangjiang Tower Park (all kinds of plant areas are about 9.8 ha), Chengdu City (104°09′30.42″ E, 30°63′18.89″ N). Fifty plants were surveyed, and disease incidence was recorded as approximately 30%. Initially, chlorotic necrotic patches appeared on the culms, and gradually the patches became white, expanded to both ends, and encircled the whole culm with black edge and conidiomata, which eventually led to wilt and death. Five samples from different bamboos were collected and one of them were used for morphological observation. Five single conidia isolates were carried out on potato dextrose agar (PDA) at 25±1℃ (Chomnunti et al. 2014). Colonies were initially white and then yellowish in the center with abundant aerial mycelia. On the culm, conidiomata were dry, black, and filamentous. Conidiophores were reduced to conidiogenous cells. Conidiogenous cells were smooth, hyaline, ampulliform to doliiform. Conidia were ellipsoid to globose, dark brown, smooth and aseptate, measuring 5.2 to 9.4 × 4.4 to 7.3 μm, (=8.2 × 6.5μm, n=50). On the PDA medium, conidia were globose to subglobose, olive green to pale brown, and smooth, larger than those from the host in size, measuring 9.0 to 18 × 7.5 to 9.5 μm ( =36.6 × 18.8 μm, n=50). These asexual structures were extremely similar to Apiospora locuta-pollinis (F. Liu & L. Cai) X.G. Tian & Tibpromma (Zhao et al. 2018). DNA was extracted from the representative strain (SICAUCC 22-0036), and the internal transcribed spacer (ITS), translation elongation factor 1-alpha (tef1-α), beta-tubulin (tub2), 28S large subunit rDNA (LSU) were amplified and sequenced with primers ITS1/ITS4 (White et al. 1990), EF1-728F (Carbone & Kohn 1999)/EF2 (O’Donnell et al. 1998), T1 (O’Donnell & Cigelnik 1997)/Bt2b (Glass & Donaldson 1995) and LR0R/LR5 (Rehner & Samuels 1994). The newly generated sequences were deposited in GenBank with accession nos. ON228609 (ITS), ON324018 (tef1-α), ON237657 (tub2), and ON228665 (LSU). Nucleotide blast showed 98.97%, 100% and 99.46% identities with A. locuta-pollinis (LC11683, ex-holotype) (accession nos. MF939595, MF939622, MF939616), and LSU data missing. Phylogenetic analyses using maximum likelihood showed a 92% bootstrap support value in a clade with A. locuta-pollinis (Fig 2). Eight healthy plants (2-year-old) were used for the pathogenicity test. Culms of four healthy bamboos were wounded via sterile double-edged blade and sprayed with conidial suspension (105 conidia/ml) prepared from 4-week-old cultures that were incubated on PDA at 25℃. The other four bamboos were sprayed with sterile distilled water as controls. Inoculated plants were placed in a growth chamber (25℃, 90% relative humidity, 12-h photoperiod). About 60 days later, necrotic patches similar to those observed in the field were found on the inoculated culms, and no symptoms were observed on the controls. The pathogen was reisolated from the diseased culms with identical morphology as previously described. To our knowledge, this is the first report of culm blight on P. aureosulcata 'Spectabilis' caused by A. locuta-pollinis. The risk of this pathogen needs further evaluation, and effective control measures should be taken.

Juglans sigillata Dode (Iron walnut) is mostly distributed in southwestern China, and valued for wood and nuts (Feng et al. 2018). In April 2020, we surveyed a walnut garden located in Baisha Town, Wanyuan City, (Sichuan, China), where brown spot symptoms were observed on leaves of ten trees among of 100 plants, and this disease can result in a reduced growth potential when trees are severely infected. Necrotic and subcircular lesions with conidiamata were observed on diseased leaves. Symptomatic leaves were collected and taken back to the laboratory forfurther analysis. Using the single spore isolation technique developed by Chomnunti et al. (2014), five isolates were grown from the infected leaves on Potato Dextrose Agar medium (PDA). The five isolates had similar colony morphology, which was initially white, suborbicular, gradually turning yellowish with black spots, developing fluffy aerial mycelium. Morphological characteristics were examined using light microscopy on the PDA. Conidiogenous cells were subcylindrical to cylindrical, or ampulliform, hyaline, rarely branched. Macroconidia were lunate, reniform, hyaline, 1-3-septate, mostly 1-septate, distinctly constricted at the septum, the basal cell was bluntly rounded, the apical cell had an acute end, and the basal cell was equal to or larger than the apical cell, measuring 22 to 40.5 × 2.5 to 8.3 μm (mean = 32 × 6.2 μm, n = 50). Microconidia were botuliform, or subfusiform, hyaline, both ends were rounded, straight or curved, aseptate, and measured 10 to 28.5 × 1.9 to 3.7 μm (mean= 17.2 × 2.7 μm, n = 20). A multilocus approach was conducted for precise identification of a representative isolate SICAUCC 20-0012. The internal transcribed spacer regions (ITS), guanine nucleotide-binding protein subunit beta gene (MS204), and translation elongation factor 1-alpha (tef1-α) of isolate SICAUCC 20-0012 were amplified and sequenced as described by Sogonov et al. (2008) and Walker et al. (2012a). GenBank Accession Nos. for ITS, MS204, and tef1-α are MW250303, MW246773, and MW246775, respectively. Phylogenetic analyses showed 100% support with Ophiognomonia leptostyla (Fr.) Sogonov, and the morphology was consistent with the asexual stage of O. leptostyla documented by Walker et al. (2012b). To test Koch's postulates, five healthy plants of J. sigillata (2- to 3-year-old) with 5-8 leaves per plant were inoculated with conidial suspensions (104 conidia/mL) after wounded with a small pin as described by Desai et al. (2019), and the same number of healthy plants were wounded and sprayed with sterile distilled water as controls. Plants were sprayed regularly with distilled water every day and placed in a growth chamber at 25℃ with a 12-h fluorescent light/dark regimen. After 15 days, typical brown spot symptoms developed on inoculated leaves, but not on the controls. The fungus O. leptostyla was reisolated from the lesion as described above but not from non-inoculated leaves. O. leptostyla has been reported on some walnut trees; for example: J. ailantifolia, J. californica, J. cinerea, J. hindsii, J. major, J. mandshurica, J. nigra, and J. regia (Farr & Rossman 2020). However, to our knowledge, this is the first report of O. leptostyla causing brown leaf spot on J. sigillata. J. sigillata is an economically important tree in southwest China, and fungicide treatments should be considered to prevent the spread of this fungus before it becomes more widespread. Chunlin Yang, Yu Deng, and Feihu Wang contributed equally to this work. This research was supported by the Key Research and Development Project of Sichuan Province (2021YFYZ0032).