Scope 5

Photographs of fresh stromata were taken using an ultra-depth of field stereomicroscope (digital microscope system Keyence VHX-7000) to illustrate the macrostructures. Sections were made using a stereomicroscope (Leica S9i) and mounted in water or a rehydrated 5% KOH solution. Photomicrographs of perithecia, asci, ascospores, conidiophores, conidia, and phialides were taken with a compound light microscope (Zeiss Scope 5 with color camera AxioCam 208) to observe the morphological characteristics. All measurements of the observed structures were made with ZEN2 (blue edition) software.

In the preliminary screening, a 5 mm-diameter disc of mycelium was taken from the margin of the fungal isolates, transferred to the center of a Petri dish containing 1% WA, and then incubated in the dark at 25 °C for 2 weeks. After the incubation period, Petri dishes were inoculated with a 1 ml nematode suspension containing 1000 nematodes. The nematode suspension was divided into 4-5 drops and spread uniformly inside the periphery of the fungal colonies. Plates without fungi served as controls. The morphology and the average size of the hyphal rings were observed and recorded. Four replicates of each isolate were utilized. The number of nematodes captured in the hyphal rings was counted using a stereomicroscope (Leica S9i, Germany). Microscopic images were captured with a compound light microscope (Zeiss Scope 5) equipped with an AxioCam 208 color camera.

A 5 mm-diameter disc of mycelium was removed from the margins of the fungal isolates, placed to the center of a petri dish containing 1% water agar (WA) and cultured for two weeks in the dark at 25°C. Petri dishes were infected with a 1 mL nematode slurry containing 1000 nematodes after the incubation period. The nematode suspension was separated into 4-5 drops and evenly distributed across the fungal colonies periphery. Control plates were made without fungus. Each strain was tested in four replicates. A compound light microscope (Zeiss Scope 5) with an AxioCam 208 color camera was used to acquire microscopic images.

From 2021 to 2022, 94 samples were collected from orchards in six locations of Guizhou Province (Congjiang, Kaiyang, Longli, Luodian, Nayong and Wengan counties), including 10 host species (cherry, loquat, passionfruit, pitaya, plum, pomegranate, Rosa roxburghii, shaddock, tangerines and walnut). To obtain pure cultures, leaf surfaces were disinfected according to Zhang et al. [21 (link)]. Abundant conidia were observed on the surface of the leaf spots examined using a dissecting microscope. Single conidia were picked off the leaves with a sterilized needle and placed on a drip board containing sterilized water. After 12 h, the germination of conidium was observed, and they were then transferred to potato dextrose agar (PDA) and incubated at room temperature (28 °C) for 10 days. Morphological characteristics of the fungi were observed and photographed using a compound light microscope (Zeiss Scope 5) with an attached camera (AxioCam 208 color). All new taxa were registered in the Index Fungorum database (www.indexfungorum.org) (accessed on 13 October 2022). Dried holotype specimens were conserved in the Herbarium of the Department of Plant Pathology, Agricultural College, Guizhou University (HGUP), while cultures were conserved in the Departmental Culture Collection (GUCC).

Photographs of the ascomata were taken using a stereomicroscope (Keyence VHX-7000 digital microscope). Observations and photomicrographs were made from material mounted in lactic acid (60%) using a compound light microscope (Zeiss Scope 5) equipped with an AxioCam 208 colour camera with interference contrast optics. All measurements were made with ZEN2 (blue edition) and images used for figures were processed with Adobe Photoshop version 2022 software.