Digital sight ds ri2

Species identification was based on morphological characteristics of the ascomata or conidiomata produced on infected host materials. The macro-morphological characteristics including structure and size of stromata; the size, color, and shape of discs; number and diameter of ostioles per disc; presence and absence of conceptacle were determined under a Leica stereomicroscope (M205). The micro-morphological characteristics including size and shape of conidiophores and conidia were determined under a Nikon Eclipse 80i microscope equipped with a Nikon digital sight DS-Ri2 high-definition color camera with differential interference contrast (DIC). Over 10 ascomata/conidiomata were sectioned, and 10 asci and 30 ascospores/conidia were selected randomly for measurement. Colony morphology and growth rates were recorded and colony colors were described after 1 or 2 weeks according to the color charts of Rayner (1970 ). Adobe Bridge CS v.6 and Adobe Photoshop CS v.5 were used for the manual editing. Taxonomic novelties and descriptions were deposited in MycoBank (Crous et al., 2004 ).

To assess conidiation, three agar blocks (5 mm in diameter) carrying mycelia were introduced into 100 ml of liquid CMC medium. The suspension was shaken at 150 rpm for 4 days, conidia were filtered with one layer of miracloth, and the concentration of conidia was determined using a hemocytometer. The experiments were performed three times with 10 counting repetitions. Cell nuclei were stained with 1 μg/ml DAPI in the dark, observed at a magnification of ×600 under a fluorescence microscope (Nikon Eclipse Ti-S; Nikon, Tokyo, Japan) connected to a digital camera (Nikon Digital Sight DS-Ri2; Nikon, Tokyo, Japan), and analyzed using NIS-elements 4.5 imaging software (Nikon, Tokyo, Japan). For conidial germination assays, conidia were resuspended to 5 × 10⁴ conidia/ml in sterile water, and 50-μl droplets of the suspensions were placed on glass coverslips (Fisher Scientific) and incubated at 25°C for 3 h.

For the isolates isolated from diseased branches, the fruiting bodies on the specimens corresponding to the isolates were used for morphological observation. For the isolates isolated from leaf spots, reproductive structures formed on PDA were used for morphological observation. The structure and size of conidiomata were photographed using the Leica stereomicroscope (M205 FA) (Leica Microsystems, Wetzlar, Germany). Over 30 conidiomata were sectioned, and 50 conidia were selected randomly to measure their lengths and widths using a Nikon Eclipse 80 i microscope (Nikon Corporation, Tokyo, Japan) equipped with a Nikon digital sight DS-Ri2 high-definition color camera with differential interference contrast (DIC). Colony color on PDA were described according to the color charts of Rayner [59 ].