D5100 camera

A thorough morphological investigation was conducted following the techniques described in Khonsanit et al. (2020) . The samples and culture plates were photographed using a digital Nikon D5100 camera. Colony characteristics and microscopic measurements of phialides and conidia (length and width) were done after growth on potato dextrose agar medium (PDA, 20 g Difco potato dextrose agar, 1 L distilled water) and incubation under white light/dark cycles at room temperature; 30 phialides and conidia were measured for each strain. To statistically test the difference between the genetic groups within each of B.asiatica and B. bassiana, five strains for each genetic group were examined for the width and the length of conidia and phialides (Table S3). The data were analysed with one-factor ANOVA for testing difference between species, as well as between the genetic groups within each species.

The characteristics of the sori and teliospores were examined on infected plants. Pictures of sori were taken using a Nikon D5100 camera. Teliospore characteristics were studied using a compound microscope (LM; BX60F; Olympus optical Co. Ltd., Japan) equipped with ProgRes C5 camera (JENOPTIK, Germany) and CapturePro software; and a scanning electron microscope (SEM; Hitachi SU6600) at 5.0 kV at the Clemson University Electron Microscopy Facility. For LM, teliospores were mounted in lactic acid (85–90 %, VWR, International, LLC) (Savchenko et al. 2014 (link)) and examined at 1 000 × magnification. The diameters of 30 teliospores, oriented in plane view so that they appeared globose, were measured from each sample collection. The colors of the sori and the teliospores were described according to Rayner (1970) . For SEM examination, teliospores were dusted on double-sided adhesive carbon tape, mounted on aluminum stubs, and sputter-coated with platinum using a Cressington sputter-coater (ca. 30 nm in 6 min).

The invasion of transfected cells in monolayer, anoikis resistant and reseeding conditions were assayed in modified Boyden chambers as described previously, with certain modifications (16 (link)). Briefly, for the invasion assay, 8-μm pore membrane filters were placed in Transwell chambers (Corning, Inc.) and the upper chamber was coated with 30 μg Matrigel (BD Biosciences) at 37°C in an incubator overnight. A suspension of 2×10⁵ cells from each culture condition was added to the upper chamber (200 μl/chamber), and DMEM was added to the lower chamber (500 μl/chamber). The chamber was incubated at 37°C for 24 h in a humidified atmosphere of 5% CO₂. Subsequently, the cells on the upper surface were removed with a cotton swab, and the cells were fixed with 30% methanol, followed by staining with 0.5% crystal violet in 20% methanol, both steps were performed for 15 min at room temperature. Finally, stained cells were counted in 5 randomly selected fields, using an inverted microscope (Eclipse TS100; Nikon Corporation) and then imaged using an attached D5100 camera (Nikon Corporation) (magnification, x100).