Zen 2 core

The initiation of fruiting body development was quantified after 3 days of incubation on BMM-covered microscope slides. Ascogonia and protoperithecia were counted separately within an area of 0.5 cm² located 1 cm behind the growth front. An Axio Imager M1 (Zeiss) with a SpectraX LED lamp (Lumencor) and a Photometrix Cool SnapHQ camera (Roper Scientific, Martinsried, Germany) was used for quantification. Experiments were repeated for three biological replicats per strain. For assessment of fruiting body formation, images of strains were taken after 7 days of incubation in Petri dishes on solid BMM with a Zeiss Stemi 2000-C binocular, using an AxioCam ERc5s with the software ZEN 2 core (version 2.5, Zeiss, Jena, Germany).

Stained sections were analyzed in bright field microscopy to measure tubular diameter (at least 50 seminiferous tubules per section – Microscope Software Zen 2 core, ZEISS, Germany) and assess histological changes in testicular parenchyma. Changes in tubules were classified into four scores (Score I: highest damage to Score IV: full normality). Score I: complete absence of Sertoli and germ cells inside the tubules; Score II: presence of Sertoli cells only (containing or not abnormalities – e.g., vacuolar degeneration, blebbing, etc.); Score III: presence of normal Sertoli cells and abnormal germ cells; and Score IV: presence of Sertoli and germ cells without morphological alterations.

Fractured charpy v-notch samples were placed in the microscope (AX10, ZEISS, GER) below the 10× (Epiplan 10×/0.25 HD M27, ZEISS, GER) lens where it was imaged (Axiocam 105 color, ZEISS, GER) and saved by software (ZEN 2 core, ZEISS, GER).