This research explored the diversity and evolutionary relationships of puffball mushrooms in Israel, discovering six previously unknown species in the region. The study combined traditional microscopic examination with modern DNA analysis to better understand how these fungi are related to each other. This work is important for everyday life in several ways: • Helps mushroom enthusiasts and experts better identify potentially edible puffball species in Israel • Contributes to understanding biodiversity conservation in Middle Eastern ecosystems • Advances knowledge of fungi that have shown potential medicinal properties including anti-cancer effects • Improves our ability to track and monitor fungal species distributions as climate changes • Provides foundation for future research into useful compounds these fungi might produce