Techniques for locating and analyzing subterranean Lycopodium and Diphasiastrum gametophytes in the field

Homosporous club mosses have an archaic life cycle, alternating two locationally, nutritionally, and physiologically independent generations. The sexual generation of club mosses-the gametophytes (or prothallia)-are among the least researched botanical subjects. The gametophytes are responsible for not only sexual reproduction, but also the determination of recruitment of the new sporophyte generation, species habitat selection, migration, and evolution. Researchers often fail to find juvenile club moss populations and thus do not discover subterranean long-lived achlorophyllous gametophytes. To date, the gametophytes of most club moss species remain undiscovered in nature and are not scientifically documented. Almost all researchers who have previously located subterranean club moss gametophytes declared that their first find was due to luck and that subsequently the researcher's intuition plays the most important role; however, intuition and good luck are not scientific methods. In our review, we combine our knowledge with data available in the literature and discuss the following questions using a methodical approach: (1) How can we locate a subterranean club moss gametophyte population? (2) How can we extract the gametophytes? and (3) What new knowledge about club moss population development can be gained by analyzing juvenile club moss populations?