Among the different secondary structures that DNA can adopt, G-quadruplex is a noncanonical form that has recently started to garner attention about the possible layers of regulation they could introduce in cellular processes. Here, we outline how the presence of G-quadruplexes can be probed in legumes and other plant genomes. This chapter describes various in silico approaches that can be utilized to identify putative G-quadruplex forming sequences (GQSes) and validate their formation through in vitro experimental approaches. © Springer Science+Business Media, LLC, part of Springer Nature 2020.