Sporeling regeneration and ex situ growth of Isoëtes cangae (Isoetaceae): Initial steps towards the conservation of a rare Amazonian quillwort

Isoëtes L. is a nearly cosmopolitan rhizomorphic lycopsid genus fundamental to disentangling plant evolution because of its singular position in the Plantae kingdom. A threatened Amazonian species recently described is I. cangae, inhabiting an oligotrophic upland lake over an iron-rich region facing substantial environmental alterations. In this study, we aim to optimize a reproduction protocol that can assist in the conservation efforts of Isoëtes species in tropical areas. We observed an average of 126 megaspores or 318,000 microspores per sporangia in field-collected plants. We obtained an elevated percentage of sporelings (63%) from an in vitro mixed culture of mega- and microspores. In addition, we recovered a large number of sporelings around adult plants after cultivating them over quartzite sand and distilled water. The megaspore nutrient reserve ensured longstanding sporeling growth and development in these environmental conditions. However, the sporeling growth rate increased when they were planted over their original lake sediments or over an organic substrate, with nonsignificant differences between the substrates. An increase in temperature also increased the sporeling growth rate. Together with the very low sporeling mortality, these results suggest that I. cangae is a resilient species able to grow and colonize environments with low trophic characteristics with a range of substrates and temperature regimes. The protocols established in this study could be useful for the ex situ propagation and growth of a large number of sporelings, representing critical steps towards the population rescue and conservation of tropical Isoëtes species.