Microencapsulation with the usage of sodium alginate: A promising method for preserving stonewort (Characeae, Charophyta) oospores to support laboratory and field experiments

This work proposes the use of a biomaterial sodium alginate for microencapsulating stonewort oospores. The microencapsulation method has been adopted and optimized to improve research work carried out on the oospores. A detailed protocol for the production of microcapsules with stonewort oospores has been presented, with particular attention paid to the critical stages and factors interfering with the efficient production process. The advantages and disadvantages of this method have also been indicated. Microcapsules with oospores can be successfully used in various laboratory (observation of the germination phases) and field experiments (reintroduction of rare species of stonewort). Alginate microcapsule may consist of water taken directly from the habitat of the stonewort, a medium, as well as specific additives (antibiotics or fungicides). The hardness of the encasement is regulated by the concentration of the sodium alginate. The optimum alginate concentration for oospores is between 2 and 3%. Encasements produced with less than 2% sodium alginate concentration do not form circular microcapsules during gelation. On the other hand, concentration >3% may limit the oospores' ability to germinate (higher density of the microcapsule). In such a dense environment, the germinating primary cells may encounter difficulties in outgrowing the microcapsule. It has been shown that alginate encasement creates an effective protective layer for the oospores inside it. The oospore is protected from mechanical damage as well as from direct exposure to the external environment; especially from pathogens and other microorganisms that can inhibit its germination and limit the growth of the protonemal and primary rhizoidal cell. The main application of oospore microencapsulation using sodium alginate is facilitating further mechanical manipulations of oospores, which by nature are very small structures (max. length 1 mm), while a microcapsule with an oospore embedded inside it can have a diameter from 3 to 5 mm (as required).