In vitro multiplication of agave tobala (Agave potatorum Zucc.) using Ebb-and-Flow bioreactor

Abstract

Agave tobala (Agave potatorum Zucc.) is one of the most important wild species to produce a Mexican alcoholic beverage called mezcal. Unfortunately, natural propagation is not sufficient to fulfill the demands. The objective of this study was to develop a protocol for the commercial micropropagation of A. potatorum using a temporary immersion system (TIS). The effect of liquid medium using an Ebb-and-Flow bioreactor and semi-solid medium as a control treatment was compared. In TIS different immersion frequencies, medium volume per explant, and additional air supply were evaluated. In all experiments, the hyperhydricity percentage, number of shoots per explant, as well as shoot length, and total chlorophyll content were recorded at 60 d of culture. In addition, the survival rate during acclimatization was evaluated at 60 d. The results show that the temporary immersion system with an immersion of 2 min every 12 h, with a culture medium volume of 50 mL and an air supply frequency of 2 min every 12 h obtained the best development parameters compared to semi-solid medium, with 29.60 shoots per explant, an average shoot length of 2.01 cm, and an hyperhydricity percentage of 3.33. The highest chlorophyll content was observed in semi-solid culture media and temporary immersion every 12 and 16 h. Regarding medium volume per explant, the highest chlorophyll contents were found in the volumes of 100, 75 and 50 mL per explant. For air supply, the frequencies of 8 and 4 h had a higher chlorophyll content. For acclimatization, no differences in percent survival were found between the culture systems evaluated. In conclusion, this study shows that the use of Ebb-and-Flow bioreactor is a suitable alternative for large-scale micropropagation of A. potatorum.