Development of control strategies for bacteria and fungi associated with a micropropagated new cultivar of orange-fleshed sweet potato (Ipomoea batatas cv. Agrosavia–Aurora)

Abstract

AbstractVegetative propagation of sweet potato lead to the accumulation of diseases from generation to generation, which represents a threat to both productivity and conservation of genetic resources. In vitro techniques can help to overcome phytosanitary problems by applying plant material cleaning strategies. The objective of this study was to develop in vitro micropropagation strategies for the production of high-quality plant material of an orange-fleshed variety of sweet potato recently released in Colombia. Molecular identification of contaminating microorganisms was performed by sequencing the 16S rRNA gene for bacteria and ITS for fungi. Five disinfection protocols were evaluated, three of which were previously developed for sweet potato and included disinfection with 0.5, 1, and 2% sodium hypochlorite respectively, while two protocols are proposed in this work and included washing with povidone-iodine, disinfection with sodium hypochlorite 2%; one of these two new protocols also contains acetic acid and quaternary ammonium. For the evaluation of the viability of in vitro plants after disinfection, they were acclimatized in a greenhouse, reintroduced, and a molecular testing by PCR of 16S rRNA gene and ITS was carried out to verify the phytosanitary status of the material. The contaminating microorganisms found were filamentous fungi of the genera Fusarium, Sarocladium, Cladosporium and Aspergillus, yeasts of the genera Pseudozyma and Moesziomyces, and the actinobacterium Curtobacterium sp. The results indicated that washing with povidone-iodine and disinfection with 2% sodium hypochlorite, acetic acid and quaternary ammonium was the most efficient disinfection protocol, reducing the number of contaminated cultures by up to 10% and eradicating 70% of contaminants. The in vitro plants established in the greenhouse remained healthy and, after reintroduction, the molecular test for bacteria and fungi was negative. These results allowed the generation of an optimized protocol that can be incorporated into the in vitro micropropagation process to generate contamination-free sweet potato seeds.