Characterization of biotic contaminants in aquatic lace plant tissue culture and development of a protocol for mitigation

Aquatic plant cultures are a critical component of agriculture and research. Despite this, innovation in the maintenance of sterile aquatic plant cultures, more specifically submerged aquatic cultures, is lacking, and propagators must use exacting prophylactic measures to alleviate contamination problems. The goal of this project was to identify contaminants in a lace plant ( Aponogeton madagascariensis) culture and use this information to develop an efficient cleansing protocol. Bacterial genera Paenibacillus, Pseudomonas, and Bacillus, as well as the fungal genus Meyerozyma, were consistently identified in contaminated lace plants. Three cleansing protocols were tested for effectiveness in reducing contamination without compromising plant growth and development. The most effective method consisted of two steps: surface sterilization of corm with 1% sodium hypochlorite, and media supplemented with 0.1% broad-spectrum biocide Plant Preservative Mixture™. This two-step method offered a cost-effective and straightforward broad-spectrum sterilization procedure for lace plant cultures that minimized contamination while supporting normal plant growth and development. This protocol may be effective for minimizing contamination in other aquatic plant cultures containing multiple types of microbes. To the best of our knowledge, this is the first paper to report an efficient protocol to minimize contaminants in submerged aquatic plant cultures.