The Use of Sodium Hypochlorite and Plant Preservative Mixture Significantly Reduces Seed-Borne Pathogen Contamination When Establishing In Vitro Cultures of Wheat (Triticum aestivum L.) Seeds
Anita Bošnjak Mihovilović, Snježana Kereša, Boris Lazarević, Snježana Pintarić, Katarina Martinko, Zvjezdana Marković, Katarina Turkalj, Ivanka Habuš Jerčić
Abstract
In vitro plants that are free of pathogens are crucial for biotechnological breeding methods. The present study investigates the effects of sterilization with sodium hypochlorite (NaClO) and the addition of Plant Preservative Mixture (PPMTM) to the growth medium on pathogen elimination, germination, and seedling development of the winter wheat (Triticum aestivum L.) variety Mara. The sterilization treatments differed in the duration of seed sterilization in 4% NaClO and the PPM concentration added to the growth medium. Pathogenic fungi of the genera Aspergillus, Cladosporium, Penicillium, and Bipolaris were completely eliminated when the seeds were sterilized in NaClO and placed on growth media with the addition of PPM. Extending the duration of the sterilization treatment with NaClO to 50 min reduced Fusarium contamination, while the interaction between the 50 min sterilization treatment with NaClO and the addition of PPM to the growth medium reduced Alternaria contamination. Our results suggest that PPM could complement sterilization procedures with NaClO in the introduction of highly infected wheat seeds in vitro. Seed germination was not affected by sterilization with NaClO or by the addition of PPM. However, PPM at a concentration of 4 mL L−1 had a negative effect on seedling development.