Development of an Effective Sterilization Protocol for Plant Tissue Culture Studies in Superfruit Aronia [Aronia melanocarpa (Michaux) Elliot]

Year 2024, Volume: 7 Issue: 6, 679 - 685, 15.11.2024

Çağlar Kaya , Şehnaz Özatay

https://doi.org/10.47115/bsagriculture.1548432

Abstract

Effective sterilization protocols are crucial for a successful tissue culture study in Aronia. These protocols directly influence contamination rates, shoot health, and root development. In this context, the study aims is to develop an effective sterilization protocol for plant tissue culture studies in Aronia [Aronia melanocarpa (Michaux) Elliot], commonly known as the "superfruit." In the study, the Nero Aronia variety shoot tips were used as material. The sterilized shoot tips were transferred to the respective plant tissue culture media in a randomized parcels trial pattern with three replicates, each containing three explants per replicate. Various concentrations and combinations of sterilizing agents, such as sodium hypochlorite (NaOCl), hydrogen peroxide (H2O2), mercuric chloride (HgCl2), and ethanol (C2H5OH), were evaluated to determine their effectiveness in maintaining tissue health and reducing contamination. Twelve protocols were developed, incorporating different concentrations of these chemicals. The data were subjected to statistical analysis using the SAS software package (LSD0.05-Proc GLM). Analysis of variance (ANOVA) was performed to evaluate differences among sterilization treatments for each parameter. The results showed that the combination of 5% NaOCl and 3% H2O2 (10 min each) provided the lowest average contamination rate 0.0%, the highest average number uncontaminated explants 9.0 pieces, shoot length 3.0 cm and root length 2.5 cm demonstrating the sterilization efficiency of this combination. On the other hand, protocols containing HgCl2, especially at higher concentrations, resulted in impaired root development. High ethanol concentrations also contributed to effective sterilization, with the combination of 7% NaOCl (10 min) and 80% ethanol (5 min) yielding a low contamination rate (22.0%) and preserving tissue health. This study emphasizes balancing sterilization protocols between effective contamination control and tissue viability. The findings are expected to benefit the improvement and development of tissue culture techniques for Aronia and similar species, providing a basis for further research on effective sterilization practices, currently limited in Aronia tissue culture.

Keywords

Aronia [Aronia melanocarpa (Michaux) Elliot], in vitro, Sterilization, Contamination rate, Shoot length, Root length

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