Document Type : Research Article
Authors
1
الهیه فاز دو انتهای بلور زیتون همت 5 شرقی قطعه 2301
2
University of Zanjan
3
Faculty member, Department of Horticultural Sciences, Faculty of Agriculture, Zanjan University
10.22067/jhs.2024.86540.1322
Abstract
The effect of time and different concentrations of sodium hypochlorite, carbendazim fungicide and mercuric chloride in inhibiting the contamination of inverted tulip bulbs (Fritillaria spp.) in tissue culture environment
Abstract
Introduction: Success in tissue culture technique, especially in bulbous plants, depends on the microbial contamination control during in vitro culture. Decontamination is considered as a fundamental challenge in the technique of cell, tissue and plant organ culture. Although there are various methods for this purpose, the development of disinfection methods specific to each species is considered an important factor in the establishment and success of the tissue culture system. Applying different treatments can control the microbial contamination and consequently increase the percentage of explant survival.
Materials and Methods: This study aimed to investigate the effect of time and different concentrations of sodium hypochlorite, carbendazim fungicide and mercuric chloride in inhibiting the contamination of inverted tulip bulbs (Fritillaria spp.) in tissue culture environment. So, an experiment was done as a completely randomized design at four replications in the biotechnology laboratory of Zanjan University during 2023. The experimental treatments consisted of 0.1% fungicide at different times (30, 25, 35 and 40 minutes), 5 levels of sodium hypochlorite (1, 1.5, 2, 2.5 and 3%) at different times (7, 9, 10, 12 and 15 minutes), 70% alcohol with time (60 and 90 seconds) and mercury chloride (0.1 and 0.2%). Bulbs that collected from nature were transferred to the Biotechnology Laboratory of the Faculty of Agriculture, Department of Horticulture, Zanjan University and kept them in the refrigerator at 4°C for two weeks. After this time, the bulbs were washed with detergent and then remained in water for 30 minutes. Then it was disinfected by using the above-mentioned treatments: carbendazim 0.1%, alcohol 70%, sodium hypochlorite, and finally using mercury chloride. It should be noted that at the end of each step, Fritillaria bulbs were washed with sterile distilled water. Also, after the end of the disinfection treatments, the desired explants were cultured in MS basic culture medium. The statistical analysis of this experiment was carried out using SAS software, version 9.1. And the differences between mean values were compared using Duncan’s multiple range test method at the 5% significant level (p < 0.05).
Results and Discussion: The results of the present study showed that, despite the fact that the lowest percentage of contamination was achieved in 40 minutes of carbendazim fungicide, 70% alcohol in 90 seconds, 3% concentration and 15 minutes of sodium hypochlorite and 0.2% mercury chloride concentration. But the survival rate of explants decreased to the lowest level and caused the explants to turn brown. According to the results, the best treatment with 80% decontamination and high survival rate, 30 minutes of fungicide, 60 seconds of 70% alcohol, 2% and 2.5% sodium hypochlorite for 12 and 10 minutes, respectively. Mercuric chloride was 0.1% for 7 minutes. Therefore, it is suggested to use the mentioned treatments in in vitro cultivation conditions to reduce the contamination and micropropagation of the bulbs of Fritillaria Spp.
Decontamination is considered as a fundamental challenge in the technique of cell, tissue and plant organ culture. Therefore,the present study the effect of Carbendazim fungicide ./1 mg L -1 at different times (30, 25, 35 and 40 minutes), 5 levels of hypochlorite sodium (1, 1.5, 2, 2.5 and 3), at different times (12, 15, 10, 9 and 7 minutes), alcohol 70% with time (1.5 and 1minutes) and mercuric chloride (duration of 7 minutes with concentrations of 1.5 and 1 mg L -1) for survival and remove contamination in onions fritillaria was executed. The results of the present study showed that although the lowest percentage of contamination was achieved in 40 minutes of carbendazim fungicide, 70% alcohol in 1.5 minutes, 3% concentration and 15 minutes of sodium hypochlorite and 1.5% concentration of mercury chloride, but the survival rate of explants It decreased to the lowest level and caused the explants to turn brown. However, the best treatment with 80% decontamination and high survival rate, 30 minutes of fungicide, one minute of 70% alcohol, concentration of 3% and time of 9 minutes of sodium hypochlorite and concentration of 0.1 mg/liter with time of 7 minutes was mercuric chloride.
Keyword: Extinction, In vitro, Mercuric chloride, Disinfection, Fritillaria.
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