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

A. B. Mihovilović, S. Kereša, Boris Lazarević, S. Pintarić, Katarina Martinko, Zvjezdana Marković, Katarina Turkalj, I. Jerčić
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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.
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在建立小麦(Triticum aestivum L.)种子的体外培养基时使用次氯酸钠和植物防腐剂混合物可显著减少种子传播的病原体污染
无病原体的离体植物对生物技术育种方法至关重要。本研究调查了用次氯酸钠(NaClO)灭菌和在生长培养基中添加植物防腐剂混合物(PPMTM)对冬小麦(Triticum aestivum L.)品种马拉的病原体消除、发芽和幼苗发育的影响。灭菌处理的不同之处在于种子在 4%NaClO 中的灭菌持续时间和添加到生长培养基中的 PPM 浓度。当种子在 NaClO 中灭菌并置于添加了 PPM 的生长培养基上时,曲霉属、帚霉属、青霉属和双孢蘑菇属的病原真菌被完全消灭。将用 NaClO 消毒处理的时间延长至 50 分钟可减少镰刀菌的污染,而用 NaClO 消毒处理 50 分钟与在生长培养基中添加 PPM 之间的相互作用可减少交替孢霉的污染。我们的研究结果表明,PPM 可作为 NaClO 灭菌程序的补充,用于体外引入高度感染的小麦种子。用 NaClO 消毒或添加 PPM 都不会影响种子发芽。然而,浓度为 4 mL L-1 的 PPM 对幼苗的发育有负面影响。
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