Introduction of Salicornia europaea L into in vitro culture

M. F. Koryazhkina, N. A. Dmitrieva, Е. V. Trizno, A. B. Sediki, A. M. Utesheva, E. S. Skorobogatova
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Abstract

The goal of the research was to develop a technology for introducing Salicornia europaea L.into in vitro culture. Methods of cultivating meristems and callus cultures were studied. To cultivate meristems, the tip of the apical bud were used as an explant. Fragments of stems and leaves were used toobtain callus tissue. To study the influence of various factors on germination, seeds were soaked in sterile tap water and in solutions of gibberellin, cytokinin, auxin and NaCl, and were also subjected to cold stratification (independently and with subsequent placement in Knop’s agarized medium). Salicornia seeds were sterilized with various antiseptics: 70% alcohol, 10% aqueous solution of sodium hypochlorite («Belizna»), amoxicillinand 3% hydrogen peroxide. The ability of the culture to form callus was studied in MS medium. As a result, it was determined that the highest germination of seeds was observed in Knop medium after treating theseeds with a suspension of green algae of the Scenedesmus genus, as well as after preliminary cold stratification, and slightly less after treating the seeds with a solution of NaCl and gibberellic acid. The most effective method of seed sterilization turned out to be treatment with alcohol followed by treatment with sodium hypochlorite. A comparative analysis of seed germination in filter paper in Petri dishes, Knop agar medium, Murashige and Skoog (hormone-free) was carried out. The ability of S. europaea L. to form callusin MS medium with phytohormones was assessed. Conclusion. To improve germination, it is recommended to subject the seeds to cold stratification. To obtain aseptic explants quickly, it is recommended to germinate the seeds in the Knop nutrient medium, having previously sterilized them with alcohol, then with sodiumhypochlorite, followed by washing with distilled water. The most suitable type of microclonal propagationfor S. europaea L. is the production of callus tissue followed by induction of organogenesis or embryogenesis.
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将欧洲盐生植物引入体外培养
该研究的目标是开发一种将欧洲沙利筋(Salicornia europaea L.)引入离体培养的技术。研究了培养分生组织和胼胝体的方法。培养分生组织时,使用顶芽的顶端作为外植体。茎和叶的碎片用于获得胼胝组织。为了研究各种因素对萌发的影响,将种子浸泡在无菌自来水以及赤霉素、细胞分裂素、辅助素和氯化钠溶液中,并进行低温层积(独立层积,然后放入克诺普琼脂培养基中)。莎草种子用不同的防腐剂进行消毒:70% 的酒精、10% 的次氯酸钠水溶液("Belizna")、阿莫西林和 3% 的过氧化氢。在 MS 培养基中研究了培养物形成茧的能力。结果表明,用 Scenedesmus 属绿藻悬浮液处理这些种子并进行初步冷藏后,在 Knop 培养基中观察到的种子萌发率最高,而用氯化钠和赤霉素溶液处理种子后,萌发率稍低。最有效的种子消毒方法是先用酒精处理,再用次氯酸钠处理。对培养皿滤纸、克诺普琼脂培养基、穆拉希和斯库格(无激素)培养基中种子的萌发进行了比较分析。评估了 S. europaea L. 在含有植物激素的 MS 培养基中形成胼胝体的能力。结论。为提高发芽率,建议对种子进行低温层积处理。为快速获得无菌外植体,建议先用酒精消毒,再用次氯酸钠消毒,然后用蒸馏水清洗,最后在克诺普营养培养基中催芽。最适合 S. europaea L.的微克隆繁殖类型是先产生胼胝组织,然后诱导器官发生或胚胎发生。
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