РАЗРАБОТКА РЕЖИМА СТЕРИЛИЗАЦИИ ВЗРОСЛОГО И ЮВЕНИЛЬНОГО БИОМАТЕРИАЛА ДУБА ЧЕРЕШЧАТОГО (QUERCUS ROBUR L.) ДЛЯ ВВЕДЕНИЯ В КУЛЬТУРУ ТКАНЕЙ
Authors: Краснов В.Г. , Сергеев Р.В. , Романов Е.М. , Тимаков А.А. 
Published: February 2025

В работе представлены результаты экспериментального исследования по получению стерильных морфогенных эксплантов из взрослого дерева и ювенильного материала дуба черешчатого. Максимальное количество стерильных эксплантов –76,7 % – получено при обработке почек 5 % раствором гипохлорита кальция в течение 15 минут. При использовании Лизоформина 3000 лучший результат показало использование 10 % раствора в течение 10 минут. При этом 86,7 % эксплантов были стерильные и морфогенные. Последующее увеличение концентрации данного стерилизатора приводило к гибели эксплантов. Максимальное количество здоровых эксплантов выявлено при стерилизации ювенильного материала дуба черешчатого в течение четырёх минут в 3 % растворе Лизоформина 3000.

Introduction. With the Quercus robur L. propagation in vitro, it is possible to obtain healthier seedlings that retain the properties of the mother plus trees. It will facilitate the forest seed plantations establishment and, together with other measures, will ensure the preservation and increase of productivity of oak plantations in the European part of the country. The study is aimed at developing of an optimal sterilization regime for obtaining the morphogenic plantations when introducing the different-age English oak seedlings in vitro. The buds of adult and juvenile English oak trees grown in the containers under laboratory conditions are the object of the study. Material and methods. Sterilization of the buds of a 50-year-old tree. The buds cut from the branches were sterilized in the solution of detergent for 10 minutes. Next, the material was immersed in 70 % ethanol for 1 minute, sterilized in a solution of calcium hypochlorite at a concentration of 0.5; 1; 1.5 % (option I) and in a solution of Lysoformin 3000 at a concentration of 5, 10, 15 % for 5, 10, 15 minutes (option II). After sterilization, the explants were cleaned off bud scales and planted into a growing medium. The WPM growing medium was used to introduce the tissue of buds of trees into the culture. Before sterilization, 6-benzylaminopurine and naphthyl acetic acid were added to the medium. Sterilization of juvenile material. Green cuttings of two month seedlings of English oak, grown in laboratory conditions, were used. They were cut into 15-20 mm segments, soaked in Fairy water solution for 20 minutes, and washed in the running water for 30-40 minutes. Then, these segments were placed for 30-40 seconds in 70 % ethanol and transferred to a sterilizing 3 % solution of Lysoformin 3000 in accordance with the experimental options: 1, 2, 3, 4, 5 and 6 min. The explants were washed three times in sterile distilled water, the ends where the plant cells were damaged by the sterilizing agent were removed and placed into the MS nutrient medium. The subsequent development of explants of all variants in the medium occurred at 21-22 oC, 1800 Lux illumination, and photoperiod of 16/8. Results. The use of the solution of calcium hypochlorite showed the best results (76.7 % of sterile explants) at a concentration of 1.5 % for 15 minutes. Lysoformin 3000 showed the best result (86.7 % of sterile and morphogenic explants) at 10 % concentration for 10 minutes. In two months, the percentage of healthy plants varied from 71.4 to 90.0 %. The maximum number of healthy explants was detected during the sterilization for 4 minutes in a 3% solution of Lysoformin 3000. Conclusions. The best results in the sterilization of buds of an adult English oak tree (Quercus robur L.) were found when using the calcium hypochlorite at a concentration of 1.5% for 15 minutes and 10.0 % Lysoformin 3000 for 10 minutes. As for the juvenile material, the maximum number of healthy explants was obtained by means of sterilization in a 3% solution of Lysofermin 3000 for 4 minutes. The obtained results can be used to develop the technology of propagation and growing of English oak seedlings in vitro.