METHOD FOR MULTI-STAGE SURFACE STERILIZATION OF UPPER PARTS OF GENERATIVE SHOOTS OF BETA VULGARIS L. BEFORE INTRODUCTION INTO IN VITRO CULTURE Russian patent published in 2023 - IPC A01H4/00 

FIELD: biotechnology.

SUBSTANCE: invention relates to a method of surface sterilization of the upper parts of the generative shoots of Beta vulgaris selected in the field in order to obtain an aseptic culture of valuable specimens and increase the efficiency of sugar beet clonal micropropagation. The method includes multi-stage sterilization of the upper parts of the central inflorescences of Beta vulgaris. The selection of plant material 7–10 cm long is carried out. The pre-treatment of the material is carried out in a soap solution, followed by cutting the explants into pieces 2–2.5 cm in size. The first stage of sterilization is carried out in a NaOCl solution (0.008–0.011%) with the addition of Tween 20 (2 drops per 1 liter of sterilizing solution) for 15–20 minutes. At the second stage, the explants are treated in an antibiotic solution (tetracycline 0.008–0.011%, ceftriaxone 0.020–0.030%) for 15–20 minutes. The third stage is the main treatment in a solution of a chlorine-containing compound — NaOCl (0.8–1.1%) with the addition of Tween 20 (2 drops per 1 liter of sterilizing solution) for 40 minutes. After each stage of sterilization, the explants are washed with sterile water. All solutions are prepared on the basis of sterile water, each stage of sterilization and washing is carried out on an orbital shaker with moderate shaking intensity to prevent mechanical damage to the plant material.

EFFECT: improvement of the quality of the process of sterilization of the upper parts of generative shoots of Beta vulgaris by reducing contamination by microorganisms and fungi while maintaining a high level of viability of explants when introduced into in vitro culture.

1 cl, 5 dwg, 1 tbl