Abstract
The aim of research was to study the effect of a method of protoplast isolation on Hippophae rhamnoides cell viability. The main objectives were the release of protoplasts H. rhamnoides from callus tissue by a combination of mechanical and enzymatic effects, as well as evaluation of the general state of isolated cells after isolation and purification. The objects of studies were mesophyll cells and leaf mesophyll explants introduced in culture in vitro. It is assumed that considerable damage of cells in the process of mechanical-enzymatic treatment is not only a result of mechanical impact, but also an effect of the release of metabolic products (polyphenolic components) of cells and their decay. Also protoplasts are dedifferentiated and simplify the process of differentiation in various breeding and biotechnological experiments. As a result of the work, an optimized method of H. rhamnoides protoplast isolation with decreased levels of cell damage is set up. It is revealed that important factors for high yield of viable protoplasts are: proper concentration of D-mannite as an osmoticum; proper concentration of cellulase R10 and macerozime R10; use of sodium tiosulfate as an antioxidante; proper time of incubation of leaf explants in medium with enzymes.
Highlights
The aim of research was to study the effect of a method of protoplast isolation on Hippophae rhamnoides cell viability
The main objectives were the release of protoplasts H. rhamnoides from callus tissue by a combination of mechanical and enzymatic effects, as well as evaluation of the general state of isolated cells after isolation and purification
The objects of studies were mesophyll cells and leaf mesophyll explants introduced in culture in vitro
Summary
PREPARATION OF PROTOPLASTS OF SEA BUCKTHORN (HIPPOPHAE RHAMNOIDES L.) Целью исследований стало изучение влияния способа выделения протопластов на жизнеспособность интактных клеток Hippophae rhamnoides L. Протопласты каллусов являются дедифференцированными и позволяют упростить процесс специализации клеток при проведении различных селекционных и биотехнологических процессов. В результате был установлен оптимальный метод выделения протопластов H. rhamnoides со сниженным уровнем повреждения клеток.
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