PREPARATION OF PROTOPLASTS OF SEA BUCKTHORN (HIPPOPHAE RHAMNOIDES L.)

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 stat...

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Main Authors: M. V. Skaptsov, S. V. Smirnov, M. G. Kutsev
Format: Article
Language:English
Published: Altay State University 2015-04-01
Series:Turczaninowia
Online Access:http://journal.asu.ru/index.php/tur/article/view/704
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author M. V. Skaptsov
S. V. Smirnov
M. G. Kutsev
author_facet M. V. Skaptsov
S. V. Smirnov
M. G. Kutsev
author_sort M. V. Skaptsov
collection DOAJ
description 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.
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spelling doaj.art-82d5dc3965db460994a9fcd144c2663f2022-12-22T00:50:38ZengAltay State UniversityTurczaninowia1560-72591560-72672015-04-0116310.14258/turczaninowia.16.3.20471PREPARATION OF PROTOPLASTS OF SEA BUCKTHORN (HIPPOPHAE RHAMNOIDES L.)M. V. Skaptsov0S. V. Smirnov1M. G. Kutsev2Altai State University, Barnaul, RussiaAltai State University, Barnaul, RussiaAltai State University, Barnaul, RussiaThe 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.http://journal.asu.ru/index.php/tur/article/view/704
spellingShingle M. V. Skaptsov
S. V. Smirnov
M. G. Kutsev
PREPARATION OF PROTOPLASTS OF SEA BUCKTHORN (HIPPOPHAE RHAMNOIDES L.)
Turczaninowia
title PREPARATION OF PROTOPLASTS OF SEA BUCKTHORN (HIPPOPHAE RHAMNOIDES L.)
title_full PREPARATION OF PROTOPLASTS OF SEA BUCKTHORN (HIPPOPHAE RHAMNOIDES L.)
title_fullStr PREPARATION OF PROTOPLASTS OF SEA BUCKTHORN (HIPPOPHAE RHAMNOIDES L.)
title_full_unstemmed PREPARATION OF PROTOPLASTS OF SEA BUCKTHORN (HIPPOPHAE RHAMNOIDES L.)
title_short PREPARATION OF PROTOPLASTS OF SEA BUCKTHORN (HIPPOPHAE RHAMNOIDES L.)
title_sort preparation of protoplasts of sea buckthorn hippophae rhamnoides l
url http://journal.asu.ru/index.php/tur/article/view/704
work_keys_str_mv AT mvskaptsov preparationofprotoplastsofseabuckthornhippophaerhamnoidesl
AT svsmirnov preparationofprotoplastsofseabuckthornhippophaerhamnoidesl
AT mgkutsev preparationofprotoplastsofseabuckthornhippophaerhamnoidesl