In vitro DIRECT SHOOT REGENERATION FROM Rhodiola rosea L. LEAF EXPLANTS

Wild plant species are of great interest as a source of pharmacologically valuable compounds but a great number of them are endemic and/or endangered ones. Modern plant biotechnology can provide reliable methods for their utilization without disturbing natural populations. In vitro culture methods f...

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Main Authors: Matvieieva N., Belokurova V., Ratushniak Y., Shcherbak N., Kuchuk M.
Format: Article
Language:English
Published: National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry 2023-06-01
Series:Biotechnologia Acta
Subjects:
Online Access:https://biotechnology.kiev.ua/images/BTA/2023/3_2023/Matvieieva_3_2023.pdf
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author Matvieieva N.
Belokurova V.
Ratushniak Y.
Shcherbak N.
Kuchuk M.
author_facet Matvieieva N.
Belokurova V.
Ratushniak Y.
Shcherbak N.
Kuchuk M.
author_sort Matvieieva N.
collection DOAJ
description Wild plant species are of great interest as a source of pharmacologically valuable compounds but a great number of them are endemic and/or endangered ones. Modern plant biotechnology can provide reliable methods for their utilization without disturbing natural populations. In vitro culture methods for Rhodiola species are being intensively developed to include them into various biotechnological programmes. Aim. Development of a protocol for direct Rhodiola rosea L. plant regeneration from leaf explants. Methods. The leaves of R. rosea aseptically growing plants were used as the explants. Several variants of Murashige and Skoog (1962) agar-solidified culture medium supplemented with different combinations of auxins (1-naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D)) and cytokinins (kinetin and 6-benzylaminopurine (BAP)) were estimated as potential regeneration-inducing media. Regeneration frequency was calculated as the percentage of leaves that produced shoots. Results. The use of MS medium supplemented with 2.5 mg/l BAP and 1.0 mg/l 2,4-D allowed inducing shoot formation with 100% frequency. An increase in the 2,4-D content up to 2.5 mg/l and decrease in BAP content to 1.0 mg/l resulted in decreasing of the regeneration frequency to 62.5%. Regeneration frequency was 25% and 62%, respectively, on the media containing 1.0 mg/l kinetin + 2.5 mg/l 2,4-D and 2.5 mg/l kinetin + 1.0 mg/l 2,4-D. Conclusions. R. rosea leaf explants have demonstrated high regeneration capacity with using the studied combinations of plant growth regulators. MS medium supplemented with 2.5 mg/l BAP and 1.0 mg/l 2,4-D allowed inducing shoot regeneration in leaf explants with the frequency of 100%. The frequency of regeneration was lower in the case of substitution of BAP for kinetin. The other types of morphogenesis (formation of adventitious roots and/or callus) were also observed.
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spelling doaj.art-39fb206345104d03805ff283f17451552023-12-13T16:00:02ZengNational Academy of Sciences of Ukraine, Palladin Institute of BiochemistryBiotechnologia Acta2410-77512410-776X2023-06-01163455010.15407/biotech16.03.045In vitro DIRECT SHOOT REGENERATION FROM Rhodiola rosea L. LEAF EXPLANTS Matvieieva N.0 Belokurova V.1 Ratushniak Y.2Shcherbak N.3Kuchuk M.4Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, KyivInstitute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, KyivInstitute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, KyivInstitute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, KyivInstitute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, KyivWild plant species are of great interest as a source of pharmacologically valuable compounds but a great number of them are endemic and/or endangered ones. Modern plant biotechnology can provide reliable methods for their utilization without disturbing natural populations. In vitro culture methods for Rhodiola species are being intensively developed to include them into various biotechnological programmes. Aim. Development of a protocol for direct Rhodiola rosea L. plant regeneration from leaf explants. Methods. The leaves of R. rosea aseptically growing plants were used as the explants. Several variants of Murashige and Skoog (1962) agar-solidified culture medium supplemented with different combinations of auxins (1-naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D)) and cytokinins (kinetin and 6-benzylaminopurine (BAP)) were estimated as potential regeneration-inducing media. Regeneration frequency was calculated as the percentage of leaves that produced shoots. Results. The use of MS medium supplemented with 2.5 mg/l BAP and 1.0 mg/l 2,4-D allowed inducing shoot formation with 100% frequency. An increase in the 2,4-D content up to 2.5 mg/l and decrease in BAP content to 1.0 mg/l resulted in decreasing of the regeneration frequency to 62.5%. Regeneration frequency was 25% and 62%, respectively, on the media containing 1.0 mg/l kinetin + 2.5 mg/l 2,4-D and 2.5 mg/l kinetin + 1.0 mg/l 2,4-D. Conclusions. R. rosea leaf explants have demonstrated high regeneration capacity with using the studied combinations of plant growth regulators. MS medium supplemented with 2.5 mg/l BAP and 1.0 mg/l 2,4-D allowed inducing shoot regeneration in leaf explants with the frequency of 100%. The frequency of regeneration was lower in the case of substitution of BAP for kinetin. The other types of morphogenesis (formation of adventitious roots and/or callus) were also observed.https://biotechnology.kiev.ua/images/BTA/2023/3_2023/Matvieieva_3_2023.pdfrhodiola rosea l.leaf explantsshoot regenerationgrowth regulators.
spellingShingle Matvieieva N.
Belokurova V.
Ratushniak Y.
Shcherbak N.
Kuchuk M.
In vitro DIRECT SHOOT REGENERATION FROM Rhodiola rosea L. LEAF EXPLANTS
Biotechnologia Acta
rhodiola rosea l.
leaf explants
shoot regeneration
growth regulators.
title In vitro DIRECT SHOOT REGENERATION FROM Rhodiola rosea L. LEAF EXPLANTS
title_full In vitro DIRECT SHOOT REGENERATION FROM Rhodiola rosea L. LEAF EXPLANTS
title_fullStr In vitro DIRECT SHOOT REGENERATION FROM Rhodiola rosea L. LEAF EXPLANTS
title_full_unstemmed In vitro DIRECT SHOOT REGENERATION FROM Rhodiola rosea L. LEAF EXPLANTS
title_short In vitro DIRECT SHOOT REGENERATION FROM Rhodiola rosea L. LEAF EXPLANTS
title_sort in vitro direct shoot regeneration from rhodiola rosea l leaf explants
topic rhodiola rosea l.
leaf explants
shoot regeneration
growth regulators.
url https://biotechnology.kiev.ua/images/BTA/2023/3_2023/Matvieieva_3_2023.pdf
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AT ratushniaky invitrodirectshootregenerationfromrhodiolarosealleafexplants
AT shcherbakn invitrodirectshootregenerationfromrhodiolarosealleafexplants
AT kuchukm invitrodirectshootregenerationfromrhodiolarosealleafexplants