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Regeneration of Genetically Stable Plants from in Vitro Vitrified Leaves of Different Carnation Cultivars

This study was conducted to investigate the efficacy of shoot regeneration from different leaf types (normal leaves and vitrified leaves) from three different carnation cultivars ‘Kumbuyl’, ‘Denev’, and ‘Jinju’ using different combinations of 3-indole butyric acid (IBA) and thidiazuron (TDZ) concent...

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Autores principales: Thu, Ho Thi Minh, Naing, Aung Htay, Jeong, Hui Yeong, Kim, Chang Kil
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7465169/
https://www.ncbi.nlm.nih.gov/pubmed/32731438
http://dx.doi.org/10.3390/plants9080950
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author Thu, Ho Thi Minh
Naing, Aung Htay
Jeong, Hui Yeong
Kim, Chang Kil
author_facet Thu, Ho Thi Minh
Naing, Aung Htay
Jeong, Hui Yeong
Kim, Chang Kil
author_sort Thu, Ho Thi Minh
collection PubMed
description This study was conducted to investigate the efficacy of shoot regeneration from different leaf types (normal leaves and vitrified leaves) from three different carnation cultivars ‘Kumbuyl’, ‘Denev’, and ‘Jinju’ using different combinations of 3-indole butyric acid (IBA) and thidiazuron (TDZ) concentrations. The shoot tips cultured on Murashige and Skoog (MS) basal media (Type 1 media) produced normal leaves, while those cultured-on media supplemented with plant growth regulators and/or vitamin (Type 2 media and Type 3 media) produced vitrified leaves for all cultivars. Culture of normal leaf segments on MS medium containing different combinations of IBA and TDZ concentrations induced callus in all treatments; however, the callus was unable to induce shoots and finally became necrotic. In contrast, no callus induction was observed in the control (hormone-free treatment). When vitrified leaf segments underwent the same treatments, shoots were induced from the vitrified leaves (derived from Type 2 media) but were unhealthy and gradually died, whereas those induced from Type 3 media were vitrified and healthy. The optimal combination for the best shoot regeneration and number of shoots per explants varied depending on the genotypes used. The vitrified shoots induced from the leaves of Type 3 media transformed into normal shoots and survived well under greenhouse conditions. According to the results of random amplified polymorphic DNA (RAPD) analysis, the banding patterns of twelve primers that were detected in vitrified leaf-induced normalized shoots were identical to those of normal in vitro grown plants, indicating that no genetic variation had occurred during the procedure. Taken together, this study indicates that vitrified leaves can be used for shoot regeneration of recalcitrant carnation cultivars, regardless of the genotypes and types of vitrified leaves. However, as the number of shoots per explants was still low, further investigation is warranted to obtain a more efficient shoot regeneration protocol for genetic transformation of the cultivars.
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spelling pubmed-74651692020-09-04 Regeneration of Genetically Stable Plants from in Vitro Vitrified Leaves of Different Carnation Cultivars Thu, Ho Thi Minh Naing, Aung Htay Jeong, Hui Yeong Kim, Chang Kil Plants (Basel) Article This study was conducted to investigate the efficacy of shoot regeneration from different leaf types (normal leaves and vitrified leaves) from three different carnation cultivars ‘Kumbuyl’, ‘Denev’, and ‘Jinju’ using different combinations of 3-indole butyric acid (IBA) and thidiazuron (TDZ) concentrations. The shoot tips cultured on Murashige and Skoog (MS) basal media (Type 1 media) produced normal leaves, while those cultured-on media supplemented with plant growth regulators and/or vitamin (Type 2 media and Type 3 media) produced vitrified leaves for all cultivars. Culture of normal leaf segments on MS medium containing different combinations of IBA and TDZ concentrations induced callus in all treatments; however, the callus was unable to induce shoots and finally became necrotic. In contrast, no callus induction was observed in the control (hormone-free treatment). When vitrified leaf segments underwent the same treatments, shoots were induced from the vitrified leaves (derived from Type 2 media) but were unhealthy and gradually died, whereas those induced from Type 3 media were vitrified and healthy. The optimal combination for the best shoot regeneration and number of shoots per explants varied depending on the genotypes used. The vitrified shoots induced from the leaves of Type 3 media transformed into normal shoots and survived well under greenhouse conditions. According to the results of random amplified polymorphic DNA (RAPD) analysis, the banding patterns of twelve primers that were detected in vitrified leaf-induced normalized shoots were identical to those of normal in vitro grown plants, indicating that no genetic variation had occurred during the procedure. Taken together, this study indicates that vitrified leaves can be used for shoot regeneration of recalcitrant carnation cultivars, regardless of the genotypes and types of vitrified leaves. However, as the number of shoots per explants was still low, further investigation is warranted to obtain a more efficient shoot regeneration protocol for genetic transformation of the cultivars. MDPI 2020-07-28 /pmc/articles/PMC7465169/ /pubmed/32731438 http://dx.doi.org/10.3390/plants9080950 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Thu, Ho Thi Minh
Naing, Aung Htay
Jeong, Hui Yeong
Kim, Chang Kil
Regeneration of Genetically Stable Plants from in Vitro Vitrified Leaves of Different Carnation Cultivars
title Regeneration of Genetically Stable Plants from in Vitro Vitrified Leaves of Different Carnation Cultivars
title_full Regeneration of Genetically Stable Plants from in Vitro Vitrified Leaves of Different Carnation Cultivars
title_fullStr Regeneration of Genetically Stable Plants from in Vitro Vitrified Leaves of Different Carnation Cultivars
title_full_unstemmed Regeneration of Genetically Stable Plants from in Vitro Vitrified Leaves of Different Carnation Cultivars
title_short Regeneration of Genetically Stable Plants from in Vitro Vitrified Leaves of Different Carnation Cultivars
title_sort regeneration of genetically stable plants from in vitro vitrified leaves of different carnation cultivars
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7465169/
https://www.ncbi.nlm.nih.gov/pubmed/32731438
http://dx.doi.org/10.3390/plants9080950
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