Abstract

The present study is intended to analyze the occurrence of potent, low produce, naturally occurring stilbenes in protoplasts of wild species and hybrids of Dendrobium. The wild species selected for the study was Dendrobium ovatum, endemic to Western Ghats of India. Protoplasts were isolated from leaves and tepal tissues of all the species and were cultured purely to generate homofusants and cross-cultured to raise heterofusants. Phytochemical composition of protoplast culture with atypical and pure microcolonies was performed using mass spectrometry. Enzyme cocktail of 4% pectinase together with 2% cellulase displayed the highest competence for protoplast isolations. Maximum protoplast density of 30.11 × 104/ml was obtained from D. ovatum leaves in 2 h. Subcellular features such as the presence of partially formed cell wall, the position of the nucleus, chloroplast density, colony existence, and integrity of the plasma membrane were analyzed. Among the pure and cross-cultured protoplasts, the number of heterofusants and homofusants formed were enumerated. The spectral feature extraction of the mass spectrometry indicated the presence of five phenolic marker compounds, viz., tristin, confusarin, gigantol, moscatilin, and resveratrol, some of them in pure and others in assorted protoplast cultures raised from Dendrobium leaves and tepals. The study demonstrated that protoplast fusion technique enabled phytochemical assemblage in vitro as stilbenes tend to get restricted either in a tissue or species specific manner. This is the first report showing the presence of resveratrol, moscatilin, tristin, gigantol, and confusarin in wild and hybrid species from cultured Dendrobium protoplasts in vitro.

Original languageEnglish
Pages (from-to)1517-1528
Number of pages12
JournalProtoplasma
Volume254
Issue number4
DOIs
Publication statusPublished - 01-07-2017

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Dendrobium
Protoplasts
Phytochemicals
coculture
Coculture Techniques
protoplasts
phytopharmaceuticals
stilbenes
Stilbenes
resveratrol
Mass Spectrometry
mass spectrometry
leaves
protoplast fusion
Polygalacturonase
In Vitro Techniques
polygalacturonase
Cellulase
endo-1,4-beta-glucanase
Chloroplasts

All Science Journal Classification (ASJC) codes

  • Plant Science
  • Cell Biology

Cite this

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title = "Dendrobium protoplast co-culture promotes phytochemical assemblage in vitro",
abstract = "The present study is intended to analyze the occurrence of potent, low produce, naturally occurring stilbenes in protoplasts of wild species and hybrids of Dendrobium. The wild species selected for the study was Dendrobium ovatum, endemic to Western Ghats of India. Protoplasts were isolated from leaves and tepal tissues of all the species and were cultured purely to generate homofusants and cross-cultured to raise heterofusants. Phytochemical composition of protoplast culture with atypical and pure microcolonies was performed using mass spectrometry. Enzyme cocktail of 4{\%} pectinase together with 2{\%} cellulase displayed the highest competence for protoplast isolations. Maximum protoplast density of 30.11 × 104/ml was obtained from D. ovatum leaves in 2 h. Subcellular features such as the presence of partially formed cell wall, the position of the nucleus, chloroplast density, colony existence, and integrity of the plasma membrane were analyzed. Among the pure and cross-cultured protoplasts, the number of heterofusants and homofusants formed were enumerated. The spectral feature extraction of the mass spectrometry indicated the presence of five phenolic marker compounds, viz., tristin, confusarin, gigantol, moscatilin, and resveratrol, some of them in pure and others in assorted protoplast cultures raised from Dendrobium leaves and tepals. The study demonstrated that protoplast fusion technique enabled phytochemical assemblage in vitro as stilbenes tend to get restricted either in a tissue or species specific manner. This is the first report showing the presence of resveratrol, moscatilin, tristin, gigantol, and confusarin in wild and hybrid species from cultured Dendrobium protoplasts in vitro.",
author = "Abitha Thomas and Ipsita Pujari and Vasudeep Shetty and Joshi, {Manjunath B.} and Rai, {Padmalatha S.} and Kapaettu Satyamoorthy and Babu, {Vidhu Sankar}",
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language = "English",
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pages = "1517--1528",
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Dendrobium protoplast co-culture promotes phytochemical assemblage in vitro. / Thomas, Abitha; Pujari, Ipsita; Shetty, Vasudeep; Joshi, Manjunath B.; Rai, Padmalatha S.; Satyamoorthy, Kapaettu; Babu, Vidhu Sankar.

In: Protoplasma, Vol. 254, No. 4, 01.07.2017, p. 1517-1528.

Research output: Contribution to journalArticle

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T1 - Dendrobium protoplast co-culture promotes phytochemical assemblage in vitro

AU - Thomas, Abitha

AU - Pujari, Ipsita

AU - Shetty, Vasudeep

AU - Joshi, Manjunath B.

AU - Rai, Padmalatha S.

AU - Satyamoorthy, Kapaettu

AU - Babu, Vidhu Sankar

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N2 - The present study is intended to analyze the occurrence of potent, low produce, naturally occurring stilbenes in protoplasts of wild species and hybrids of Dendrobium. The wild species selected for the study was Dendrobium ovatum, endemic to Western Ghats of India. Protoplasts were isolated from leaves and tepal tissues of all the species and were cultured purely to generate homofusants and cross-cultured to raise heterofusants. Phytochemical composition of protoplast culture with atypical and pure microcolonies was performed using mass spectrometry. Enzyme cocktail of 4% pectinase together with 2% cellulase displayed the highest competence for protoplast isolations. Maximum protoplast density of 30.11 × 104/ml was obtained from D. ovatum leaves in 2 h. Subcellular features such as the presence of partially formed cell wall, the position of the nucleus, chloroplast density, colony existence, and integrity of the plasma membrane were analyzed. Among the pure and cross-cultured protoplasts, the number of heterofusants and homofusants formed were enumerated. The spectral feature extraction of the mass spectrometry indicated the presence of five phenolic marker compounds, viz., tristin, confusarin, gigantol, moscatilin, and resveratrol, some of them in pure and others in assorted protoplast cultures raised from Dendrobium leaves and tepals. The study demonstrated that protoplast fusion technique enabled phytochemical assemblage in vitro as stilbenes tend to get restricted either in a tissue or species specific manner. This is the first report showing the presence of resveratrol, moscatilin, tristin, gigantol, and confusarin in wild and hybrid species from cultured Dendrobium protoplasts in vitro.

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