Dendrobium protoplast co-culture promotes phytochemical assemblage in vitro

Thomas, Abitha and Pujari, Ipsita and Shetty, Vasudeep and Joshi, Manjunath B and Rai, Padmalatha S and Satyamoorthy, K and Babu, Vidhu Sankar (2015) Dendrobium protoplast co-culture promotes phytochemical assemblage in vitro. Protoplasma, 254 (4). pp. 1-12. ISSN 0033-183X

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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 heterfusants and homofusants formed were enumerated. The spectral feature extraction of the mass spectrometry indicated the presence of five phenolicmarker 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.

Item Type: Article
Uncontrolled Keywords: Dendrobium; FACS; fusants; mass spectrometry; protoplas; stilbene.
Subjects: Life Sciences > MLSC Manipal
Depositing User: KMC Library
Date Deposited: 27 Feb 2017 11:31
Last Modified: 27 Feb 2017 11:31
URI: http://eprints.manipal.edu/id/eprint/148351

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