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.
authors.affiliation: Department of Plant Sciences, School of Life Sciences, Planetarium Complex, Manipal University, Manipal Udupi, Karnataka, 576 104, India.
authors.forename: Abitha
authors.initials: A
authors.lastname: Thomas
authors.affiliation: Department of Plant Sciences, School of Life Sciences, Planetarium Complex, Manipal University, Manipal Udupi, Karnataka, 576 104, India.
authors.forename: Ipsita
authors.initials: I
authors.lastname: Pujari
authors.affiliation: Department of Plant Sciences, School of Life Sciences, Planetarium Complex, Manipal University, Manipal Udupi, Karnataka, 576 104, India.
authors.forename: Vasudeep
authors.initials: V
authors.lastname: Shetty
authors.affiliation: Department of Ageing Research, School of Life Sciences, Planetarium Complex, Manipal University, Manipal Udupi, Karnataka, 576 104, India.
authors.forename: Manjunath B
authors.initials: MB
authors.lastname: Joshi
authors.affiliation: Department of Biotechnology, School of Life Sciences, Planetarium Complex, Manipal University, Manipal Udupi, Karnataka, 576 104, India.
authors.forename: Padmalatha S
authors.initials: PS
authors.lastname: Rai
authors.affiliation: Department of Cell and Molecular Biology, School of Life Sciences, Planetarium Complex, Manipal University, Manipal Udupi, Karnataka, 576 104, India.
authors.forename: Kapaettu
authors.initials: K
authors.lastname: Satyamoorthy
authors.affiliation: Department of Plant Sciences, School of Life Sciences, Planetarium Complex, Manipal University, Manipal Udupi, Karnataka, 576 104, India. vidhu.sankar@manipal.edu.
authors.forename: Vidhu Sankar
authors.initials: VS
authors.lastname: Babu
mesh_terms: D002478:Cells, Cultured; D018920:Coculture Techniques; D031667:Dendrobium / Q000166:cytology* / Q000378:metabolism; D010636:Phenols / Q000378:metabolism*; D064209:Phytochemicals / Q000378:metabolism; D018515:Plant Leaves / Q000166:cytology / Q000378:metabolism; D011523:Protoplasts / Q000166:cytology; D013267:Stilbenes / Q000378:metabolism
chemical_list: D010636:Phenols; D064209:Phytochemicals; D013267:Stilbenes
keywords: Dendrobium; FACS; Fusants; Mass spectrometry; Protoplast; Stilbene