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A high-throughput flow cytometry system for early screening of in vitro made polyploids in Dendrobium hybrids

TitoloA high-throughput flow cytometry system for early screening of in vitro made polyploids in Dendrobium hybrids
Tipo di pubblicazioneArticolo su Rivista peer-reviewed
Anno di Pubblicazione2018
AutoriGrosso, V., Farina Anna, Giorgi Debora, Nardi Luca, Diretto Gianfranco, and Lucretti Sergio
RivistaPlant Cell, Tissue and Organ Culture
Volume132
Paginazione57-70
ISSN01676857
Parole chiaveAnti-tumoral activity, Antioxidants, Biomolecules, colchicine, Cycle Value, Diagnosis, Flow cytometry, Flow cytometry systems, High resolution mass spectrometry, International markets, International trade, LC-HRMS, liquid chromatography, Mass Spectrometry, Metabolites, Plants (botany), screening, Secondary metabolites, Stochastic systems
Abstract

Orchids of the genus Dendrobium hold a high economical value in the international markets both as pot plants or cut flowers, and for the production of some metabolites with antioxidant and anti-tumoral activities. Manipulating ploidy levels of Dendrobium species is one of the possible methods to develop new varieties with increased ornamental value and a higher production of secondary metabolites. In this work, we present a new and fast flow cytometry approach to obtain and select Dendrobium phalaenopsis × Dendrobium loddigesii polyploids, through an early in vitro screening on protocorm like bodies (PLBs) after antimitotic treatment. Our approach allows the identification of the best time of treatment on control PLBs and the assessment of best conditions for polyploid recovery just one month after treatments, by using Cycle Value. We were able to discard about the two-third of the unchanged material by drastically reducing the explants to work with and the corresponding costs. Different conditions, regarding concentrations and exposition time, were tested using colchicine or amiprophos-methyl (APM). A high polyploids recovery, up to 80%, were obtained with both antimitotic agents, and those materials were further characterized by liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS), to identify independent polyploids explants with increased levels in high-value molecules as shihunidine and hircinol, together with stochastic events and genotype-specific metabolite fluctuations. © 2017, Springer Science+Business Media B.V.

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URLhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85029766111&doi=10.1007%2fs11240-017-1310-8&partnerID=40&md5=58619e297b243c1da097444290d4d645
DOI10.1007/s11240-017-1310-8
Citation KeyGrosso201857