A Protocol for the Induction of Polyploids in Phalaenopsis Orchids by In Vitro Method Without Using Anti-microtubule Agents

  • Wen-Huei Chen
  • Ching-Yan Tang
Part of the Springer Protocols Handbooks book series (SPH)


Polyploidy plays an important role in the breeding of many crops as well as horticultural plants of the world. It is also important for the variety improvement of Phalaenopsis orchids. Most of the commercial orchids are tetraploids. However, most of the wild species of Phalaenopsis are diploid. Thus, a barrier to hybridization between these two groups of varieties limits the source of germplasm available for the breeding programs. Hence, a simple technique to scale up the ploidy level of diploid species is needed. Traditionally, chemical induction using anti-microtubule agents such as colchicine or oryzalin is used for polyploid induction. However, there are several disadvantages to these methods, such as complicated procedures, problems of toxicity to plants, and the occurrence of unfavorable chimeras. Search for alternate procedures might give better result to achieve the same objective. Due to the occurrence of endopolyploid cells in the tissues of protocorms and PLBs of the orchid, there is a chance to regenerate polyploid plants from these cells. A protocol for dissecting protocorms or PLBs in tissue culture without using anti-microtubule agents and the use of flow cytometry with 4,6-diamidino-2-phenylindole (DAPI) staining is described in details in this chapter. This is a simple, effective, and reliable technique to produce large numbers of polyploid plants in Phalaenopsis orchids. It might have a great impact on new variety development of the orchid in the future.

Key words

Chromosome doubling DAPI Endopolyploidy Flow cytometry In vitro methods Phalaenopsis orchids Polyploidy induction Protocorm Protocorm-like body 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Wen-Huei Chen
    • 1
  • Ching-Yan Tang
    • 2
  1. 1.Orchid Research and Development Center, National Cheng Kung UniversityTainanRepublic of China
  2. 2.Department of Life SciencesNational University of KaohsiungKaohsiungRepublic of China

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