Recent Protocols on Genetic Transformation of Orchid Species

  • Chia-Wen Li
  • Ming-Tsair ChanEmail author
Part of the Springer Protocols Handbooks book series (SPH)


Gene transfer technology is an important tool for accelerating the process of breeding. Agrobacterium-mediated genetic transformation method has been applied to commercial orchid species, such as those in the genera Cattleya, Cymbidium, Dendrobium, Oncidium, Phalaenopsis, and Vanda. This chapter describes an efficient selection and plant regeneration protocol for Agrobacterium-mediated transformation of a model orchid, Erycina pusilla. Selection of suitable experimental materials, generation and propagation of explants, and components of the medium used in the serials of transformation procedures are detailed and compared with other orchid transformation protocols. In addition, some tips for improving the transformation efficiency are also included.

Key words

Agrobacterium tumefaciens Erycina pusilla Oncidium orchid Phalaenopsis Protocorm-like body Transformation 



This work was supported by grants from Academia Sinica and the National Science Council of the Republic of China.


  1. 1.
    Chin DP, Mishiba K, Mii M (2007) Agrobacterium-mediated transformation of protocorm-like bodies in Cymbidium. Plant Cell Rep 26:735–743CrossRefGoogle Scholar
  2. 2.
    Teixeira da Silva JA, Dobránszki J, Cardoso JC, Chandler SF, Zeng S (2016) Methods for genetic transformation in Dendrobium. Plant Cell Rep 35:483–504CrossRefGoogle Scholar
  3. 3.
    Li C-W, Liao C-H, Huang X, Chan M-T (2015) Orchids (Oncidium and Phalaenopsis). In: Wang K (ed) Agrobacterium protocols: volume 2. Humana Press, Totowa, NJ, pp 331–346Google Scholar
  4. 4.
    Mishiba K, Chin DP, Mii M (2005) Agrobacterium-mediated transformation of Phalaenopsis by targeting protocorms at an early stage after germination. Plant Cell Rep 24:297–303CrossRefGoogle Scholar
  5. 5.
    Phlaetita W, Chin DP, Tokuhara K, Nakamura I, Masahiro M (2015) Agrobacterium-mediated transformation of protocorm-like bodies in Dendrobium formidible ‘Ugusu’. Plant Biotechnol 32:225–231CrossRefGoogle Scholar
  6. 6.
    Shrestha BR, Chin DP, Tokuhara K, Mii M (2010) Agrobacterium-mediated transformation of Vanda using protocorm-like bodies. Asia-Pacific J Molec Biol Biotechnol 18:225–228Google Scholar
  7. 7.
    Zhang L, Chin DP, Mii M (2010) Agrobacterium-mediated transformation of protocorm-like bodies in Cattleya. Plant Cell Tiss Org Cult 103:41–47CrossRefGoogle Scholar
  8. 8.
    Chai D, Lee SM, Ng JH, Yu H (2007) L-methionine sulfoximine as a novel selection agent for genetic transformation of orchids. J Biotechnol 131:466–472CrossRefGoogle Scholar
  9. 9.
    Semiarti E, Indrianto A, Purwantoro A, Machida Y, Machida C (2011) Agrobacterium-mediated transformation of Indonesian orchids for micropropagation. In: INTECH Open Access Publisher, pp 215–240Google Scholar
  10. 10.
    Chan YL, Lin K-H, Liao L-J, Chen W-H, Chan M-T (2005) Gene stacking in Phalaenopsis orchid enhances dual tolerance to pathogen attack. Transgenic Res 14:279–288CrossRefGoogle Scholar
  11. 11.
    You SJ, Liau C-H, Huang H-E, Feng T-Y, Prasad V, Hsiao H-H, J-C L, Chan M-T (2003) Sweet pepper ferredoxin-like protein (pflp) gene as a novel selection marker for orchid transformation. Planta 217:60–65PubMedGoogle Scholar
  12. 12.
    Lee S-H, Li C-W, Liau C-H, Chang P-Y, Lin C-S, Chan M-T (2015) Establishment of an Agrobacterium-mediated genetic transformation procedure for the experimental model orchid Erycina pusilla. Plant Cell Tiss Org Cult 120:211–220CrossRefGoogle Scholar
  13. 13.
    Qin X, Liu Y, Mao S, Li T, Wu H, Chu C, Wang Y (2011) Genetic transformation of lipid transfer protein encoding gene in Phalaenopsis amabilis to enhance cold resistance. Euphytica 177:33–43CrossRefGoogle Scholar
  14. 14.
    Pan IC, Liao D-C, Wu F-H, Daniell H, Singh ND, Chang C, Shih M-C, Chan M-T, Lin C-S (2012) Complete chloroplast genome sequence of an orchid model plant candidate: Erycina pusilla apply in tropical Oncidium breeding. PLoS One 7:e34738CrossRefGoogle Scholar
  15. 15.
    Gnasekaran P, Antony JJJ, Uddain J, Subramanian S (2014) Agrobacterium-mediated transformation of the recalcitrant Vanda Kasem’s Delight orchid with higher efficiency. Sci Worl J 2014:583934Google Scholar
  16. 16.
    Tokuhara K, Mii M (1993) Micropropagation of Phalaenopsis and Doritaenopsis by culturing shoot tips of flower stalk buds. Plant Cell Rep 13:7–11CrossRefGoogle Scholar
  17. 17.
    Porebski S, Bailey LG, Baum BR (1997) Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Molec Biol Reprod 15:8–15CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of Life ScienceFu-Jen Catholic UniversityNew Taipei CityRepublic of China
  2. 2.Agricultural Biotechnology Research Center, Academia SinicaTainanRepublic of China

Personalised recommendations