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Hsing HX, Lin YJ, Tong CG, et al. Efficient and heritable transformation of Phalaenopsis orchids. Bot Stud. 2016;57(1):30doi: 10.1186/s40529-016-0146-6.
Hsing, H. X., Lin, Y. J., Tong, C. G., Li, M. J., Chen, Y. J., & Ko, S. S. (2016). Efficient and heritable transformation of Phalaenopsis orchids. Botanical studies, 57(1), 30. https://doi.org/10.1186/s40529-016-0146-6
Hsing, Hong-Xian, et al. "Efficient and heritable transformation of Phalaenopsis orchids." Botanical studies vol. 57,1 (2016): 30. doi: https://doi.org/10.1186/s40529-016-0146-6
Hsing HX, Lin YJ, Tong CG, Li MJ, Chen YJ, Ko SS. Efficient and heritable transformation of Phalaenopsis orchids. Bot Stud. 2016 Dec;57(1):30. doi: 10.1186/s40529-016-0146-6. Epub 2016 Oct 20. PMID: 28597440; PMCID: PMC5430590.
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Bot Stud. 2016 Dec;57(1):30. doi: 10.1186/s40529-016-0146-6. Epub 2016 Oct 20.

Efficient and heritable transformation of Phalaenopsis orchids.

Botanical studies

Hong-Xian Hsing, Yi-Jyun Lin, Chii-Gong Tong, Min-Jeng Li, Yun-Jin Chen, Swee-Suak Ko

Affiliations

  1. Academia Sinica Biotechnology Center in Southern Taiwan, Tainan, 741, Taiwan.
  2. Agricultural Biotechnology Research Center, Academia Sinica, Taipei, 115, Taiwan.
  3. Academia Sinica Biotechnology Center in Southern Taiwan, Tainan, 741, Taiwan. [email protected].
  4. Agricultural Biotechnology Research Center, Academia Sinica, Taipei, 115, Taiwan. [email protected].

PMID: 28597440 PMCID: PMC5430590 DOI: 10.1186/s40529-016-0146-6

Abstract

BACKGROUND: Phalaenopsis orchid (Phal. orchid) is visually attractive and it is important economic floriculture species. Phal. orchids have many unique biological features. However, investigation of these features and validation on their biological functions are limited due to the lack of an efficient transformation method.

RESULTS: We developed a heritable and efficient Agrobacterium- mediated transformation using protocorms derived from tetraploid or diploid Phal. orchids. A T-DNA vector construct containing eGFP driven by ubiquitin promoter was subjected to transformation. An approximate 1.2-5.2 % transformation rate was achieved. Genomic PCR confirmed that hygromycin selection marker, HptII gene and target gene eGFP were integrated into the orchid genome. Southern blotting indicated a low T-DNA insertion number in the orchid genome of the transformants. Western blot confirmed the expression of eGFP protein in the transgenic orchids. Furthermore, the GFP signal was detected in the transgenic orchids under microscopy. After backcrossing the pollinia of the transgenic plants to four different Phal. orchid varieties, the BC1 progenies showed hygromycin resistance and all surviving BC1 seedlings were HptII positive in PCR and expressed GFP protein as shown by western blot.

CONCLUSIONS: This study demonstrated a stable transformation system was generated for Phal. orchids. This useful transformation protocol enables functional genomics studies and molecular breeding.

Keywords: Agrobacterium tumefaciens; Hygromycin selection; Phalaenopsis aphrodite; Protocorm; Transformation; eGFP

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