Efficient and heritable transformation of Phalaenopsis orchids

Autor: Yi-Jyun Lin, Chii-Gong Tong, Min-Jeng Li, Hong-Xian Hsing, Swee-Suak Ko, Yun-Jin Chen
Rok vydání: 2016
Předmět:
Zdroj: Botanical Studies
ISSN: 1999-3110
DOI: 10.1186/s40529-016-0146-6
Popis: 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. Electronic supplementary material The online version of this article (doi:10.1186/s40529-016-0146-6) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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