Establishment of an Agrobacterium-mediated genetic transformation procedure for the experimental model orchid Erycina pusilla

Shu Hong Lee, Chia Wen Li, Chia Hui Liau, Pao Yi Chang, Li Jen Liao, Choun Sea Lin, Ming Tsair Chan

研究成果: Article同行評審

16 引文 斯高帕斯(Scopus)

摘要

The establishment of a genetic transformation system for Erycina pusilla (E. pusilla) would be a major step for orchid research and help establish the species as a potential experimental model. Here we describe the development of an efficient selection and plant regeneration protocol for Agrobacterium-mediated transformation of E. pusilla protocorms. Plants were transformed with a binary plasmid containing the Arabidopsis thaliana gene Methionine sulfoxide reductase B7 (MSRB7) using the A. tumefaciens strain EHA105. Explants were selected on hygromycin-containing New Dogashima medium (Tokuhara and Mii in Plant Cell Rep 13(1):7–11, 1993) with 6-benzylaminopurine and α-naphthaleneacetic acid for 5 months to diminish the ratio of false positive explants, and it was determined that 3 month-old E. pusilla protocorms represented the optimal stage for transformation. Positive transformants were confirmed by Southern blot and PCR analyses, and by methyl viologen tolerance assays. Since E. pusilla has a shorter life cycle than most other orchids, self-pollinated T1 progeny was obtained within only 18 months. In addition, the results indicated that overexpression of MSRB genes can be used to avoid the effects of abiotic stress in economically important plants such as orchids.

原文English
頁(從 - 到)211-220
頁數10
期刊Plant Cell, Tissue and Organ Culture
120
發行號1
DOIs
出版狀態Published - 2015 一月

All Science Journal Classification (ASJC) codes

  • 園藝科學

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