title: Orchids (Oncidium and Phalaenopsis).
abstract: This chapter describes an efficient and reproducible method for large-scale propagation of Oncidium and Phalaenopsis protocorm-like bodies (PLBs) using floral stalk sections and seeds, respectively. The propagated PLBs can be used for Agrobacterium-mediated transformation. An advanced transformation system for Oncidium and Phalaenopsis orchids has been established. This protocol demonstrates that the time during which the PLBs are cocultivated with Agrobacterium is the key to promoting transformation efficiency. Modified DNA and RNA extraction methods are also provided to diminish polysaccharide contamination and to improve the quality for further molecular analysis.
authors.affiliation: Department of Biotechnology, TransWorld University, Yunlin County 640, Douliu, Taiwan, s43306jw@gmail.com.
authors.forename: Chia-Wen
authors.initials: CW
authors.lastname: Li
authors.forename: Chia-Hui
authors.initials: CH
authors.lastname: Liao
authors.forename: Xia
authors.initials: X
authors.lastname: Huang
authors.forename: Ming-Tsair
authors.initials: MT
authors.lastname: Chan
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000254:growth & development; D015139:Blotting, Southern; D018920:Coculture Techniques; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification; D005818:Genetic Engineering / Q000379:methods*; D029595:Orchidaceae / Q000166:cytology / Q000235:genetics* / Q000254:growth & development*; D018749:RNA, Plant / Q000235:genetics / Q000302:isolation & purification; D012038:Regeneration; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014170:Transformation, Genetic
chemical_list: D018744:DNA, Plant; D018749:RNA, Plant