Agrobacterium-mediated transformation of Cymbidium sinensis.
- Author:
Li XIE
;
Fen WANG
;
Ruizhen ZENG
;
Herong GUO
;
Yuliang ZHOU
;
Zhisheng ZHANG
- Publication Type:Journal Article
- MeSH:
Agrobacterium;
Coculture Techniques;
Genetic Engineering;
Glucuronidase;
Orchidaceae;
genetics;
Plants, Genetically Modified;
genetics;
Polymerase Chain Reaction;
Transformation, Genetic
- From:
Chinese Journal of Biotechnology
2015;31(4):542-551
- CountryChina
- Language:Chinese
-
Abstract:
Genetic transformation is an effective method to improve breeding objective traits of orchids. However, there is little information about genetic transformation of Cymbidium sinensis. Rhizomes from shoot-tip culture of C. sinensis cv. 'Qijianbaimo' were used to establish a practical transformation protocol of C. sinensis. Pre-culture time, concentration and treating methods of acetosyringone, concentration of infection bacteria fluid (OD600), infection time, and co-culture time had significant effects on β-glucuronidase (GUS) transient expression rate of C. sinensis cv. 'Qijianbaimo' rhizome. The GUS transient expression rate of rhizome was the highest (11.67%) when rhizomes pre-cultured for 39 d were soaked in bacterium suspension (OD600 = 0.9) supplemented with 200 μmol/L acetosyringone for 35 min, followed by culturing on co-culture medium supplemented with 200 μmol/L acetosyringone for 7 d. Under this transformation conditions, 3 transgenic plantlets, confirmed by GUS histochemical assay and PCR, were obtained from 400 regenerated plantlets, and the genetic transformation rate was 0.75%. This proved that it was feasible to create new cultivars by the use of Agrobacterium-mediated genetic transformation in C. sinense.
