Agrobacterium tumefaciens-Mediated genetic transformation in tea (Camellia sinensis [L.] O. Kuntze) View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2004-06

AUTHORS

S. Joseph Lopez, R. Raj Kumar, P. K. Pius, N. Muraleedharan

ABSTRACT

We have developed a system to produce transgenic plants in tea (Camelia sinensis [L.] O. Kuntze) viaAgrobacterium tumefaciens-mediated transformation of embryogenic calli. Cotyledon-derived embryogenic callus cultures were cocultivated with anA. tumefaciens strain (AGL 1) harboring a binary vector carrying the hygromycin phosphotransferase (hpt II), glucuronidase (uid A), and green fluorescent protein (GFP) genes in the tDNA region. Following cocultivation, embryogenic calli were cultured in medium containing 500 mg/L carbenicillin for 1 wk and cultured on an antibiotic selection medium containing 75 mg/L hygromycin for 8–10 wk. Hygromycin-resistant somatic embryos were selected. The highest production efficiency of hygromycin-resistant calli occurred with cocultivation for 6–7 d in the presence of 400 μM acetosyringone (AS). Hygromycin-resistant somatic embryos developed into complete plantlets in regeneration medium containing half-strength Murashige and Skoog (MS) salts with 1 mg/L benzyl amino purine (BAP) and 9 mg/L giberellic acid (GA3). Transformants were subjected to GFP expression analysis, β-glucuronidase (GUS) histochemical assay, PCR analysis, and Southern hybridization to confirm gene integration. More... »

PAGES

201-202

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf02772730

DOI

http://dx.doi.org/10.1007/bf02772730

DIMENSIONS

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