Stable genetic transformation of castor (Ricinus communis L.) via Agrobacterium tumefaciens-mediated gene transfer using embryo axes from mature seeds

• Published in: Plant cell reports Vol. 23; no. 12; pp. 803 - 810
• Main Authors: Sujatha, M, Sailaja, M
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.03.2005; Springer Nature B.V
• Subjects: Agrobacterium tumefaciens; Agrobacterium tumefaciens - genetics; Anti-Bacterial Agents - pharmacology; Biological and medical sciences; Biotechnology; Castor; castor beans; Cefotaxime - pharmacology; Cinnamates - pharmacology; embryo (plant); Embryos; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Plant - drug effects; Gene Expression Regulation, Plant - genetics; Gene Transfer Techniques; Genetic engineering; Genetic Engineering - methods; Genetic technics; genetic transformation; genetic vectors; Genetic Vectors - genetics; Genome, Plant; Hygromycin B - analogs & derivatives; Hygromycin B - pharmacology; Methods. Procedures. Technologies; Microbiology; micropropagation; oil crops; plant regeneration; Plant Shoots - drug effects; Plant Shoots - genetics; Plants, Genetically Modified - embryology; Plants, Genetically Modified - genetics; Ricinus - drug effects; Ricinus - embryology; Ricinus - genetics; Ricinus communis; seed maturation; seeds; Seeds - drug effects; Seeds - embryology; Seeds - genetics; Transformation, Genetic - genetics; Transgenic animals and transgenic plants; Transgenic plants; Embryo; Vegetals; Genetic transformation; Castor; Agrobacterium tumefaciens; Meristem; Meristem-based transformation; Genetic transfer; Dicotyledones; Angiospermae; Bacteria; Euphorbiaceae; Spermatophyta; Transgenic plant; Rhizobiaceae; Ricinus communis
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-004-0898-4

A protocol for the transformation of castor embryo axes using the pCAMBIA vector 1304 in disarmed Agrobacterium tumefaciens strain EHA105 is presented. Co-cultivated explants were initially subjected to expansion and proliferation on MS medium with 0.5 mg l-1 TDZ followed by three cycles of selection on medium with 0.5 mg l-1 BA and increasing concentrations of hygromycin (20-40-60 mg l-1). Selected shoot clusters were transferred to medium with 0.5 mg l-1 BA for proliferation and 0.2 mg l-1 BA for shoot elongation. Elongated shoots were rooted on half-strength MS medium with 2.0 mg l-1 NAA. The presence and stable integration of the hpt gene was confirmed through PCR, RT-PCR, PCR-Southern blot, sequence analysis, Southern blot analysis and PCR analysis of progeny. Southern blot analysis of the primary transformants showed single copy integration and progeny analysis revealed monogenic inheritance of the introduced gene. This paper reports the first successful attempt at producing transgenic castor.