Transformation of radish (Raphanus sativus L.) via sonication and vacuum infiltration of germinated seeds with Agrobacterium harboring a group 3 LEA gene from B. napus

• DOI: 10.1007/s00299-005-0973-5
• Publication Date: 2005-04-20T08:36:18Z
• Authors: Byong-Jin Park; Zaochang Liu; Akira Kanno; Toshiaki Kameya
• Keywords: 0106 biological sciences; Base Sequence; Brassica napus; Germination; Genes, Plant; Plants, Genetically Modified; Polymerase Chain Reaction; 01 natural sciences; Raphanus; Blotting, Southern; Transformation, Genetic; DNA Primers; Rhizobium

A protocol for producing transgenic radish (Raphanus sativus) was obtained by using both ultrasonic and vacuum infiltration assisted, Agrobacterium-mediated transformation. The Agrobacterium strain LBA4404 contained the binary vector pBI121-LEA (late embyogenesis abundant), which carried a Group 3 LEA gene, from Brassica napus. Among six combinations, Agrobacterium-mediated transformation assisted by a combination of 5-min sonication with 5-min vacuum infiltration resulted in the highest transformation frequency. The existence, integration and expression of transferred LEA gene in transgenic T(1) plants were confirmed by PCR, genomic Southern and Western blot analysis. Transgenic radish demonstrated better growth performance than non-transformed control plants under osmotic and salt stress conditions. Accumulation of Group 3 LEA protein in the vegetative tissue of transgenic radish conferred increased tolerance to water deficit and salt stress.