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.
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.
Park
Byong-Jin
BJ
Department of Environmental Life Sciences, Graduate school of Life Sciences, Tohoku University, 2-1-1 Katahira, Aobaku, Sendai 980-8577, Japan.
Liu
Zaochang
Z
Kanno
Akira
A
Kameya
Toshiaki
T
D001483:Base Sequence; D015139:Blotting, Southern; D029688:Brassica napus / Q000235:genetics*; D017931:DNA Primers; D017343:Genes, Plant; D018525:Germination; D030821:Plants, Genetically Modified / Q000196:embryology / Q000235:genetics / Q000382:microbiology; D016133:Polymerase Chain Reaction; D031224:Raphanus / Q000196:embryology / Q000235:genetics* / Q000382:microbiology; D012231:Rhizobium / Q000235:genetics / Q000502:physiology*; D014170:Transformation, Genetic
D017931:DNA Primers