A simple shoot multiplication procedure using internode explants, and its application for particle bombardment and Agrobacterium-mediated transformation in watercress

• Published in: Journal of plant research Vol. 122; no. 4; pp. 455 - 463
• Main Authors: Ogita, S.(Toyama Prefectural Univ., Imizu (Japan)), Usui, M, Shibutani, N, Kato, Y
• Format: Journal Article
• Language: English
• Published: Japan Springer Japan 01.07.2009; Springer Nature B.V
• Subjects: ADN; AGROBACTERIUM; Biolistics - methods; Biomedical and Life Sciences; Brassicaceae - cytology; Brassicaceae - genetics; Brassicaceae - microbiology; BROTES; Cell culture; Cell division; Confocal microscopy; CULTIVO IN VITRO; CULTURE IN VITRO; DNA; ENTRE NOEUD; EXPLANT; EXPLANTES; EXPLANTS; Gene Expression Profiling; Genetic engineering; GENETIC TRANSFORMATION; Grasses; Green Fluorescent Proteins - genetics; IN VITRO CULTURE; INTERNODES; INTERNODIOS; Life Sciences; Nasturtium officinale; NUCLEOTIDE SEQUENCE; Onions - genetics; Organogenesis; Organogenesis - drug effects; particle bombardment; Plant Biochemistry; Plant Ecology; Plant Growth Regulators - pharmacology; Plant Physiology; Plant Sciences; Plant Shoots - cytology; Plant Shoots - drug effects; Plant Shoots - embryology; Plant Shoots - growth & development; Plant Stems - cytology; Plant Stems - drug effects; Plant Stems - embryology; Plant Stems - growth & development; PLANTAS TRANSGENICAS; PLANTE TRANSGENIQUE; Plants, Genetically Modified; POUSSE; Regular Paper; Rhizobium - metabolism; SECUENCIA NUCLEOTIDICA; SEQUENCE NUCLEOTIDIQUE; SHOOTS; Studies; thidiazuron; TRANSFORMACION GENETICA; Transformation; TRANSFORMATION GENETIQUE; Transformation, Genetic - drug effects; TRANSGENIC PLANTS; Multiple shoot; Green florescent protein; Watercress; Particle bombardment
• ISSN: 0918-9440; 1618-0860
• DOI: 10.1007/s10265-009-0226-8

A shoot multiplication system derived from internode explants was investigated with the aim of improving genetic characteristics of watercress (Nasturtium officinale R. Br.). Internodes of ca. 1 cm excised from in vitro stock shoot culture were placed on half-strength Murashige and Skoog (MS) medium supplemented with 3 microM 2,4-dichlorophenoxyacetic acid as a pre-treatment. Laser scanning microscopy indicated clearly that the first sign of meristematic cell division could be seen after 1-2 days of pre-culture, and meristematic tissues multiplied along the vascular cambium of the internode segment during 7 days of culture. Multiple shoots could be obtained from more than 90% of the pre-treated explants when they were subsequently transferred to MS medium supplemented with 1 microM thidiazuron for 3 weeks. These findings indicate that pre-treatment of the internodes for 7 days promoted their capacity for organogenesis. Using this pre-treatment, frequent generation of transgenic watercress plants was achieved by adapting particle bombardment and Agrobacterium-mediated transformation techniques with a construct expressing a synthetic green florescent protein gene.