Agrobacterium-mediated transformation of Eruca sativa

作者：Slater Susan M H^*; Keller W A; Scoles G

来源：Plant Cell, Tissue and Organ Culture, 2011, 106(2): 253-260.

DOI：10.1007/s11240-010-9915-1

• Agrobacterium tumefaciens
• Brassicaceae
• Eruca sativa

An Agrobacterium tumefaciens-mediated transformation system was developed for Eruca sativa (eruca). Hypocotyl explants were co-cultivated with bacterial cells carrying a plasmid harboring a uidA:nptII fusion gene along a phosphinothricin acetyl transferase (PAT) gene cassette, for a period of 2 days. These were grown on a high cytokinin/auxin medium containing 5.0 mg l(-1) 6-benzyladenine (BA), 1.0 mg l(-1) indole-3-acetic acid (IAA), and 0.1 mg l(-1) alpha-naphthaleneacetic acid (NAA). Explants were then transferred to a lower cytokinin/auxin medium containing 2.0 mg l(-1) BA and 0.1 mg l(-1) NAA along with 5.0 mg l(-1) silver nitrate and 300 mg l(-1) TimentinA (R). Upon transfer to a selection medium containing either 20 mg l(-1) kanamycin or 2 mg l(-1) L-phosphinothricin (L-ppt), shoot regenerants were observed. Expression of the transgenes in putative transformants was confirmed using a histochemical GUS assay. Presence of the PAT transgene in GUS-positive T0 plants was confirmed by Southern blot analysis. Moreover, spot tests of T1 seedlings were conducted using the L-ppt herbicide. A transformation frequency of 1.1% was obtained with more than 60% of transgenic lines containing single copies of the transgenes.

• 出版日期2011-8