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Transgenic yellow lupin (Lupinus luteus)
H Li1, S J Wylie1, M G K Jones1
1Western Australian State Agricultural Biotechnology Centre (SABC) and Centre for Legumes in Mediterranean Agriculture (CLIMA), School of Biological Sciences and Biotechnology, Division of Science and Engineering, Murdoch University, Perth, W.A. 6150, Australia e-mail: lihua@central.murdoch.edu.au Fax: +61-8-9360-6303, , , , , , AU.
Abstract:
Transgenic yellow lupin (Lupinus luteus L.) plants have been generated by meristem co-cultivation with Agrobacterium tumefaciens. The binary plasmid pPZBNIa contains the bar gene under the control of a CaMV 35 S promoter. The transformation method involves inoculation of embryonic axis explants with A. tumefaciens, flooding the meristem with glufosinate, and initial culture on non-selective medium. Shoots were transferred to culture medium containing 20 mg/l glufosinate. Following subculture, shoots were grafted onto non-transgenic narrow-leafed lupin (L. angustifolius L.) seedling rootstocks, or rooted in vitro. The overall transformation efficiency, as determined at the T1 generation, was 0.05%-0.75%. The transgenic nature of plants grown to the T6 generation was confirmed by phosphinothricin acetyl transferase, PCR and Southern analyses.
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