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Transgenic plant production from leaf discs of Moricandia arvensis using Agrobacterium tumefaciens

Author:
Rashid, H., Toriyama, K., Hinata, K.
Source:
Plant cell reports 1996 v.15 no.11 pp. 799-803
ISSN:
0721-7714
Subject:
Moricandia arvensis, genetic transformation, leaves, gene transfer, Agrobacterium tumefaciens, transgenic plants, cell suspension culture, shoots, callus, internodes, stems, genetic vectors, genes, kanamycin, hygromycin B, beta-glucuronidase, flowering, enzyme activity, culture media, benzyladenine, dose response, methodology, genetic techniques and protocols, explants, naphthaleneacetic acid
Abstract:
A high frequency shoot regeneration (80%) was developed from callus of leaf discs and stem internodes of Moricandia arvensis. Leaf discs were shown to be a preferable starting material for transformation experiments. Agrobacterium tumefaciens strain GV3101/pMP90 used in this study contained a binary vector with genes for kanamycin resistance, hygromycin resistance and beta-glucuronidase (GUS). Maximum transformation efficiency (10.3%) was achieved by using kanamycin at the rate of 200 mg/l as a selection agent. Presence of tobacco suspension culture during co-cultivation and a pre-selection period of seven days after co-cultivation was essential for successful transformation. Transgenic plants grew to maturity and exhibited flowering in a glasshouse. GUS activity was evident in all parts of leaf and the presence of GUS gene in plant genome was confirmed by PCR analysis.
Agid:
1436721