Premium
Transformation of Brassica napus by using the aadA gene as selectable marker and inheritance studies of the marker genes
Author(s) -
Schröder Mia,
Dixelius Christina,
Råhlén Lennart,
Glimelius Kristina
Publication year - 1994
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1994.tb06652.x
Subject(s) - biology , selectable marker , genetics , gene , marker gene , southern blot , transformation (genetics) , agrobacterium tumefaciens , locus (genetics) , genetic marker , brassica , dna , microbiology and biotechnology , botany
An Agrobacterium tumefaciens ‐mediated transformation system for Brassica napus has been improved. We investigated several marker genes for transformation of Brassica napus , and the aadA gene, which confers resistance to streptomycin and spectinomycin, was found to be the most suitable. Forty‐three out of 193 putative transformants in the T 1 generation were investigated by Southern blot analysis. Transformants containing a range of 1 to 10 integrated T‐DNA copies per genome were found. Total DNA from 35 plants showed hybridisation to both the aadA and the nptll marker gene probes, from 5 plants only to one marker gene probe and from 3 plants DNA did not hybridise to any of the gene probes. Furthermore, more complex integration patterns such as direct repeated copies of the T‐DNA, both as tandem and inverted copies, were observed. Inheritance of the marker genes in the T 2 generation was studied in 37 of the plants. This revealed that 22% of the plants that contained both marker genes, segregated as one single locus (3:1) for both genes, while 46% of the plants gave a segregation pattern corresponding to one T‐DNA locus for at least one of the marker genes. Moreover, these inheritance patterns appeared to be more or less independent of the number of genes seen in the Southern blot analysis of the T, generation. In this study we show that the introduced marker genes are inherited by the T; generation in a less predictable way than was earlier reported for B. napus .