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High‐efficiency transformation of Arabidopsis thaliana with a selectable marker gene regulated by the T‐DNA 1′ promoter
Author(s) -
Mengiste Tesfaye,
Amedeo Paolo,
Paszkowski Jerzy
Publication year - 1997
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1997.12040945.x
Subject(s) - selectable marker , arabidopsis thaliana , transformation (genetics) , gene , genetics , biology , dna , arabidopsis , computational biology , microbiology and biotechnology , mutant
Summary Two selectable marker genes harbouring the bar coding region but differing in their promoters were compared in an Arabidopsis thaliana transformation assay using in planta infiltration with Agrobacterium tumefaciens . Surprisingly, in four Arabidopsis ecotypes examined, the 1′ promoter from the right T‐DNA was superior to the most commonly used 35S promoter of cauliflower mosaic virus (CaMV). The ecotype Wassilewskija gave the highest transformation frequencies, with an average of between 5.3 and 6.3 % of the seedlings subjected to the selection. This is approximately 30‐fold higher than previously reported results. Analysis of T‐DNA integration patterns in single transformed plants or pooled populations revealed independent T‐DNA integration events in each case. Results show that the 1′ promoter is an attractive alternative to the 35S promoter for the generation of T‐DNA insertion lines. The 1′ promoter may be especially beneficial for the secondary transformation of transgenic strains containing the 35S promoter to exclude homology‐mediated gene silencing.