A facile method for screening for phosphinothricin (PPT)-resistant transgenic wheats

Citation
S. Rasco-gaunt et al., A facile method for screening for phosphinothricin (PPT)-resistant transgenic wheats, MOL BREED, 5(3), 1999, pp. 255-262
Citations number
29
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Plant Sciences","Animal & Plant Sciences
Journal title
MOLECULAR BREEDING
ISSN journal
1380-3743 → ACNP
Volume
5
Issue
3
Year of publication
1999
Pages
255 - 262
Database
ISI
SICI code
1380-3743(1999)5:3<255:AFMFSF>2.0.ZU;2-N
Abstract
A bioassay was developed for identifying transgenic wheat plants based on D e Block et al.'s [8] ammonium-multiwell assay which allows qualitative and quantitative evaluation of the expression of the enzyme phosphinothricin ac etyl transferase (PAT). Important parameters in the assay are the use of yo ung leaf tissues, short incubation period (6 h) and a high light intensity during incubation (250 mu mol s(-1) m(-2)). The assay is quick and results are obtained within a day. Ammonium measurements based on a colourimetric ( modified Berthelot) reaction are conducted using sodium salicylate to avoid the use of phenol. Results of the assay show high correlation with Basta l eaf painting tests and polymerase chain reaction (PCR) results. Thus, the a ssay may be used as a facile screen for bar-expressing transgenic cereals.