Repositorium

What is a repositorium?

The repositorium is a searchable database that provides data on relevant articles from journals, company web pages and web pages of governmental agencies about studies/applications of genome-editing in model plants and agricultural crops in the period January 1996 to May 2018. Search options are article type, technique, plant, traits or free text. The repositorium is based on the systematic map of Dominik Modrzejewski et al., published in the journal environmental evidence. (Download article PDF).

Chimeric RNA/DNA oligonucleotide-based site-specific modification of the tobacco acetolactate syntase gene


Typ / Jahr

Journal Article / 2003

Autoren

Kochevenko, Andrej; Willmitzer, Lothar

Abstract

Single amino acid substitutions at either of two crucial positions in acetolactate synthase (ALS) result in a chlorsulfuron-insensitive form of this enzyme and, as a consequence, a herbicide-resistant phenotype. Here, we describe the successful in vivo targeting of endogenous tobacco (Nicotiana tabacum) ALS genes using chimeric RNA/DNA and all-DNA oligonucleotides at two different locations. Similar number of conversion events with two different chimeras indicates the absence of restricting influence of genomic target sequence on the gene repair in tobacco. Chlorsulfuron-resistant plants were regenerated from calli after mesophyll protoplast electroporation or leaf tissue particle bombardment with these specifically constructed chimeras. Sequence analysis and enzyme assays proved the resulting alterations to ALS at both DNA and protein levels. Furthermore, foliar application of chlorsulfuron confirmed the development of resistant phenotypes. Lines with proline-196-alanine, threonine, glutamine, or serine substitutions or with tryptophan-573-leucine substitutions were highly resistant at both cellular and whole plant levels, whereas lines with proline-196-leucine substitutions were less resistant. The stability of these modifications was demonstrated by the continuous growth of calli on chlorsulfuron-containing medium and by the transmission of herbicide resistance to progeny in a Mendelian manner. Ability of haploid state to promote chimera-mediated conversions is discussed.

Keywords
Acetolactate Synthase/genetics/metabolism; Amino Acid Sequence; Amino Acid Substitution; Base Sequence; DNA, Plant/chemistry/genetics; DNA/genetics; Drug Resistance/genetics; Gene Targeting/methods; Molecular Sequence Data; Mutagenesis, Site-Directed; Oligonucleotides/genetics; phenotype; RNA/genetics; Sequence Analysis, DNA; Sequence Homology, Nucleic Acid; Sulfonamides; Tobacco/drug effects/enzymology/genetics; Triazines/pharmacology
Periodical
Plant physiology
Periodical Number
1
Page range
174–184
Volume
132
DOI
10.1104/pp.102.016857

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
209 Kochevenko, Andrej
Germany
nicotiana tabacum Oligonucleotide-directed mutagenesis
ALS
herbicide tolerance
PM
Basic research
Basic research