Generation and characterization of soybean and marker‐free tobacco plastid transformants over‐expressing a bacterial 4‐hydroxyphenylpyruvate dioxygenase which provides strong herbicide tolerance
- 28 November 2006
- journal article
- Published by Wiley in Plant Biotechnology Journal
- Vol. 5 (1), 118-133
- https://doi.org/10.1111/j.1467-7652.2006.00226.x
Abstract
Plant 4-hydroxyphenylpyruvate dioxygenase (HPPD) is part of the biosynthetic pathway leading to plastoquinone and vitamin E. This enzyme is also the molecular target of various new bleaching herbicides for which genetically engineered tolerant crops are being developed. We have expressed a sensitive bacterial hppd gene from Pseudomonas fluorescens in plastid transformants of tobacco and soybean and characterized in detail the recombinant lines. HPPD accumulates to approximately 5% of total soluble protein in transgenic chloroplasts of both species. As a result, the soybean and tobacco plastid transformants acquire a strong herbicide tolerance, performing better than nuclear transformants. In contrast, the over-expression of HPPD has no significant impact on the vitamin E content of leaves or seeds, quantitatively or qualitatively. A new strategy is presented and exemplified in tobacco which allows the rapid generation of antibiotic marker-free plastid transformants containing the herbicide tolerance gene only. This work reports, for the first time, the plastome engineering for herbicide tolerance in a major agronomic crop, and a technology leading to marker-free lines for this trait.Keywords
This publication has 53 references indexed in Scilit:
- Plastid marker‐gene excision by transiently expressed CRE recombinaseThe Plant Journal, 2006
- Patterns of Protein Oxidation in Arabidopsis Seeds and during GerminationPlant Physiology, 2005
- Plastid-Expressed Betaine Aldehyde Dehydrogenase Gene in Carrot Cultured Cells, Roots, and Leaves Confers Enhanced Salt TolerancePlant Physiology, 2004
- Generation of marker-free plastid transformants using a transiently cointegrated selection geneNature Biotechnology, 2004
- Persistence of Unselected Transgenic DNA during a Plastid Transformation and Segregation Approach to Herbicide ResistancePlant Physiology, 2003
- A chloroplast transgenic approach to hyper‐express and purify Human Serum Albumin, a protein highly susceptible to proteolytic degradationPlant Biotechnology Journal, 2003
- Complete Sequence of the Maize Chloroplast Genome: Gene Content, Hotspots of Divergence and Fine Tuning of Genetic Information by Transcript EditingJournal of Molecular Biology, 1995
- Translation of psbA mRNA is regulated by light via the 5′‐untranslated region in tobacco plastidsThe Plant Journal, 1994
- Amino Acid Biosynthesis Inhibitors As HerbicidesAnnual Review of Biochemistry, 1988
- Why do chloroplasts and mitochondria contain so many copies of their genome?BioEssays, 1987