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Plant-Parasitic Nematode-Protected and Herbicide Tolerant Soybean
BASF Plant Science GmbH
The soy (Glycine max) has been modified for plant
parasitic nematode resistance and herbicide tolerance. The soy
contains Bacillus thuringiensis Cry14Ab1 for resistance to
nematode plant parasites, such as soybean cyst nematode. The
protein is related to other crystal proteins and thought to act on
the lining of the intestine of nematodes to confer resistance. For
herbicide tolerance, a Pseudomonas fluorescens
4-hydroxyphenylpyruvate dioxygenase (HPPD) gene with point
mutations in the C-terminal to allow for and enhance tolerance to
HPPD-inhibitor herbicides, such as isoxaflutole and mesotrione.
The term Recipient organism refers to an organism (either already modified or non-modified) that was subjected to genetic modification, whereas Parental organisms refers to those that were involved in cross breeding or cell fusion.
- Agrobacterium-mediated DNA transfer
Polyubiquitin10 gene promoter
4-Hydroxyphenylpyruvate dioxygenase gene
Optimized Transit Peptide
CaMV Enhanced 35S promoter
Two gene expression cassettes were inserted in to the soy
(Glycine max) genome. The gene cassettes are present in
the counter-clockwise orientation.
Transcription of Bacillus thuringiensis cry14Ab1 is under
control of the Arabidopsis thaliana ubiquitin 10 promoter
and the Cauliflower mosaic virus (CaMV) 35S terminator.
The ubiquitin 10 promoter promotes expression of cry14Ab1
at high levels throughout all plant tissues.
Transcription of Pseudomonas fluorescens
4-hydroxyphenylpyruvate dioxygenase 9 (hppdPf-4Pa) is
under control of a CaMV 35S enhanced promoter and terminator.
At the 5' end of the transcript contains a synthetic optimized
peptide from Zea mays and Helianthus annuus and a
Tobacco etch virus (TEV) leader sequence. The transit
peptide targets the protein to the chloroplast of the cells and the
TEV leader sequence enhances translation of the transcript. The
enhanced promoter contains duplicated enhancer sequences and
promotes high levels of expression.
- Sequencing analysis indicated a single T-DNA insertion without
- A 21 basepair sequence corresponding to ORIpVS1 vector backbone
sequence was inserted into the soy genome. This sequence does not
correspond to the antibiotic resistance gene.
- The T-DNA insertion resulted in a 63 basepair deletion in the soy
- Bioinformatic analysis suggested the T-DNA was inserted into the
3' untranslated region of putative BON1-associated protein 1-like
protein on chromosome 7.
- The coding sequence of hppdPf-4Pa contains changes that
result in the following amino acid substitutions in the HPPD
-- glutamic acid residue was changed for a proline residue at
-- a glycine residue for a tryptophan residue at position
-- a lysine residue for an alanine residue at position 339; and
-- an alanine residue for a glutamine residue at position 340.
- Resistance to diseases and pests
- Resistance to herbicides
- Tolerance to isoxaflutole
- Tolerance to mesotrione
4-Hydroxyphenylpyruvate dioxygenase (HPPD) is a iron
(II)-dependent non-heme oxygenase that catalyzes the conversion of
4-hydroxyphenylpyruvate to homogentisate in the tyrosine catabolism
pathway. Herbicides inhibiting HPPD prevent the breakdown of
tyrosine resulting in stunted growth due to excess tyrosine and
oxidative damage because the precursors for the photosynthetic
transport chain and antioxidative systems (tocopherols and
carotenoids) are not being produced.