GMO molecular characterization details

GMO detailed information

GMO:
5715
Alias:

Cibus Canola Event 5715, GE canola, GE rape, gene editing, Herbicide tolerant canola
UID:
Tradename:

SU Canola
Companies/Institutions:
Cibus US
Species:
Brassica napus
(canola, oilseed rape, rapeseed)
Traits:
EU authorisation:

no Reference materials could be found
No results found.
no Literature reference could be found

General Information

Description:

The canola event 5715 was developed by conventional breeding of two herbicide tolerant mutants.

Brassica napus carries two complete genomes designated "A" and "C". Each genome has an acetohydroxyacid synthase (ahas) gene (ahas1 gene in the C genome and ahas3 in the A genome) coding for an AHAS enzyme.

Event 5715 is a cross between Cibus canola line BnALS-57 (donor of the single nucleotide mutation in the ahas1 gene of the C genome) and the commercially available Clearfield canola variety SP Cougar CL (donor of the single nucleotide mutation in the ahas3 gene of the A genome).

BnALS-57 was selected as a mutant variant of the wild-type parental line BN2. Mutants of BN2 were generated using an oligonucleotide-directed mutagenesis (ODM) approach known as the Rapid Trait Development System™ (RTDS™). No changes were observed in the promoter regions or locations that would affect endogenous gene expression. This was confirmed by sequencing a 2500-base pair (bp) region upstream of the AHAS1 gene which was determined to be identical with GenBank accession Z11524. The developer hypothesized that the single nucleotide mutation was the result of a spontaneous somaclonal variation that occurred during the tissue culture process, and not due to the specific oligonucleotide used in the RTDS protocol.

Being a crossing product of BnALS-57 and SP Cougar CL, 5715 possesses a specific single nucleotide mutation in the ahas1 and ahas3 gene, respectively. Both the ahas1 and ahas3 genes encode a protein subunit (i.e., a monomer) of an acetohydroxyacid synthase (ahas) enzyme. The single nucleotide mutation in the subunits of each of the ahas enzymes results in conformational changes in their configuration, reducing the binding affinity of imidazolinone and sulfonylurea herbicides, thus conferring herbicide tolerance to the canola plant.

DNA sequencing was used to confirm homozygosity of the single nucleotide mutation in both the ahas1 and ahas3 genes. Furthermore, plants from an advanced breeding generation of Cibus canola event 5715 demonstrated consistent imidazolinone and sulfonylurea herbicide tolerance, phenotypically confirming the stable inheritance of the trait.

Links regarding to this GMO:

BCH http://bch.cbd.int/database/record.shtml?documentid=110268,

Health Canada http://www.canada.ca/en/health-canada/services/food-nutrition/genetically-modified-foods-other-novel-foods/approved-products/novel-food-information-cibus-canola-event-5715-imidazolinone-sulfonylurea-herbicide-tolerant.html

Transformation / Modification technique:

Oligonucleotide-directed mutagenesis (ODM)

Inserts / Modification

Order	Name	Donor	Size (bp)	Position	Definition	Trait	Comment
	CS-ahas1_genome edited-HOST	Brassica napus		-	Acetohydroxy acid synthase 1 gene	ALS/AHAS inhibitor tolerance	mutation introduced through genome editing resutling in a tryptophan to leucine amino acid change at position 574 (aa)
	CS-ahas3-HOST	Brassica napus		-	Acetohydroxy acid synthase 3 gene	ALS/AHAS inhibitor tolerance	gene of Clearfield rape contains a mutation resutling in a tryptophan to leucine amino acid change at position 574 (not genome edited)

Acc.No.	Reference	Sequence	Size (bp)	Annotation
	patent documents	No

Detection Methods

7 results

Name	Type	Target	Description	Validation	Standardisation	Verification
QT-TAX-FatA primer1/FatA primer2	taxon-specific	Brassica napus	Quantitative real-time PCR (TaqMan) method for the detection of oilseed rape (Monsanto, 2004, A recommended procedure for real-time quantitative TaqMan PCR for Roundup Ready canola RT73). Estimated number of target copies per haploid genome (Jacchia et al. 2018): 2	in-house validation	none	1
QT-TAX-BN-012	taxon-specific	Brassica napus	Quantitative real-time PCR (TaqMan) method for detection of oilseed rape cruciferin A (CruA) gene (EU-RL GMFF GMOMETHODS database as of March 06, 2014). Estimated number of target copies per haploid genome (Jacchia et al. 2018): 2	ring trial validation	EU reference method	1
QT-TAX-BN-003	taxon-specific	Brassica napus	Quantitative real-time PCR (TaqMan) method for detection of oilseed rape cruciferin A gene (Jacchia et al. 2019) (EU-RL GMFF GMOMETHODS database as of January 15, 2020)	ring trial validation	EU reference method	1
QT-TAX-BN-002	taxon-specific	Brassica napus	Quantitative real-time PCR (TaqMan) method for detection of oilseed rape cruciferin storage protein (EU-RL GMFF GMOMETHODS database as of January 27, 2014). Estimated number of target copies per haploid genome (Jacchia et al. 2018): 4	ring trial validation	EU reference method	1
QT-TAX-BN-001	taxon-specific	Brassica napus	Quantitative real-time PCR (TaqMan) method for detection of oilseed rape acyl-ACP thioesterase (FatA) gene (EU-RL GMFF GMOMETHODS database as of January 23, 2014) Estimated number of target copies per haploid genome (Jacchia et al. 2018): 1	ring trial validation	EU reference method	1
QL-TAX-BN-004	taxon-specific	Brassica napus	Qualitative conventional PCR method for detection of oilseed rape high mobility group protein I/Y gene (Pan et al., 2007).	ring trial validation	unknown	1
QL-TAX-BN-003	taxon-specific	Brassica napus	Qualitative real-time PCR (SYBRGreen) method for detection of oilseed rape cruciferin A gene(Barbau-Piednoir et al., 2014)	ring trial validation	unknown	1

Name:	edited_5715
Comment:	The 5715 event is the crossing product of BnALS-57 (genome edited line) containing the PM2 single point mutation in the ahas1 gene (genome C) and Clearfield rape (commercially available line obtained using conventional breeding techniques) which contains the PM2 single point mutation in the ahas3 gene (genome A). The PM2 mutation consists of a G to T substitution. As a result, codon 574 reads TTG instead of TGG and codes for leucine instead of tryptophan in the ahas protein.
Map:

European GMO Initiative for a Unified Database System

GMO detailed information

GMO:

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UID:

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Traits:

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General Information

Links regarding to this GMO:

Inserts / Modification

Transformation Vectors

Detection Methods

Reference Materials

Authorisation

Literature reference