SYN-IR6Ø4-5 × DAS-Ø15Ø7-1 × SYN-Ø53Ø7-1 × MON-ØØØ21-9 - Herbicide tolerant, insect resistant maize | BCH-LMO-SCBD-115543 | Living Modified Organism | Biosafety Clearing-House

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Living Modified Organism (LMO)
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Decisions on the LMO Risk Assessments  
last updated: 22 Apr 2020
Living Modified Organism identity
The image below identifies the LMO through its unique identifier, trade name and a link to this page of the BCH. Click on it to download a larger image on your computer. For help on how to use it go to the LMO quick-links page.
Herbicide tolerant, insect resistant maize
EN
MIR604 x TC1507 x 5307 x GA21
Yes
SYN-IR6Ø4-5 × DAS-Ø15Ø7-1 × SYN-Ø53Ø7-1 × MON-ØØØ21-9
The modified maize event was a result of cross-breeding modified parental lines and demonstrates herbicide tolerance and insect resistance. For Lepidoptera resistance, the maize expresses Bacillus thuringiensis Cry1F. For Coleoptera resistance, the maize expresses B. thuringiensis mCry3A and eCry3.1Ab. In addition to the insecticidal proteins, the maize also expresses Streptomyces viridochromogenes phosphinothricin N-acetyltransferase for glufosinate tolerance and a modified native Zea mays 5-enolpyruvylshikimate-3-phosphate synthase for tolerance to glyphosate. A selectable marker, Escherichia coli phosphomannose isomerase, is also present and was used during the transformation of the parental line using mannose selection.
EN
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.
  • BCH-ORGA-SCBD-246-6 Organism Zea mays (Maize, Corn, MAIZE)
    Crops
  • BCH-LMO-SCBD-15105-12 Living Modified Organism SYN-IR6Ø4-5 - Agrisure™ RW Rootworm-Protected maize
    Resistance to diseases and pests (Insects, Coleoptera (beetles))
  • BCH-LMO-SCBD-14841-13 Living Modified Organism DAS-Ø15Ø7-1 - Herculex™ I maize
    Dow AgroSciences, Pioneer Hi-Bred International Inc. | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths)), Resistance to herbicides (Glufosinate)
  • BCH-LMO-SCBD-104791-4 Living Modified Organism SYN-Ø53Ø7-1 - Agrisure® Duracade™ Maize
    Syngenta Seeds GmbH | Resistance to diseases and pests (Insects, Coleoptera (beetles), Western corn rootworm (Diabrotica virgifera), Northern corn rootworm (Diabrotica barberi))
  • BCH-LMO-SCBD-14794-18 Living Modified Organism MON-ØØØ21-9 - Roundup Ready™ maize
    Monsanto | Resistance to herbicides (Glyphosate)
EN
Characteristics of the modification process
pZM26; PHI8999A; pSYN12274; pDPG434
EN
  • Cross breeding
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
  • BCH-GENE-SCBD-103881-2 Metallothionein-like gene promoter | Zea mays (Maize, Corn, MAIZE)
    Promoter
  • BCH-GENE-SCBD-43634-3 mCry3A | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Coleoptera (beetles), Western corn rootworm (Diabrotica virgifera))
  • BCH-GENE-SCBD-100269-8 Nopaline Synthase Gene Terminator | Agrobacterium tumefaciens (Agrobacterium)
    Terminator
  • BCH-GENE-SCBD-100362-7 Ubiquitin gene promoter | Zea mays (Maize, Corn, MAIZE)
    Promoter
  • BCH-GENE-SCBD-103627-5 Ubiquitin Intron 1 | Zea mays (Maize, Corn, MAIZE)
    Intron
  • BCH-GENE-SCBD-15003-7 Phosphomannose Isomerase gene | Escherichia coli (ECOLX)
    Protein coding sequence | Mannose tolerance,Selectable marker genes and reporter genes
  • BCH-GENE-SCBD-14987-8 Cry1F | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-GENE-SCBD-100363-5 ORF25 PolyA Terminator sequence | Agrobacterium tumefaciens (Agrobacterium)
    Terminator
  • BCH-GENE-SCBD-100287-7 CaMV 35S promoter | Cauliflower mosaic virus (CaMV)
    Promoter
  • BCH-GENE-SCBD-15002-4 Phosphinothricin N-acetyltransferase gene | Streptomyces viridochromogenes (STRVR)
    Protein coding sequence | Resistance to herbicides (Glufosinate)
  • BCH-GENE-SCBD-100290-6 CaMV 35S terminator | Cauliflower mosaic virus (CaMV)
    Terminator
  • BCH-GENE-SCBD-104788-2 Cestrum Yellow Leaf Curling Virus promoter | Cestrum yellow leaf curling virus (CYLCV)
    Promoter
  • BCH-GENE-SCBD-104789-2 eCry3.1Ab | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Coleoptera (beetles), Western corn rootworm (Diabrotica virgifera), Northern corn rootworm (Diabrotica barberi))
  • BCH-GENE-SCBD-100364-5 Rice actin 1 gene promoter | Oryza sativa (Rice, ORYSA)
    Promoter
  • BCH-GENE-SCBD-101419-4 Optimized Transit Peptide
    Transit signal
  • BCH-GENE-SCBD-46333-8 5-enolpyruvylshikimate-3-phosphate synthase | Zea mays (Maize, Corn, MAIZE)
    Protein coding sequence | Resistance to herbicides (Glyphosate)
DNA insert from MIR604 vector pZM26
The parental plant contains two expression cassettes: (i) modified Cry3a (mcry3a) originally from Bacillus thuringiensis and (ii) phosphomannose isomerase (pmi) from Escherichia coli.

Expression mcry3a is under control of a Zea mays metallothionein-like gene promoter and an Agrobacterium tumefaciens nopaline synthase (nos) terminator. Transcription of pmi is under the control of Z. mays ubiquitin gene promoter and an A. tumefaciens nos terminator. The transcript initially also contains an intron from Z. mays ubiquitin-1 to enhance gene expression.

Note:
- mcry3a was originally obtained from the native cry3A gene, but was modified to enhance gene expression in maize. The synthetic version of the protein (mCry3a) contains the same amino acid sequences of the native version, except for the modified serine-protease recognition site.
- The following changes in the pmi occurred: the valine at position 61 has been substituted by alanine (V61A) and glutamine at position 210 has been substituted by histidine (Q210H). Please note no apparent change of function occurred.
- Southern blot and qPCR analysis indicated that a single insertion of both expression cassettes occurred and there was no integration of the vector backbone.

DNA insert from TC1507 vector PHI8999A
DNA fragment PHI8999A contains two adjacent plant gene expression cassettes for Bacillus thuringiensis cry1F and Streptomyces viridochromogenes pat.

Transcription of cry1F is directed by the promoter and first exon and intron of the maize (Zea mays) ubiquitin gene and terminates at the Agrobacterium tumefaciens ORF25 terminator.

Transcription of the pat gene commences from the Cauliflower Mosaic Virus (CaMV) 35S promoter and ends at the CaMV 35S terminator.

Note:
- The coding sequence of both genes has been optimized to achieve a high level of expression in maize.
- The sequences of the complete cry1F and pat are identical to those in the original plasmid.
- The Cry1F protein includes the F604K (phenylalanine to lysine at position 604) amino acid substitution, which was introduced to create a specific restriction site for cloning purposes.

DNA insert from 5307 vector pSYN12274
The DNA insert contains two gene cassettes for an engineered chimeric protein eCry3.1Ab and an Escherichia coli phosphomannose isomerase (pmi).

Transcription of ecry3.1Ab is under control of a Cestrum Yellow Leaf Curling Virus promoter and an Agrobacterium tumefaciens nopaline synthase (nos) terminator. Transcription of pmi is under control of a Zea mays ubiquitin gene promoter and a nos terminator. The promoter contains the first intron of the ubiquitin gene, which will be initially included in the mRNA before splicing and for enhancing expression of pmi. Transcription is expected to be constitutive under both promoters and result in elevated levels of transgene expression.

Note:
- eCry3.1Ab is a result of a fusion of the 5′ end (Domain I, Domain II and 15 amino acids of Domain III) of a modified Cry3A gene (mcry3A) and the 3′ end (Domain III and Variable Region 6) of a synthetic Cry1Ab gene. The sequences were sourced from Bacillus thuringiesis.
- Southern blot analysis indicated that the parental line contains a single insertion of the vector and there was no integration of the vector backbone.
- Sequencing analysis indicated that the right and left T-DNA borders were truncated.

DNA insert from GA21 vector pDPG434
Transcription of Zea mays modified 5-enolpyruvylshikimate-3-phosphate synthase (mepsps) commences from the Oryza sativa (rice) actin 1 promoter and terminates at the Agrobacterium tumefaciens nopaline synthase terminator. The transcribed elements (from 5’ to 3’) are expected to be as follows: first intron of rice actin 1, a synthetic transit peptide and mepsps. Transcription of mepsps is expected to occur constitutively due to the rice actin promoter. Gene expression is additionally enhanced by the rice actin intron. Post-translation, the optimized transit peptide targets mEPSPS to the chloroplasts.

Note:
- The coding sequence of mepsps was obtained through site-directed mutagenesis to create a modified version of the native enzyme to confer glyphosate tolerance with similar enzymatic function.
- The Rice Actin 1 promoter contains a portion of the first intron of the Actin 1 and thus corresponds to the 5’ end of the gene.
- The optimized transit peptide was derived from maize and sunflower (Helianthus sp.) ribulose 1,5 –bisphosphate carboxylase oxygenase sequences.
- Southern blot analysis indicated that an insert containing three complete tandem copies of the insert and one incomplete copy were inserted into the parental genome. The incomplete copy contains rice actin promoter, the optimized transit peptide and a truncated mepsps sequence without the nos 3’ untranslated region (as uncovered by sequence analysis).
- Sequencing analysis indicated a truncated rice actin promoter in the 5’ end of the insertion event, only containing 148 bp of the promoter region.
- The modified maize expresses only the full-length mEPSPS protein.
EN
LMO characteristics
EN
  • Food
  • Feed
Detection method(s)
EN
Additional Information
EN