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Living Modified Organism (LMO)
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BCH-LMO-SCBD-269704-1   |   PDF   |   Print   |  
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Decisions on the LMO Risk Assessments  
last updated: 30 May 2024
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 × MON89034 × 5307 × GA21
Yes
SYN-IR6Ø4-5 × MON-89Ø34-3 × SYN-Ø53Ø7-1 × MON-ØØØ21-9
The maize (Zea mays) was produced through crossing modified parental lines for resistance to insect pests and tolerance to herbicides. For protection from Lepidoptera insects, the maize expresses Bacillus thuringiensis Cry1A.105 and Cry2Ab2 proteins, which have a pore-forming mode of action that selectively damages the midgut epithelium line of feeding larvae. For resistance to Coleoptera pests, the modified maize expresses B. thuringiensis eCry3.1Ab and mCry3A, which also have a pore forming mode of action. For tolerance to glyphosate, the maize expresses a modified maize 5-enolpyruvylshikimate-3-phosphate, which has a low binding affinity for the herbicidal compound and allows for the continued synthesis of aromatic amino acids through the shikimate pathway. In addition, the modified maize also contains gene cassettes for Escherichia coli phosphomannose isomerase, which was used as a selectable marker during transformation by allowing for the transformed maize plants to tolerate the presence of mannose.
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-43773-18 Living Modified Organism MON-89Ø34-3 - YieldGard™ VT Pro™
    Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • 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; PV-ZMIR245; pSYN12274; pDPG434
EN
  • Cross breeding
 
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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-100366-6 CaMV Enhanced 35S promoter | Cauliflower mosaic virus (CaMV)
    Promoter
  • BCH-GENE-SCBD-100354-6 5' untranslated leader from chlorophyll a/b-binding protein | Triticum aestivum (Wheat)
    Leader sequence
  • BCH-GENE-SCBD-100355-6 Rice actin 1, intron | Oryza sativa (Rice, ORYSA)
    Intron
  • BCH-GENE-SCBD-43771-9 Cry1A.105 | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-GENE-SCBD-100356-6 Heat shock protein 17.3 terminator | Triticum aestivum (Wheat)
    Terminator
  • BCH-GENE-SCBD-101507-5 FMV 34S promoter | Figwort mosaic virus (Figwort mottle virus, FMV, CMoVb)
    Promoter
  • BCH-GENE-SCBD-100359-7 Hsp70 intron | Zea mays (Maize, Corn, MAIZE)
    Intron
  • BCH-GENE-SCBD-100360-4 Transit peptide and first intron of Rubisco SSU | Zea mays (Maize, Corn, MAIZE)
    Transit signal
  • BCH-GENE-SCBD-14988-7 Cry2Ab2 | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • 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-101419-5 Optimized chloroplast transit peptide | Zea mays (Maize, Corn, MAIZE) | Helianthus annuus (Sunflower, HELAN)
    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 (SYN-IR6Ø4-5) 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 MON89034 (MON-89Ø34-3) vector PV-ZMIR245:
Maize line MON89034 expresses two Bt-toxins encoded by Bacillus thuringiensis cry1A.105  and cry2Ab2.

Transcription of cry1A.105 begins at the Cauliflower mosaic virus (CaMV) Enhanced 35S promoter and finishes at the wheat (Triticum aestivum) wheat heat shock protein 17.3 terminator. The transcript initially includes (5' to 3'): wheat 5' untranslated leader from the chlorophyll a/b-binding protein, Oryza sativa (rice) actin 1 intron and cry1A.105. The wheat 5' untranslated leader sequence and the rice intron enhance the expression of cry1A.105.

Transcription of cry2Ab2 commences from the Figwort mosaic virus (FMV) 34S promoter and terminates at the Agrobacterium tumefaciens nopaline synthase (nos) terminator. The transcript initially includes (5' to 3'): maize heat shock protein 70 (hsp70) intron, maize transit peptide and first intron from the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase and cry2Ab32. The hsp70 regulates and enhances gene expression, while the transit peptide targets Cry2Ab2 to the chloroplast.
 
Note:
  • The viral promoters are expected to be constitutively active and promote high levels of transcription.
  • The coding sequence of cry2Ab2 was codon-optimized for expression within plant systems.
  • A second T-DNA insertion (containing CaMV 35S promoter, Escherichia coli neomycin phosphotransferase and A. tumefaciens nos  terminator) was initially inserted into the genome for kanamycin selection during transformation. However, once transformants were regenerated, the selectable marker was bred out of the parental line using convention breeding techniques.
  •  Southern blot analyses indicated a single copy of the cry1A.105 and the cry2Ab2 cassettes. No backbone plasmid DNA or nptII sequences were detected. PCR and DNA sequence analyses provided the complete DNA sequence of the insert and confirmed the organization of the elements within the insert. Furthermore, sequence analysis indicated that MON89034 no longer has the duplicated enhancer elements compared to the original e35S promoter in PV-ZMIR245, possibly due to a recombination event that resulted in its deletion.

DNA insert from 5307 (SYN-Ø53Ø7-1) 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 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 (MON-ØØØ21-9) vector pDPG434
The DNA insert from the GA21 genome contains one gene cassette: Zea mays modified 5-enolpyruvylshikimate-3-phosphate synthase (mepsps).

Transcription of 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 annuus) 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.

For more information, kindly refer to the parental LMO records.
EN
LMO characteristics
EN
  • Feed
  • Food
Additional Information
EN
Records referencing this document Show in search
Record type Field Record(s)
Country's Decision or any other Communication LMO identification 1
Risk Assessment generated by a regulatory process Living modified organism(s) 1

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