MON-89Ø34-3 × SYN-IR162-4 × MON-ØØ6Ø3-6 × DAS-4Ø278-9 - Herbicide-tolerant, insect-resistant maize | BCH-LMO-SCBD-265847 | 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: 16 Nov 2023
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
MON89034 × MIR162 × NK603 × DAS40278
Yes
MON-89Ø34-3 × SYN-IR162-4 × MON-ØØ6Ø3-6 × DAS-4Ø278-9
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 Cry1A.105, Cry2Ab2 and VIP3Aa20 proteins, which have a pore-forming mode of action that selectively damages the midgut epithelium line of feeding larvae. For tolerance to glyphosate, the maize expresses Agrobacterium tumefaciens 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. For tolerance to aryloxyphenoxypropionate and 2,4-dichlorophenoxyacetic acid herbicides (acetyl‐CoA carboxylase inhibitors), the modified maize expresses Sphingobium herbicidovorans aryloxyalkanoate dioxygenase, which cleaves 2,4-dichlorophenoxyacetic acid into non-herbicidal dichlorophenol and glyoxylate, as well as inactivates aryloxyphenoxypropionate. In addition, the modified maize also contains a gene cassette for Escherichia coli phosphomannose isomerase, which was used as a selectable marker during transformation by allowing for the transformed maize plants to use mannose as a carbon source.
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-43773-18 Living Modified Organism MON-89Ø34-3 - YieldGard™ VT Pro™
    Monsanto Company | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-LMO-SCBD-100885-13 Living Modified Organism SYN-IR162-4 - Agrisure™ Viptera maize
    Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-LMO-SCBD-14776-17 Living Modified Organism MON-ØØ6Ø3-6 - Roundup Ready™ maize
    Monsanto | Resistance to herbicides (Glyphosate)
  • BCH-LMO-SCBD-104814-1 Living Modified Organism DAS-4Ø278-9 - Enlist™ Maize
    Dow AgroSciences GmbH | Resistance to herbicides, Tolerance to 2,4-Dichlorophenoxyacetic acid, Tolerance to aryloxyphenoxypropionate
EN
Characteristics of the modification process
PV-ZMIR245; pNOV1300; PV-ZMGT32; pDAS1740
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-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-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-100887-5 Vegetative insecticidal protein 3Aa20 | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-GENE-SCBD-101406-4 Phosphoenolpyruvate carboxylase, intron 9 | Zea mays (Maize, Corn, MAIZE)
    Intron
  • BCH-GENE-SCBD-100290-6 CaMV 35S terminator | Cauliflower mosaic virus (CaMV)
    Terminator
  • 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-100364-5 Rice actin 1 gene promoter | Oryza sativa (Rice, ORYSA)
    Promoter
  • BCH-GENE-SCBD-100365-6 Chloroplast transit peptide 2 | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)
    Transit signal
  • BCH-GENE-SCBD-14979-7 5-enolpyruvylshikimate-3-phosphate synthase gene | Agrobacterium tumefaciens (Agrobacterium)
    Protein coding sequence | Resistance to herbicides (Glyphosate)
  • BCH-GENE-SCBD-104795-4 RB7 matrix attachment region | Nicotiana tabacum (Tobacco, TOBAC )
    Enhancer
  • BCH-GENE-SCBD-104812-3 Aryloxyalkanoate dioxygenase gene | Sphingobium herbicidovorans (SPHHE)
    Protein coding sequence | Resistance to herbicides,Tolerance to 2,4-Dichlorophenoxyacetic acid,Tolerance to aryloxyphenoxypropionate
  • BCH-GENE-SCBD-104813-4 Per5 3' Untranslated Region | Zea mays (Maize, Corn, MAIZE)
    Terminator
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 MIR162 (SYN-IR162-4) vector pNOV1300
The parental genome contains two gene cassettes: a variant of the native Bacillus thuringiensis vegetative insecticidal protein 3Aa (vip3Aa20) and Escherichia coli phosphomannose isomerase.
 
 Transcription of vip3Aa20 commences at the Zea mays ubiquitin promoter and then transcribes vip3Aa20 followed by intron 9 of Z. mays phosphoenolpyruvate carboxylase, before terminating at the Cauliflower mosaic virus 35S terminator. Higher levels of expression are expected due to the promoter and the enhancement by the intron.

The pmi coding sequence is under the control of Z. mays ubiquitin promoter and transcription terminates at the Agrobacterium tumefaciens nopaline synthase gene (nos) terminator.

Note:
  • Southern blot analyses demonstrated that the T-DNA insert contains: (i) single copies of a vip3Aa20 gene and a pmi gene; (ii) two copies of the maize ubiquitin promoter; (iii) one copy of the nos terminator; and (iv) no backbone sequences from transformation plasmid pNOV1300.
  • In the parental MIR162 maize, a variant of the native B. thuringiensis vegetative insecticidal protein 3Aa (vip3Aa20), named vip3Aa19, which has codon changes that result in a single M129I amino acid substitution was inserted into the transformation cassette. During the transformation process an additional DNA mutation resulted in a K284Q amino acid substitution.
 
DNA insert from NK603 (MON-ØØ6Ø3-6) vector PV-ZMGT32
The plant expression plasmid vector, PV-ZMGT32 contains two adjacent plant gene expression cassettes each containing a single copy of the Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps). In the first (5' end) expression cassette, the cp4-epsps gene is under the transcriptional regulation of an Oryza sativa actin promoter and an A. tumefaciens nopaline synthase gene (nos) terminator. An O. sativa actin intron is also present in the transcript for enhanced expression of the coding sequence. The second cassette consists of another cp4-epsps gene regulated by a Cauliflower mosaic virus enhanced 35S promoter (containing a duplicated enhancer region) and a nos terminator. Similarly, an intron from the Zea mays heat shock protein 70 (hsp70) was included for enhancing expression of the coding sequence. Both promoters of the gene cassettes are expected to promoter high levels of transcription.

Note:
- The parental NK603 line contained a single, intact insertion containing both cp4-epsps gene cassettes.
- Due to a restriction digest prior to particle bombardment, the vector backbone, containing Escherichia coli neomycin phosphotransferase II and origin of replication, were not incorporated into the parental genome.

DNA insert from DAS40278 (DAS-4Ø278-9) vector pDAS1740
The parental LMO was generated using the Whiskers-mediated transformation method. Sphingobium herbicidovorans aryloxyalkanoate dioxygenase-1 (aad-1) is under the control of Zea mays ubiquitin gene promoter and Z. mays root preferential cationic peroxidase terminator. Elevated levels of transcription are expected to occur due to the constitutive nature of the ubiquitin promoter.

Note:
  • The aad-1 coding sequence was optimized for expression in the plant.
  • Southern blot analysis indicated that a single complete copy of the transformation cassette was stably integrated into the host genome at a single locus.
  • No integration of the vector backbone occurred.
 
For more information, kindly refer to the parental LMO records.
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LMO characteristics
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  • Feed
  • Food
Additional Information
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