MON-89Ø34-3 × SYN-IR162-4 - Insect resistant maize | BCH-LMO-SCBD-115664 | Living Modified Organism | Biosafety Clearing-House

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Living Modified Organism (LMO)

Decisions on the LMO Risk Assessments  
last updated: 31 Jul 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.
Insect resistant maize
EN
MON89034 x MIR162
Yes
MON-89Ø34-3 × SYN-IR162-4
The modified maize was produced through the cross breeding of two modified parental lines for Lepidoptera resistance through the expression of Bacillus thuringiensis Cry1A.105, Cry2Ab2 and Vegetative insecticidal protein 3Aa20. The modified maize also contains a selectable, Escherichia coli phosphomannose isomerase, for mannose selection during parental transformation.
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))
EN
Characteristics of the modification process
PV-ZMIR245; pNOV1300
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-101415-9 Ti plasmid left border repeat | Agrobacterium tumefaciens (Agrobacterium)
    Plasmid vector
  • 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-101416-6 Ti plasmid right border repeat | Agrobacterium tumefaciens (Agrobacterium)
    Plasmid vector
  • 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
DNA insert from MON89034 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) 35S 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 MON 89034 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 vector pNOV1300
In the parental MIR162 maize, a variant of the native B. thuringiensis vegetative insecticidal protein 3Aa (vip3Aa), termed vip3Aa20, was inserted into the transformation cassette. Transcription of vip3Aa20 commences at the Z. mays ubiquitin gene promoter and then transcribes vip3Aa20 followed by intron 9 of Z. mays phosphoenolpyruvate carboxylase, before terminating at the CaMV 35S terminator. The intron enhances expression of the transgene.

A second expression cassette, containing E. coli phosphomannose isomerase (pmi), was also inserted into the parental genome. The gene is under the control of another ubiquitin promoter and transcription terminates at the Agrobacterium tumefaciens nopaline synthase (nos) terminator.

Note:
- Southern blot analyses demonstrated that the T-DNA insert contains: (i) single copies of vip3Aa20 and 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.
- vip3Aa20 is a variant of the native vip3Aa, which has codon changes that result in M129I (methionine to isoleucine at position 129) and K284Q (lysine to glutamine at position 284) amino acid substitutions.
EN
LMO characteristics
EN
  • Food
  • Feed
Additional Information
EN
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