MON-89Ø34-3 × MON-87411-9 × DAS-4Ø278-9 - Insect-resistant, herbicide-tolerant maize | BCH-LMO-SCBD-265635 | 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 Oct 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.
Insect-resistant, herbicide-tolerant maize
EN
MON89034 × MON87411 × DAS40278
Yes
MON-89Ø34-3 × MON-87411-9 × 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 Bacillus thuringiensis Cry1A.105 and Cry2Ab2 protein, which have a pore-forming mode of action that selectively damages the midgut epithelium line of feeding larvae. For protection from Coleoptera insects, the maize expresses B. thuringiensis Cry3Bb1, which also has a pore-forming mode of action in the midgut epithelium lining. In addition, the maize contains an RNA interference cassette that specifically targets Diabrotica virgifera virgifera Snf7, an essential cellular component of endosomal sorting complex required for transport and thus confers resistance to this pest. 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. Further, the maize expresses Sphingobium herbicidovorans aryloxyalkanoate dioxygenase, which cleaves 2,4-dichlorophenoxyacetic acid into non-herbicidal dichlorophenol and glyoxylate, as well as inactivates aryloxyphenoxypropionate herbicides (acetyl‐CoA carboxylase inhibitors).
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-108881-1 Living Modified Organism MON-87411-9 - Maize modified for herbicide tolerance and insect resistance
    Monsanto | Resistance to diseases and pests (Insects, Coleoptera (beetles), Western corn rootworm (Diabrotica virgifera), Northern corn rootworm (Diabrotica barberi)), 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; PV-ZMIR10871; 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-108875-2 Snf7 coding sequence | Diabrotica virgifera virgifera (Western corn rootworm, DIAVI)
    Protein coding sequence | Resistance to diseases and pests (Insects, Coleoptera (beetles), Western corn rootworm (Diabrotica virgifera))
  • BCH-GENE-SCBD-101877-5 rbcS-E9 gene terminator | Pisum sativum (Garden pea, PEA)
    Terminator
  • BCH-GENE-SCBD-108876-1 pIIG gene promoter | Zea mays (Maize, Corn, MAIZE)
    Promoter
  • BCH-GENE-SCBD-14993-5 Cry3Bb1 | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Coleoptera (beetles))
  • BCH-GENE-SCBD-108877-1 Alpha Tubulin 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-108880-1 Alpha Tubulin Gene terminator | Oryza sativa (Rice, ORYSA)
    Terminator
  • BCH-GENE-SCBD-104795-4 RB7 matrix attachment region | Nicotiana tabacum (Tobacco, TOBAC )
    Enhancer
  • BCH-GENE-SCBD-100362-7 Ubiquitin gene promoter | Zea mays (Maize, Corn, MAIZE)
    Promoter
  • 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 MON87411 (MON-87411-9) vector PV-ZMIR10871
The MON87411 genome contains three cassettes: an RNA interference (RNAi) cassette targeting Diabrotica virgifera virgifera, Bacillus thuringiensis cry3Bb1 and Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps).

Transcription of the RNAi cassette commences from the Cauliflower mosaic virus 35S enhanced promoter and terminates at the Pisum sativum ribulose bisphosphate carboxylase small chain 2 terminator. The transcript initially contains a Zea mays heat shock protein 70 intron, which contributes to enhanced expression in vegetative tissues of the plant, and two partial coding sequences of the D. virgifera virgifera Snf7p gene, which encodes the SNF7 subunit of the ESCRT-III complex. The two Snf7p sequences are in an inverted orientation, separated by a 150-nucleotide intervening sequence, which allows base pairing between the inverted sequences and hairpin RNA formation post-transcription, which then triggers an RNAi response. Due to RNAi processing, small interfering RNA molecules (roughly 21-23 nucleotides in length) will be produced and thus no translation into protein will occur from this cassette.

Transcription of the cry3Bb1 is under control of the Z. mays physical impedance induced protein promoter and Triticum aestivum (wheat) heat shock protein 17.3 terminator. The transcript also contains a wheat 5' untranslated leader from chlorophyll a/b-binding protein and Oryza sativa actin 1 intron for enhanced expression of the transgene. Expression of cp4-epsps is under control of an O. sativa alpha tubulin promoter and terminator. The transcript additionally contains Arabidopsis thaliana chloroplast targeting peptide 2 to sequester the protein to the chloroplast.
 
Note:
  • Sequencing, PCR and bioinformatic analyses indicate that a single, intact insertions of the three gene cassettes occurred in the parental line.
  • No plasmid backbone was detected.

DNA insert from DAS40278 (DAS-4Ø278-9) vector pDAS1740
The 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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