MON-87427-7 × MON-95379-3 × MON-87411-9 × MON-87419-8 - Insect-protected, herbicide-tolerant maize | BCH-LMO-SCBD-260556 | Living Modified Organism | Biosafety Clearing-House

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Living Modified Organism (LMO)

Decisions on the LMO Risk Assessments  
last updated: 07 Jun 2022
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-protected, herbicide-tolerant maize
EN
MON87427 × MON95379 × MON87411 × MON87419
MON-87427-7 × MON-95379-3 × MON-87411-9 × MON-87419-8
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 synthetic Cry1B.868 and Cry1Da_7 proteins, which have pore-forming mode of action that is independent of the receptors that other Bt toxins interact with. For protection from Coleoptera insects, the maize expresses Bacillus thuringiensis Cry3Bb1, which has a pore-forming mode of action. 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. For tolerance to glufosinate, the maize expresses Streptomyces viridochromogenes phosphinothricin N-acetyltransferase, which inactivates phosphinothricin through acetylation. 
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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-104758-3 Living Modified Organism MON-87427-7 - Maize modified for tissue selective glyphosate tolerance
    Resistance to herbicides - Glyphosate
  • BCH-LMO-SCBD-258895-1 Living Modified Organism MON-95379-3 - Insect-protected maize
    Bayer CropScience Deutschland GmbH | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths), Cotton bollworm (Helicoverpa spp.), European corn borer (Ostrinia nubilalis), Fall armyworm (Spodoptera frugiperda))
  • 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-111531-4 Living Modified Organism MON-87419-8 - Dicamba and Glufosinate Tolerant Maize
    Resistance to herbicides - Glufosinate
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Characteristics of the modification process
PV-ZMAP1043; PV-ZMIR522223; PV-ZMIR10871; PV-ZMHT507801
EN
  • Cross breeding
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
DNA insert from MON87427 PV-ZMAP1043
The MON87427 genome contains one gene cassette Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps). Transcription of cp4-epsps commences from the Cauliflower mosaic virus (CaMV) enhanced 35S promoter and ends at the A. tumefaciens nopaline synthase (nos) gene terminator. The transcript contains a Zea mays heat shock protein 70 (hsp70) intron, Arabidopsis thaliana N-terminal chloroplast transit peptide sequence, and cp4-epsps.  The CaMV enhanced 35S promoter-hsp70 combination promotes gene expression in female and vegetative tissues, but not in male reproductive tissues (pollen microspores and tapetum).

Note:
  • Southern blot analyses indicate that a single copy of the T-DNA was inserted at a single site in the parental maize genome and no plasmid vector backbone sequences were detected to have been integrated. DNA sequencing analyses further indicated that the expected T-DNA sequences were integrated.
  • The cp4-epsps coding sequence is the codon optimized coding sequence of the aroA gene from Agrobacterium sp. strain CP4 encoding CP4 EPSPS.
  • The expression of cp4-epsps from the MON87411 parental genome is expected to overcome the tissue specific expression from the MON87427 genome.


DNA insert from MON95379 vector PV-ZMIR522223
The MON95379 genome contains two gene cassettes: synthetic cry1B.868 and synthetic cry1Da_7.

The cry1B.868 coding sequence is under control of a Zea mays ubiquitin promoter and an Oryza sativa lipid transfer-like protein terminator. The promoter contains the promoter, leader and intron sequences from the maize ubiquitin gene. High levels of transcription are expected in all tissues due to the constitutive nature of the promoter.

The cry1Da_7 coding sequence is under control of a Setaria italica promoter and an O. sativa gos2 terminator. The first intron of the rice actin 15 gene was also included and likely improves expression of the gene. 

Note:
  • Both cry1B.868 and cry1Da_7 sequences are derived from Bacillus thuringiensis sequences. Refer to the genetic element records for more information.
  • During development of the modified maize, a c4-epsps cassette (rice tubulin A terminator; cp4-epsps; Arabidopsis thaliana chloroplast transit peptide 2; and rice tubulin A promoter) was removed using Cre-lox excision. The T-DNA right border was also truncated (lost) during transformation.
  • Next-generation sequencing indicated that a single, intact copy of the the intended DNA insertion was present in the parental genome. No backbone or other unexpected sequences were detected.


DNA insert from MON87411 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 MON87419 vector PV-ZMHT507801
The MON87419 genome contains two gene cassettes:  Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (pat) and Stenotrophomonas maltophilia dicamba monooxygenase (dmo).

The pat coding sequence is under the control of the Andropogon gerardii ubiquitin  promoter and the Oryza sativa alpha-amylase/trypsin inhibitor terminator. The transcript includes the A. gerardii 5' untranslated leader sequence and an intron from ubiquitin before (5') coding sequence of pat.

The dmo coding sequence is under control of the Peanut chlorotic streak caulimovirus (PC1SV) full-length transcript promoter and the Triticum aestivum (wheat) heat shock protein 17 terminator. The transcript produced contains a wheat chlorophyll a/b-binding 5' untranslated leader sequence (for improved gene expression), an O. sativa actin 1 untranslated region and intron (for improved gene expression), the untranslated  and targeting region of Petunia hybrida chloroplast transit peptide 4 (for chloroplast targeting of the protein) and dmo.

Note:
  • Originally, the plasmid vector contained two T-DNA elements that were inserted during the initial transformation event: one containing the dmo and pat expression cassettes, and a second containing an Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps) expression cassette. The cp4-epsps expression cassette is regulated by the O sativa actin 1 promoter and 5′ untranslated leader, O. sativa intron, the Arabidopsis thaliana chloroplast targeting peptide 2 targeting sequence, and the A. tumefaciens nopaline synthase  3′ untranslated region. Subsequent traditional breeding, segregation, selection, and screening were used to isolate those plants that contain the dmo and pat expression cassettes (T-DNA I) and do not contain the cp4-epsps expression cassette (T-DNA II).
  • Molecular characterization of MON87419 indicated that a single copy of T-DNA I was integrated into the maize genome at a single intact locus that includes all expected elements within the insert, with the exception of incomplete Right and Left Border sequences. These analyses also showed no PV-ZMHT507801 backbone elements or T-DNA II sequences were present in the event.


For more information, kindly refer to the parental LMO records.
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LMO characteristics
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