MON-87427-7 × MON-8746Ø-4 × DAS-59122-7 - Drought-tolerant, herbicide-tolerant, insect-resistant maize | BCH-LMO-SCBD-116309 | 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: 05 Oct 2021
Living Modified Organism identity
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Drought-tolerant, herbicide-tolerant, insect-resistant maize
EN
MON87427 × MON87460 × 59122
Yes
MON-87427-7 × MON-8746Ø-4 × DAS-59122-7
The maize (Zea mays) was produced through cross breeding of modified parental maize lines for drought tolerance, herbicide tolerance and insect resistance. For abiotic tolerance, the maize expresses Bacillus subtillus cold shock protein to enhance natural abiotic (drought) stress responses. For herbicide tolerance, the maize expresses Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (glyphosate tolerance - enzyme variant) and Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (glufosinate tolerance - enzymatic inactivation). The expression of cp4-epsps is restricted to female and vegetative tissues. For Coleoptera resistance, the maize expresses B. thuringiensis Cry34Ab1 and Cry35Ab1. Additionally, the maize contains an Escherichia coli neomycin phosphotransferase II cassette for kanamycin selection, which was used during transformation of a parental line.
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-104758-3 Living Modified Organism MON-87427-7 - Maize modified for tissue selective glyphosate tolerance
    Monsanto | Resistance to herbicides (Glyphosate)
  • BCH-LMO-SCBD-103066-6 Living Modified Organism MON-8746Ø-4 - Droughtgard™ Maize
    Resistance to antibiotics (Kanamycin, Neomycin), Tolerance to abiotic stress (Cold / Heat, Drought)
  • BCH-LMO-SCBD-15165-13 Living Modified Organism DAS-59122-7 - Herculex™ RW Rootworm Protection maize
    Pioneer Hi-Bred International Inc. | Resistance to diseases and pests (Insects, Coleoptera (beetles)), Resistance to herbicides (Glufosinate)
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Characteristics of the modification process
PV-ZMAP1043; PV-ZMAP595; PHP17662
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  • 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
Transcription of 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps) from Agrobacterium tumefaciens 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.

DNA insert from MON87460 vector PV-ZMAP595
The T-DNA insert contains the following gene cassettes: Bacillus subtillus cold shock protein (cspB) and Escherichia coli neomycin phosphotransferase II (nptII). 

Transcription of cspB is under control of the Oryza sativa actin 1 promoter and Agrobacterium tumefaciens transcript 7 gene 3' untranslated region. The transcript initially contains an O. sativa actin 1 intron for enhanced gene expression of cspB. The sequence is removed (spliced) prior to protein translation. Constitutive expression of cspB is expected due to the actin promoter.

Transcription of nptII is under control of the Cauliflower mosaic virus (CaMV) 35S promoter and A. tumefaciens nopaline synthase terminator. High levels of transcription are expected due to the CaMV promoter.

Note:
- The coding sequence of cspB has been codon optimized for optimal expression within plant cells.
- Southern blot analysis indicated that no vector backbone sequences were inserted into the parental genome
- Southern blot analysis indicated that the parental genome contains a single insertion
- Sequencing analyses confirm the Southern blot analyses.
- A 22 base pair deletion of genomic DNA at the insert-to-plant DNA junction occurred.
- loxP sites can be found in the parental genome and could potentially allow for the excision of the nptII cassette by CRE recombinase.

DNA insert from 59122 vector PHP17662:
Transcription of Bacillus thuringiensis cry34Ab1 starts at Zea mays ubiquitin gene promoter and terminates at the Solanum tuberosum proteinase inhibitor II gene terminator. Transcription of B. thuringiensis cry35Ab1 commences from the (Triticum aestivum (wheat) peroxidase gene promoter and stops at another S. tuberosum proteinase inhibitor II gene terminator.

Note:
- The coding sequence of cry34Ab1 and cry35Ab1  has been adapted to the codon usage in maize as to achieve optimal expression in planta.
- The cry34Ab1 and cry35Ab1 were cloned from B. thuringiensis strain PS149B1.
- Sequence analysis of 59122 done by the European Food Safety Authority indicated that this LMO contains one complete copy of the T-DNA of PHP17662 without internal rearrangements. All three gene cassettes, cry34Ab1, cry35Ab1 and pat, are intact within the transgenic event. The DNA sequences of the genes in 59122 are identical to those in the original plasmid except for two nucleotide differences in the wheat peroxidise promoter. At the 5' T-DNA end a deletion of 22 bp is observed and at the 3' T-DNA end a deletion of 25 bp is observed. The absence of vector backbone in maize 59122 was also demonstrated.

For more information, kindly refer to the parental LMO records.
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LMO characteristics
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  • Food
  • Feed
Additional Information
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Records referencing this document Show in search
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Laboratory for detection and identification of LMOs LMO(s) detectable by the laboratory 1
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