MON-8746Ø-4 × MON-89Ø34-3 × DAS-Ø15Ø7-1 - Drought-tolerant, Glufosinate-tolerant, Lepidoptera-resistant maize | BCH-LMO-SCBD-116313 | Living Modified Organism | Biosafety Clearing-House


Living Modified Organism (LMO)

Decisions on the LMO Risk Assessments  
last updated: 06 Oct 2021
Living Modified Organism identity
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Drought-tolerant, Glufosinate-tolerant, Lepidoptera-resistant maize
MON87460 × MON89034 × TC1507
MON-8746Ø-4 × MON-89Ø34-3 × DAS-Ø15Ø7-1
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 Streptomyces viridochromogenes phosphinothricin N-acetyltransferase, which inactivates glufosinate herbicides through acetylation. For Lepidoptera tolerance, the maize expresses Bacillus thuringiensis Cry1A.105, Cry1F and Cry2Ab2.  Additionally, the maize contains an Escherichia coli neomycin phosphotransferase II cassette for kanamycin selection, which was used during transformation of a parental line.
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)
  • BCH-LMO-SCBD-103066-6 Living Modified Organism MON-8746Ø-4 - Droughtgard™ Maize
    Resistance to antibiotics - Kanamycin Tolerance to abiotic stress - Cold / Heat, Drought
  • BCH-LMO-SCBD-43773-18 Living Modified Organism MON-89Ø34-3 - YieldGard™ VT Pro™
    Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths)
  • BCH-LMO-SCBD-14841-13 Living Modified Organism DAS-Ø15Ø7-1 - Herculex™ I maize
    Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths)), Resistance to herbicides (Glufosinate)
Characteristics of the modification process
PV-ZMAP595; PV-ZMIR245; PHI8999A
  • Cross breeding
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
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.

- 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 MON89034 vector PV-ZMIR245
Two insecticidal protein expression cassettes were inserted into the genome. Bacillus thuringiensis cry1A.105 expression is under the control of the CaMV 35S enhanced promoter, which first transcribes wheat (Triticum aestivum) 5' untranslated region of the chlorophyll a/b-binding protein (cab) and a rice actin 1 intron before transcribing cry1A.105. Transcription terminates at the wheat heat shock protein 17.3 terminator. Expression of the B. thuringiensis cry2Ab2 starts at the Figwort mosaic virus 34S promoter, which transcribes the Zea mays heat shock protein 70 (hsp70), then the Z. mays transit peptide and the cry2Ab2 coding sequence, before terminating at the nos terminator.

- The Cry2Ab2 coding sequence was modified for optimal expression in plants.
- South blot analysis confirmed that single insertions of both cry2Ab2 and cry1A.105, as well as no vector backbone were present and in the parent.
- A deletion removed the duplicated enhancer elements compared to the original CaMV 35S enhanced promoter in PV-ZMIR245.
- The selectable marker, nptII, cassette was bred out of the parental line and became not associated with this transformation event.

DNA insert from TC1507 vector PHI8999A
DNA fragment PHI8999A contains two adjacent plant gene expression cassettes for Bacillus thuringiensis cry1F and Streptomyces viridochromogenes pat.

Transcription of cry1F is directed by the promoter and first exon and intron of the maize (Zea mays) ubiquitin gene and terminates at the Agrobacterium tumefaciens ORF25 terminator.

Transcription of the pat gene commences from the Cauliflower mosaic virus (CaMV) 35S promoter and ends at the CaMV 35S terminator.

- The coding sequence of both genes has been optimized to achieve a high level of expression in maize.
- The sequences of the complete cry1F and pat are identical to those in the original plasmid.
- The CRY1F protein includes the F604K (phenylalanine to lysine at position 604) amino acid substitution, which was introduced to create a specific restriction site for cloning purposes.

For more information, kindly refer to the parental LMO records.
LMO characteristics
  • Food
  • Feed
Additional Information
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