MON-87427-7 × MON-8746Ø-4 × MON-ØØ6Ø3-6 - Herbicide-tolerant, drought-tolerant maize | BCH-LMO-SCBD-116168 | Living Modified Organism | Biosafety Clearing-House


Living Modified Organism (LMO)

Decisions on the LMO Risk Assessments  
published: 05 Jul 2021 last updated: 01 Oct 2021
Living Modified Organism identity
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Herbicide-tolerant, drought-tolerant maize
MON87427 × MON87460 × NK603
MON-87427-7 × MON-8746Ø-4 × MON-ØØ6Ø3-6
The modified maize (Zea mays) was produced through the cross breeding of modified parental lines for herbicide tolerance and drought tolerance. For herbicide tolerance, the maize expresses Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase, a variant of an endogenous enzyme that confers for glyphosate tolerance. For drought tolerance, the maize expresses Bacillus subtilis cold shock protein, which binds RNA and maintains cellular functions under water-limited conditions (improvement of natural abiotic stress responses). The maize also contains an Escherichia coli neomycin phosphotransferase II cassette, which allowed for kanamycin selection during transformation of some of the parental lines.
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-104758-3 Living Modified Organism MON-87427-7 - Maize modified for tissue selective glyphosate tolerance
    Resistance to herbicides - Glyphosate
  • 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-14776-17 Living Modified Organism MON-ØØ6Ø3-6 - Roundup Ready™ maize
    Resistance to herbicides - Glyphosate
Characteristics of the modification process
  • 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).

- 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: Bacillius 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 NK603 vector PV-ZMGT32
The plant expression plasmid vector, PV-ZMGT32 contains two adjacent plant gene expression cassettes each containing a single copy of the cp4-epsps. In the first (5' end) expression cassette, the cp4-epsps gene is under the transcriptional regulation of an Oryza sativa actin promoter and a nos terminator. An O. sativa actin intron is also present in the transcript for enhanced expression of the coding sequence. The second cassette consists of another cp4-epsps gene regulated by an CaMV enhanced 35S promoter (containing a duplicated enhancer region) and a nos terminator. Similarly, an intron from the maize heat shock protein 70 (hsp70) was included for enhancing expression of the coding sequence. Both promoters of the gene cassettes are expected to promoter high levels of transcription.

- The parental NK603 line contained a single, intact insertion containing both cp4-epsps gene cassettes.
- Due to restriction digest prior to particle bombardment, the vector backbone, containing E. coli neomycin phosphotransferase II and origin of replication, were not incorporated into the parental genome.

Kindly refer to the parental LMO records for more information.
LMO characteristics
  • Food
  • Feed
Additional Information
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Country's Decision or any other Communication Living modified organism(s) 1
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