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Modified Organism
SYN-IR162-4 × MON-89Ø34-3 × MON-ØØØ21-9 - Herbicide tolerant, insect resistant maize
Record information and status
Record ID
Date of creation
2020-06-05 20:56 UTC (austein.mcloughlin@cbd.int)
Date of publication
2020-06-05 20:56 UTC (austein.mcloughlin@cbd.int)

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.

LMO name
Herbicide tolerant, insect resistant maize
Transformation event
MIR162 × MON89034 × GA21
Unique identifier
SYN-IR162-4 × MON-89Ø34-3 × MON-ØØØ21-9
800 North Lindbergh Blvd.
St. Louis, MO
United States of America, 63167
Phone:+ 1 314 694-1000
Fax:+1 314 694-3080
The modified maize event was a result of cross-breeding modified parental lines and demonstrates herbicide tolerance and insect resistance. For Lepidoptera resistance, the maize expresses Bacillus thuringiensis Vegetative insecticidal protein 3Aa20, Cry1A.105 and Cry2Ab2. In addition to the insecticidal proteins, the maize expresses a modified native Zea mays 5-enolpyruvylshikimate-3-phosphate synthase for tolerance to glyphosate. A selectable marker, Escherichia coli phosphomannose isomerase, is also present and was used during the transformation of the parental line using mannose selection.
Recipient Organism or Parental Organisms
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.
Zea mays - Maize, Corn, MAIZE
SYN-IR162-4 - Agrisure™ Viptera maize
Mannose tolerance Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths) Selectable marker genes and reporter genes
Show detection method(s)
MON-89Ø34-3 - YieldGard™ VT Pro™
Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths)
Show detection method(s)
MON-ØØØ21-9 - Roundup Ready™ maize
Resistance to herbicides - Glyphosate
Show detection method(s)
Characteristics of the transformation process
pNOV1300; PV-ZMIR245; pDPG434
Techniques used for the modification
  • Cross breeding
Genetic elements construct
Ubiquitin gene promoter
1.99 Kb
Vegetative insecticidal protein 3Aa20
2.37 Kb
Phosphoenolpyruvate carboxylase, intron 9
0.11 Kb
CaMV 35S terminator
0.07 Kb
Ubiquitin gene promoter
1.99 Kb
Phosphomannose Isomerase gene
1.18 Kb
Nopaline Synthase Gene Terminator
0.25 Kb
Ti plasmid left border repeat
0.24 Kb
CaMV Enhanced 35S promoter
0.30 Kb
5' untranslated leader from chlorophyll a/b-binding protein
0.06 Kb
Rice actin 1, intron
0.48 Kb
3.53 Kb
Heat shock protein 17.3 terminator
0.21 Kb
FMV 34S promoter
0.56 Kb
Hsp70 intron
0.80 Kb
Transit peptide and first intron of Rubisco SSU
0.40 Kb
1.91 Kb
Nopaline Synthase Gene Terminator
0.25 Kb
Ti plasmid right border repeat
0.23 Kb
Rice actin 1 gene promoter
1.37 Kb
Optimized Transit Peptide
0.37 Kb
5-enolpyruvylshikimate-3-phosphate synthase
1.34 Kb
Nopaline Synthase Gene Terminator
0.24 Kb
Further details
Notes regarding the genetic elements introduced or modified in this LMO
DNA insert from MIR162 vector pNOV1300
In the parental MIR162 maize, a variant of the native B. thuringiensis vegetative insecticidal protein 3Aa (vip3Aa), termed vip3Aa20, was inserted into the transformation cassette. Transcription of vip3Aa20 commences at the Z. mays ubiquitin gene promoter and then transcribes vip3Aa20 followed by intron 9 of Z. mays phosphoenolpyruvate carboxylase, before terminating at the CaMV 35S terminator. The intron enhances expression of the transgene.

A second expression cassette, containing E. coli phosphomannose isomerase (pmi), was also inserted into the parental genome. The gene is under the control of another ubiquitin promoter and transcription terminates at the Agrobacterium tumefaciens nopaline synthase (nos) terminator.

- Southern blot analyses demonstrated that the T-DNA insert contains: (i) single copies of vip3Aa20 and pmi gene; (ii) two copies of the maize ubiquitin promoter; (iii) one copy of the nos terminator; and iv) no backbone sequences from transformation plasmid pNOV1300.
- Vip3Aa20 is a variant of the native Vip3Aa, which has codon changes that result in M129I (methionine to isoleucine at position 129) and K284Q (lysine to glutamine at position 284) amino acid substitutions.

DNA insert from MON89034 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) 35S 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.

- 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 MON 89034 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 GA21 vector pDPG434
Transcription of Zea mays modified 5-enolpyruvylshikimate-3-phosphate synthase (mepsps) commences from the Oryza sativa (rice) actin 1 promoter and terminates at the Agrobacterium tumefaciens nopaline synthase terminator. The transcribed elements (from 5' to 3') are expected to be as follows: first intron of rice actin 1, a synthetic transit peptide and mepsps. Transcription of mepsps is expected to occur constitutively due to the rice actin promoter. Gene expression is additionally enhanced by the rice actin intron. Post-translation, the optimized transit peptide targets mEPSPS to the chloroplasts.

- The coding sequence of mepsps was obtained through site-directed mutagenesis to create a modified version of the native enzyme to confer glyphosate tolerance with similar enzymatic function.
- The Rice Actin 1 promoter contains a portion of the first intron of the Actin 1 and thus corresponds to the 5' end of the gene.
- The optimized transit peptide was derived from maize and sunflower (Helianthus sp.) ribulose 1,5 -bisphosphate carboxylase oxygenase sequences.
- Southern blot analysis indicated that an insert containing three complete tandem copies of the insert and one incomplete copy were inserted into the parental genome. The incomplete copy contains rice actin promoter, the optimized transit peptide and a truncated mepsps sequence without the nos 3' untranslated region (as uncovered by sequence analysis).
- Sequencing analysis indicated a truncated rice actin promoter in the 5' end of the insertion event, only containing 148 bp of the promoter region.
- The modified maize expresses only the full-length mEPSPS protein.
LMO characteristics
Modified traits
  • Selectable marker genes and reporter genes
  • Tolerance to mannose
Common use(s)
  • Food
  • Feed
Additional Information
Other relevant website address or attached documents