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Modified Organism
MON-89Ø34-3 × DAS-59122-7 × DAS-4Ø278-9 - Herbicide tolerance, insect resistant maize
Record information and status
Record ID
115525
Status
Published
Date of creation
2020-04-16 20:16 UTC (austein.mcloughlin@cbd.int)
Date of publication
2020-04-16 20:16 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 tolerance, insect resistant maize
Transformation event
MON89034 x 59122 x DAS40278
Unique identifier
MON-89Ø34-3 × DAS-59122-7 × DAS-4Ø278-9
Developer(s)
Description
The modified maize was produced by crossing modified parental lines to result in a line with herbicide tolerance and insect resistance. For Lepidoptera resistance, the maize expresses Bacillus thuringiensis Cry1A.105 and Cry2Ab2. For Coleoptera resistance, the maize expresses B. thuringiensis Cry34Ab1 and Cry35Ab1. In addition to the insecticidal proteins, the maize also expresses Streptomyces viridochromogenes phosphinothricin N-acetyltransferase for glufosinate tolerance and Sphingobium herbicidovorans aryloxyalkanoate dioxygenase for tolerance to 2,4-dichlorophenoxyacetic acid and aryloxyphenoxypropionate acetyl coenzyme A carboxylase inhibitors.
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
MON-89Ø34-3 - YieldGard™ VT Pro™
Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths)
Show detection method(s)
DAS-59122-7 - Herculex™ RW Rootworm Protection maize
Resistance to diseases and pests - Insects - Coleoptera (beetles) Resistance to herbicides - Glufosinate
Show detection method(s)
DAS-4Ø278-9 - Enlist™ Maize
Dow AgroSciences GmbH Resistance to herbicides Tolerance to 2,4-Dichlorophenoxyacetic acid Tolerance to aryloxyphenoxypropionate
Show detection method(s)
Characteristics of the transformation process
Vector
PV-ZMIR245; PHP17662; pDAS1740
Techniques used for the modification
  • Cross breeding
Genetic elements construct
 
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
 
 
Cry1A.105
3.53 Kb
 
 
Heat shock protein 17.3 terminator
0.21 Kb
 
 
FMV 35S promoter
0.56 Kb
 
 
Hsp70 intron
0.80 Kb
 
 
Transit peptide and first intron of Rubisco SSU
0.40 Kb
 
 
Cry2Ab2
1.91 Kb
 
 
Nopaline Synthase Gene Terminator
0.25 Kb
 
 
Ti plasmid right border repeat
0.23 Kb
 
 
Ubiquitin gene promoter
1.99 Kb
 
 
Cry34Ab1
0.37 Kb
 
 
Proteinase inhibitor II gene terminator
0.32 Kb
 
 
Peroxidase gene promoter
1.30 Kb
 
 
Cry35Ab1
1.15 Kb
 
 
Proteinase inhibitor II gene terminator
0.32 Kb
 
 
CaMV 35S promoter
0.55 Kb
 
 
Phosphinothricin N-acetyltransferase gene
0.55 Kb
 
 
CaMV 35S terminator
0.20 Kb
 
 
RB7 matrix attachment region
1.17 Kb
 
 
Ubiquitin gene promoter
1.99 Kb
 
 
Aryloxyalkanoate dioxygenase gene
0.89 Kb
 
 
Per5 3' Untranslated Region
0.37 Kb
 
 
RB7 matrix attachment region
1.17 Kb
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
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.

Note:
- 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 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.

DNA insert from DAS40278 vector pDAS1740:
The LMO was generated using the Whiskers-mediated transformation method. Sphingobium herbicidovorans aryloxyalkanoate dioxygenase-1 (aad-1)  is under the control of Zea mays ubiquitin gene promoter and Z. mays root preferential cationic peroxidase terminator. The aad-1 coding sequence was optimized for expression in the plant.

Note:
- Southern blot analysis indicated that a single complete copy of the transformation cassette was stably integrated into the host genome at a single locus
- No integration of the vector backbone occurred.
LMO characteristics
Modified traits
  • Tolerance to 2,4-Dichlorophenoxyacetic acid
  • Tolerance to aryloxyphenoxypropionate
Additional Information
Other relevant website address or attached documents