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Modified Organism
MON-89Ø34-3 × MON-88Ø17-3 × DAS-4Ø278-9 - Herbicide tolerant, insect resistant maize
Record information and status
Record ID
115524
Status
Published
Date of creation
2020-04-16 19:55 UTC (austein.mcloughlin@cbd.int)
Date of publication
2020-04-16 19:55 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
MON89034 x MON88017 x DAS40278
Unique identifier
MON-89Ø34-3 × MON-88Ø17-3 × DAS-4Ø278-9
Developer(s)
Dow AgroSciences GmbH
Dow AgroSciences GmbH
Truderinger Straße 15
München, Bayern
Germany, 81677
Phone:+ 49 89 - 4 55 33 - 0
Fax:+ 49 89 - 4 55 33 - 111
Email:DowAgroSciencesD@dow.com
Url:http://www.dowagro.com/de
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 Cry 3Bb1. In addition to the insecticidal proteins, the maize also expresses Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase for tolerance to glyphosate 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)
MON-88Ø17-3 - YieldGard™ VT™ Rootworm/RR2™ Maize
Resistance to diseases and pests - Insects - Coleoptera (beetles) Resistance to herbicides - Glyphosate
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; PV-ZMIR39; 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
 
 
Rice actin 1 gene promoter
0.93 Kb
 
 
Rice actin 1, intron
0.46 Kb
 
 
Chloroplast transit peptide 2
0.23 Kb
 
 
5-enolpyruvylshikimate-3-phosphate synthase gene
0.26 Kb
 
 
Nopaline Synthase Gene Terminator
0.26 Kb
 
 
CaMV Enhanced 35S promoter
0.61 Kb
 
 
5' untranslated leader from chlorophyll a/b-binding protein
0.07 Kb
 
 
Rice actin 1, intron
0.46 Kb
 
 
Cry3Bb1
1.96 Kb
 
 
Heat shock protein 17.3 terminator
0.23 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 MON88017 vector PV-ZMIR39:
Maize line 88017 contains A. tumefaciens 5-enolpyruvylshikimate-3-phosphate (epsps) and B. thuringiensis cry3Bb1 .

Transcription of epsps starts from the Oryza sativa (rice) Actin 1 promoter and terminates at the A. tumefaciens nos  terminator. The transcript initially includes (from 5' to 3'): rice actin 1 intron for enhanced gene expression, Arabidopsis thaliana chloroplast transit peptide 2 and epsps. The A. thaliana transit peptide 2 targets the EPSPS protein to the chloroplast of the plant's cell.

Transcription of cry3Bb1 commences from the Cauliflower Mosaic Virus (CaMV) 35S enhanced promoter and terminates at the wheat heat shock protein 17.3 terminator. The transcript initially includes (from 5' to 3'): wheat 5' untranslated leader from chlorophyll a/b-binding, rice Actin 1 intron and cry3Bb1. The wheat untranslated leader and the rice actin intron regulate and enhance expression of the downstream cry3Bb1 element.

Note:
- The wild-type cry3Bb1 coding sequence was modified to encode the following six specific amino acid substitutions, resulting in the synthetic cry3Bb1 coding sequence present in the vector:
-- 2A (alanine insertion at position 2)
-- H232R (histidine to arginine substitution at position 223)
-- S312L (serine to leucine substitution at position 312)
-- N314T (asparagine to threonine substitution at 314)
-- E318K (glutamic acid to lysine substitution at position 318)
-- Q349R (glutamine to arginine substitution at position 349).
- Molecular analyses of MON88017 confirmed that single copies of the epsps and cry3Bb1 genes are integrated at a single locus in the corn genome with all expression elements intact and no plasmid bacterial backbone present. Plasmid PV-ZMIR39 contains the left and right transfer DNA (T-DNA) border sequences that facilitate transformation.

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