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Modified Organism
SYN-IR162-4 × SYN-IR6Ø4-5 × SYN-Ø53Ø7-1 × MON-ØØØ21-9 - Glyphosate tolerant, insect resistant maize
Record information and status
Record ID
115541
Status
Published
Date of creation
2020-04-22 15:36 UTC (austein.mcloughlin@cbd.int)
Date of publication
2020-04-22 15:36 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
Glyphosate tolerant, insect resistant maize
Transformation event
MIR162 x MIR604 x 5307 x GA21
Unique identifier
SYN-IR162-4 × SYN-IR6Ø4-5 × SYN-Ø53Ø7-1 × MON-ØØØ21-9
Developer(s)
Syngenta Seeds GmbH
Syngenta Seeds GmbH
Zum Knipkenbach 20
Bad Salzuflen
Germany, 32107
Phone:+49 52 22 5308-0
Fax:+49 52 22 5308-12
Email:info.seeds@syngenta.com
Url:Syngenta Seeds Germany
Description
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. For Coleoptera resistance, the maize expresses B. thuringiensis mCry3A and eCry3.1Ab. In addition to the insecticidal proteins, the maize also 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)
SYN-IR6Ø4-5 - Agrisure™ RW Rootworm-Protected maize
Mannose tolerance Resistance to diseases and pests - Insects - Coleoptera (beetles) - Western corn rootworm (Diabrotica virgifera) Selectable marker genes and reporter genes
Show detection method(s)
SYN-Ø53Ø7-1 - Agrisure® Duracade™ Maize
Mannose tolerance Resistance to diseases and pests - Insects - Coleoptera (beetles) - Western corn rootworm (Diabrotica virgifera), Northern corn rootworm (Diabrotica barberi) Selectable marker genes and reporter genes
MON-ØØØ21-9 - Roundup Ready™ maize
Resistance to herbicides - Glyphosate
Show detection method(s)
Characteristics of the transformation process
Vector
pNOV1300; pZM26; pSYN12274; 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
 
 
Metallothionein-like gene promoter
2.56 Kb
 
 
mCry3A
1.80 Kb
 
 
Nopaline Synthase Gene Terminator
0.25 Kb
 
 
Ubiquitin gene promoter
0.98 Kb
 
 
Ubiquitin Intron 1
1.01 Kb
 
 
Phosphomannose Isomerase gene
1.18 Kb
 
 
Nopaline Synthase Gene Terminator
0.25 Kb
 
 
Cestrum Yellow Leaf Curling Virus promoter
0.35 Kb
 
 
eCry3.1Ab
1.98 Kb
 
 
Nopaline Synthase Gene Terminator
0.25 Kb
 
 
Ubiquitin gene promoter
1.99 Kb
 
 
Phosphomannose Isomerase gene
1.18 Kb
 
 
Nopaline Synthase Gene Terminator
0.25 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.

Note:
- 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 MIR604 vector pZM26
The parental plant contains two expression cassettes: (i) modified Cry3a (mCry3a) originally from Bacillus thuringiensis and (ii) phosphomannose isomerase (pmi) from Escherichia coli.

Expression mCry3a is under control of a Zea mays metallothionein-like gene promoter and an Agrobacterium tumefaciens nopaline synthase (nos) terminator. Transcription of pmi is under the control of Z. mays ubiquitin gene promoter and an A. tumefaciens nos terminator. The transcript initially also contains an intron from Z. mays ubiquitin-1 to enhance gene expression.

Note:
- mCry3a was originally obtained from the native cry3a gene, but was modified to enhance gene expression in maize. The synthetic version of the protein (mCry3a) contains the same amino acid sequences of the native version, except for the modified serine-protease recognition site.
- The following changes in the pmi occurred: the valine at position 61 has been substituted by alanine (V61A) and glutamine at position 210 has been substituted by histidine (Q210H). Please note no apparent change of function occurred.
- Southern blot and qPCR analysis indicated that a single insertion of both expression cassettes occurred and there was no integration of the vector backbone.

DNA insert from 5307 vector pSYN12274
The DNA insert contains two gene cassettes for an engineered chimeric protein eCry3.1Ab and an Escherichia coli phosphomannose isomerase (pmi).

Transcription of eCry3.1Ab is under control of a Cestrum Yellow Leaf Curling Virus promoter and an Agrobacterium tumefaciens nopaline synthase (nos) terminator. Transcription of pmi is under control of a Zea mays ubiquitin gene promoter and a nos terminator. The promoter contains the first intron of the ubiquitin gene, which will be initially included in the mRNA before splicing and for enhancing expression of pmi. Transcription is expected to be constitutive under both promoters and result in elevated levels of transgene expression.

Note:
- eCry3.1Ab is a result of a fusion of the 5′ end (Domain I, Domain II and 15 amino acids of Domain III) of a modified Cry3A gene (mcry3A) and the 3′ end (Domain III and Variable Region 6) of a synthetic Cry1Ab gene. The sequences were sourced from Bacillus thuringiesis.
- Southern blot analysis indicated that the parental line contains a single insertion of the vector and there was no integration of the vector backbone.
- Sequencing analysis indicated that the right and left T-DNA borders were truncated.

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 Cry1Ab, Vegetative insecticidal protein 3Aa20, and Cry1F. For Coleoptera resistance, the maize expresses B. thuringiensis mCry3A. In addition to the insecticidal proteins, the maize also expresses Streptomyces viridochromogenes phosphinothricin N-acetyltransferase for glufosinate tolerance and 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.

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.

Note:
- 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