SYN-E3272-5 × SYN-BTØ11-1 × DAS-59122-7 × SYN-IR6Ø4-5 × DAS-Ø15Ø7-1 × MON-ØØØ21-9 - Insect-resistant, herbicide-tolerant, amylase producing maize | BCH-LMO-SCBD-260417 | Living Modified Organism | Biosafety Clearing-House
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Living Modified Organism (LMO)
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Decisions on the LMO Risk Assessments  
last updated: 20 May 2022
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.
Insect-resistant, herbicide-tolerant, amylase producing maize
EN
3272 × Bt11 × 59122 × MIR604 × TC1507 × GA21
Yes
SYN-E3272-5 × SYN-BTØ11-1 × DAS-59122-7 × SYN-IR6Ø4-5 × DAS-Ø15Ø7-1 × MON-ØØØ21-9
The maize (Zea mays) was produced by crossing modified parental lines for tolerance to herbicides, resistance to insect pests and improved use in biofuel. For tolerance to Lepidoptera pests, the maize expresses Bacillus thuringiensis Cry1Ab and Cry1F. For resistance to Coleoptera pests, the maize expresses B. thuringiensis modified Cry3A, Cry34Ab1 and Cry35Ab1. For tolerance to glufinosate, the maize expresses Streptomyces viridochromogenes phosphinothricin N-acetyaltransferase. For tolerance to glyphosate, the maize expresses a modified maize 5-enolpyruvylshikimate-3-phosphate synthase, which does not bind the herbicidal compound with high affinity. For improved use in downstream biofuel applications, the maize expresses a chimeric Thermococcales sp. alpha amylase, which has increased thermostability and starch hydrolysis activity during the high temperatures in dry-grind ethanol production from maize. The maize also contains Escherichia coli phosphomannose isomerase, which was used as a selectable marker during transformation of the parental lines.
EN
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)
    Crops
  • BCH-LMO-SCBD-15109-9 Living Modified Organism SYN-E3272-5 - Enogen™ Maize
    Use in industrial applications (Biofuel production)
  • BCH-LMO-SCBD-14797-15 Living Modified Organism SYN-BTØ11-1 - YieldGard™ maize
    Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths), European corn borer (Ostrinia nubilalis)), Resistance to herbicides (Glufosinate)
  • BCH-LMO-SCBD-15165-13 Living Modified Organism DAS-59122-7 - Herculex™ RW Rootworm Protection maize
    Pioneer Hi-Bred International Inc. | Resistance to diseases and pests (Insects, Coleoptera (beetles)), Resistance to herbicides (Glufosinate)
  • BCH-LMO-SCBD-15105-12 Living Modified Organism SYN-IR6Ø4-5 - Agrisure™ RW Rootworm-Protected maize
    Resistance to diseases and pests (Insects, Coleoptera (beetles))
  • BCH-LMO-SCBD-14841-13 Living Modified Organism DAS-Ø15Ø7-1 - Herculex™ I maize
    Dow AgroSciences, Pioneer Hi-Bred International Inc. | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths)), Resistance to herbicides (Glufosinate)
  • BCH-LMO-SCBD-14794-18 Living Modified Organism MON-ØØØ21-9 - Roundup Ready™ maize
    Monsanto | Resistance to herbicides (Glyphosate)
EN
Characteristics of the modification process
pNOV7013; pZO1502; PHP17662; pZM26; PHI8999A; pDPG434
EN
  • Cross breeding
 
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Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
DNA insert from 3272 vector
The DNA insert in the 3272 genome contains two gene cassettes: Thermococcales sp. alpha amylase (amy797E) and Escherichia coli phosphomannose isomerase (pmi).

The coding sequence of amy797E is under control of the Zea mays (maize) 27kD gamma-zein promoter and Cauliflower mosaic virus 35S terminator. A maize phosphoenolpyruvate carboxylase intron 9 was included to enhance expression of amy797E. A synthetic SEKDEL endoplasmic retention signal was also included to retain the AMY797E protein in the endoplasmic reticulum of cells. Due to the endosperm (seed)-specific nature of the promoter, expression of AMY797E is expected in the endosperm of maize kernels only.

The pmi coding sequence is under control of a maize ubiquitin promoter and Agrobacteirum tumefaciens nopaline synthase terminator. The first intron of the maize ubiquitin sequence was included to enhance expression of the pmi sequence.

Note:
  • The amy797E is a chimeric, thermostable protein that consists of three thermostable amylase enzymes. The sequence was then further codon optimized for expression in maize.
  • Southern blot analysis indicated that the 3272 genome contains a single insertion without vector backbone sequences or re-arrangements.

DNA insert from Bt11 vector pZO1502
The DNA insert from the Bt11 genome contains two gene cassettes: Bacillus thuringiensis cry1Ab and Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (pat).

Transcription of cry1Ab gene is under control of the Cauliflower mosaic virus (CaMV) 35S promoter and the Agrobacterium tumefaciens nopaline synthase (nos) terminator. The transcript initially contains Zea mays alcohol dehydrogenase 1 intron 6, which enhance gene expression.

Transcription of pat is under control of the CaMV 35S promoter and nos terminator. The transcript initially contains Zea mays alcohol dehydrogenase 1 intron 2 to enhance gene expression.

Note:
  • The CaMV promoter associated with cry1Ab, was isolated from the CM1841 strain of CaMV using DdeI restriction digestion. However, the DdeI sites were converted into SacI sites.
  • The CaMV promoter associated with pat was isolated from the Cabb-S strain of CaMV (AluI to DdeI fragment) and subsequently modified.
  • The cry1Ab coding sequence encodes a truncated version corresponding to the N-terminal 615 amino acids of the full length Cry1Ab protein.
  • The cloning of pat did not result in any amino acid sequence changes in the parental line.
  • The nos terminator corresponds to the 423 to 678 basepairs of the nopaline synthase gene in A. tumefaciens.


DNA insert from 59122 vector PHP17662:
The DNA insert from the 59122 genome has three gene cassettes: Bacillus thuringiensis cry34Ab1, Bacillus thuringiensis cry35Ab1 and Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (pat).

Transcription of cry34Ab1 starts at Zea mays ubiquitin gene promoter and terminates at the Solanum tuberosum proteinase inhibitor II gene terminator.

Transcription of cry35Ab1 commences from the (Triticum aestivum peroxidase gene promoter and stops at another S. tuberosum proteinase inhibitor II gene terminator.

The pat coding sequence is under control of a CaMV 35S promoter and terminator. Due to the constitutive nature of the promoter, high levels of expression are expected.

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 peroxidase 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 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 TC1507 vector PHI8999A
DNA fragment PHI8999A contains two adjacent plant gene expression cassettes for Bacillus thuringiensis cry1F and Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (pat).

Transcription of cry1F is directed by the promoter and first exon and intron of the maize (Zea mays) ubiquitin gene and terminates at the Agrobacterium tumefaciens ORF25 terminator.

Transcription of the pat gene commences from the CaMV 35S promoter and ends at the CaMV 35S terminator.

Note:
  • The coding sequence of both genes has been optimized to achieve a high level of expression in maize.
  • The sequences of the complete cry1F and pat are identical to those in the original plasmid.
  • The Cry1F protein includes the F604K (phenylalanine to lysine at position 604) amino acid substitution, which was introduced to create a specific restriction site for cloning purposes.


DNA insert from GA21 vector pDPG434
The DNA insert from the GA21 genome contains one gene cassette: Zea mays modified 5-enolpyruvylshikimate-3-phosphate synthase (mepsps).

Transcription of 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.


Kindly refer to the parental records for more information.
EN
LMO characteristics
EN
  • Biofuel
Detection method(s)
EN
Additional Information
EN
Records referencing this document Show in search
Record type Field Record(s)
Laboratory for detection and identification of LMOs LMO(s) detectable by the laboratory 1

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