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Living Modified Organism (LMO)
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Decisions on the LMO Risk Assessments  
last updated: 10 Jul 2026
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 × MIR162 × GA21
Yes
SYN-E3272-5 × SYN-BTØ11-1 × SYN-IR162-4 × MON-ØØØ21-9
The maize (Zea mays) was produced through cross‑breeding of modified parental varieties SYN-E3272-5, SYN-BTØ11-1, SYN-IR162-4, and MON-ØØØ21-9.
Maize SYN-E3272-5 was modified for optimized bioethanol production. The LM maize also includes a selectable marker. To achieve this trait, the maize expresses a thermostable alpha-amylase encoded by the chimeric amy797E gene, derived from alpha-amylase gene segments originating from strains of the order Thermococcales (thermostable bacteria), which remains active under high temperature to catalyze the hydrolysis of starch into dextrins and optimize the starch liquefaction process. For the transformants selection marker, the maize expresses the manA gene from Escherichia coli, also known as pmi, which encodes for phosphomannose isomerase. which allows mannose utilization.
Maize SYN-BTØ11-1 was modified for resistance to the Lepidoptera insect European corn borer (Ostrinia nubilalis) and tolerance to glufosinate-ammonium herbicides. To achieve resistance to the European corn borer, the maize expresses cry1Ab gene from Bacillus thuringiensis subsp. kurstaki, which have a pore forming mode-of-action in the epithelial lining of feeding larvae; cry1Ab produces a crystal delta-endotoxin that binds to specific midgut receptors in the insect larva, forming pores that disrupt osmotic balance and cause fatal cell lysis. To achieve resistance to glufosinate, the maize expresses the gene from Streptomyces viridochromogenes encoding phosphinothricin N-acetyltransferase encoding gene, which inactivates the active compound L-phosphinothricin through acetylation of the primary amino group.
Maize SYN-IR162-4 was modified for resistance against lepidopteran insect pests including fall armyworm (Spodoptera frugiperda), true armyworm (Pseudaletia unipuncta), beet armyworm (Spodoptera exigua), corn earworm (Helicoverpa zea), black cutworm (Agrotis ipsilon), western bean cutworm (Striacosta albicosta). The LM maize was also modified to express a selectable marker. To achieve resistance against lepidopteran insects, the maize expresses the vegetative insecticidal protein gene (vip3Aa20), a variant of the native vip3Aa gene isolated from Bacillus thuringiensis strain AB88, which is secreted during the vegetative growth phase of the bacterium; Vip3Aa20 is a vegetative insecticidal protein that binds to specific midgut receptors distinct from those recognized by Cry proteins, disrupts the integrity of epithelial cell membranes, and ultimately causes gut paralysis and death of the feeding larvae. For the selection marker, the maize expresses the manA gene from Escherichia coli, which encodes for phosphomannose isomerase.
Maize MON-ØØØ21-9 was modified for tolerance to glyphosate-containing herbicides. To achieve glyphosate tolerance, the maize expresses the modified enzyme 5-enolpyruvylshikimate-3-phosphate synthase, encoded by the mepsps gene. The isolated endogenous maize epsps gene was modified through site-directed mutagenesis, such that its encoded enzyme was insensitive to inactivation by glyphosate, and inserted into the inbred AT maize variety. The modified maize line permits farmers to use glyphosate-containing herbicides for weed control in the cultivation of maize.
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-14797-16 Living Modified Organism SYN-BTØ11-1 - Agrisure™ CB/LL
    Syngenta | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths), European corn borer (Ostrinia nubilalis)), Resistance to herbicides (Glufosinate)
  • BCH-LMO-SCBD-100885-13 Living Modified Organism SYN-IR162-4 - Agrisure™ Viptera maize
    Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-LMO-SCBD-14794-18 Living Modified Organism MON-ØØØ21-9 - Roundup Ready™ maize
    Monsanto | Resistance to herbicides (Glyphosate)
AT variety (for event GA21)
EN
Characteristics of the modification process
pNOV7013; pZO1502 derived from pUC18; pNOV1300; pDPG434
EN
  • Cross breeding
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
  • BCH-GENE-SCBD-14985-12 Cry1Ab | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-GENE-SCBD-103867-2 Alcohol dehydrogenase 1, intron 2 | Zea mays (Maize, Corn, MAIZE)
    Intron
  • BCH-GENE-SCBD-101406-4 Phosphoenolpyruvate carboxylase, intron 9 | Zea mays (Maize, Corn, MAIZE)
    Intron
  • BCH-GENE-SCBD-103622-5 27kD gamma-zein Promoter | Zea mays (Maize, Corn, MAIZE)
    Promoter
  • BCH-GENE-SCBD-103625-3 Alcohol dehydrogenase 1, intron 6 | Zea mays (Maize, Corn, MAIZE)
    Intron
  • BCH-GENE-SCBD-102033-4 SEKDEL ER retention signal
    Transit signal
  • BCH-GENE-SCBD-100887-5 Vegetative insecticidal protein 3Aa20 | Bacillus thuringiensis (Bt, Bacillus, BACTU)
    Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
  • BCH-GENE-SCBD-14966-7 amy797E alpha amylase | Thermococcales spp. (Thermococcus)
    Protein coding sequence | Thermostable alpha-amylase,Use in industrial applications (Biofuel production)
  • BCH-GENE-SCBD-46333-8 5-enolpyruvylshikimate-3-phosphate synthase | Zea mays (Maize, Corn, MAIZE)
    Protein coding sequence | Resistance to herbicides (Glyphosate)
  • BCH-GENE-SCBD-101419-5 Optimized chloroplast transit peptide | Zea mays (Maize, Corn, MAIZE) | Helianthus annuus (Sunflower, HELAN)
    Transit signal
  • BCH-GENE-SCBD-103627-5 Ubiquitin Intron 1 | Zea mays (Maize, Corn, MAIZE)
    Intron
  • BCH-GENE-SCBD-15003-7 Phosphomannose Isomerase gene | Escherichia coli (ECOLX)
    Protein coding sequence | Mannose tolerance,Selectable marker genes and reporter genes
  • BCH-GENE-SCBD-100364-5 Rice actin 1 gene promoter | Oryza sativa (Rice, ORYSA)
    Promoter
  • BCH-GENE-SCBD-100355-6 Rice actin 1, intron | Oryza sativa (Rice, ORYSA)
    Intron
  • BCH-GENE-SCBD-15002-5 Phosphinothricin N-acetyltransferase | Streptomyces viridochromogenes (STRVR)
    Protein coding sequence | Resistance to herbicides (Glufosinate)
  • BCH-GENE-SCBD-100287-7 CaMV 35S promoter | Cauliflower mosaic virus (CaMV)
    Promoter
  • BCH-GENE-SCBD-100290-6 CaMV 35S terminator | Cauliflower mosaic virus (CaMV)
    Terminator
  • BCH-GENE-SCBD-100269-8 Nopaline Synthase Gene Terminator | Agrobacterium tumefaciens (Agrobacterium)
    Terminator
  • BCH-GENE-SCBD-100362-7 Ubiquitin gene promoter | Zea mays (Maize, Corn, MAIZE)
    Promoter
DNA insert from SYN-E3272-5 (vector pNOV7013)
The transforming plasmid carried a T-DNA sequence comprising two gene expression cassettes: modified thermostable alpha-amylase gene (amy797E) from the archaeal order Thermococcales and phosphomannose isomerase (pmi) gene from Escherichia coli.
(1) The amy797E gene is regulated by the Zea mays 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.
(2) The pmi gene is regulated by the polyubiquitin promoter from Zea mays, and the nopaline synthase terminator from Agrobacterium tumefaciens. The Z. mays ubiquitin intron (intron 1) was also included to enhance expression of the pmi sequence. High levels of transcription are expected due to the strong constitutive promoter accompanied by intron-mediated enhancement.
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.
  • After cleavage of the N-terminal signal peptide, the mature AMY797E protein consists of 441 amino acids with a molecular weight of approximately 50.2 kDa. 
  • Southern blot analysis indicated that the 3272 genome contains a single insertion without vector backbone sequences or re-arrangements.The chimeric amy797E gene was generated to combine the best features of three thermostable amylase enzymes and the coding region of the amy797E gene was modified for the preferred codon usage for maize.

DNA insert from SYN-BTØ11-1 (vector pZO1502)
The DNA insert from maize SYN-BTØ11-1 contained two genetic cassettes expressing the cry1Ab gene from Bacillus thuringiensis subsp. kurstaki and the phosphinothricin N-acetyltransferase (PAT) encoding gene from Streptomyces viridochromogenes. Both cassettes were introduced by particle acceleration (biolistic) transformation.
(1) The cry1Ab gene is under the control of the 35S Cauliflower mosaic virus promoter and the Agrobacterium tumefaciens nopaline synthase gene terminator. An alcohol dehydrogenase 1 intron (intron 6) from Zea mays was included to enhance expression of the cry1Ab sequence from Bacillus thuringiensis. High levels of transcription are expected due to the strong constitutive promoter accompanied by intron-mediated enhancement.
(2) The pat gene is regulated by the 35S promoter from Cauliflower mosaic virus and the nopaline synthase gene terminator from Agrobacterium tumefaciens. The Zea mays alcohol dehydrogenase 1 intron (intron 2) from was also included to enhance expression of the phosphinothricin N-acetyltransferase sequence from Streptomyces viridochromogenes. High levels of transcription are expected due to the strong constitutive promoter accompanied by intron-mediated enhancement.
Note
  • The plasmid carrying the DNA transfer was digested by restriction enzyme NotI and only the fragments containing the two expression cassettes were inserted into the LM maize. The AMPr gene encoding ampicillin resistance and an origin of replication were not inserted.
  • Southern blot analysis confirmed the absence of unwanted DNA fragments in the transformant generations.

DNA insert from SYN-IR162-4 (vector pNOV1300)
The transforming plasmid for maize SYN-IR162-4 contained a transfer DNA (T-DNA) region comprising two genetic cassettes: one expressing the vegetative insecticidal protein gene, vip3Aa20, from Bacillus thuringiensis for the modified resistant trait and one expressing phosphomannose isomerase (PMI) as a selection marker.
(1) The vip3Aa20 gene is regulated by the ubiquitin gene promoter from Zea mays and the 35S Cauliflower mosaic virus terminator. The Zea mays phosphoenolpyruvate carboxylase intron 9 (iPEPC9) from Z. mays was also included to enhance expression of the Vip3Aa20 sequence from Bacillus thuringiensis. High levels of transcription are expected due to the strong constitutive promoter accompanied by intron-mediated enhancement.
(2) The pmi gene was used as a selectable marker for transformants and is under the control of the Z. mays ubiquitin gene promoter and the nopaline synthase gene terminator from Agrobacterium tumefaciens.
Note
  • The variant of the native B. thuringiensis Vip3Aa, named vip3Aa19, has codon changes that result in a single M129I amino acid substitution was inserted into the transformation cassette. During the transformation process an additional DNA mutation resulted in a K284Q amino acid substitution. This final form was designated the name Vip3Aa20.
  • The pNOV1300 vector region between the left and right border sequences, which included the vip3Aa19 and pmi gene expression cassettes, was inserted into the maize genome during transformation.
  • Southern blot analyses demonstrated that the T-DNA insert contains: i) single copies of a vip3Aa20 gene and a pmi gene; ii) two copies of the ZmUbiInt promoter; iii) one copy of the NOS terminator; and iv) no backbone sequences from transformation plasmid pNOV1300.

DNA insert from MON-ØØØ21-9 (vector pDPG434)
The DNA insert from maize MON-ØØØ21-9 contained one genetic cassette expressing the modified Zea mays 5-enolpyruvylshikimate-3-phosphate synthase (mepsps).
The mepsps gene is under the control of the Oryza sativa (rice) actin 1 promoter and the Agrobacterium tumefaciens nopaline synthase terminator. A rice actin 1 intron was included to enhance expression of the mepsps sequence, and a synthetic optimized chloroplast transit peptide, derived from maize and sunflower (Helianthus sp.) ribulose-1,5-bisphosphate carboxylase oxygenase sequences, to direct the translated mEPSPS protein to the chloroplast. High levels of transcription are expected due to the strong constitutive promoter accompanied by intron-mediated enhancement.
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.
  • Southern blot analysis indicated that the transformed DNA integrated into the host genome at a single site. Analysis further indicated that no sequences from the vector backbone were integrated into the host genome.
  • 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 - Sequencing analysis indicated a truncated rice actin promoter in the 5’ end of the insertion event, only containing the last 148bp of the 3’ end of the rice actin promoter including the rice actin intron. 
  • 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). 
    • One set of analysis indicated that two complete copies and one partial copy of the transformation cassette were integrated at a single site into the host genome. The partial copy was composed of the rice actin promoter and the EPSPS coding sequence but not the nos terminator element.
  • The modified maize expresses only the full-length mEPSPS protein.
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LMO characteristics
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  • Biofuel
  • Feed
  • Food
Additional Information
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