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Modified Organism
MON-87427-7 × MON-89Ø34-3 × MON-ØØ81Ø-6 × SYN-IR162-4 × MON-87411-9 × MON-87419-8 - Herbicide tolerant, insect resistant maize
Record information and status
Record ID
115691
Status
Published
Date of creation
2020-08-25 20:50 UTC (austein.mcloughlin@cbd.int)
Date of publication
2020-08-25 20:50 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
MON87427 × MON89034 × MON810 × MIR162 × MON87411 × MON87419
Unique identifier
MON-87427-7 × MON-89Ø34-3 × MON-ØØ81Ø-6 × SYN-IR162-4 × MON-87411-9 × MON-87419-8
Developer(s)
Description
The maize has been produced through cross breeding of modified parental lines to confer tolerance to herbicides and resistance to insects. For Lepidoptera resistance, the modified maize expresses Bacillus thuringiensis Cry1Ab, Cry1A.105, Cry2Ab2 and Vegetative insecticidal protein 3Aa20. For Coleoptera resistance, the maize expresses B. thuringiensis Cry3Bb1. Additionally, the maize contains an RNA interference cassette targeting Diabrotica virgifera virgifera DvSnf7, an essential cellular component of endosomal sorting complex required for transport, for protection against the pest. For tolerance to glyphosate, the maize expresses Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase, a variant of an endogenous gene, which prevents the herbicide interference of the shikimate pathway (aromatic amino acid biosynthesis). For tolerance to glufinosate, the modified maize expresses Streptomyces viridochromogenes phosphinothricin N-acetyltransferase, which inactivates phosphinothricin (glufinosate) by transferring an acetyl group from acetyl-CoA. For diacamba tolerance, the maize expresses Stenotrophomonas maltophilia dicamba monooxygenase, which catalyzes the oxygen demethylation of diacamba. The modified maize also expresses Escherichia coli phosphomannose isomerase for mannose selection during transformation.
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-87427-7 - Maize modified for tissue selective glyphosate tolerance
Resistance to herbicides - Glyphosate
Show detection method(s)
MON-89Ø34-3 - YieldGard™ VT Pro™
Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths)
Show detection method(s)
MON-ØØ81Ø-6 - YieldGard™ maize
Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths)
Show detection method(s)
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)
MON-87411-9 - Maize modified for herbicide tolerance and insect resistance
Monsanto Resistance to diseases and pests - Insects - Coleoptera (beetles) - Western corn rootworm (Diabrotica virgifera) Resistance to herbicides - Glyphosate
MON-87419-8 - Dicamba and Glufosinate Tolerant Maize
Resistance to herbicides - Glufosinate
Characteristics of the transformation process
Vector
PV-ZMAP1043; PV-ZMIR245; PV-ZMBK07 and PV-ZMGT10; pNOV1300; PV-ZMIR10871; PV-ZMHT507801
Techniques used for the modification
  • Cross breeding
Genetic elements construct
 
CaMV Enhanced 35S promoter
0.62 Kb
 
 
Hsp70 intron
0.80 Kb
 
 
Chloroplast transit peptide 2
0.23 Kb
 
 
5-enolpyruvylshikimate-3-phosphate synthase gene
1.37 Kb
 
 
Nopaline Synthase Gene Terminator
0.25 Kb
 
 
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
 
 
CaMV Enhanced 35S promoter
0.61 Kb
 
 
Hsp70 intron
0.80 Kb
 
 
Cry1Ab
3.46 Kb
 
 
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
 
 
CaMV Enhanced 35S promoter
0.62 Kb
 
 
Hsp70 intron
0.80 Kb
 
 
Snf7 coding sequence
0.24 Kb
 
 
Snf7 coding sequence
0.24 Kb
 
 
rbcS-E9 gene terminator
0.63 Kb
 
 
pIIG gene promoter
0.95 Kb
 
 
5' untranslated leader from chlorophyll a/b-binding protein
0.06 Kb
 
 
Rice actin 1, intron
0.48 Kb
 
 
Cry3Bb1
1.96 Kb
 
 
Heat shock protein 17.3 terminator
0.21 Kb
 
 
Alpha Tubulin Gene promoter
2.18 Kb
 
 
Chloroplast transit peptide 2
0.23 Kb
 
 
5-enolpyruvylshikimate-3-phosphate synthase gene
1.37 Kb
 
 
Alpha Tubulin Gene terminator
0.58 Kb
 
 
Ubiquitin gene promoter
1.64 Kb
 
 
Ubiquitin leader sequence
0.10 Kb
 
 
Ubiquitin Intron Sequence
1.04 Kb
 
 
Phosphinothricin N-acetyltransferase gene
0.55 Kb
 
 
RA5B gene terminator
0.21 Kb
 
 
PC1SV Promoter
0.43 Kb
 
 
5' untranslated leader from chlorophyll a/b-binding protein
0.06 Kb
 
 
Rice actin 1, intron
0.48 Kb
 
 
Chloroplast Transit Peptide 4
0.22 Kb
 
 
Dicamba monooxygenase gene
1.02 Kb
 
 
Heat shock protein 17.3 terminator
0.21 Kb
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
DNA insert from PV-ZMAP1043
Transcription of  Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (epsps) commences from the Cauliflower mosaic virus (CaMV) enhanced 35S promoter and terminates at the A. tumefaciens nopaline synthase (nos) terminator. The transcript contains a Zea mays heat shock protein 70 (hsp70) intron, an Arabidopsis thaliana N-terminal chloroplast transit peptide sequence for chloroplast targeting of the protein and epsps. The CaMV enhanced 35S promoter-hsp70 combination promotes gene expression in female and vegetative tissues, but not in male reproductive tissues (pollen microspores and tapetum).

Note:
- Southern blot analyses indicate that a single copy of the T-DNA was inserted at a single site in the parental maize genome and no plasmid vector backbone sequences were detected to have been integrated. DNA sequencing analyses further indicated that the expected T-DNA sequences were integrated.
-The epsps coding sequence is the codon optimized coding sequence of the aroA gene from Agrobacterium sp. strain CP4 encoding EPSPS.


DNA insert from PV-ZMIR245
Two insecticidal protein expression cassettes were inserted into the genome. Bacillus thuringiensis cry1A.105 expression is under the control of the CaMV 35S enhanced promoter, which first transcribes wheat (Triticum aestivum) 5' untranslated region of the chlorophyll a/b-binding protein (cab) and a rice actin 1 intron before transcribing cry1A.105. Transcription terminates at the wheat heat shock protein 17.3 terminator. Expression of the B. thuringiensis cry2Ab2 starts at the Figwort mosaic virus (FMV) promoter, which transcribes the Zea mays heat shock protein 70 (hsp70), then the Z. mays transit peptide and the cry2Ab2 coding sequence, before terminating at the nos terminator.

Note:
- The Cry2Ab2 coding sequence was modified for optimal expression in plants.
- South blot analysis confirmed that single insertions of both cry2Ab2 and cry1A.105, as well as no vector backbone were present and in the parent.
- A deletion removed the duplicated enhancer elements compared to the original CaMV e35S promoter in PV-ZMIR245.
- The selectable marker, nphII, cassette was bred out of the parental line and became not associated with this transformation event.


DNA insert from PV-ZMBK07
A partial insert containing Bacillus thuringiensis cry1Ab was inserted into the parental maize genome. Transcription is directed from the Cauliflower Mosaic Virus 35S enhanced promoter. The transcript contains a Zea mays heat shock protein 70 (ZmHsp70) intron and the coding sequence of cry1Ab. ZmHsp70 enhances expression of cry1Ab.

Note:
- The coding sequence of cry1Ab has been codon optimized for expression in plants. The codon optimization did not result in any changes to the amino acid sequence relative to the native sequence.
- Southern blot analysis indicated that a single partial insert is found within the parental genome.
- Southern blot analysis did not detect the presence of the Escherichia coli neomycin phosphotransferase II gene nor any DNA from plasmid PVZMGT10 (containing genes for glyphosate tolerance - cp4 epsps).
- ELISA protein analysis and feeding assays indicated expression of Cry1Ab.


DNA insert from pNOV1300
In the parental MIR162 maize, a variant of the native B. thuringiensis vegetative insecticidal protein 3Aa (vip3Aa20), named vip3Aa19, which 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. 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. A second expression cassette, containing the E. coli phosphomannose isomerase gene, 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 gene (nos) terminator.

Note:
- 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 PV-ZMHT507801
The Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (pat) gene is under the control of the Andropogon gerardii ubiquitin  promoter and the Oryza sativa alpha-amylase/trypsin inhibitor terminator. The transcript includes the A. gerardii 5' untranslated leader sequence and an intron from ubiquitin before (5') coding sequence of pat.

The Stenotrophomonas maltophilia dicamba monooxygenase (dmo) gene is under control of the Peanut chlorotic streak caulimovirus (PC1SV) full-length transcript promoter and the Triticum aestivum (wheat) heat shock protein 17 terminator. The transcript produced contains a wheat chlorophyll a/b-binding 5' untranslated leader sequence (for improved gene expression), an O. sativa actin 1 untranslated region and intron (for improved gene expression), the untranslated  and targeting region of Petunia hybrida chloroplast transit peptide 4 (for chloroplast targeting of the protein) and dmo.

Note:
- Originally, the plasmid vector contained two T-DNA elements that were inserted during the initial transformation event: one containing the dmo and pat expression cassettes, and a second containing an Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps) expression cassette. The cp4-epsps expression cassette is regulated by the O sativa actin 1 promoter and 5′ untranslated leader, O. sativa intron, the Arabidopsis thaliana chloroplast targeting peptide 2 targeting sequence, and the A. tumefaciens nopaline synthase  3′ untranslated region. Subsequent traditional breeding, segregation, selection, and screening were used to isolate those plants that contain the dmo and pat expression cassettes (T-DNA I) and do not contain the cp4-epsps expression cassette (T-DNA II).
- Molecular characterization of MON87419 indicated that a single copy of T-DNA I was integrated into the maize genome at a single intact locus that includes all expected elements within the insert, with the exception of incomplete Right and Left Border sequences. These analyses also showed no PV-ZMHT507801 backbone elements or T-DNA II sequences were present in the event.


DNA insert from PV-ZMIR10871:
The MON87411 genome contains an RNA interference (RNAi) cassette targeting Diabrotica virgifera virgifera, a Bacillus thuringiensis Cry3Bb1 cassette and an Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (epsps) cassette.

Transcription of the RNAi cassette commences from the Cauliflower mosaic virus 35S enhanced promoter and terminates at the Pisum sativum ribulose bisphosphate carboxylase small chain 2 terminator. The transcript initially contains a Zea mays heat shock protein 70 intron, which contributes to enhanced expression in vegetative tissues of the plant, and two partial coding sequences of the D. virgifera virgifera Snf7p gene, which encodes the SNF7 subunit of the ESCRT-III complex. The two Snf7p sequences are in an inverted orientation, separated by a 150 nucleotide intervening sequence, which allows base pairing between the inverted sequences and hairpin RNA formation post-transcription, which then triggers an RNAi response. Due to RNAi processing, small interfering RNA molecules (roughly 21-23 nucleotides in length) will be produced and thus no translation into protein will occur from this cassette.

Transcription of the cry3Bb1 is under control of the Z. mays physical impedance induced protein promoter and Triticum aestivum (wheat) heat shock protein 17.3 terminator. The transcript also contains a wheat 5' untranslated leader from chlorophyll a/b-binding protein and Oryza sativa actin 1 intron for enhanced expression of the transgene. Expression of epsps is under control of an O. sativa alpha tubulin promoter and terminator. The transcript additionally contains Arabidopsis thaliana chloroplast targeting peptide 2 to sequester the protein to the chloroplast.

Note:
- Sequencing, PCR and bioinformatic analyses indicate that a single, intact insertions of the three gene cassettes occurred in the parental line.
- No plasmid backbone was detected.


For more information, kindly refer to the parental line records.
LMO characteristics
Modified traits
  • Selectable marker genes and reporter genes
  • Tolerance to mannose
  • Resistance to Diabrotica virgifera virgifera
Common use(s)
  • Food
  • Feed

Records referencing this document (2)
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2record(s) found
Country's Decision or any other Communication1 record
Risk Assessment1 record