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Modified Organism
MON-87427-7 × MON-8746Ø-4 × MON-89Ø34-3 × SYN-IR162-4 × MON-ØØ6Ø3-6 - Maize modified for herbicide tolerance, insect resistance and drought tolerance
Record information and status
Record ID
115672
Status
Published
Date of creation
2020-08-06 20:23 UTC (austein.mcloughlin@cbd.int)
Date of publication
2020-08-06 20:23 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
Maize modified for herbicide tolerance, insect resistance and drought tolerance
Transformation event
MON87427 × MON87460 × MON89034 × MIR162 × NK603
Unique identifier
MON-87427-7 × MON-8746Ø-4 × MON-89Ø34-3 × SYN-IR162-4 × MON-ØØ6Ø3-6
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
Bayer CropScience
Bayer CropScience Deutschland GmbH
Bayer CropScience AG
Alfred-Nobel-Str. 50
40789 Monheim am Rhein
Monheim am Rhein
Germany, 40789
Phone:+49 21 73 - 38-0
Url:Bayer CropScience
Description
The modified maize (Zea mays) was produced through the cross breeding of modified parental lines for herbicide tolerance, insect resistance and drought tolerance. For Lepidoptera resistance, the maize expresses Bacillus thuringiensis  Cry1A.105 and Cry2Ab2. For herbicide tolerance, the maize expresses Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase, a variant of an endogenous enzyme that confers for glyphosate tolerance. For drought tolerance, the maize expresses Bacillus subtilis cold shock protein, which binds RNA and maintains cellular functions under water-limited conditions (improvement of natural abiotic stress responses). The maize also contains an Escherichia coli neomycin phosphotransferase II cassette, which allowed for kanamycin selection, and E. coli phoshomannose isomerase, which allowed for mannose selection, during transformation of some of the parental lines.
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-8746Ø-4 - Droughtgard™ Maize
Resistance to antibiotics - Kanamycin Tolerance to abiotic stress - Cold / Heat, Drought
Show detection method(s)
MON-89Ø34-3 - YieldGard™ VT Pro™
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-ØØ6Ø3-6 - Roundup Ready™ maize
Resistance to herbicides - Glyphosate
Show detection method(s)
Characteristics of the transformation process
Vector
PV-ZMAP1043; PV-ZMAP595; PV-ZMIR245; pNOV1300; PV-ZMGT32
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 right border repeat
0.36 Kb
 
 
Rice actin 1 gene promoter
0.92 Kb
 
 
Rice actin 1, intron
0.48 Kb
 
 
Cold shock protein gene
0.20 Kb
 
 
Transcript 7 gene 3' untranslated region
0.51 Kb
 
 
loxP recombination site
0.03 Kb
 
 
CaMV 35S promoter
0.29 Kb
 
 
Neomycin Phosphotransferase II
0.79 Kb
 
 
Nopaline Synthase Gene Terminator
0.25 Kb
 
 
loxP recombination site
0.03 Kb
 
 
Ti plasmid left border repeat
0.44 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
 
 
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
 
 
Phosphinothricin N-acetyltransferase gene
1.18 Kb
 
 
Nopaline Synthase Gene Terminator
0.25 Kb
 
 
Rice actin 1 gene promoter
0.80 Kb
 
 
Rice actin 1, intron
0.60 Kb
 
 
Chloroplast transit peptide 2
0.20 Kb
 
 
5-enolpyruvylshikimate-3-phosphate synthase gene
1.40 Kb
 
 
Nopaline Synthase Gene Terminator
0.30 Kb
 
 
CaMV Enhanced 35S promoter
0.60 Kb
 
 
Hsp70 intron
0.80 Kb
 
 
Chloroplast transit peptide 2
0.20 Kb
 
 
5-enolpyruvylshikimate-3-phosphate synthase gene
1.40 Kb
 
 
Nopaline Synthase Gene Terminator
0.30 Kb
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
DNA insert from PV-ZMAP1043
Transcription of 5-enolpyruvylshikimate-3-phosphate synthase (cp4 epsps) from Agrobacterium tumefaciens commences from the Cauliflower mosaic virus (CaMV) enhanced 35S promoter and ends at the A. tumefaciens nopaline synthase (nos) gene terminator. The transcript contains a Zea mays heat shock protein 70 (hsp70) intron, Arabidopsis thaliana N-terminal chloroplast transit peptide sequence, and cp4 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 cp4 epsps coding sequence is the codon optimized coding sequence of the aroA gene from Agrobacterium sp. strain CP4 encoding CP4 EPSPS.


DNA insert from PV-ZMAP595
The T-DNA insert contains the following gene cassettes: Bacillius subtillus cold shock protein (cspB) and Escherichia coli neomycin phosphotransferase II (nptII). 

Transcription of cspB is under control of the Oryza sativa actin 1 promoter and Agrobacterium tumefaciens trasncript 7 gene 3' untranslated region. The transcript initially contains an O. sativa actin 1 intron for enhanced gene expression of cspB. The sequence is removed (spliced) prior to protein translation. Constitutive expression of cspB is expected due to the actin promoter.

Transcription of nptII is under control of the Cauliflower Mosaic Virus (CaMV) 35S promoter and A. tumefaciens nopaline synthase terminator. High levels of transcription are expected due to the CaMV promoter.

Note:
- The coding seuquence of cspB has been codon optimized for optimal expression within plant cells.
- Southern blot analysis indicated that no vector backbone sequences were inserted into the parental genome
- Southern blot analysis indicated that the parental genome contains a single insertion
- Sequencing analyses confirm the Southern blot analyses.
- A 22 base pair deletion of genomic DNA at the insert-to-plant DNA junction occurred.
- loxP sites can be found in the parental genome and could potentially allow for the excision of the nptII cassette by CRE recombinase.


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 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-ZMGT32
The plant expression plasmid vector, PV-ZMGT32 contains two adjacent plant gene expression cassettes each containing a single copy of the cp4 epsps. In the first (5' end) expression cassette, the cp4 epsps gene is under the transcriptional regulation of an Oryza sativa actin promoter and a nos terminator. An O. sativa actin intron is also present in the transcript for enhanced expression of the coding sequence. The second cassette consists of another cp4 epsps gene regulated by an CaMV enhanced 35S promoter (containing a duplicated enhancer region) and a nos terminator. Similarly, an intron from the maize heat shock protein 70 (hsp70) was included for enhancing expression of the coding sequence. Both promoters of the gene cassettes are expected to promoter high levels of transcription.

Note:
- The parental NK603 line contained a single, intact insertion containing both cp4 epsps gene cassettes.
- Due to restriction digest prior to particle bombardment, the vector backbone, containing E. coli neomycin phosphotransferase II and origin of replication, were not incorporated into the parental genome.


For more information, kindly refer to the parental LMO records.
LMO characteristics
Modified traits
  • Selectable marker genes and reporter genes
Common use(s)
  • Food
  • Feed
Additional Information
Other relevant website address or attached documents