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Modified Organism
MON-ØØ81Ø-6 × MON-8746Ø-4 - Insect resistant, Drought tolerant maize (Water efficient maize for Africa)
Record information and status
Record ID
115607
Status
Published
Date of creation
2020-06-17 16:13 UTC (austein.mcloughlin@cbd.int)
Date of publication
2020-06-17 16:13 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
Insect resistant, Drought tolerant maize (Water efficient maize for Africa)
Transformation event
MON810 × MON87460
Unique identifier
MON-ØØ81Ø-6 × MON-8746Ø-4
Developer(s)
Dr. Murenga Mwimali
Principal Scientist/Maize Breeder
Biotechnology
Kenya Agricultural and Livestock Research Organization/Monsanto Kenya/CIMMYT/AATF
Nairobi
Kenya
Phone:(+254) (722 915 500)
Email:mwimali@yahoo.co.uk
Description
The  modified maize was produced through the crossing of two modified parental lines (MON810 and MON87460) to achieve insect resistance and drought tolerance. For resistance to Lepidoptera pests, the maize expresses Bacillus thuringiensis Cry1Ab, which causes pore formation in the midgut epithelial cells of larvae and results in loss of gut lining integrity and eventual death. 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 during transformation of the MON87460 parental line.
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-ØØ81Ø-6 - YieldGard™ maize
Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths)
Show detection method(s)
MON-8746Ø-4 - Droughtgard™ Maize
Resistance to antibiotics - Kanamycin Tolerance to abiotic stress - Cold / Heat, Drought
Show detection method(s)
Characteristics of the transformation process
Vector
PV-ZMBK07 and PV-ZMAP595
Techniques used for the modification
  • Cross breeding
Genetic elements construct
 
CaMV Enhanced 35S promoter
0.61 Kb
 
 
Hsp70 intron
0.80 Kb
 
 
Cry1Ab
3.46 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
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
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 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.

For more information, kindly refer to the parental records.
LMO characteristics
Modified traits
  • Selectable marker genes and reporter genes
Common use(s)
  • Food
  • Feed
Additional Information
Additional Information
The Water Efficient Maize for Africa (WEMA) project is a public-private partnership led by the African Agricultural Technology Foundation (AATF) to address the effects of drought on farmers. The project partners have been developing new drought-tolerant African maize varieties since 2008, incorporating conventional advanced plant breeding and biotechnology.