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Modified Organism
OKA-NBØØ3-1 - Arctic™ Fuji Apple
Record information and status
Record ID
115072
Status
Published
Date of creation
2019-07-30 14:02 UTC (austein.mcloughlin@cbd.int)
Date of publication
2019-07-30 14:02 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
Arctic™ Fuji Apple
Transformation event
NF872
Unique identifier
OKA-NBØØ3-1
Developer(s)
Neal Carter
Okanagan Specialty Fruits Inc.
PO Box 1533
Summerland, BC
Canada, V0H 1Z0
Description
The modified Fuji apple uses RNA interference to suppress polyphenol oxidase activity, which results in a reduction in browning of the fruit. Enzymatic browning of the apple can cause changes to the nutrition and flavour profile of the fruit, as well as resulting in losses post-harvest. The modified apples did not show differences in nutrition, agronomic traits or pest susceptibility compared to the unmodified variety.
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.
Malus domestica - Apple, MALDO
Point of collection or acquisition of the recipient organism
Commercial apple cultivar Fuji
Related LMOs
OKA-NBØØ1-8 - Arctic™ "Golden Delicious" Apple
Neal Carter Resistance to antibiotics - Kanamycin
OKA-NBØØ2-9 - Arctic™ "Granny Smith" Apple
Neal Carter Resistance to antibiotics - Kanamycin
Characteristics of the transformation process
Vector
pGEN-03
Techniques used for the modification
  • Agrobacterium-mediated DNA transfer
Genetic elements construct
 
Nopaline Synthase Gene Promoter
0.31 Kb
 
 
Neomycin Phosphotransferase II
0.98 Kb
 
 
Nopaline Synthase Gene Terminator
0.26 Kb
 
 
CaMV 35S promoter
0.70 Kb
 
 
5' Untranslated Leader of AMV RNA4
0.00 Kb
 
 
PPO Suppression Transgene
1.82 Kb
 
 
Nopaline Synthase Gene Terminator
0.26 Kb
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
The Arctic Fuji apple contains two introduced gene cassettes: Escherichia coli neomycin phosphotransferase II (nptII) and polyphenol oxidase (PPO) supression cassettes.

Transcription of NPTII commences from the Agrobacterium tumefaciens nopaline synthase gene (nos) promoter and terminates at the nos terminator. The gene is used as a selection marker for the selecting transformants.

The apple (Malus domestica) PPO suppression construct is under the control of the Cauliflower Mosaic Virus (CaMV) 35S promoter, the 5' untranslated leader sequence from the Alfalfa Mosaic Virus RNA4, and the nos terminator. The CaMV promoter contains a duplicated enhancer. The construct contains segments for the following apple PPO genes: PPO2 (394 bp), GPO2 (457bp), APO5 (457 bp), and pSR7 (453 bp). The transgene suppressor is placed in the sense orientation and is expected to form double stranded RNA (dsRNA) within the apple cells.

Note:
- Whole genome sequencing confirmed that three insertions had occurred on chromosomes 3, 13, and 17
- Three T-DNAs were inserted on chromosome 3, two T-DNAs were inserted on chromosome 13, and a single T-DNA was inserted on chromosome 147.
- One of the inserts (on chromosome 3) contained 1400 bp of the vector backbone, however, none of the functional elements were present
- The insertion on chromosome 17 contained a partial CaMV promoter, which is in reverse orientation (pointed towards the apple genome) and does not contain 127 bp of the CaMV 35S core promoter, the proximal or the medial regions.
LMO characteristics
Modified traits
How the expression of the gene(s) was affected
Transcription of the PPO transgene supression construct results in a transcript with homology to the following endogenous genes: PPO2, GPO2, APO5, and pSR7. Due to the homologous sequences, the transcripts can base pair to form dsRNA structures, which trigger an RNA interference response. These structures are recognized by the cell and processed into small interfering RNAs, which target transcripts with sequence homology for degradation. The resulting destruction of the endogenous mRNAs results in reduced levels of the PPO protein accumulation in the plastids and thus decreased PPO activity in the transformed apples.
Common use(s)
  • Food
Detection method(s)
External link(s)
Additional Information