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Modified Organism
FDP-ØØ114-5 - Pineapple modified for altered color and reduced ripening
Record information and status
Record ID
111940
Status
Published
Date of creation
2017-05-29 15:07 UTC (dina.abdelhakim@cbd.int)
Date of publication
2017-05-29 15:07 UTC (dina.abdelhakim@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
Pineapple modified for altered color and reduced ripening
Transformation event
EF2-114
Unique identifier
FDP-ØØ114-5
Developer(s)
Del Monte Fresh Produce Company
241 Sevilla Avenue
Coral Gables, Florida
United States of America, 33134
Description
Pineapple was genetically modified to alter the color of the fruit from yellow to pink through accumulation of lycopene in place of β-carotene, and to alter flower control.

The modification of fruit colour was carried out through the insertion of a gene coding for phytoene synthase, which is involved in a key step in lycopene synthesis and two RNAi constructs that were designed to suppress endogenous expression of each of lycopene β-cyclase and lycopene ε-cyclase which convert lycopene to other carotenoids.

Additionally the suppression of flowering was carried out through the insertion of an RNAi construct that is intended to suppress endogenous expression of 1-aminocyclopropane-1-carboxylic acid synthase, the penultimate step in ethylene biosynthesis.
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.
Ananas comosus - Pineapple, ANACO
Point of collection or acquisition of the recipient organism
Variety: MD2
Characteristics of the transformation process
Vector
pHCW.T-7 and pHCWflACC3ʹ-2
Techniques used for the modification
  • Agrobacterium-mediated DNA transfer
Genetic elements construct
 
Epoxide hydrolase promoter
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Tetrameric ubiquitin gene promoter
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Acetohydroxy acid synthase gene
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Acetohydroxy acid synthase gene terminator
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Bromelain inhibitor gene promoter
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Phytoene synthase gene
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Tetrameric ubiquitin gene terminator
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Bromelain inhibitor gene promoter
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Lycopene beta cyclase gene
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Light-inducible tissue-specific LS1 intron 2
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Lycopene beta cyclase gene
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Tetrameric ubiquitin gene terminator
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Bromelain inhibitor gene promoter
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Lycopene epsilon-cyclase gene
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Light-inducible tissue-specific LS1 intron 2
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Lycopene epsilon-cyclase gene
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Tetrameric ubiquitin gene terminator
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Tetrameric ubiquitin gene promoter
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1-aminocyclopropane-1-carboxylic acid synthase gene
0.00 Kb
 
 
Light-inducible tissue-specific LS1 intron 2
0.00 Kb
 
 
1-aminocyclopropane-1-carboxylic acid synthase gene
0.00 Kb
 
 
Tetrameric ubiquitin gene terminator
0.00 Kb
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
DNA insert from vector pHCW.T-7

Vector pHCW.T-7 contains four expression cassettes:

The SuRBHra expression cassette, intended to function as a plant selectable marker, which is composed of:
* A constitutive fusion promoter EHS-Ubpp, comprising the promoter/5ʹ untranslated region (UTR) of the epoxide hydrolase (EHS) gene and the promoter/5ʹ UTR of the tetrameric ubiquitin (Ubi) gene, both isolated from pineapple
* SuRBHra, a mutant acetolactate synthase (ALS) gene from tobacco (Nicotiana tabacum)
* A transcription terminator sequence corresponding to the 3ʹ untranslated region of the tobacco ALS gene

The BRIp-PSY-Ubpter expression cassette intended to express the phytoene synthase protein, which is composed of:
* A modified promoter derived from the pineapple BRI gene
* The tangerine (Citrus unshiu) phytoene synthase (PSY) gene
* The transcription terminator sequence corresponding to the 3ʹ UTR and flanking region of the pineapple Ubi gene

The BRIp-bLcy RNAi-Ubpter expression cassette, intended to suppress expression of the pineapple lycopene β-cyclase protein, which is composed of:
* A modified promoter derived from the pineapple BRI gene
* A partial coding sequence of pineapple bLcy in the sense orientation,
* The intron 2 sequence of the potato (Solanum tuberosum) ST-LSI gene,
* A partial coding sequence of pineapple bLcy in the anti-sense orientation
* The transcription terminator sequence corresponding to the 3ʹ UTR and flanking region of the pineapple Ubi gene

The BRIp-eLcy RNAi-Ubpter expression cassette, intended to suppress expression of the pineapple lycopene ε-cyclase protein, which is composed of:
* A modified promoter derived from the pineapple BRI gene
* A partial coding sequence of pineapple eLcy in the sense orientation
* The intron 2 sequence of the potato (S. tuberosum) ST-LSI gene
* * A partial coding sequence of pineapple eLcy in the anti-sense orientation
The transcription terminator sequence corresponding to the 3ʹ UTR and flanking region of the pineapple Ubi gene

DNA insert from vector pHCWflACC3ʹ-2

Vector pHCWflACC3ʹ-2 contains two expression cassettes:

The SuRBHra expression cassette described above, intended as a plant selectable marker

The Ubpp-flACC3ʹ RNAi-Upbter expression cassette, intended to suppress expression of 1-aminocyclopropane-1-carboxylic acid synthase (ACS), which is composed of:
* A promoter sequence corresponding to the promoter/5ʹ region of the pineapple Ubi gene
* A partial coding sequence of pineapple meristem ACS gene flACC3ʹ in the sense orientation
* The intron 2 sequence of the potato (S. tuberosum) ST-LSI gene
* A partial coding sequence of pineapple meristem ACS gene flACC3ʹ in the anti-sense orientation
* The transcription terminator sequence corresponding to the 3ʹ UTR and flanking region of the pineapple Ubi gene.

Characteristics of the introduced DNA

Southern blot analysis was carried out to determine the characteristics and stability of the introduced DNA. Studies provided data that was consistent with four complete integrations of the pHCW.T-7 T-DNA, one irregular integration of the pHCWflACC3ʹ-2 T-DNA, and three partial integrations of the pHCWflACC3ʹ-2 right border region.

Furthermore, the data indicated that at least one intact copy of pHCW.T-7 is present in the EF2-114 genome, in addition to rearranged copies of pHCW.T-7, the exact structure of which could not be completely elucidated. In addition the data supports that no intact copies of pHCWflACC3ʹ-2 are present in EF2-114, and that extensive rearrangements of pHCWflACC3ʹ-2 genetic elements had occurred.

Copy number analysis was consistent with the integration of four copies of the pHCW.T-7 plasmid, of which two were complete. There was also evidence of one partial integration of the pHCWflACC3ʹ-2 plasmid.
LMO characteristics
Modified traits
Common use(s)
  • Food
Additional Information