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Modified Organism
SHD-06101-3 - Rose modified for purple colouration (OS10036101)
Record information and status
Record ID
115387
Status
Published
Date of creation
2020-02-12 06:35 UTC (schandler@florigene.com.au)
Date of publication
2020-02-12 15:53 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
Rose modified for purple colouration (OS10036101)
Transformation event
10036101
Unique identifier
SHD-06101-3
Developer(s)
Dr Yukihisa Katsumoto
Principal Researcher
Research Institute
Suntory Holdings Limited (SHD)
1-1-1 Wakayamadai, Shimamoto-cho
Mishima-gun, Osaka
Japan, 618-8503
Phone:+81 75 962 9132
Fax:+81 75 962 3791
Email:Yoshikazu_Tanaka@suntory.co.jp,Yukihisa_Katsumoto@suntory.co.jp
Description
Rose OS10036101 flowers have a mauve colour due to the biosynthesis of the anthocyanin pigment delphinidin, which is not produced in non-transgenic rose. Viola sp. flavonoid 3', 5'-hydroxylase (F3'5'H) and iris dihydroflavonol reductase (DFR) were inserted to produce this blue-coloured delphinidin anthocyanin pigment. F3'5'H directs the production of an intermediate dihydromyricetin, which is then utilized as a substrate by iris DFR to produce delphinidin. A third gene gene cassette silences the expression of the endogenous rose DRF, which is responsible for the production of cyanidin, through the production of  double stranded RNA to cause RNA interference and thus prevent cyanidin production. The intermediate metabolites are then used for delphinidin biosynthesis instead. The rose additionally contains a selectable marker (kanamycin resistance), E. coli neomycin phosphotransferase II, for the selection of transformants during cloning.

A related LMO( http://bch.cbd.int/database/record.shtml?documentid=115379) has been generated using the same transformation vector and parent variety. That transformation event differs from SHD-06101-3 in the concentration of delphnidin in petals, resulting in a slightly different shade of flower colour. There are also slight differences in petal number and stem length.
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.
Rosa hybrida - Rose, ROSHC
Related LMOs
SHD-03701-6 - Rose modified for mauve colouration (Rose OS10033701)
Dr Yukihisa Katsumoto Changes in quality and/or metabolite content - Pigmentation / Coloration, Flavonoids (e.g. anthocyanin) Resistance to antibiotics - Kanamycin
Show detection method(s)
Characteristics of the transformation process
Vector
pSFL1003
Techniques used for the modification
  • Agrobacterium-mediated DNA transfer
Genetic elements construct
 
Nopaline Synthase Gene Promoter
0.32 Kb
 
 
Neomycin Phosphotransferase II
0.97 Kb
 
 
Nopaline Synthase Gene Terminator
0.26 Kb
 
 
CaMV Enhanced 35S promoter
0.76 Kb
 
 
Dihydroflavonol-4-reductase
1.26 Kb
 
 
D8 gene terminator
0.83 Kb
 
 
CaMV Enhanced 35S promoter
0.75 Kb
 
 
Dihydroflavonol-4-reductase
0.20 Kb
 
 
Dihydroflavonol-4-reductase
0.10 Kb
 
 
Mannopine synthase gene terminator
0.69 Kb
 
 
CaMV Enhanced 35S promoter
0.75 Kb
 
 
Flavonoid 3’, 5’-hydroxylase gene
1.78 Kb
 
 
Nopaline Synthase Gene Terminator
0.28 Kb
 
Further details
Notes regarding the genetic elements introduced or modified in this LMO
Iris DFR
Transcription of the Iris sp. dihydroflavonol-4-reductase (DFR) is under the control of a Cauliflower Mosaic Virus (CaMV) 35S enhanced promoter and a Petunia hybrid phospholipid transfer protein homologue (D8) terminator. Transcription is expected to be constitutive and at high levels due to the CaMV 35S enhanced promoter.

RNAi cassette
The RNA interference (RNAi) cassette was designed to target the endogenous dihydroflavonol-4-reductase (DFR) gene. Transcription begins from the CaMV 35S enhanced promoter and terminates at an Agrobacterium tumefaciens mannopine synthase terminator. The transcript is expected to contain two complementary sections of DFR, which are expected to form a hairpin RNA (hpRNA) to trigger an RNAi response. Thus, no protein is expected to be produced from this transcript (see "Other gene(s) whose expression was affected by the transformation").

Pansy F3'5'H
The Viola sp. flavonoid 3', 5'-hydroxylase (F3'5'H) gene is under the transcriptional control of a CaMV 35S enhanced promoter and an A. tumefaciens nopaline synthase terminator. Transcription is expected to be constitutive, occurring at elevated levels due to the CaMV 35S enhanced promoter.

Selectable marker
Transcription of the Escherichia coli neomycin phosphotransferase II gene is under the control of the A. tumefaciens nopaline synthase gene promoter and terminator.

Note:
- Iris sp. dihydroflavonol-4-reductase sequence corresponds to the cDNA of the transcript
- Viola sp. flavonoid 3', 5'-hydroxylase sequence corresponds to the cDNA of the transcript
- None of the elements have been modified from the sequences registered in the BCH genetic element registry.

For more details regarding the vector, kindly refer to the supplementary information provided.
LMO characteristics
Modified traits
  • Selectable marker genes and reporter genes
How the expression of the gene(s) was affected
After transcription of the sense and anti-sense segments of the Rosa hybrida dihydroflavonol reductase (DFR), the transcript is expected to form a hairpin structure due to base pairing. This gives the transcript double strandedness, which elicits an RNAi response. The hpRNA complexes with DICER, which processes the hpRNA into small interfering RNA (siRNA). These siRNA then associate with ARGONAUTE proteins to form the RNA induced silencing complex, which actively targets the endogenous DFR transcripts for degradation and results in a reduction of DFR expression.
Common use(s)
  • Ornamental
Detection method(s)
Additional information
The event may be identified by southern analysis.
Additional Information
Additional Information
More deails of the vector are provided in the attached word document.
Relevant literature;

Flower colour and cytochromes P450
Yoshikazu Tanaka and Filippa Brugliera
Transactions of the Royal Society of London B: Biological Sciences 368;1612: 20120432 (2013)

Katsumoto, Y., Fukuchi-Mizutani, M., Fukui, Y., Brugliera, F., Holton, T. A., Karan, M. and Tanaka, Y. (2007). Engineering of the rose flavonoid biosynthetic pathway successfully generated blue-hued flowers accumulating delphinidin. Plant and Cell Physiology 48; 1589-1600.