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Living Modified Organism
(LMO)
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.
Disease-resistant, low-reducing sugars and reduced browning potato
EN
BG25
Yes
SPS-ØBG25-7
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Person:SPS International5369 West Irving StreetBoise, Idaho
83706, United States of AmericaPhone: +1 (208) 780-6066,Fax:Email: simplot@simplot.com,Website:Related OrganizationJ.R. Simplot Company ()5369 West Irving StreetBoise, Idaho
83706, United States of AmericaPhone: +1 (208) 780-6066,Fax:Email: simplot@simplot.com,Website: -
Organization:J.R. Simplot Company ()Private sector (business and industry)5369 West Irving StreetBoise, ID
83706, United States of AmericaPhone: +1 (208) 780-6066,Fax:Email:Website:
Solanum tuberosum was modified to confer tolerance to late blight disease caused by the fungus Phytophthora infestans and resistance to Potato Virus Y as well as lower levels of reducing sugars and a reduction of polyphenol oxidase. The LM potato was developed to provide durable protection against late blight and Potato virus Y, while also reducing the levels of reducing sugars and polyphenol oxidase activity that contribute to darkening and black spot bruising in tubers. To achieve tolerance to late blight disease, BG25 expresses three resistance (R) proteins, VNT1, AMR3, and BLB2, encoded by the genes Rpi-vnt1, Rpi-amr3, and Rpi-blb2, derived from wild Solanum species, S. venturii, S. americanum, and S. bulbocastanum, respectively. Transcription of these genes produces a Coiled-Coil Nucleotide-Binding Leucine-Rich Repeat (CC-NB-LRR) resistance protein that recognizes specific Phytophthora infestans effector proteins and activates defense pathways. To confer resistance to Potato virus Y, the LM potato expresses inverted repeat sequences of the viral coat protein gene (PVY-CP) derived from Potato virus Y, which trigger the plant’s RNA interference (RNAi) pathway and block viral replication of the viral coat protein. To confer the low-reducing sugar trait, BG25 expresses inverted repeat sequences from endogenous Solanum tuberosum var. Ranger Russet potato genes that silence vacuolar invertase (VInv) gene, thereby blocking the conversion of sucrose into glucose and fructose and lowering their accumulation during storage. To confer the reduced browning trait, the LM potato expresses inverted repeat DNA sequence from the wild edible potato, Solanum verrucosum, that silence the polyphenol oxidase gene, blocking the conversion of colourless polyphenols in the plant tissue to coloured quinones which produce dark melanin pigments. For selection of putative transformants during the transformation stage, a modified acetolactate synthase gene (StmAls) from S. tuberosum was also incorporated as a selectable marker, conferring tolerance to herbicide imidazolinone.
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.
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BCH-ORGA-SCBD-12106-6 Organism Solanum tuberosum (Potato, SOLTU)Crops
Russet Burbank
EN
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SPS-ØØF10-7 - Innate™ Ranger Russet Potato| J.R. Simplot Company | Changes in quality and/or metabolite content (Pigmentation / Coloration, Protein and amino acids)
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SPS-ØØØZ6-5 - Elevate potato| J.R. Simplot Company | Changes in quality and/or metabolite content (Pigmentation / Coloration, Protein and amino acids), Reduced acrylamide potential, Reduced enzymatic browning, Resistance to diseases and pests (Fungi), Resistance to Phytophthora infestans
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SPS-ØØX17-5 - Innate® Acclimate Ranger Russet Potato| J.R. Simplot Company | Changes in quality and/or metabolite content (Carbohydrates, Pigmentation / Coloration, Protein and amino acids), Resistance to diseases and pests (Fungi)
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SPS-ØØE56-7 - Low acrylamide potential, reduced browning Russet potato| J.R. Simplot Company | Changes in quality and/or metabolite content (Carbohydrates, Pigmentation / Coloration, Protein and amino acids)
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SPS-ØØØW8-4 - Innate™ Russet Burbank Potato| J.R. Simplot Company | Changes in physiology and/or production, Changes in quality and/or metabolite content (Carbohydrates, Pigmentation / Coloration, Protein and amino acids), Resistance to diseases and pests
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SPS-ØØF37-7 - Innate™ Ranger Russet Potato| J.R. Simplot Company | Changes in quality and/or metabolite content (Pigmentation / Coloration, Protein and amino acids)
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SPS-ØØE24-2 - Innate™ Russet Burbank Potato| J.R. Simplot Company | Changes in quality and/or metabolite content (Pigmentation / Coloration, Protein and amino acids)
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SPS-ØØE12-8 - Innate™ Russet Burbank Potato| J.R. Simplot Company | Changes in quality and/or metabolite content (Pigmentation / Coloration, Protein and amino acids)
pSIM4363
EN
- Agrobacterium-mediated DNA transfer
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Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
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BCH-GENE-SCBD-260835-2 Ubiquitin 7 promoter | Solanum tuberosum (Potato, SOLTU)Promoter
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BCH-GENE-SCBD-260836-3 Ubiquitin 7 intron | Solanum tuberosum (Potato, SOLTU)Intron
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BCH-GENE-SCBD-111058-1 Ubiquitin-ribosomal protein gene terminator | Solanum tuberosum (Potato, SOLTU)Terminator
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BCH-GENE-SCBD-260837-2 Ubiquitin 7 exon 1 | Solanum tuberosum (Potato, SOLTU)Protein coding sequence | Enhanced expression
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BCH-GENE-SCBD-101415-9 Ti plasmid left border repeat | Agrobacterium tumefaciens (Agrobacterium)Plasmid vector
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BCH-GENE-SCBD-280571-3 Acetolactate synthase | Solanum tuberosum (Potato, SOLTU)Protein coding sequence | Resistance to herbicides (Imidazolinone, Sulfonylurea)
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BCH-GENE-SCBD-109063-2 Phytophthora infestans Resistance gene 1 terminator | Solanum venturii (Wild Potato, SOLVN)Terminator
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BCH-GENE-SCBD-102155-7 Phytophthora infestans Resistance gene 1 | Solanum venturii (Wild Potato, SOLVN)Protein coding sequence | Resistance to diseases and pests (Fungi)
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BCH-GENE-SCBD-109062-2 Phytophthora infestans Resistance gene 1 Promoter | Solanum venturii (Wild Potato, SOLVN)Promoter
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BCH-GENE-SCBD-280720-1 Phytophthora Resistance gene 3 Promoter | Solanum americanum (American black nightshade, Black nightshade berries)Promoter
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BCH-GENE-SCBD-280721-1 Phytophthora Resistance gene 3 | Solanum americanum (American black nightshade, Black nightshade berries)Protein coding sequence | Resistance to diseases and pests (Fungi)
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BCH-GENE-SCBD-280722-1 Phytophthora Resistance gene 3 Terminator | Solanum americanum (American black nightshade, Black nightshade berries)Terminator
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BCH-GENE-SCBD-103775-2 Phytophthora infestans Resistance gene 2 Promoter | Solanum bulbocastaneum (Nightshade, Ornamental nightshade)Promoter
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BCH-GENE-SCBD-41318-4 Phytophthora infestans Resistance gene 2 | Solanum bulbocastaneum (Nightshade, Ornamental nightshade)Protein coding sequence | Resistance to diseases and pests (Phytophthora infestans resistance)
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BCH-GENE-SCBD-103776-2 Phytophthora infestans Resistance gene 2 Terminator | Solanum bulbocastaneum (Nightshade, Ornamental nightshade)Terminator
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BCH-GENE-SCBD-14997-6 Granule bound starch synthase gene promoter | Solanum tuberosum (Potato, SOLTU)Promoter
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BCH-GENE-SCBD-109064-2 Vacuolar invertase gene | Solanum tuberosum (Potato, SOLTU)Protein coding sequence | Changes in quality and/or metabolite content (Carbohydrates)
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BCH-GENE-SCBD-106424-3 Polyphenol oxidase 5 gene | Solanum verrucosum (SOLVR)Protein coding sequence | Changes in quality and/or metabolite content (Pigmentation / Coloration)
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BCH-GENE-SCBD-115073-1 Spacer sequence | Solanum tuberosum (Potato, SOLTU)Spacer sequence
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BCH-GENE-SCBD-106420-1 ADP glucose pyrophosphorylase gene promoter | Solanum tuberosum (Potato, SOLTU)Promoter
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BCH-GENE-SCBD-15020-3 PVY coat protein | Potato virus Y (PVY)Protein coding sequence | Resistance to diseases and pests (Viruses, Potato virus Y (PVY))
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BCH-GENE-SCBD-115074-1 Spacer sequence | Solanum tuberosum (Potato, SOLTU)Spacer sequence
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BCH-GENE-SCBD-101416-7 Ti plasmid right border repeat | Agrobacterium tumefaciens (Agrobacterium)Plasmid vector
Information on the inserted DNA sequences
The transforming plasmid pSIM4363 carried a transfer DNA (T-DNA) comprising of six cassettes which contain a modified acetolactate synthase (StmAls) gene from Solanum tuberosum, Resistance-genes (R-genes): Rpi-vnt1 from Solanum venturii, Rpi-amr3 from Solanum americanum and Rpi-blb2 from Solanum bulbocastanum, RNA interference (RNAi) cassettes with inverted repeat DNA sequences from Solanum tuberosum, Solanum verrucosum targeting vacuolar invertase (VInv) and polyphenol oxidase (Ppo), and from potato plants infected with PVY strain N targeting Potato virus Y coat protein (PVY-CP), respectively.
(1) Selectable marker
Transcription of the modified acetolactate synthase (StmALS) coding sequence was directed by the polyubiquitin promoter (pUbi7) from Solanum tuberosum var. Ranger Russet which includes a gene intron StUbi7 and monomer from S. tuberosum that enhance transcriptional activity. The intron enhances expression of StmALS and is spliced out before translation, and the ubiquitin monomer is expressed as an N-terminal fusion peptide with the StmALS protein then cleaved off. The ubiquitin terminator (tUbi3) from Solanum tuberosum was used for transcript termination and polyadenylation. Due to the constitutive nature of the polyubiquitin promoter, moderate to high levels of expression are expected in most plant tissues. This cassette was used only for selection and was not intended as a commercial trait.
R-gene cassettes (2), (3) and (4)
These genes encode intracellular immune receptors that enable the potato to recognize specific Phytophthora infestans effector proteins and activate immune response in the LM potato. Expression is localized in leaf and tuber tissues and inducible upon pathogen infection, providing protection against late blight.
Transcription of the modified acetolactate synthase (StmALS) coding sequence was directed by the polyubiquitin promoter (pUbi7) from Solanum tuberosum var. Ranger Russet which includes a gene intron StUbi7 and monomer from S. tuberosum that enhance transcriptional activity. The intron enhances expression of StmALS and is spliced out before translation, and the ubiquitin monomer is expressed as an N-terminal fusion peptide with the StmALS protein then cleaved off. The ubiquitin terminator (tUbi3) from Solanum tuberosum was used for transcript termination and polyadenylation. Due to the constitutive nature of the polyubiquitin promoter, moderate to high levels of expression are expected in most plant tissues. This cassette was used only for selection and was not intended as a commercial trait.
R-gene cassettes (2), (3) and (4)
These genes encode intracellular immune receptors that enable the potato to recognize specific Phytophthora infestans effector proteins and activate immune response in the LM potato. Expression is localized in leaf and tuber tissues and inducible upon pathogen infection, providing protection against late blight.
(2) The Rpi-vnt1 gene from Solanum venturii was regulated by its native promoter and terminator sequences (pVnt1 and tVnt1).
(3) The Rpi-amr3 gene expression from Solanum americanum was under the control of its native pAmr3 promoter and tAmr3 terminator.
(4) The Rpi-blb2 gene from Solanum bulbocastanum was transcribed under the control of the native pBlb2 promoter and tBlb2 terminator.
RNAi cassettes (5) and (6)
The modified potato contains two RNAi cassettes, one designed to target vacuolar invertase (VInv) and polyphenol oxidase (Ppo) and another targeting Potato virus Y coat protein (PVY-CP). The coding sequences from these cassettes contain two complementary (inverted) sequences with homology to the target gene or transcript, separated by an intron. Upon transcription, the intron functions as a loop and the sequences base pair due to complementarity (sense-antisense orientations), forming a hairpin RNA structure (hpRNA). The hpRNA acts as double stranded RNA (dsRNA) which will trigger an RNAi response in the cell, which will process the hpRNA into small interfering RNA. Due to the processing of the hairpin RNA, no translation is expected to occur from these gene cassettes.
RNAi cassettes (5) and (6)
The modified potato contains two RNAi cassettes, one designed to target vacuolar invertase (VInv) and polyphenol oxidase (Ppo) and another targeting Potato virus Y coat protein (PVY-CP). The coding sequences from these cassettes contain two complementary (inverted) sequences with homology to the target gene or transcript, separated by an intron. Upon transcription, the intron functions as a loop and the sequences base pair due to complementarity (sense-antisense orientations), forming a hairpin RNA structure (hpRNA). The hpRNA acts as double stranded RNA (dsRNA) which will trigger an RNAi response in the cell, which will process the hpRNA into small interfering RNA. Due to the processing of the hairpin RNA, no translation is expected to occur from these gene cassettes.
(5) Transcription of the RNAi cassette targeting vacuolar invertase (VInv) and polyphenol oxidase (Ppo) was directed by the granule-bound starch synthase (pGbss) and ADP-glucose pyrophosphorylase (pAgp) promoters from S. tuberosum, both of which are tuber-active and ensure expression in starch-accumulating tissues. The cassette contains sense and antisense fragments of VInv from S. tuberosum and the 3′ untranslated region (UTR) of Ppo from S. verrucosum separated by an intron from Gbss. The results in silencing of endogenous VInv and Ppo mRNA, which reduces glucose and fructose accumulation during storage and minimizes black spot bruising.
(6) The RNA interference cassette targeting the Potato virus Y coat protein (PVY-CP) is regulated by the polyubiquitin promoter (pUbi7) from S. tuberosum, which includes an intron and ubiquitin monomer that enhance transcriptional activity, and terminated by the ubiquitin terminator (tUbi3). The cassette contains sense and antisense fragments of the PVY coat protein gene separated by an intron derived from Gbss. The promoter’s constitutive activity allows expression in multiple tissues, preventing systemic viral replication and conferring durable PVY protection.
The right and left border regions of pSIM4363
The right border region of the plasmid contains a 25 base pair border repeat recognized by the Agrobacterium VirD1/VirD2 endonuclease complex, accompanied by an adjacent buffer sequence of 161 base pairs. This region serves as the primary cleavage site to release the single-stranded T-DNA from the plasmid. This occurs with the VirD2 enzyme introducing a site-specific nick and remaining attached to the 5′ end of the newly created T-strand. The buffer sequence supports the primary cleavage by providing structural stability and ensuring precise excision and, thus, accurate initiation of DNA transfer during integration into the host genome (GenBank: AY566555). Using the remaining intact strand of the plasmid as a template, Agrobacterium’s replication machinery begins synthesizing a new DNA strand, and the VirD2–T-strand complex is then displaced toward the left border. The left border region consists of a 25 base pair border repeat together with a 162 base pair buffer sequence is also recognized by the VirD1/VirD2 complex. It functions as the termination site for T-DNA processing, marking the endpoint of the single-stranded DNA. When VirD2 reaches the left border, it performs a second, precise cut that releases the entire T-DNA region from the plasmid as a single-stranded molecule. The presence of the buffer region helps reduce the effects of truncations or minimize loss of sequence during transfer and integration, ensuring that the complete T-DNA cassette is successfully incorporated into the plant genome (GenBank: AY566555).
Notes:
- For more information on RNAi, kindly refer to the "Please describe how the expression of the gene(s) was affected" section of this record below;
- Illumina Next Generation Sequencing, Sanger sequencing and droplet digital PCR analyses confirmed absence of backbone sequences (a small number of reads mapped to backbone sequences due to the presence of two endogenous potato sequences in the plasmid backbone) and identified a single intact T-DNA insertion (29,957 bp) at a single genomic locus on chromosome 12;
- All cassettes maintained expected order and orientation (minor truncations only at border sequences due to double-stranded break repair mechanisms);
- ddPCR and PCR analyses on leaf-derived genomic DNA demonstrated heritable stability across generations;
- Bioinformatic screening detected no expressed ORFs or the three R-genes with homology to known allergens or toxins;
- Reverse transcription quantitative real-time PCR verified the expected RNA transcript levels of Vlnv, Ppo and the three R-genes in tuber tissues, immunoblot analysis to confirm the expression level of StmALS gene in leaf and tuber tissues, and a PVY resistance assay to confirm the absence of viral CP RNA transcript levels.
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BCH-GENE-SCBD-109064-2 Vacuolar invertase gene | Solanum tuberosum (Potato, SOLTU)Protein coding sequence | Changes in quality and/or metabolite content (Carbohydrates)
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BCH-GENE-SCBD-106424-3 Polyphenol oxidase 5 gene | Solanum verrucosum (SOLVR)Protein coding sequence | Changes in quality and/or metabolite content (Pigmentation / Coloration)
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BCH-GENE-SCBD-15020-3 PVY coat protein | Potato virus Y (PVY)Protein coding sequence | Resistance to diseases and pests (Viruses, Potato virus Y (PVY))
RNA interference
In RNA interference (RNAi), two complementary (inverted) sequences with homology to a target gene or transcript are used to trigger gene silencing. An RNAi response is an anti-viral response triggered by the recognition of dsRNA. Host DICER recognizes dsRNA, cleaving the dsRNA into small interfering RNA (siRNA), roughly 21-23 bp long (size is host dependent). The siRNA is then bound by ARGONAUTE family proteins, which unwind the duplex, leaving a single strand of the siRNA and activating the RNA-induced silencing complex (RISC) complex. The RISC complex targets transcripts with homology to the siRNA and degrades them.
Upon transcription of the two vacuolar invertase and polyphenol oxidase, and Potato virus Y coat protein RNAi suppression cassettes, a hairpin RNA will be formed with double stranded secondary structure and then an RNA interference response will be triggered. The hairpin RNA will complex with DICER, which will cleave the hairpin RNA into small interfering RNA of 21 to 24 nucleotides in length. Following this, these double stranded small interfering RNAs will then associate with ARGONAUTE, which will unwind the duplex, leaving one strand associated with ARGONAUTE. This protein-RNA complex is now the activated RISC, which will then use the remaining strand of siRNA as a guide to target messenger RNA (mRNA) that has sequence homology to it. Thus, the cellular response is directed at mRNA encoding full-length vacuolar invertase, polyphenol oxidase and Potato virus Y coat protein, resulting in degradation of the mRNA, a reduction in the overall translation of the protein, and silencing of the expression of the endogenous genes.
In RNA interference (RNAi), two complementary (inverted) sequences with homology to a target gene or transcript are used to trigger gene silencing. An RNAi response is an anti-viral response triggered by the recognition of dsRNA. Host DICER recognizes dsRNA, cleaving the dsRNA into small interfering RNA (siRNA), roughly 21-23 bp long (size is host dependent). The siRNA is then bound by ARGONAUTE family proteins, which unwind the duplex, leaving a single strand of the siRNA and activating the RNA-induced silencing complex (RISC) complex. The RISC complex targets transcripts with homology to the siRNA and degrades them.
Upon transcription of the two vacuolar invertase and polyphenol oxidase, and Potato virus Y coat protein RNAi suppression cassettes, a hairpin RNA will be formed with double stranded secondary structure and then an RNA interference response will be triggered. The hairpin RNA will complex with DICER, which will cleave the hairpin RNA into small interfering RNA of 21 to 24 nucleotides in length. Following this, these double stranded small interfering RNAs will then associate with ARGONAUTE, which will unwind the duplex, leaving one strand associated with ARGONAUTE. This protein-RNA complex is now the activated RISC, which will then use the remaining strand of siRNA as a guide to target messenger RNA (mRNA) that has sequence homology to it. Thus, the cellular response is directed at mRNA encoding full-length vacuolar invertase, polyphenol oxidase and Potato virus Y coat protein, resulting in degradation of the mRNA, a reduction in the overall translation of the protein, and silencing of the expression of the endogenous genes.
- Food
EN
EN
- EUginius: BG25 potato - detailed information (SPS-ØBG25-7) [URL] [ English ]
- OECD BioTrack Product Database: SPS-ØBG25-7 [URL] [ English ]
- FSANZ: A1289 Safety assessment - Food derived from disease-resistant, low-reducing sugars and reduced browning potato line BG25 [PDF] [ English ]
- FSANZ: A1289 - Food derived from disease-resistant, low-reducing sugars and reduced browning potato line BG25 [URL] [ English ]
- FAO: SPS-ØBG25-7 details [URL] [ English ]
| Record type | Field | Record(s) | |
|---|---|---|---|
| Risk Assessment generated by a regulatory process | Living modified organism(s) | 1 | |
| Country's Decision or any other Communication | LMO identification | 1 | |
