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Living Modified Organism
(LMO)
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Poplar with modified lignin
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pHG8-CAD4
No
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Person:Prof.dr. Wout Boerjan, Departement PlantensysteembiologieTechnologiepark 927Gent,
B-9052, BelgiumPhone: +32 9 331 38 81,Fax:Email: wout.boerjan@psb.vib-ugent.be,Related OrganizationVIB-UGent ()Academic or research instituteTechnologiepark 927Gent,
B-9052, BelgiumPhone: +32 9 331 38 81,Fax:Email: wout.boerjan@psb.vib-ugent.be,
The genetically modified trees have an altered lignin composition resulting from the downregulation of the cinnamyl alcohol dehydrogenase (CAD) enzyme through RNA interference. CAD catalyzes the last step in the monolignol synthesis. The remaining CAD activity in the modified trees is about 15% of that in wildtype trees. The altered lignin composition has a positive effect on the ease with which the lignin can be broken down to gain access to the valuable sugar content in the cellulose and hemicellulose in the wood of the trees. A lowered CAD activity has been found to naturally occur in the U.S. in loblolly pine (Pinus taeda) and in wild black poplar in Europe with comparable effects on the wood composition.
The modified trees also carry the NPT-II selection marker gene allowing an easy selection of transformed plants using kanamycin.
EN
The modified trees also carry the NPT-II selection marker gene allowing an easy selection of transformed plants using kanamycin.
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-101269-4 Organism Populus tremula x Populus alba (Gray Poplar)Trees
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pHellsgate8
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- Agrobacterium-mediated DNA transfer
0.346 kb
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1.024 kb
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0.766 kb
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1.024 kb
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0.706 kb
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0.184 kb
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0.822 kb
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0.253 kb
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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-15001-5 Neomycin Phosphotransferase II | Escherichia coli (ECOLX)Protein coding sequence | Resistance to antibiotics (Kanamycin)
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BCH-GENE-SCBD-115133-1 Cinnamyl alcohol dehydrogenase | Populus tremula x Populus alba (Gray Poplar)Double-stranded RNA
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BCH-GENE-SCBD-100287-7 CaMV 35S promoter | Cauliflower mosaic virus (CaMV)Promoter
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BCH-GENE-SCBD-103123-6 Pyruvate orthophosphate dikinase, Intron 3 | Flaveria trinervia (Clustered Yellowtops, speedyweed, flaveria, yellow twinstem)Intron
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BCH-GENE-SCBD-100271-5 Octopine Synthase Gene Terminator | Agrobacterium tumefaciens (Agrobacterium)Terminator
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BCH-GENE-SCBD-100270-6 Nopaline Synthase Gene Promoter | Agrobacterium tumefaciens (Agrobacterium)Promoter
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BCH-GENE-SCBD-100269-8 Nopaline Synthase Gene Terminator | Agrobacterium tumefaciens (Agrobacterium)Terminator
RNA interference cassette:
Transcription of the hairpin RNA (HpRNA) is under control of the Cauliflower Mosaic Virus (CaMV) 35S promoter and the Agrobacterium tumefaciens octopine synthase terminator. The transcript is expected to contain an inverted repeat of the Populus tremula x Populus alba cinnamyl alcohol dehydrogenase (CAD) gene separated by a segment sourced from Flaveria trinervia pyruvate orthophosphate dikinase intron 3, which is essential to form a loop that allows the inverted repeats to base pair and form the RNA secondary structure after transcription. Then the hpRNA is sufficient to trigger an RNAi response. Due to the RNAi response, the hpRNA is processed into small interfering RNA (siRNA) and thus no translation to protein is expected to occur.
Selectable marker:
The pHellsgate8 vector also contains a selectable marker for kanamycin resistance. The Escherichia coli neomycin phosphotransferase II gene is under transcriptional control of the A. tumefaciens nopaline synthase promoter and terminator.
EN
Transcription of the hairpin RNA (HpRNA) is under control of the Cauliflower Mosaic Virus (CaMV) 35S promoter and the Agrobacterium tumefaciens octopine synthase terminator. The transcript is expected to contain an inverted repeat of the Populus tremula x Populus alba cinnamyl alcohol dehydrogenase (CAD) gene separated by a segment sourced from Flaveria trinervia pyruvate orthophosphate dikinase intron 3, which is essential to form a loop that allows the inverted repeats to base pair and form the RNA secondary structure after transcription. Then the hpRNA is sufficient to trigger an RNAi response. Due to the RNAi response, the hpRNA is processed into small interfering RNA (siRNA) and thus no translation to protein is expected to occur.
Selectable marker:
The pHellsgate8 vector also contains a selectable marker for kanamycin resistance. The Escherichia coli neomycin phosphotransferase II gene is under transcriptional control of the A. tumefaciens nopaline synthase promoter and terminator.
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BCH-GENE-SCBD-115133-1 Cinnamyl alcohol dehydrogenase | Populus tremula x Populus alba (Gray Poplar)Double-stranded RNA
Post-transcription, the inverted repeat segments of the poplar cinnamyl alcohol dehydrogenase (CAD) basepair to form a double stranded region separated a loop derived from the Flaveria trinervia pyruvate orthophosphate dikinase intron 3. The double strandedness of the hpRNA is recognized by host cell RNA interference machinery; DICER processes the hpRNA into siRNA molecules (21-24 base pairs in length), which then associate with ARGONAUTE to form the RISC complex. The siRNA are then used to guide host machinery to degrade transcripts with sequence homology. Thus, the host machinery targets the endogenous CAD transcripts, resulting in silencing of CAD gene expression.
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- Research
- Timber
- Fiber/textile
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Three lines have been created with the same genetics: pHG8-CAD4, pHG8-CAD19 and pHG8-CAD24. Thus, the transformation event listed above represents a placeholder and will be updated should more information become available.
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