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Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.).
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This investigation reports a protocol for transfer and expression of foreign chimeric genes in loblolly pine (Pinus taeda L.). Transformation was achieved by co-cultivation of mature zygotic embryos with Agrobacterium tumefaciens strain LBA4404 which harbored a binary vector (pBI121) including genes
Additional virulence genes and sonication enhance Agrobacterium tumefaciens-mediated loblolly pine transformation.
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Additional virulence (vir) genes in Agrobacterium tumefaciens and sonication were investigated for their impact on transformation efficiency in loblolly pine (Pinus taeda L.). Mature zygotic embryos of loblolly pine were co-cultivated with disarmed A. tumefaciens strain EHA105 containing either
Growth and differentiation of transgenic callus regulated by phytohormones and antibiotics in transformation of loblolly pine.
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Mature zygotic embryos of loblolly pine (Pinus taeda L.) were transformed by Agrobacterium tumefaciens strain LBA 4404 harbouring the plasmid pBI121 which carried the selectable marker gene, neomycin phosphotransferase II (npt II) controlled by the promoter of the nopaline synthase gene, and the
Seasonal changes in the transient expression of a 35S CaMV-GUS gene construct introduced into Scots pine buds.
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Seasonal changes in the transient expression of Beta-glucuronidase gene (GUS) driven by a constitutive 35S CaMV-promoter in Scots pine (Pinus sylvestris L.) buds were studied by the microprojectile DNA-delivery method. Buds were collected from 5-, 15- and 50-year-old trees. In buds from all age
Differential responses of the promoters from nearly identical paralogs of loblolly pine (Pinus taeda L.) ACC oxidase to biotic and abiotic stresses in transgenic Arabidopsis thaliana.
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Promoters from an ACC oxidase gene (PtACO1) and its nearly identical paralog (NIP) (PtACO2) of loblolly pine (Pinus taeda L.) were recovered from genomic DNA using PCR amplification. Transgenic Arabidopsis plants harboring genetic constructs from which beta-glucuronidase (GUS) expression was driven
Expression analysis of four Pinus radiata male cone promoters in the heterologous host Arabidopsis.
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Four male cone-specific promoters were isolated from the genome of Pinus radiata D. Don, fused to the beta-glucuronidase (GUS) reporter gene and analysed in the heterologous host Arabidopsis thaliana (L.) Heynh. The temporal and spatial activities of the promoters PrCHS1, PrLTP2, PrMC2 and PrMALE1
High-efficiency Agrobacterium-mediated transformation of Norway spruce (Picea abies) and loblolly pine (Pinus taeda).
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Agrobacterium-mediated gene transfer is the method of choice for many plant biotechnology laboratories; however, large-scale use of this organism in conifer transformation has been limited by difficult propagation of explant material, selection efficiencies and low transformation frequency. We have
Regeneration of transgenic loblolly pine (Pinus taeda L.) from zygotic embryos transformed with Agrobacterium tumefaciens.
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Embryos of 24 open-pollinated families of loblolly pine (Pinus teade L.) were used as explants to conduct in vitro regeneration. Then, Agrobacterium tumefaciens strain GV3101 harboring the plasmid pPCV6NFHygGUSINT was used to transform mature zygotic embryos of seven families of loblolly pine. The
The promoter of a cytosolic glutamine synthetase gene from the conifer Pinus sylvestris is active in cotyledons of germinating seeds and light-regulated in transgenic Arabidopsis thaliana.
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We have isolated and characterized a genomic clone encoding Scots pine (Pinus sylvestris) cytosolic glutamine synthetase (GS). The clone contains the 5' end half of the gene including part of the coding region and 980 bp upstream of the translation initiation codon. The major transcription start
A promoter from the loblolly pine PtNIP1;1 gene directs expression in an early-embryogenesis and suspensor-specific fashion.
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The PtNIP1;1 gene encodes an aquaglyceroporin that is expressed early in embryogenesis and appears to be expressed preferentially in the suspensor [V.T. Ciavatta et al. (2001) Plant Physiol 127:211-224]. An 899-bp fragment 5' to the PtNIP1;1 open reading frame (NIP(-899)) was cloned from loblolly
Transient expression from microprojectile-mediated DNA transfer in pinus taeda.
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Transfer of plasmid DNA to Pinus taeda L. (loblolly pine) cotyledon cells by microprojectile bombardment has been demonstrated using beta-glucuronidase (GUS). GUS histochemical staining indicated active enzyme in localized centers (blue spots) 24 hours after bombardment. GUS expression declined
Promoters for pregenomic RNA of banana streak badnavirus are active for transgene expression in monocot and dicot plants.
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Two putative promoters from Australian banana streak badnavirus (BSV) isolates were analysed for activity in different plant species. In transient expression systems the My (2105 bp) and Cv (1322 bp) fragments were both shown to have promoter activity in a wide range of plant species including
Sequence analysis and functional characterization of the promoter of the Picea glauca Cinnamyl Alcohol Dehydrogenase gene in transgenic white spruce plants.
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The enzyme Cinnamyl Alcohol Dehydrogenase (CAD) catalyses the last step of lignin monomer synthesis, and is considered as a molecular marker of cell wall lignification in different plants species. Here, we report the isolation and analysis of 5' flanking genomic DNA regions upstream to the CAD gene,
Stable Agrobacterium-mediated transformation of maritime pine based on kanamycin selection.
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An efficient transformation protocol based on kanamycin selection was developed for Agrobacterium-mediated transformation of maritime pine embryonal masses. The binary vector pBINUbiGUSint, which contained neomycin phosphotransferase II (nptII) as a selectable marker gene and β -glucuronidase (uidA)
Transgenic Pinus radiata from Agrobacterium tumefaciens-mediated transformation of cotyledons.
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A method for Agrobacterium tumefaciens-mediated transformation of Pinus radiata cotyledon explants was developed using commercially available open-pollinated seed. Pinus radiata is the most widely planted commercial conifer species in the Southern Hemisphere. Reports on transformation of this
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