Insect-resistant transgenic Pinus radiata

Grace, Lynette J.; Charity, Julia A.; Gresham, Belinda A.; Kay, Nod; Walter, Christian

doi:10.1007/s00299-004-0912-x

Cited by 60 publications

(16 citation statements)

References 38 publications

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“…to infect a diverse group of plant species has been well documented, expression of transgenes still remains difficult in the gymnosperms, even with newly developed super-virulent and ternary Agrobacterium strains [118][119][120]. In the case of these recalcitrant conifers, particle bombardment-mediated transformation has been utilized to introduce both linear and plasmid DNA constructs, with numerous reports of both transient and stable transformation in many of these gymnosperm species [121][122][123][124]. However, due to the non-receptive nature of gymnosperms to regenerate after transgene introduction, and the possibility of fragmented or multiple copy transformation events leading to transgene silencing, these recalcitrant conifers have lagged behind when compared to genetic engineering of agricultural crops.…”

Section: Transformation Of Taxus Sppmentioning

confidence: 99%

Advancements in the Understanding of Paclitaxel Metabolism in Tissue Culture

Vongpaseuth¹,

Roberts

2007

CPB

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Paclitaxel is a potent chemotherapeutic agent approved in the treatment of a variety of cancers, and under evaluation for the treatment of Alzheimer's and heart disease. Originally isolated from Taxus brevifolia, this highly substituted ring diterpenoid belongs to a family of plant secondary metabolites known as taxoids. Paclitaxel is currently supplied through both a semi-synthetic process and plant cell culture. Taxus spp. cell culture offers the potential to produce large amounts of paclitaxel and related taxoids, although variability in accumulation and low yields represent key limitations. Thus, intense efforts have been put forth towards understanding Taxus spp. metabolism to increase paclitaxel accumulation in cell culture. While elicitation and environmental optimization have provided some success in increasing paclitaxel accumulation in vitro, understanding metabolism of paclitaxel on the molecular level is essential for process optimization. Utilizing direct and indirect molecular techniques, a further understanding of paclitaxel biosynthesis has been gained, though knowledge into other aspects of paclitaxel global metabolism, such as regulation, transport, and degradation is lacking. Taxus spp. cell cultures are highly heterogeneous, displaying significant cell-cell variability in growth and paclitaxel accumulation. Information gathered on culture subpopulations as well as putative transcriptional bottlenecks in paclitaxel biosynthesis, coupled with successful transformation of Taxus spp. will allow for the targeted metabolic engineering of Taxus spp. or other model organisms for paclitaxel accumulation to ensure future supply of this important pharmaceutical.

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Section: Transformation Of Taxus Sppmentioning

confidence: 99%

Advancements in the Understanding of Paclitaxel Metabolism in Tissue Culture

Vongpaseuth¹,

Roberts

2007

CPB

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“…In particular, success with gymnosperms have been mixed (9). In the case of these recalcitrant conifers, such as Picea glauca, P. mariana, P. abies, Pinus radiata, P. pinaster, and Larix laricina, particle bombardment-mediated transformation has been utilized to introduce both linear and plasmid DNA constructs, with numerous reports of both transient and stable transformation (10)(11)(12)(13). However, because of the nonreceptive nature of gymnosperms to regenerate after transgene introduction and the possibility of fragmented or multiple copy transformation events leading to transgene silencing, metabolic engineering of these recalcitrant conifers has lagged compared to genetic engineering of agricultural crops.…”

Section: Introductionmentioning

confidence: 99%

Development of a Particle Bombardment-Mediated Transient Transformation System for Taxus spp. Cells in Culture

et al. 2007

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In developing and developed nations, plant cell culture systems are used to supply desirable compounds in lieu of chemical synthesis or natural extraction. When plant cell culture systems are unable to meet commercial demand, metabolic engineering offers a method to increase yields. However, to benefit from metabolic engineering approaches, effective transient transformation methods are required to rapidly identify and characterize key regulatory genes before intensive, time-consuming stable transformation efforts can proceed. This paper describes a particle bombardment-mediated transient transformation system for Taxus spp. in cell culture. Optimal parameters were established for the T. cuspidata cell line P991 and the T. canadensis cell line CO93D, resulting in reliable, efficient, transient expression of the firefly luciferase gene under control of the constitutive CaMV 35S promoter. Multiple bombardments and larger gold microcarriers (1.6 vs 1.0 microm in diameter) were particularly effective in increasing luciferase activity and in reducing variation among replicates. This particle bombardment-mediated transformation system was also shown to be capable of transiently expressing the DsRed and beta-glucuronidase reporter genes under the control of the maize ubiquitin and CaMV 35S promoters, respectively. With the ability to transiently transform Taxus spp. cell cultures using a variety of promoters and reporters, characterization of genes related to paclitaxel accumulation in culture can now proceed.

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“…In most of conifer species, the embryogenic system for genetic transformation is obtained from EM derived from immature zygotic embryos and they served as the best starting material for the successful transformation (Klimaszewska et al, 1997(Klimaszewska et al, , 2001Nigro et al, 2004;Grace et al, 2005;Tereso et al, 2006;Tang et al, 2007).…”

Section: Agrobacterium-mediated Transformationmentioning

confidence: 99%

Agrobacterium-mediated Transformation via Somatic Embryogenesis System in Korean fir (Abies koreana Wil.), A Korean Native Conifer

Lee¹,

Moon²,

Park

2014

Korean Journal of Plant Resources

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-This study was conducted to establish an efficient transformation system by using somatic embryogenesis in an important Korean native conifer, Korean fir (Abies koreana). Embryogenic masses were induced from mature zygotic embryos of the Korean fir on Schenk and Hildebrandt medium, which was supplemented with thidiazuron. For genetic transformation, the embryogenic masses were co-cultivated with a disarmed Agrobacterium tumefaciens strain C58/pMP90 containing the plasmid vector pBIV10 or LBA4404 containing the plasmid vector MP90. Both vectors contain the kanamycin resistance and beta-glucuronidase (GUS) reporter genes. A total of 48 lines of embryogenic masses were selected on mLV medium containing 50 μg/mL of kanamycin after 4 weeks of culture, following 3 days of co-cultivation with A. tumefaciens strain C58/pMP90 carrying pBIV10 (none of the lines was cultivated with strain LBA4404 carrying MP90). Quantitative real-time PCR was performed, and high levels of GUS transcripts were observed in the 48 putative transgenic lines; however, the control (non-transgenic line) showed negative results. Results of histochemical staining showed that the expression of the GUS reporter gene was observed in somatic embryos that developed from the embryogenic masses of all 48 lines. Stably transformed cultures were successfully produced by co-cultivation with A. tumefaciens strain C58/pMP90 carrying pBIV10 in Korean fir. Here, we have reported an Agrobacterium-mediated gene transfer protocol via somatic embryogenesis that may be helpful in developing breeding and conservation strategies for the Korean fir.

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Insect-resistant transgenic Pinus radiata

Cited by 60 publications

References 38 publications

Advancements in the Understanding of Paclitaxel Metabolism in Tissue Culture

Advancements in the Understanding of Paclitaxel Metabolism in Tissue Culture

Development of a Particle Bombardment-Mediated Transient Transformation System for Taxus spp. Cells in Culture

Agrobacterium-mediated Transformation via Somatic Embryogenesis System in Korean fir (Abies koreana Wil.), A Korean Native Conifer

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