Transgene integration patterns and expression levels in transgenic tissue lines of Picea mariana, P. glauca and P. abies

Klimaszewska, Krystyna; Lachance, Denis; Bernier‐Cardou, Michèle; Rutledge, Robert G.

doi:10.1007/s00299-003-0626-5

Cited by 22 publications

(12 citation statements)

References 23 publications

(34 reference statements)

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“…So far, stable transformation of embryogenic tissues by Agrobacterium has been achieved in hybrid larch (Levée et al 1997), Picea abies (Wenck et al 1999), Picea glauca (Le et al 2001;Klimaszewska et al 2003), Chamaecyparis obtusa (Taniguchi et al 2005), Pinus radiata (Cerda et al 2002), Pinus strobus (Levée et al 1999), Picea mariana (Klimaszewska et al 2001).…”

Section: Introductionmentioning

confidence: 98%

Agrobacterium-mediated transformation of embryogenic tissues of hybrid firs (Abies spp.) and regeneration of transgenic emblings

et al. 2009

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A genetic transformation system has been developed for selected embryogenic cell lines of hybrids Abies alba x A. cephalonica (cell lines AC2, AC78) and Abies alba x A. numidica (cell line AN72) using Agrobacterium tumefaciens. The cell lines were derived from immature or mature zygotic embryos on DCR medium containing BA (1 mg l(-1)). The T-DNA of plant transformation vector contained the beta-glucuronidase reporter gene under the control of double dCaMV 35S promoter and the neomycin phosphotransferase selection marker gene driven by the nos promoter. The regeneration of putative transformed tissues started approximately 1 week after transfer to the selection medium containing 10 mg geneticin l(-1). GUS activity was detected in most of the geneticin-resistant sub-lines AN72, AC2 and AC78, and the transgenic nature of embryogenic cell lines was confirmed by PCR approach. Plantlet regeneration from PCR-positive embryogenic tissues has been obtained as well. The presence of both gus and nptII genes was confirmed in 11 out of 36 analysed emblings.

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Section: Introductionmentioning

confidence: 98%

Agrobacterium-mediated transformation of embryogenic tissues of hybrid firs (Abies spp.) and regeneration of transgenic emblings

et al. 2009

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“…Agrobacterium-based system has been widely applied to different tissues of several pine species (Levé e et al 1999;Wenck et al 1999;Ló pez et al 2000;Trontin et al 2002;Tereso et al 2003;Tang et al 2001;Tang and Newton 2004;Grant et al 2004), but transformation remains highly dependent on the genotype. Somatic embryogenesis (SE) from immature zygotic embryos has been the most commonly used method for regeneration of transformed conifer plants (Levé e et al 1997(Levé e et al 1999Wenck et al 1999;Klimaszewska et al 2001Klimaszewska et al 2003. In P. pinaster, SE was previously achieved (Bercetche and Pâ ques 1995;Lelu et al 1999;Miguel et al 2004) and used in the genetic transformation of French genotypes (Trontin et al 2002).…”

Section: Introductionmentioning

confidence: 98%

Stable Agrobacterium-mediated transformation of embryogenic tissues from Pinus pinaster Portuguese genotypes

Tereso¹,

Miguel²,

Zoglauer

et al. 2006

Plant Growth Regul

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Protocols for genetic transformation of maritime pine (Pinus pinaster Sol. ex Aiton) embryogenic tissues were developed using the Agrobacterium C58pMP90/pPCV6NFGUS. This is the first report of Agrobacterium-mediated T-DNA integration in P. pinaster confirmed by Southern blot analysis. The omission of casein hydrolysate from culture medium during cocultivation and subsequent subculture was crucial to control Agrobacterium growth. Two different transformation protocols were compared: (1) bacterial drops were spread over embryogenic clumps; (2) a mixture of bacterial and embryogenic cell suspensions was plated on filter paper.The highest frequency of transformation (22 independent transformed lines per g fresh weight, for embryogenic clone 31/668/00) was obtained with Protocol 2. The same basic procedure allowed transformation of embryogenic cell suspensions, which was dependent on subculture age. From 52 hygromycin-resistant independent lines obtained, 47 showed stable uidA gene expression and were PCR-positive for uidA gene and 42 for hpt gene. No residual Agrobacterium was detected in the transformed lines. Transgene integration was achieved using both protocols, as confirmed by Southern hybridization. From 38 (90%) transformed lines successfully cryopreserved and recovered, 71% regrown replicates have maintained the frequency of cell aggregates and early-formed embryos with uidA expression. Maturation of 44 transformed lines gave rise to 3 mature somatic embryos, each one coming from a different transformed line. Our results show the high potential of Protocol 2 for application to different culture systems.

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“…Such an approach to the problem of somatic embryogenesis in coniferous trees was understandable, and the positive results achieved at each stage of culture are now the basis for more advanced studies of this process, already at the molecular level (Stasolla et al 2002;BishopHurley et al 2003;Silveira et al 2004;Lippert et al 2005;Peña and Séguin 2001;Klimaszewska et al 2003).…”

Section: Introductionmentioning

confidence: 99%

Somatic embryogenesis of selected spruce species (Picea abies, P. omorika, P. pungens 'Glauca' and P. breweriana)

Hazubska-Przybył

Bojarczuk

2011

Acta Soc Bot Pol

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Somatic embryogenesis was studied in four spruce species (Picea abies, P. omorika, P. pungens Glauca and P. breweriana) to determine if this method can be used for in vitro propagation of coniferous trees. The highest frequency of initiation of embryogenic tissue was obtained when mature zygotic embryos were used as explants. It ranged then from 10.8% (P. breweriana) to 23.75% (P. omorika and P. pungens Glauca). The frequency of embryogenic tissue initiation was strongly affected by medium composition, i.e. addition of appropriate auxins (2,4-D, NAA, Picloram) and sucrose concentration (10-20 g·l -1 ). A lower frequency was obtained in Picea omorika (10%) when megagametophytes (endosperms with immature zygotic embryos) were used as explants. No embryogenic tissue was produced from hypocotyls, cotyledons and needles. A satisfactory frequency was achieved with the use of somatic embryos of Picea abies (30%). The proliferation of embryogenic cell lines of spruces was affected by medium type. The experiments resulted in production of somatic plantlets of P. abies and P. omorika. This enables the application of this method of spruce micropropagation for genetic and breeding research or for nursery production.

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Transgene integration patterns and expression levels in transgenic tissue lines of Picea mariana, P. glauca and P. abies

Cited by 22 publications

References 23 publications

Agrobacterium-mediated transformation of embryogenic tissues of hybrid firs (Abies spp.) and regeneration of transgenic emblings

Agrobacterium-mediated transformation of embryogenic tissues of hybrid firs (Abies spp.) and regeneration of transgenic emblings

Stable Agrobacterium-mediated transformation of embryogenic tissues from Pinus pinaster Portuguese genotypes

Somatic embryogenesis of selected spruce species (Picea abies, P. omorika, P. pungens 'Glauca' and P. breweriana)

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