Agrobacterium tumefaciens-mediated transformation of hybrid larch (Larix kaempferi T L. decidua) and transgenic plant regenerationn

Levée, Valérie; Lelu, Marie-Anne; Jouanin, Lise; Cornu, D.; Pilate, Gilles

doi:10.1007/s002990050301

Cited by 103 publications

(54 citation statements)

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“…The addition of 100 M coniferyl alcohol during the cocultivation step increased the induction frequency of kanamycin resistant calli. Similar results have been reported by Levee et al [9]. Coniferyl alcohol is a phenolic compound that is released by the wounded plant cells and is a virulence inducer similar to acetosyringone [19] and syringaldehyde.…”

Section: Discussionsupporting

confidence: 89%

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Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.)

Tang

2001

Cell Res

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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 for b-glucuronidase (GUS) and neomycin phosphotransferase (NPTII). Factors influencing transgene expression including seed sources of loblolly pine, concentration of bacteria, and the wounding procedures of target explants were investigated. The expression of foreign gene was confirmed by the ability of mature zygotic embryos to produce calli in the presence of kanamycin, by histochemical assays of GUS activity, by PCR analysis, and by Southern blot. The successful expression of the GUS gene in different families of loblolly pine suggests that this transformation system is probably useful for the production of the genetically modified conifers.

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Section: Discussionsupporting

confidence: 89%

“…However, successful transfer of foreign genes into conifers by Agrobacterium and the regeneration of transgenic plants have been achieved with larch (Larix deciduas) [8], hybrid larch (L. KaempfericL. Decidua) [9], Norway spruce (Picea abies) [10], and white pine (Pinus strobes) [2].…”

Section: Introductionmentioning

confidence: 99%

Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.)

Tang

2001

Cell Res

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“…Similar conclusions were obtained during Agrobacterium-mediated transformation of hybrid larch ESM with 4 out of 7 genotypes transformed with very contrasted efficiencies [29]. Such results suggested that our Agrobacterium procedure may be applicable to a wide range of selected genotypes after identification of main variation sources (e.g.…”

Section: Discussionsupporting

confidence: 71%

“…Considering acetosyringone concentration in the cocultivation, up to 200 µM did not lead to an increase in transformation yields. In contrast, transformation was found to be influenced by the presence of this plant elicitor in Picea abies and Pinus taeda (25-50 µM, [49]), Pinus strobus (100 µM, [30]) and hybrid larch (100 µM, [29]). …”

Section: Discussionmentioning

confidence: 99%

“…Such a promising tissue culture system undeniably provides a good target for stable genetic transformation of conifers such as larch [23,29], spruce [10,48,49], and pine [6,30,47] and offers new prospects for rapid and efficient introduction of desirable traits, mostly unknown (e.g. pests and herbicide tolerance) or with low heritability (e.g.…”

Section: Introductionmentioning

confidence: 99%

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Towards genetic engineering of maritime pine (Pinus pinaster Ait.)

Trontin¹,

Harvengt²,

Garin³

et al. 2002

Ann. For. Sci.

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-Using our improved protocols for somatic embryogenesis in Pinus pinaster, transgenic tissues and plantlets were recovered after microprojectile bombardment (biolistic) or cocultivation of embryonal-suspensor masses (ESM) with Agrobacterium tumefaciens. Transformation experiments were carried out with selectable hpt gene (hygromycin B resistance) and reporter gus gene (β-glucuronidase activity). With both methods, hygromycin was shown to be an effective selective agent of transformed cells within 4-19 weeks. The mean number of hygromycin-resistant lines expressing gus per gram ESM subjected to DNA transfer, ranged from 7.0 to 8.5 using biolistic and 0 to 67.3 during Agrobacterium experiments. Mature somatic embryos obtained from some transformed lines were converted into plantlets and grown in the greenhouse. The whole process (from transformation to plant acclimatisation) could be completed within only 12 months. The transgenic state of ESM, somatic embryos and plants was confirmed by histochemical GUS assays and molecular methods.

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Plant Transformation Technology: Agrobacterium ‐Mediated Transformation

Komari

Ishida

Hiei

2004

Handbook of Plant Biotechnology

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The soil phytopathogen Agrobacterium tumefaciens induces tumours, known as crown galls, mainly on dicotyledonous plants. Such tumours are generated by a complex, multi‐step transformation process. Another species, A. rhizogenes , causes hairy root disease in higher plants via an identical process. Agrobacterium has been utilised for the transfer of genes to dicotyledonous plants. Now, monocotyledonous plants are routinely transformed by Agrobacterium despite the fact that these plants, including important cereals, were thought until recently to be outside the range of this technology. Most of the common transformation methods heavily depend on tissue culture technology and we refer to such methods as in vitro transformation. The only case of a routine tissue culture free method is the ‘ in planta ’ transformation of Arabidopsis thaliana . Numerous critical factors are involved in both approaches. In transformation in vitro , key factors include choice of vectors and bacterial strains, types of plant tissues to be infected, procedures of preparation the tissues, protocols of infection and co‐cultivation, methods for subsequent culture and selection of transformed cells, antibiotics to remove infecting bacteria, techniques for regeneration of transgenic plants and genotypes of plants. It is our opinion that the type and quality of the starting material is the most important one among them. The capacity to serve as a host plant for crown gall disease is no longer a prerequisite for a host of vector systems based on Agrobacterium . The real prerequisite for in vitro transformation is the availability of technology for dedifferentiation of tissues and regeneration of plants in a given species. On the other hand, the biological processes involved in in planta transformation are yet to be elucidated. Recent evidence suggested that ovules are the primary target, and further basic understanding is likely to help extend the number of species transformed by the method. The plant species that are routinely transformed by Agrobacterium are expanding rapidly. Some gymnosperms, several forest trees and fruit trees, various legumes, and cereal and non‐cereal monocotyledons, which were once considered very recalcitrant, are now in a long list of transformable plants. Arabidopsis , tobacco and rice are the top three species that were transformed during the last two years. Although it is only half a dozen years since the current procedure of rice transformation mediated by Agrobacterium was published, the economic importance and the accumulation of genomic information has made rice the species of focus in plant biotechnology. The advantages of Agrobacterium ‐mediated transformation include the transfer of pieces of DNA with defined ends and minimal rearrangement, the transfer of relatively large segments of DNA, the integration of small numbers of copies of genes into plant chromosomes and the high quality and fertility of transgenic plants. However, transformation does not always produce such ‘clean’ events. Formation of repeats of transgenes, certain rearrangements, integration of non‐target DNA segments and unstable expression of transgenes are among the complications. Although the majority of transgenic plants usually appear ‘good’ in a particular test, accumulation of ‘dropped’ plants is significant after multiple rounds of characterisation and screening. Therefore, further improvement in each of steps is highly desired. Application of the gene transfer mediated by Agrobacterium is further expanding. Transient expression of genes delivered by Agrobacterium is now a useful tool in the study of promoters and gene function. Vectors specifically designed to carry very large segments of DNA have been developed and extensively tested. Insertional mutagenesis by DNA transferred by Agrobacterium is a routine technique in genomics study in Arabidopsis and rice. Methods for targeted integration of transgenes to genomes of higher plants have been drawing considerable attention. Various technologies for the production of ‘selection marker free’ transgenic plants are now in place for better public acceptance of biotechnology products.

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Agrobacterium tumefaciens-mediated transformation of hybrid larch (Larix kaempferi T L. decidua) and transgenic plant regenerationn

Cited by 103 publications

References 20 publications

Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.)

Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.)

Towards genetic engineering of maritime pine (Pinus pinaster Ait.)

Plant Transformation Technology: Agrobacterium ‐Mediated Transformation

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