Transformation of Brassica napus and Brassica oleracea Using Agrobacterium tumefaciens and the Expression of the bar and neo Genes in the Transgenic Plants

Block, Marc De; Brouwer, Dirk De; Tenning, Paul

doi:10.1104/pp.91.2.694

Cited by 399 publications

(217 citation statements)

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“…The oligonucleotide used had the sequence 5'-CACATAGCAAAC-CATGGAAACAAG-3'. Subsequently, the modified HindIII fragment was excised and used to replace the corresponding unmodified fragment of pAT2S lBG, resulting in pAT2S IC 11. pAT2S1BG contains the 3.6 kb genomic BglII fragment also used in the construction of pTAD7 (see above) (12). These two fragments were cloned into the unique BglII site (made blunt through S 1 nuclease treatment) of the binary vector pGSC 1 703A, resulting in pTAD3.…”

Section: Ptad3mentioning

confidence: 99%

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Expression and Processing of an Arabidopsis 2S Albumin in Transgenic Tobacco

Clercq

Vandewiele²,

Rycke³

et al. 1990

Plant Physiol.

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2S albumin seed storage proteins undergo a complex series of posttranslational proteolytic cleavages. In order to determine if this process is correctly carried out in transgenic plants, the gene AT2S1 encoding an Arabidopsis thaliana 2S albumin isoform has been expressed in transgenic tobacco. Initial experiments using a reporter gene demonstrated that the AT2S1 promoter directs seed specific expression in both transgenic tobacco and Brassica napus plants. The entire AT2S1 gene was then transferred into tobacco plants, where it showed a tissue specific and developmentally regulated expression. Arabidopsis 2S albumin accumulates up to 0.1% of the total high-salt extractable seed protein. Protein sequencing demonstrated that the amino termini of the two Arabidopsis 2S albumin subunits were correctly processed, suggesting that the protease(s) necessary for posttranslational processing of 2S albumin precursors may display common specificities among different dicot plant species. Immunocytochemical studies showed that the Arabidopsis 2S albumin is localized in the protein body matrix of tobacco endosperm and embryo. Correct processing and targeting of the 2S albumin in transgenic plants suggests that modified versions could be expressed, allowing the study of 2S albumin processing and in particular the possible roles of the processed fragments in protein stability and/or targeting.

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Section: Ptad3mentioning

confidence: 99%

“…Brassica napus cv Westar was transformed as described by De Block et al (12). In all transformation procedures kanamycin was used as selectable marker.…”

Section: Plant Transformationsmentioning

confidence: 99%

Expression and Processing of an Arabidopsis 2S Albumin in Transgenic Tobacco

Clercq

Vandewiele²,

Rycke³

et al. 1990

Plant Physiol.

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“…The Basta gene-based selection system brought about an easy and reproducible selection method and it has successfully produced numerous herbicide-resistant transformed crops e.g. oilseed rape (De Block et al 1989;Kopertekh et al 2009), sugarcane (Joyce et al 2010), cassava (Koehorst-van Putten et al 2012) and tomato (Khuong et al 2013). Unfortunately, most of the vectors that contain Basta as a plant selectable marker are gateway based vectors (e.g.…”

Section: Introductionmentioning

confidence: 99%

Novel recombinant binary vectors harbouring Basta (bar) gene as a plant selectable marker for genetic transformation of plants

Nada

2016

Physiol Mol Biol Plants

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Genetic transformation is one of the most widely used technique in crop improvement. However, most of the binary vectors used in this technique, especially cloning based, contain antibiotic genes as selection marker that raise serious consumer and environmental concerns; moreover, they could be transferred to non-target hosts with deleterious effects. Therefore, the goal of this study was reconstruction of the widely used pBI121 binary vector by substituting the harmful antibiotic selection marker gene with a less-harmful selection marker, Basta (herbicide resistance gene). The generated vectors were designated as pBI121NB and pBI121CB, in which Basta gene was expressed under the control of Nos or CaMV 35S promoter, respectively. The successful integration of the new inserts into both the vectors was confirmed by PCR, restriction digestion and sequencing. Both these vectors were used in transforming Arabidopsis, Egyptian wheat and barley varieties using LBA4404 and GV3101 Agrobacterium strains. The surfactant Tween-20 resulted in an efficient transformation and the number of Arabidopsis transformants was about 6-9 %. Soaked seeds of wheat and barley were transformed with Agrobacterium to introduce the bacteria to the growing shoot apices. The percentage of transgenic lines was around 16-17 and 14-15 % for wheat and barley, respectively. The quantitative studies presented in this work showed that both LBA4404 and GV3101 strains were suitable for transforming Egyptian wheat and barley.

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“…Plant transformation systems have been developed for many economically important species of the genus Brassica such as B. napus [5], B. oleracea [6], B. juncea [7], B. rapa [8], B. nigra [9] and B. carinata [10]. This technology enables us to obtain transgenic plants with modified agronomic traits.…”

Section: Introductionmentioning

confidence: 99%