ArticleTheScientificWorldJournal2013
Stable Agrobacterium-mediated transformation of maritime pine based on kanamycin selection.
Article in TheScientificWorldJournal, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- Genetic transformation in conifers: current status and future prospects.Forestry research · 2024Review
- Embryogenic callus induction from immature zygotic embryos and genetic transformation of Larix kaempferi 3x Larix gmelinii 9.PloS one · 2021Article
- An efficient system forPlant methods · 2020Article
- Somatic Embryogenesis in Selected Conifer TreesFrontiers in plant science · 2019Review
- A novel in vitro transformation of3 Biotech · 2017Article
- A Genetic Transformation Method for Cadmium HyperaccumulatorFrontiers in plant science · 2017Article
- An efficient plant regeneration and Agrobacterium-mediated genetic transformation of Tagetes erecta.Protoplasma · 2015Article
- Sequenced genomes and rapidly emerging technologies pave the way for conifer evolutionary developmental biology.Frontiers in plant science · 2015Review
- Evaluation on the effectiveness of 2-deoxyglucose-6-phosphate phosphatase (DOG(R)1) gene as a selectable marker for oil palm (Elaeis guineensis Jacq.) embryogenic calli transformation mediated by Agrobacterium tumefaciens.Frontiers in plant science · 2015Article
- A new high-frequency Agrobacterium-mediated transformation technique for Sesamum indicum L. using de-embryonated cotyledon as explant.Protoplasma · 2014Article
- From soil to plant, the journey of P through trophic relationships and ectomycorrhizal association.Frontiers in plant science · 2014Review
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
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) as a reporter gene, was used for transformation studies. Different factors, such as embryogenic line, bacterial strain, bacterial concentration, and coculture duration, were examined and optimized. For selection of transformants, 15 mgL(-1) kanamycin was used. The highest transformation efficiency (11.4 events per gram of fresh mass) was achieved when a vigorously growing embryonal mass (embryogenic line L01) was cocultivated with Agrobacterium strain AGL1 at the optical density (OD(600 nm)) of 0.3 for 72 h. Evidence of the stable transgene integration was obtained by polymerase chain reaction for the nptII and uidA genes and expression of the uidA gene. Maturation capacity of the transgenic lines was negatively affected by the transformation process. Induction of axillary shoots by preculturing the embryos with benzyladenine allowed overcoming the low maturation rates of some transformed lines. The transgenic embryos were germinated and the axillar shoots were rooted. Transgenic plants were transferred to potting substrate showing normal growth.
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