Stable Agrobacterium-mediated genetic transformation of London plane tree (Platanus acerifolia Willd.)

London plane tree (Platanus acerifolia Willd.) is an important tree in urban landscaping but it suffers from a number of negative traits which genetic engineering could be used to address. As with many woody species, P. acerifolia has appeared recalcitrant to genetic transformation. However, the rec...

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Bibliographic Details
Published in:Plant cell reports 2007-05, Vol.26 (5), p.641-650
Main Authors: LI, Z. N, FANG, F, LIU, G. F, BAO, M. Z
Format: Article
Language:English
Subjects:
Agrobacterium
Agrobacterium tumefaciens
Agrobacterium tumefaciens - drug effects
Agrobacterium tumefaciens - physiology
Anti-Bacterial Agents - pharmacology
Antibiotics
Bacteria
Biological and medical sciences
Biotechnology
Cultivation
Deoxyribonucleic acid
DNA
DNA, Bacterial - metabolism
Explants
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Genetic transformation
Genomes
Glucuronidase - metabolism
Immunization
Inoculation
Kanamycin
Landscaping
Leaves
Magnoliopsida - drug effects
Magnoliopsida - genetics
Magnoliopsida - microbiology
Mannitol
Methods. Procedures. Technologies
Microbiology
Molecular weight
Osmotic Pressure - drug effects
Plant Leaves - drug effects
Plant Leaves - microbiology
Plant Shoots - drug effects
Plant Shoots - physiology
Plants, Genetically Modified
Platanus
Platanus acerifolia
Regeneration - drug effects
Reporter gene
Selection, Genetic
Shoots
T-DNA
Transformation, Genetic - drug effects
Transgenic animals and transgenic plants
Transgenic plants
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Summary:London plane tree (Platanus acerifolia Willd.) is an important tree in urban landscaping but it suffers from a number of negative traits which genetic engineering could be used to address. As with many woody species, P. acerifolia has appeared recalcitrant to genetic transformation. However, the recent development of a method for regenerating shoots from P. acerifolia leaf explants suggests that such material could be a target for gene-transfer. Using an Agrobacterium tumefaciens strain in which the T-DNA carries the histochemically detected reporter gene beta-glucuronidase (GUS), we have followed the transfer of genes from Agrobacterium to leaf explants of Platanus acerifolia. Using this system, we have identified a set of inoculation and co-cultivation conditions (notably: the pre-treatment of leaf explants with 0.4 M mannitol, an inoculation period of 10 min, a bacterial OD(600) of 0.8-1.0 and a co-cultivation period of 5 days) that permit a good frequency and reliability of transient gene-transfer. Optimum levels of antibiotics for bacterial elimination and kanamycin-resistant shoot regeneration were also established. By applying these parameters, we recovered eight independent stably transformed shoots that were kanamycin-resistant and contained the nptII T-DNA gene, as confirmed by PCR analysis. Furthermore, Southern blot analysis confirmed that, in at least five of these lines, the transgene was associated with high molecular weight DNA, so indicating integration into the plant genome.
ISSN:0721-7714
1432-203X
DOI:10.1007/s00299-006-0271-x