scholarly journals New Approaches to Agrobacterium tumefaciens-Mediated Gene Transfer to Plants

10.5772/66465 ◽  
2016 ◽  
Author(s):  
Mustafa Yildiz ◽  
Murat Aycan ◽  
Sunjung Park
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Gene Transfer ◽  
New Approaches

scholarly journals Strategies to produce T-DNA free CRISPRed fruit trees via Agrobacterium tumefaciens stable gene transfer

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Lorenza Dalla Costa ◽  
Stefano Piazza ◽  
Valerio Pompili ◽  
Umberto Salvagnin ◽  
Alessandro Cestaro ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Gene Transfer ◽  
Genomic Dna ◽  
Fruit Trees ◽  
Plant Genome ◽  
Stable Gene ◽  
Binary Vectors ◽  
Exogenous Dna ◽  
Left Behind ◽  
Dicotyledonous Plants

AbstractGenome editing via CRISPR/Cas9 is a powerful technology, which has been widely applied to improve traits in cereals, vegetables and even fruit trees. For the delivery of CRISPR/Cas9 components into dicotyledonous plants, Agrobacterium tumefaciens mediated gene transfer is still the prevalent method, although editing is often accompanied by the integration of the bacterial T-DNA into the host genome. We assessed two approaches in order to achieve T-DNA excision from the plant genome, minimizing the extent of foreign DNA left behind. The first is based on the Flp/FRT system and the second on Cas9 and synthetic cleavage target sites (CTS) close to T-DNA borders, which are recognized by the sgRNA. Several grapevine and apple lines, transformed with a panel of CRISPR/SpCas9 binary vectors, were regenerated and characterized for T-DNA copy number and for the rate of targeted editing. As detected by an optimized NGS-based sequencing method, trimming at T-DNA borders occurred in 100% of the lines, impairing in most cases the excision. Another observation was the leakage activity of Cas9 which produced pierced and therefore non-functional CTS. Deletions of genomic DNA and presence of filler DNA were also noticed at the junctions between T-DNA and genomic DNA. This study proved that many factors must be considered for designing efficient binary vectors capable of minimizing the presence of exogenous DNA in CRISPRed fruit trees.

scholarly journals Integrative gene transfer in the truffle Tuber borchii by Agrobacterium tumefaciens-mediated transformation

AMB Express ◽  
2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Andrea Brenna ◽  
Barbara Montanini ◽  
Eleonora Muggiano ◽  
Marco Proietto ◽  
Patrizia Filetici ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Gene Transfer ◽  
Tuber Borchii

Development of Auxotrophic Agrobacterium tumefaciens for Gene Transfer in Plant Tissue Culture

2004 ◽  
Vol 20 (3) ◽  
pp. 890-896 ◽  
Author(s):  
J.I. Collens ◽  
D.R. Lee ◽  
A.M. Seeman ◽  
W.R. Curtis
Keyword(s):  
Tissue Culture ◽  
Agrobacterium Tumefaciens ◽  
Gene Transfer ◽  
Plant Tissue Culture ◽  
Plant Tissue

The production and uses of genetically transformed plants: concluding remarks

1993 ◽  
Vol 342 (1301) ◽  
pp. 293-294
Keyword(s):  
Gene Transfer ◽  
Crop Improvement ◽  
Plant Biotechnology ◽  
Cell Types ◽  
Crop Plants ◽  
Transformed Plants ◽  
The Past ◽  
New Approaches ◽  
Transfer Genes ◽  
Biological Methods

This meeting at which the preceding collection of papers were presented occurred on the tenth anniversary of the development of techniques which have laid the foundation for modern plant biotechnology, namely the ability to transfer genes into plants and to direct their expression in appropriate cell types. The contributors have revealed many of the exciting and potentially important applications of gene transfer technology in crop plants arising from these earlier developments, and have also described new approaches which have engendered an ever-widening scope for crop improvement through the application of molecular biological methods. In the past ten years a wide variety of crop plants have been stably transformed, and it is clear that many of the major crops can now be transformed, although with widely different efficiencies.

scholarly journals Improvement of adventitious organogenesis for regeneration of transgenic plants in blackberry

Genetika ◽  
2015 ◽  
Vol 47 (2) ◽  
pp. 599-608 ◽  
Author(s):  
Alena Gajdosová ◽  
Tatjana Vujovic ◽  
Miroslava Súkeníková ◽  
Gabriela Libiaková
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Plant Regeneration ◽  
Gene Transfer ◽  
Transgenic Plants ◽  
Plant Genome ◽  
Ms Medium ◽  
Regeneration System ◽  
Promising Technique ◽  
Rubus Fruticosus ◽  
Foreign Dna

The introduction of foreign DNA into the plant genome by Agrobacterium tumefaciens is a promising technique of targeted gene transfer which depends on good working regeneration system. The aim of the work was to elaborate the system for efficient adventitious organogenesis and transgenic plant regeneration in Rubus fruticosus L. using explants from mature plants. Regeneration of putative transgenic shoots took place from flag explants cultivated vertically on MS medium with 1 mg l-1 TDZ and 0.02 mg l-1 IBA followed by transfer on MS medium with 1 mg l-1 BAP, 0.02 mg l-1 IBA and 0.1 mg l-1 GA3 supplemented with 10-15 mg l-1 hygromycin after transformation by A. tumefaciens strain LBA 4404 carrying plasmid pCambia 1304. Four putative transgenic plants of cv. 'Cacanska Bestrna' were rooted and acclimatized.

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