Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method

@article{Zhang2006AgrobacteriummediatedTO,
  title={Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method},
  author={Xiuren Zhang and Rossana Henriques and Shih-Shun Lin and Qinglin Niu and Nam‐Hai Chua},
  journal={Nature Protocols},
  year={2006},
  volume={1},
  pages={641-646}
}
Collective efforts of several laboratories in the past two decades have resulted in the development of various methods for Agrobacterium tumefaciens–mediated transformation of Arabidopsis thaliana. Among these, the floral dip method is the most facile protocol and widely used for producing transgenic Arabidopsis plants. In this method, transformation of female gametes is accomplished by simply dipping developing Arabidopsis inflorescences for a few seconds into a 5% sucrose solution containing… 
The floral inoculating protocol: a simplified Arabidopsis thaliana transformation method modified from floral dipping
TLDR
This simplified method, without floral dipping, which requires large volumes of Agrobacterim culture, offers as efficient a transformation as previously reported protocols, and reduces the required workload, cost, time, and space.
Protocol: Streamlined sub-protocols for floral-dip transformation and selection of transformants in Arabidopsis thaliana
TLDR
Three streamlined sub-protocols are developed that find that A. thaliana can be transformed by dipping directly into an A. tumefaciens culture supplemented with surfactant, eliminating the need for media exchange to a buffered solution.
Minor modifications in obtainable Arabidopsis floral dip method enhances transformation efficiency and production of homozygous transgenic lines harboring a single copy of transgene
TLDR
Modified protocol produced ~12-14% increase in transformants and increased the percentage of homozygous and single copied stable transgenic lines in Arabidopsis thaliana.
High transformation efficiency in Arabidopsis using extremely low Agrobacterium inoculum
TLDR
The data revealed that the floral dip method still guarantees a relatively high transformation rate in the Arabidopsis thaliana Col-0 ecotype even with very low Agrobacterium inoculum, which simplifies the floral dipping protocol further, which allows transformation with small bacterial culture and enables high-throughput transformation of large numbers of constructs in parallel.
High transformation efficiency in Arabidopsis using extremely low Agrobacterium inoculum
TLDR
The data revealed that the floral dip method still guarantees relatively high transformation rate in Arabidopsis thaliana Col-0 ecotype even with very low Agrobacterium inoculum, which simplifies the floral dipping protocol further, which allows transformation with small bacterial culture and enables high-throughput transformation of large numbers of constructs in parallel.
Agrobacterium-mediated floral dip transformation of the model polyploid species Arabidopsis kamchatica
TLDR
The early development of pistils in A. kamchatica was investigated and confirmed the formation of open pistil in young flower buds (stages 8–9), which is important for allowing Agrobacterium to access female reproductive tissues.
A Simplified Floral Dip Method for Transformation of Brassica napus and B. carinata
TLDR
The results showed that the Agrobacterium-mediated floral-dip transformation can be a successful strategy to develop transgenic B. napus and B. carinata in a short time without undergoing cumbersome vacuum infiltration or tissue culture steps.
Isolation of embryo-specific mutants in Arabidopsis: plant transformation.
TLDR
A detailed description with various fine-tuned steps for these two Agrobacterium-mediated floral transformation methods in Arabidopsis is outlined, one of the most powerful tools for studying gene function.
The Status of Setaria viridis Transformation: Agrobacterium-Mediated to Floral Dip.
TLDR
While the tissue culture-based method results in a higher transformation efficiency than floral dip, implementation requires a specific technical skillset that limits availability of experienced personnel to successfully perform transformations, a lack of high quality growth chambers or greenhouses that provide the necessary optimum growing conditions would reduce an already low transformation efficiency or would not result in recovery of transgenic lines.
The Status of Setaria viridis Transformation: Agrobacterium-Mediated to Floral Dip
TLDR
An overview of transformation methods reported for S. viridis is presented in this review and it is shown that a lack of high-quality growth chambers or greenhouses that provide the necessary optimum growing conditions would reduce an already low transformation efficiency or would not result in recovery of transgenic lines.
...
1
2
3
4
5
...

References

SHOWING 1-10 OF 27 REFERENCES
Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana.
  • S. Clough, A. Bent
  • Biology
    The Plant journal : for cell and molecular biology
  • 1998
TLDR
The modified method should facilitate high-throughput transformation of Arabidopsis for efforts such as T-DNA gene tagging, positional cloning, or attempts at targeted gene replacement.
Female reproductive tissues are the primary target of Agrobacterium-mediated transformation by the Arabidopsis floral-dip method.
TLDR
The results suggest that ovules are the site of productive transformation in the floral-dip method, and suggest that Agrobacterium must be delivered to the interior of the developing gynoecium prior to locule closure if efficient transformation is to be achieved.
Floral spray transformation can efficiently generate Arabidopsis transgenic plants.
TLDR
The result indicates that the floral spray method of Agrobacterium can achieve rates of in planta transformation comparable to the vacuum-infiltration and floral dip methods.
Germ-line transformation of Arabidopsis lasiocarpa
  • B. Tague
  • Environmental Science
    Transgenic Research
  • 2004
TLDR
The suitability of the germ-line transformation method developed forArabidopsis thaliana was examined for four taxa in the Brassicaceae that have not been previously transformed: Arabidopsis griffithiana, Arabidosis lasiocarpa, Arabids petraea and Capsella bursa-pastoris.
Production of transgenic crops by the floral-dip method.
TLDR
This chapter describes the simple system that has been adopted in the routine production of transgenic radish, without requiring any knowledge of plant tissue culture, that has enabled the successful production of transformed Medicago truncatula plants.
Floral dip: agrobacterium-mediated germ line transformation.
  • S. Clough
  • Biology
    Methods in molecular biology
  • 2005
TLDR
This chapter provides detailed explanations on the floral dip protocol, a simple method to transform Arabidopsis by inoculating immature flowers with Agrobacterium tumefaciens.
The Endosperm and the Embryo of Arabidopsis thaliana are Independently Transformed through Infiltration by Agrobacterium tumefaciens
TLDR
Results indicate that mature female gametes could be the targets of T-DNA in planta transformation of Arabidopsis thaliana and that the minority of ‘co-transformed’ plantlets and endosperm can be explained by simultaneous but independent transformation events.
Genomic stability in Arabidopsis thaliana transgenic plants obtained by floral dip
TLDR
Variations were observed in callus-derived A. thaliana plants, supporting the conclusion that somaclonal variation is essentially correlated with the stress imposed by the in vitro cell culture, rather than with the integration of a foreign gene.
Agrobacterium-mediated transformation of germinating seeds of Arabidopsis thaliana: A non-tissue culture approach
TLDR
All of the kanamycin-resistant plants that were tested were found to possess NPT II activity and all of the resistant plants contained at least one copy of the T-DNA and that the majority of the plants had multiple inserts.
Development of a novel Agrobacterium-mediated transformation method to recover transgenic Brassica napus plants
TLDR
Molecular and genetic analysis revealed that single and multiple copy events were generated and that the transgenes were transmitted to the T1 and T2 progeny in a Mendelian fashion.
...
1
2
3
...