Loading...
Please wait, while we are loading the content...
Similar Documents
Floral-dip transformation of flax (Linum usitatissimum) to generate transgenic progenies with a high transformation rate.
| Content Provider | Europe PMC |
|---|---|
| Author | Bastaki, Nasmah K. Cullis, Christopher A. |
| Copyright Year | 2014 |
| Abstract | Agrobacterium-mediated plant transformation via floral-dip is a widely used technique in the field of plant transformation and has been reported to be successful for many plant species. However, flax (Linum usitatissimum) transformation by floral-dip has not been reported. The goal of this protocol is to establish that Agrobacterium and the floral-dip method can be used to generate transgenic flax. We show that this technique is simple, inexpensive, efficient, and more importantly, gives a higher transformation rate than the current available methods of flax transformation.In summary, inflorescences of flax were dipped in a solution of Agrobacterium carrying a binary vector plasmid (T-DNA fragment plus the Linum Insertion Sequence, LIS-1) for 1 - 2 min. The plants were laid flat on their side for 24 hr. Then, plants were maintained under normal growth conditions until the next treatment. The process of dipping was repeated 2 - 3 times, with approximately 10 - 14 day intervals between dipping. The T1 seeds were collected and germinated on soil. After approximately two weeks, treated progenies were tested by direct PCR; 2 - 3 leaves were used per plant plus the appropriate T-DNA primers. Positive transformants were selected and grown to maturity. The transformation rate was unexpectedly high, with 50 - 60% of the seeds from treated plants being positive transformants. This is a higher transformation rate than those reported for Arabidopsis thaliana and other plant species, using floral-dip transformation. It is also the highest, which has been reported so far, for flax transformation using other methods for transformation. |
| Journal | Journal of Visualized Experiments : JoVE |
| PubMed Central reference number | PMC4396936 |
| Issue Number | 94 |
| PubMed reference number | 25549243 |
| e-ISSN | 1940087X |
| DOI | 10.3791/52189 |
| Language | English |
| Publisher | MyJove Corporation |
| Publisher Date | 2014-12-19 |
| Access Restriction | Open |
| Rights License | This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/ Copyright © 2014, Journal of Visualized Experiments |
| Subject Keyword | Plant Biology Issue 94 Flax (Linum usitatissimum) Floral-dip Plant transformation Transgenic Agrobacterium tumefaciens Plant binary vector Direct PCR testing |
| Content Type | Text |
| Resource Type | Article |
| Subject | Immunology and Microbiology Chemical Engineering Neuroscience Biochemistry, Genetics and Molecular Biology |
