Back to Search
Start Over
Agrobacterium-mediated vacuum infiltration and floral dip transformation of rapid-cycling Brassica rapa
- Source :
- BMC Plant Biology, Vol 19, Iss 1, Pp 1-9 (2019), BMC Plant Biology
- Publication Year :
- 2019
- Publisher :
- BMC, 2019.
-
Abstract
- Background Rapid-cycling Brassica rapa (RCBr), also known as Wisconsin Fast Plants, are small robust plants with a short lifecycle that are widely used in biology teaching. RCBr have been used for decades but there are no published reports of RCBr genetic transformation. Agrobacterium-mediated vacuum infiltration has been used to transform pakchoi (Brassica rapa ssp. chinensis) and may be suitable for RCBr transformation. The floral dip transformation method, an improved version of vacuum infiltration, could make the procedure easier. Results Based on previous findings from Arabidopsis and pakchoi, plants of three different ages were inoculated with Agrobacterium. Kanamycin selection was suboptimal with RCBr; a GFP screen was used to identify candidate transformants. RCBr floral bud dissection showed that only buds with a diameter less than 1 mm carried unsealed carpels, a key point of successful floral dip transformation. Plants across a wide range of inflorescence maturities but containing these immature buds were successfully transformed, at an overall rate of 0.1% (one per 1000 T1 seeds). Transformation was successful using either vacuum infiltration or the floral dip method, as confirmed by PCR and Southern blot. Conclusion A genetic transformation system for RCBr was established in this study. This will promote development of new biology teaching tools as well as basic biology research on Brassica rapa. Electronic supplementary material The online version of this article (10.1186/s12870-019-1843-6) contains supplementary material, which is available to authorized users.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Gynoecium
Agrobacterium
Vacuum infiltration
Floral dip
Plant Science
Flowers
Biology
01 natural sciences
Polymerase Chain Reaction
03 medical and health sciences
Transformation, Genetic
Rapid cycling
Arabidopsis
lcsh:Botany
Brassica rapa
medicine
Methodology Article
fungi
food and beverages
Kanamycin
biology.organism_classification
Wisconsin fast plants
lcsh:QK1-989
Transformation (genetics)
Horticulture
Blotting, Southern
030104 developmental biology
Inflorescence
Agrobacterium-mediated transformation
Genetic Engineering
Rapid-cycling Brassica rapa
010606 plant biology & botany
medicine.drug
Subjects
Details
- Language :
- English
- ISSN :
- 14712229
- Volume :
- 19
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- BMC Plant Biology
- Accession number :
- edsair.doi.dedup.....2d6d4c23166116ab0e08ce8012fc336c
- Full Text :
- https://doi.org/10.1186/s12870-019-1843-6