Abstract
BackgroundRapid-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.ResultsBased 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.ConclusionA 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.
Highlights
Rapid-cycling Brassica rapa (RCBr), known as Wisconsin Fast Plants, are small robust plants with a short lifecycle that are widely used in biology teaching
To identify a useful method for a rapid-cycling Brassica rapa (RCBr) transformation system, kanamycin resistance and green fluorescent protein (GFP) expression were tested on RcBC 1-33, the standard Wisconsin Fast Plants line of RCBr
These results suggested that RCBr is relatively kanamycin-insensitive and that GFP would be a better selection marker than kanamycin resistance in a RCBr transformation system
Summary
Rapid-cycling Brassica rapa (RCBr), known as Wisconsin Fast Plants, are small robust plants with a short lifecycle that are widely used in biology teaching. Rapid-cycling Brassica rapa (RCBr), known as Wisconsin Fast Plants, were derived from genetic crossing among multiple faster flowering Brassica rapa [1]. Because of this origin, RCBr inherited diverse traits. The plants are petite and have a very short lifecycle of about 40 days if grown under continuous light Due to these desirable characteristics, and the development of clever and inexpensive teaching modules by the Wisconsin Fast Plants program, RCBr have been used in thousands of classrooms to educate students about biology [2]. Development of a transformation system would promote both basic research on Brassica rapa and enhanced use of RCBr as a teaching tool
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