Expression vector construction of Rubisco activase gene CsRCA and genetic transformation to cucumber
2012
A:PS
Details
Title
Expression vector construction of Rubisco activase gene CsRCA and genetic transformation to cucumber
Publication Date
2012
Call Number
A:PS
Summary
Ribulose-1,5-bisphosphate carboxylase/xygenaseRubiscois a key enzyme in the photosynthetic carbon cycle. In order to elucidate the activating mechanism of Rubisco in plantsthe Rubisco activase gene CsRCA was introduced into inbred line of cucumber with the Agrobacterium-mediated method. The transgenic plants were screened by PCRand their expression in mRNA and protein level were analysed by real-time PCR and western blot respectively. The restriction enzyme result showed that the CsRCA was inserted into the binary vector pCAMBIA1301a sense expression vector containing CsRCA gene was constructed. The resulting plasmid was introduced into cucumber inbred lines‘08-1’and seven transgenic plants which contain two CsRCA copies were obtainedThe wild type plants contain one CsRCA copy. The transformation rate was about 3.5%. The seven T0 transgenic cucumber plants showed 11.98 folds in CsRCA mRNA abundance and a significant increase in expression of protein level compared with the wild typeWTplants. CsRCA over expression led to significant increase in the pigment contentphotosynthetic ratePnsoluble sugar and starch contents in T1 transgenic plant leaves. These data indicated that stable genetic CsRCA transgenic cucumber plants were obtained with Agrobacterium-mediated method. CsRCA over expression increased the Pn and carbohydrate significantly in cucumber leaves.
Journal Citation
39(5):869-878, ACTA HORTICULTURAE SINICA
Contact Information
harvest@worldveg.org
Record Appears in
Research > Published Articles