Abstract
Capsanthin/capsorubin synthase (Ccs) gene is a key gene that regulates the synthesis of capsanthin and the development of red coloration in pepper fruits. There are three tandem repeat units in the promoter region of Ccs, but the potential effects of the number of repetitive units on the transcriptional regulation of Ccs has been unclear. In the present study, expression vectors carrying different numbers of repeat units of the Ccs promoter were constructed, and the transient expression of the β-glucuronidase (GUS) gene was used to detect differences in expression levels associated with the promoter fragments. These repeat fragments and the plant expression vector PBI121 containing the 35s CaMV promoter were ligated to form recombinant vectors that were transfected into Agrobacterium tumefaciens GV3101. A fluorescence spectrophotometer was used to analyze the expression associated with the various repeat units. It was concluded that the constructs containing at least one repeat were associated with GUS expression, though they did not differ from one another. This repeating unit likely plays a role in transcription and regulation of Ccs expression.
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Abbreviations
- Ccs :
-
Capsanthin/capsorubin synthase
- GUS :
-
β-Glucuronidase
- PL0:
-
Vector carrying full-length Ccs promoter fragments with W box and AACA motif
- PL1:
-
Vector carrying full-length Ccs promoter fragments without W box or AACA motif
- PL2:
-
Vector carrying two repeat units of the promoter fragment
- PL2-1:
-
Vector carrying one repeat unit of the promoter fragment
- PL3:
-
Vector carrying no repeat units of the promoter fragment
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Acknowledgements
The authors acknowledge the following organizations for their financial support: National Key Research and Development Program of China (No. 2016YFD0101900), National Natural Science Foundation of China (No. 31272163); Innovation Scientists and Technicians Troop Construction Projects of Henan Province (C20150054); Science and Technology Development Project of Henan Province (162102110084); Key Scientific Research Projects in Universities of Henan Province (16A210010).
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Supplementary Fig. 1–5
Schematic representation of recombinant PBI121 vectors carrying Ccs gene promoter fragments (JPEG 590 kb)
Supplementary Fig. 6
DNA extraction of pepper fruits (JPEG 17 kb)
Supplementary Table 1
Primers were used in plasmid construction (DOCX 13 kb)
Supplementary Table 2
The reaction system (DOCX 13 kb)
Supplementary Table 3
The ligase reaction system (DOCX 13 kb)
Supplementary Table 4
Histochemical staining reactants for assaying GUS activity (DOCX 13 kb)
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Tian, SL., Li, Z., Li, L. et al. Analysis of tandem repeat units of the promoter of capsanthin/capsorubin synthase (Ccs) gene in pepper fruit. Physiol Mol Biol Plants 23, 685–691 (2017). https://doi.org/10.1007/s12298-017-0437-8
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DOI: https://doi.org/10.1007/s12298-017-0437-8