Abstract
The enzyme 4-coumarate:coenzyme A ligase (4CL) plays an important role in phenylpropanoid metabolism. The 5′-upstream regions of two Sm4CL genes were isolated from danshen (Salvia miltiorrhiza Bunge) and their functions were characterized by promoter-directed GUS gene expression assay in transgenic Arabidopsis. Seedlings containing pSm4CL1 promoter:GUS fusions showed apparent GUS staining in hypocotyl and those harboring pSm4CL2 promoter:GUS fusions were clearly stained in cotyledon vasculars and roots. Mature Arabidopsis transformed with pSm4CL1 promoter:GUS exhibited GUS expression which was weak in the shoots and scarcely in roots and those modified with pSm4CL2 promoter:GUS displayed obvious GUS staining in roots, stigmatic papillae, stamens and sepal veins. Semi-quantitative RT-PCR revealed that Sm4CL2 was transcribed at the highest level in roots which was also shown to be the major accumulation site of salvianolic acid B. The results suggested that Sm4CL2 rather than Sm4CL1 might be responsible for the biosynthesis of salvianolic acid B in danshen roots.
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Abbreviations
- 4CL:
-
4-coumarate:coenzyme A ligase
- GUS:
-
β-glucuronidase
- HPLC:
-
high performance liquid chromatography
- MeJA:
-
methyl jasmonate
- MS:
-
Murashige and Skoog
- RA:
-
rosmarinic acid
- RT-PCR:
-
reverse transcription polymerase chain reaction
- SAB:
-
salvianolic acid B
- X-gluc:
-
5-bromo-4-chloro-3-indolyl-β-D-glucuronide
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Acknowledgement
This work was financially supported by National Natural Science Foundation of China (No. 30300447, No. 30973878) and Shanghai Shuguang Project (No. 06SG44). We would like to thank Professor Shouming Zhong for modifying the article, Prof. Yuke He for providing Agrobacterium tumefaciens GV3101 and Arabidopsis (Columbia) and Dr. Gonghao Jiang and Mr. Guixuan Song for technical assistance. The nucleotide sequence data reported have been submitted at the Genbank database under the accession numbers EF458149 (Sm4CL1 5’ upstream region) and EF458150 (Sm4CL2 5′ upstream region).
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Jin, X.Q., Chen, Z.W., Tan, R.H. et al. Isolation and functional analysis of 4-coumarate:coenzyme A ligase gene promoters from Salvia miltiorrhiza . Biol Plant 56, 261–268 (2012). https://doi.org/10.1007/s10535-012-0085-3
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DOI: https://doi.org/10.1007/s10535-012-0085-3