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Expression Analysis of Phenylalanine Ammonia Lyase Gene and Rosmarinic Acid Production in Salvia officinalis and Salvia virgata Shoots Under Salicylic Acid Elicitation

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Abstract

Partial fragments of phenylalanine ammonia lyase (PAL) genes were cloned and characterized from Salvia officinalis (SoPAL) and Salvia virgata (SvPAL). Different concentrations (250 and 500 μM) of exogenous salicylic acid (SA) were used when correlation between PAL expression and rosmarinic acid (RA) accumulation was compared. The results showed that the deduced cDNA sequences of the partial genes had high similarities with those of known PAL gene from other plant species. Semi-quantitative reverse transcription PCR (RT-PCR) analysis revealed that exogenous application of SA led to up-regulating of the PAL expression. Further analysis showed that in S. virgata, at higher concentration of SA, higher accumulation of RA was achieved, while in S. officinalis, the higher RA accumulation was observed at lower concentration of SA. It was concluded that there was no positive correlation between the intensity of PAL transcription and the RA accumulation in the studied species. Therefore, despite of the increase in transcription rate of the PAL at the higher concentration of SA, the lower amounts of RA were accumulated in the case of S. officinalis. Consequently, the hypothesis that PAL is the rate-determining step in RA biosynthesis is not always valid and probably some other unknown factors participate in the synthesis of phenolics.

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References

  1. Kupeli Akkol E., Goger F., Kosar M., & Can Baser K. H. (2008). Phenolic composition and biological activities of Salvia halophila and Salvia virgata from Turkey. Food Chemistry, 108, 942–949.

    Article  Google Scholar 

  2. Ebrahimabadi A. H., Mazoochi A., Jookar Kashi F., Djafari-Bidgoli Z., & Batooli B. (2010). Essential oil composition and antioxidant and antimicrobial properties of the aerial parts of Salvia eremophila Boiss. From Iran. Food and Chemical Toxicology, 48, 1371–1376.

    Article  CAS  Google Scholar 

  3. Tosun M., Ercisli S., Sengul M., Ozer H., Polat T., & Ozturk E. (2009). Antioxidant properties and total phenolic content of eight Salvia species from Turkey. Biological Research, 42, 175–181.

    Article  CAS  Google Scholar 

  4. Lu Y., & Foo Y. L. (2002). Polyphenolics of Salvia. Phytochemistry, 59, 117–140.

    Article  CAS  Google Scholar 

  5. Scarpati M. L., & Oriente G. (1958). Isolamento e costituzione dell’acido rosmarinico (dal Rosmarinus off.). La Ricerca Scientifica, 28, 2329–2333.

    CAS  Google Scholar 

  6. Weitze C., & Petersen M. (2010). Enzymes of phenylpropanoid metabolism in the important medicinal plant Melissa officinalis L. Planta, 232, 731–742.

    Article  Google Scholar 

  7. Swarup V., Ghosh J., Ghosh S., Saxena A., & Basu A. (2007). Anti viral and anti inflammatory effect of rosmarinic acid in an experimental murine model of Japanese encephalitis. Journal of Antimicrobial Chemotherapy, 51, 3367–3370.

    Article  CAS  Google Scholar 

  8. Huang B., Yi B., Duan Y., Sun L., Yu X., Guo J., & Chen W. (2008). Characterization and expression profiling of tyrosine aminotransferase gene from Salvia miltiorrhiza (Dan-shen) in rosmarinic acid biosynthesis pathway. Molecular Biology Reports, 35, 601–612.

    Article  CAS  Google Scholar 

  9. Yan Q., Shi M., Ng J., & Yong Wu J. (2006). Elicitor-induced rosmarinic acid accumulation and secondary metabolism enzyme activities in Salvia miltiorrhiza hairy roots. Plant Science, 170, 853–858.

    Article  CAS  Google Scholar 

  10. Weitzel C., & Petersen M. (2011). Cloning and characterisation of rosmarinic acid synthase from Melissa officinalis L. Phytochemistry, 72, 572–578.

    Article  CAS  Google Scholar 

  11. Berger A., Meinhard J., & Petersen M. (2006). Rosmarinic acid synthase is a new member of the superfamily of BAHD acyltransferases. Planta, 224, 1503–1510.

    Article  CAS  Google Scholar 

  12. Wen P. F., Chen J. Y., Kong W. F., Pan Q. H., Wan S. B., & Huang W. D. (2005). Salicylic acid induced the expression of phenylalanine ammonia-lyase gene in grape berry. Plant Science, 169, 928–934.

    Article  CAS  Google Scholar 

  13. Song J., & Wang Z. (2009). Molecular cloning, expression and characterization of a phenylalanine ammonia-lyase gene (SmPAL1) from Salvia miltiorrhiza. Molecular Biology Reports, 36, 939–952.

    Article  CAS  Google Scholar 

  14. Song J., & Wang Z. (2011). RNAi-mediated suppression of the phenylalanine ammonia-lyase gene in Salvia miltiorrhiza causes abnormal phenotypes and a reduction in rosmarinic acid biosynthesis. Plant Research, 124, 183–192.

    Article  CAS  Google Scholar 

  15. Razzaque A., & Ellis B. E. (1997). Rosmarinic acid production in Coleus cell cultures. Planta, 137, 287–291.

    Article  Google Scholar 

  16. Chen H., & Chen F. (2000). Effect of yeast elicitor on the secondary metabolism of Ti-transformed Salvia miltiorrhiza cell suspension cultures. Plant Cell Reports, 19, 710–717.

    Article  CAS  Google Scholar 

  17. Murashige T., & Skoog F. (1962). A revised medium for rapid growth and bioassays with tobacco cultures. Physiologia Plantarum, 15, 473–497.

    Article  CAS  Google Scholar 

  18. Perez-Tortosa V., Lopez-Orenes A., Martinez-Perez A., Ferrer M. A., & Calderon A. A. (2012). Antioxidant activity and rosmarinic acid changes in salicylic acid-treated Thymus membranaceus shoots. Food Chemistry, 130, 362–369.

    Article  CAS  Google Scholar 

  19. Dong J., Wan G., & Liang Z. (2010). Accumulation of salicylic acid-induced phenolic compounds and raised activities of secondary metabolic and antioxidative enzymes in Salvia miltiorrhiza cell culture. Biotechnology, 148, 99–104.

    CAS  Google Scholar 

  20. Jiao M., Cao R., Chen H., Hao W., & Dong J. (2012). Effects of salicylic acid on synthesis of rosmarinic acid and related enzymes in the suspension cultures of Salvia miltiorrhiza. Sheng Wu Gong Xue Bao, 28, 320–328.

    CAS  Google Scholar 

  21. Petersena M., & Simmonds M. S. J. (2003). Molecules of interest rosmarinic acid. Phytochemistry, 62, 121–125.

    Article  Google Scholar 

  22. Shah, Y. and Klessig, D.F. (1999). Salicylic acid: signal perception and transduction. In P.J.J. Hooykaas, M.A. Hall, K.R. Libbenga (Eds.), Biochemistry and Molecular Biology of Plant Hormones (pp. 513–541). Amsterdam: Elsevier Science.

    Chapter  Google Scholar 

  23. Fraissinet-Tachet L., Baltz R., Chong J., Kauffmann S., Fritig B., & Saindrenan P. (1998). Two tobacco genes induced by infection, elicitor and salicylic acid encode glucosyltransferases acting on phenylpropanoids and benzoic acid derivatives, including salicylic acid. FEBS Letters, 437, 319–323.

    Article  CAS  Google Scholar 

  24. Ding C. K., Wang C. Y., Gross K. C., & Smith D. L. (2002). Jasmonate and salicylate induce the expression of pathogenesis-related-protein genes and increase resistance to chilling injury in tomato fruit. Planta, 214, 895–901.

    Article  CAS  Google Scholar 

  25. Thulke O., & Conrath U. (1998). Salicylic acid has a dual rule in the activation of defence related genes in parsley. The Plant Journal, 14, 35–42.

    Article  CAS  Google Scholar 

  26. Mizukami H., Ogawa T., Ohashi H., & Ellis B. E. (1992). Induction of rosmarinic acid biosynthesis in Lithospermum erythrorhizon cell suspension cultures by yeast extract. Plant Cell Reports, 11, 480–483.

    Article  CAS  Google Scholar 

  27. Ogata A., Tsuruga A., Matsuno M., & Mizukami H. (2004). Elicitor induced rosmarinic acid biosynthesis in Lithospermum erythrorhizon cell suspension cultures: activities of rosmarinic acid synthase and the final two cytochrome P450-catalyzed hydroxylations. Plant Biotechnology Journal, 21, 393–396.

    Article  CAS  Google Scholar 

  28. Szabo E., Thelen A., & Petersen M. (1999). Fungal elicitor preparations and methyl jasmonate enhance rosmarinic acid accumulation in in vitro-cultures of Coleus blumei. Plant Cell Reports, 18, 485–489.

    Article  CAS  Google Scholar 

  29. Bauer N., Fulgosi H., & Jelaska S. (2011). Overexpression of phenylalanine ammonia-lyase in transgenic roots of Coleus blumei alters growth and rosmarinic acid synthesis. Food Technology and Biotechnology, 49, 24–31.

    CAS  Google Scholar 

  30. Bauer N., Kiseljak D., & Jelaska S. (2009). The effect of yeast extract and methyl jasmonate on rosmarinic acid accumulation in Coleus blumei hairy roots. Biologia Plantarum, 53, 650–656.

    Article  CAS  Google Scholar 

  31. Liu R., Hu Y., Li H., & Lin Z. (2007). Production of soybean isoflavone genistein in non-legume plants via genetically modified secondary metabolism pathway. Journal of Metabolic Engineering, 9, 1–7.

    Article  Google Scholar 

  32. Li W., Koike K., Asada Y., Yoshikawa T., & Nokaido T. (2005). Rosmarinic acid production by Coleus forskohlii hairy root cultures. Plant Cell, Tissue and Organ Culture, 80, 151–155.

    Article  CAS  Google Scholar 

  33. Zhang S., Yan Y., Wang B., Liang Z., Liu Y., Liu F., & Qi Z. (2014). Selective responses of enzymes in the two parallel pathways of rosmarinic acid biosynthetic pathway to elicitors in Salvia miltiorrhiza hairy root cultures. Journal of Bioscience and Bioengineering, 117, 645–651.

    Article  CAS  Google Scholar 

  34. Staszkow A., Swarcewicz B., Banasiak J., Muth D., Jasinski M., & Stobiecki M. (2011). LC/MS profiling of flavonoid glycoconjugates isolated from hairy roots, suspension root cell cultures and seedling roots of Medicago truncatula. Metabolomics, 7, 604–613.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors wish to express their gratitude to the Council of Shahed University for the financial supports during the course of this research. The authors also thank the Biotechnology Lab of University of Kashan for the molecular and plant tissue culture research works.

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Authors and Affiliations

  1. Department of Biology, Faculty of Basic Sciences, Shahed University, Tehran, 33191-18651, Iran

    Roghayeh sadat Ejtahed & Tayebeh Radjabian

  2. Faculty of Chemistry, Department of Biotechnology, University of Kashan, Kashan, 8731751167, Iran

    Roghayeh sadat Ejtahed & Sayed Ali Hoseini Tafreshi

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  1. Roghayeh sadat Ejtahed
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  2. Tayebeh Radjabian
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  3. Sayed Ali Hoseini Tafreshi
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Correspondence to Sayed Ali Hoseini Tafreshi.

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Ejtahed, R.s., Radjabian, T. & Hoseini Tafreshi, S.A. Expression Analysis of Phenylalanine Ammonia Lyase Gene and Rosmarinic Acid Production in Salvia officinalis and Salvia virgata Shoots Under Salicylic Acid Elicitation. Appl Biochem Biotechnol 176, 1846–1858 (2015). https://doi.org/10.1007/s12010-015-1682-3

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  • DOI: https://doi.org/10.1007/s12010-015-1682-3

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