Abstract
The aim of this study was to investigate the effect of phenylalanine ammonia lyase (PAL) and isochorismate synthase (ICS) on free salicylic acid (FSA) or total salicylic acid (TSA) content, and the effect of endogenous SA on baicalin and baicalein accumulation in Scutellaria baicalensis Georgi, respectively. We amplified partial sequences of PAL and ICS genes in Scutellaria baicalensis Georgi and silenced the two genes with virus-induced gene silence (VIGS) technique, respectively. The influence of gene silence on FSA, TSA, baicalin, and baicalein accumulation in Scutellaria baicalensis Georgi were analyzed, and these parameters were also investigated under high temperature. Results indicated that PAL silence significantly affected the FSA, ICS affected TSA content. FSA significantly affected the baicalin, rather than baicalein content. Our results along with previous studies indicated PAL and ICS were different in the regulation of FSA or TSA synthesis, and FSA and TSA were different in the regulation of baicalin and baicalein synthesis in Scutellaria baicalensis Georgi.
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Abbreviations
- SA:
-
Salicylic acid
- PAL:
-
Phenylalanine ammonia lyase
- ICS:
-
Isochorismate synthase
- FSA:
-
Free salicylic acid
- TSA:
-
Total salicylic acid
- VIGS:
-
Virus induced gene silence
- IPL:
-
Isochorismate pyruvate-lyase
- HT:
-
High temperature
References
Ali M, Yu K-W, Hahn E-J, Paek K-Y (2006) Methyl jasmonate and salicylic acid elicitation induces ginsenosides accumulation, enzymatic and non-enzymatic antioxidant in suspension culture Panax ginseng roots in bioreactors. Plant Cell Rep 25(6):613–620. doi:10.1007/s00299-005-0065-6
Dewdney J, Reuber TL, Wildermuth MC, Devoto A, Cui J, Stutius LM, Drummond EP, Ausubel FM (2000) Three unique mutants of Arabidopsis identify eds loci required for limiting growth of a biotrophic fungal pathogen. Plant J 24(2):205–218
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. J Biotechnol 148(2–3):99–104. doi:10.1016/j.jbiotec.2010.05.009
Ekengren SK, Liu Y, Schiff M, Dinesh-Kumar SP, Martin GB (2003) Two MAPK cascades, NPR1, and TGA transcription factors play a role in Pto-mediated disease resistance in tomato. Plant J 36(6):905–917
Huang J, Gu M, Lai Z, Fan B, Shi K, Zhou YH, Yu JQ, Chen Z (2010) Functional analysis of the Arabidopsis PAL gene family in plant growth, development, and response to environmental stress. Plant Physiol 153(4):1526–1538. doi:10.1104/pp.110.157370
Hwang SJ (2006) Baicalin production in transformed hairy root clones of Scutellaria baicalensis. Biotechnol Bioprocess Eng 11(2):105–109. doi:10.1007/bf02931892
Kovacik J, Klejdus B (2012) Tissue and method specificities of phenylalanine ammonia-lyase assay. J Plant Physiol 169(13):1317–1320. doi:10.1016/j.jplph.2012.04.008
Kovacik J, Gruz J, Backor M, Strnad M, Repcak M (2009) Salicylic acid-induced changes to growth and phenolic metabolism in Matricaria chamomilla plants. Plant Cell Rep 28(1):135–143. doi:10.1007/s00299-008-0627-5
Lee YS, Ju HK, Kim YJ, Lim TG, Uddin MR, Kim YB, Baek JH, Kwon SW, Lee KW, Seo HS, Park SU, Yang TJ (2013) Enhancement of anti-inflammatory activity of Aloe vera adventitious root extracts through the alteration of primary and secondary metabolites via salicylic acid elicitation. PLoS One 8(12):e82479. doi:10.1371/journal.pone.0082479
Liu Y, Schiff M, Dinesh-Kumar SP (2002) Virus-induced gene silencing in tomato. Plant J 31(6):777–786. doi:10.1046/j.1365-313X.2002.01394.x
Mahdavian K, Kalantari KM, Ghorbanli M, Torkzade M (2008) The effects of salicylic acid on pigment contents in ultraviolet radiation stressed pepper plants. Biol Plant 52(1):170–172. doi:10.1007/s10535-008-0037-0
Mauch F, Mauch-Mani B, Gaille C, Kull B, Haas D, Reimmann C (2001) Manipulation of salicylate content in Arabidopsis thaliana by the expression of an engineered bacterial salicylate synthase. Plant J 25(1):67–77
Metraux JP (2002) Recent breakthroughs in the study of salicylic acid biosynthesis. Trends Plant Sci 7(8):332–334
Nugroho LH, Verberne MC, Verpoorte R (2002) Activities of enzymes involved in the phenylpropanoid pathway in constitutively salicylic acid-producing tobacco plants. Plant Physiol Biochem 40(9):755–760. doi:10.1016/S0981-9428(02)01437-7
Ogawa D, Nakajima N, Tamaoki M, Aono M, Kubo A, Kamada H, Saji H (2007) The isochorismate pathway is negatively regulated by salicylic acid signaling in O3-exposed Arabidopsis. Planta 226(5):1277–1285. doi:10.1007/s00425-007-0556-5
Pallas JA, Paiva NL, Lamb C, Dixon RA (1996) Tobacco plants epigenetically suppressed in phenylalanine ammonia-lyase expression do not develop systemic acquired resistance in response to infection by tobacco mosaic virus. Plant J 10(2):281–293. doi:10.1046/j.1365-313X.1996.10020281.x
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29(9):e45
Poulsen CH, van der Heijden R, Verpoorte R (1991) Assay of isochorismate synthase from plant cell cultures by high performance liquid chromatography. Phytochemistry 9:2873–2876
Sadeghi M, Dehghan S, Fischer R, Wenzel U, Vilcinskas A, Kavousi HR, Rahnamaeian M (2013) Isolation and characterization of isochorismate synthase and cinnamate 4-hydroxylase during salinity stress, wounding, and salicylic acid treatment in Carthamus tinctorius. Plant Signal Behav 8(11):e27335. doi:10.4161/psb.27335
Strawn MA, Marr SK, Inoue K, Inada N, Zubieta C, Wildermuth MC (2007) Arabidopsis isochorismate synthase functional in pathogen-induced salicylate biosynthesis exhibits properties consistent with a role in diverse stress responses. J Biol Chem 282(8):5919–5933. doi:10.1074/jbc.M605193200
Su H, Jiang H, Li YP (2013) Effects of PAL and ICS on the production of total flavonoids, daidzein and puerarin in Pueraria thomsonii Benth. suspension cultures under low light stress. J Plant Biochem Biot 24(1):34–41. doi:10.1007/s13562-013-0233-7
Wang L, Cao Y, Xu H, Ye J (2006) Study on quality control and fingerprint of Chinese traditional medicine Yinhuang oral liquid. Chin J Chromatogr 24(4):367–372
Xue LJ, Guo W, Yuan Y, Anino EO, Nyamdari B, Wilson MC, Frost CJ, Chen HY, Babst BA, Harding SA, Tsai CJ (2013) Constitutively elevated salicylic acid levels alter photosynthesis and oxidative state but not growth in transgenic populus. Plant Cell 25(7):2714–2730. doi:10.1105/tpc.113.112839
Wildermuth MC, Dewdney J, Wu G, Ausubel FM (2001) Isochorismate synthase is required to synthesize salicylic acid for plant defence. Nature 414 (6863):562–565. http://www.nature.com/nature/journal/v414/n6863/suppinfo/414562a_S1.html
Yoon HJ, Kim HK, Ma C-J, Huh H (2000) Induced accumulation of triterpenoids in Scutellaria baicalensis suspension cultures using a yeast elicitor. Biotechnol Lett 22(13):1071–1075. doi:10.1023/a:1005610400511
Zhang JJ, Xu MJ (2006) Effects of nitric oxide and methyl jasmonate on the baicalin production and cell growth in suspension cultures of Scutellaria baicalensis. Chin Bull Bot 23:374–379
Zhao X, Wang J, Yuan J, Wang XL, Zhao QP, Kong PT, Zhang X (2015) NITRIC OXIDE-ASSOCIATED PROTEIN1 (AtNOA1) is essential for salicylic acid-induced root waving in Arabidopsis thaliana. New phytol 207(1):211–224. doi:10.1111/nph.13327
Zhou J, Fang L, Li X, Guo L, Huang L (2012) Jasmonic Acid (JA) Acts as a signal molecule in LaCl3-Induced Baicalin Synthesis in Scutellaria baicalensis Seedlings. Biol Trace Elem Res 148(3):392–395. doi:10.1007/s12011-012-9379-8
Acknowledgments
This work was supported by the National Natural Science Foundation of China (Grant No. 81102803) and “555 talent project” of Jiangxi Province, Jiangxi Key Laboratory of Bioprocess Engineering and Co-Innovation Center for In Vitro Diagnostic Reagents and Devices of Jiangxi Province (Grant no. 31171731, 31460447, 20142BDH80003, 2013-CXTD002).
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Communicated by A. Krolicka.
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Su, H., Yu, C., Shang, J. et al. Effects of endogenous salicylic acid synthesized through PAL and ICS pathway on baicalin and baicalein accumulation in Scutellaria baicalensis Georgi. Acta Physiol Plant 38, 178 (2016). https://doi.org/10.1007/s11738-016-2193-8
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DOI: https://doi.org/10.1007/s11738-016-2193-8