Osmotic stress affects polyamine homeostasis and phenolic content in proembryogenic liquid cell cultures of Scots pine
- 417 Downloads
Polyamines (PAs) are ubiquitous polycations involved in many physiological processes in plants, including somatic embryogenesis, cellular growth and stress reactions. In the present study, we focus on the consequences in PA metabolism caused by polyethylene glycol (PEG) in proembryogenic Scots pine (Pinus sylvestris L.) liquid cultures. The growth and viability of the cell masses and changes in PA concentrations and phenolic secondary metabolites were investigated under control, 5 and 10 % PEG treatments. The effect of osmotic stress responses was investigated at the gene expression level including stress, cell division, programmed cell death and PA-related genes and PA metabolites. Moreover, the expression of ethylene and proline biosynthesis genes and phenylalanine ammonia lyase and stilbene synthase (psSTS) was analyzed. Under osmotic stress conditions, we found a consistent pattern of endogenous PAs in Scots pine proembryogenic cells. However, accumulation of free spermine (Spm) and methyl putrescine under osmotic stress might indicate their specific role in stress protection. Expression of polyamine oxidase was down-regulated under osmotic stress, suggesting the role of PA catabolism in regulation of Spm levels. Scots pine proliferating proembryogenic cells are in a developmentally undifferentiated stage where the content of secondary metabolites is generally low. However, in the present study the total content of phenolic compounds increased but the biosynthesis of phenylpropanoids and stilbenes, generally considered as stress-protective molecules, was not affected by osmotic stress.
KeywordsOsmotic stress Scots pine Embryogenic cells Liquid cell culture Polyamines Phenols
We are grateful to Ms. Eeva Pihlajaviita, Ms. Anneli Käenmäki and Ms. Hanna Leppälammi for their skillful technical help. We thank Meeri Pearson for proofreading the manuscript. This research was funded by the Thule Institute (2010–2013) (to HH), Academy of Finland (Project 121994 to TS) and Graduate School of Forest Sciences (to RMM).
Conflict of interest
The authors declare that they have no conflict of interest.
- Cvikrová M, Binarová P, Eder J, Vágner M, Hrubcová M, Zoń J, Machácˇková I (1999) Effect of inhibition of phenylalanine ammonia-lyase activity on growth of alfalfa cell suspension culture: alterations in mitotic index, ethylene production, and contents of phenolics, cytokinins, and polyamines. Physiol Plant 107:329–337CrossRefGoogle Scholar
- Cvikrová M, Malá J, Hrubcová M, Eder J, Foretová S (2008) Induced changes in phenolic acids and stilbenes in embryogenic cell cultures of Norway spruce by culture filtrate of Ascocalyx abietina. J Plant Dis Prot 115:57–62Google Scholar
- Dutra NT, Silveira V, Azevedo IG, Gomes-Neto LR, Façanha AR, Steiner N, Guerra MP, Floh EIS, Santa-Catarina C (2013) Polyamines affect the cellular growth and structure of pro-embryogenic masses in Araucaria angustifolia embryogenic cultures through the modulation of proton pump activities and endogenous levels of polyamines. Physiol Plant 148:121–132PubMedCrossRefGoogle Scholar
- Fitzmaurice GM, Laird N, Ware JH (2004) Applied longitudinal analysis. Wiley, HobokenGoogle Scholar
- Fliegmann J, Schröder G, Schanz S, Britsch L, Schröder J (1992) Molecular analysis of chalcone synthase and dihydropinosylvin synthase from Scots pine (Pinus sylvestris), and differential regulation of these and related enzyme activities in stressed plants. Plant Mol Biol 18:489–503PubMedCrossRefGoogle Scholar
- Häggman H, Vuosku J, Sarjala T, Jokela A, Niemi K (2006) Somatic embryogenesis of pine species—from functional genomics to plantation forestry. In: Mujib A, Samaj J (eds) Somatic embryogenesis. In series: plant cell monographs, vol 2. Springer, Berlin, pp 119–140Google Scholar
- Moschou PN, Paschalidis KA, Delis ID, Andriopoulou AH, Lagiotis GD, Yamakoumakis DI, Kalliopi A, Roubelakis-Angelakis KA (2008) Spermidine exodous and oxidation in the apoplast induced by abiotic stress is responsible for H2O2 signatures that direct tolerances in tobacco. Plant Cell 20:1708–1724PubMedCentralPubMedCrossRefGoogle Scholar
- Pinheiro J, Bates D, DebRoy S, Sarkar D and the R Development Core Team (2012) nlme: linear and nonlinear mixed effects models. R package version 3.1-103Google Scholar
- R Development Core Team (2012) R: a language and environment for statistical computing. R Foundation for statistical computing, Vienna, Austria. http://www.R-project.org/. ISBN 3-900051-07-0
- Silveira V, Santa-Catarina C, Tun N, Scherer G, Handro W, Guerra M, Floh E (2006) Polyamine effects on the endogenous polyamine contents, nitric oxide release, growth and differentiation of embryogenic suspension cultures of Araucaria angustifolia (Bert.) O. Ktze. Plant Sci 171:91–98CrossRefGoogle Scholar
- Skirycz A, Claeys H, De Bodt S, Oikawa A, Shinoda S, Andriankaja M, Maleux K, Eloy NB, Coppens F, Yoo SD, Saito K, Inze D (2011) Pause-and-stop: the effects of osmotic stress on cell proliferation during early leaf development in arabidopsis and a role for ethylene signaling in cell cycle arrest. Plant Cell 23:1876–1888PubMedCentralPubMedCrossRefGoogle Scholar
- Viitala K, Potila H, Savonen E-M, Sarjala T (2011) Health from forest-antioxidative properties of endophytic fungi from Scots pine roots. Proceedings of the 7th international conference on mushroom biology and mushroom products (ICMBMP7)Google Scholar
- Vuosku J (2011) A matter of life and death—polyamine metabolism during zygotic embryogenesis of pine. PhD thesis, Acta Universitatis Ouluensis, Scientiae rerum naturalium 573, Oulu, FinlandGoogle Scholar
- Vuosku J, Sutela S, Kestilä J, Jokela A, Sarjala T, Häggman H (2015) Expression of catalase and retinoblastoma-related protein genes associated with cell death processes in Scots pine zygotic embryogenesis. BMC Plant Biol 15:88. doi: 10.1186/s12870-015-0462-0 PubMedCentralPubMedCrossRefGoogle Scholar