Murat TELLİ, Fatma PEHLİVAN KARAKAŞ, Ercan Selçuk ÜNLÜ, Günce ŞAHİN

Effects of moderate high temperature and UV-B on accumulation of withanolides and relative expression of the squalene synthase gene in Physalis peruviana

Physalis peruviana L. (Cape gooseberry) is a source for a variety of phytocompounds such as withanolides, withanone,withaferin A, and withanolide A. These withanolides are high-value drug candidates due to their various pharmacological properties.To meet the increasing demands for these compounds, plant cell technology offers a reliable alternative. Exogenous addition of elicitorsis considered the most effective strategy for enhanced production of secondary metabolites. In this study, we investigated changes inwithanolide accumulation and characterized the gene expression level changes of squalene synthase enzyme in P. peruviana shootcultures exposed to mild nonlethal heat stress (45°C for 2 and 5 h) and UV-B radiation (313 nm for 15 min and 3 h). We demonstratedsignificant changes in withanolide content with 7.86- and 12.5-fold increases for 2- and 5-hmild high-temperature exposure times,respectively. Exposure to UV-B also changed the withanolide content by 7.22- and 7-fold increases for 15 min and 3 h exposure times,respectively. The relative expression level of squalene synthase gene showed consistent results with1.80- and 10.13-fold increases inwithanolide for 2- and 5-h mild high-temperature exposure times, and 1.34- and 2.01-fold increases with 15 min and 3 h UV-B exposuretimes, respectively

PDF

___

Afreen F, Zobayed SMA, Kozai T (2005). Spectral quality and UV-B stress stimulate glycyrrhizin concentration of Glycyrrhiza uralensis in hydroponic and pot system. Plant Physiology and Biochemistry 43: 1074-1081.
Akula R, Ravishankar GA (2011). Influence of abiotic stress signals on secondary metabolites in plants. Plant Signal Behaviour 6: 1720-1731.
Binder B, Peebles C, Shanks JV, San KY (2009). The effects of UV-B stress on the production of terpenoid indole alkaloids in Catharanthus roseus hairy roots. Biotechnology Progress 25 (3): 861-865.
Chaurasiya ND, Sangwan NS, Sabir F, Misra L, Sangwan RS (2012). Withanolide biosynthesis recruits both mevalonate and DOXP pathways of isoprenogenesis in Ashwagandha Withania somnifera L. (Dunal). Plant Cell Reports 31 (10): 1889-1897.
Cirak C, Radusiene J, Stanius Z, Camas N, Caliskan O et al. (2012). Secondary metabolites of Hypericum orientale L. growing in Turkey: variation among populations and plant parts. Acta Physiologiae Plantarum 34: 1313-1320.
Eichholz I, Huyskens-Keil S, Keller A, Ulrich D, Kroh LW et al. (2011). UV-B-induced changes of volatile metabolites and phenolic compounds in blueberries (Vaccinium corymbosum L). Food Chemistry 126: 60-64.
Galli V, Messias SR, Perin CE, Borowski MJ, Bamberg LA et al. (2016). Mild salt stress improves strawberry fruit quality. Food Science and Technology 73: 693-699.
Gerhardt KE, Lampi MA, Greenberg BM (2008). The effects of far-red light on plant growth and flavonoid accumulation in Brassica napusin the presence of ultraviolet B radiation. Photochemistry and Photobiology 84: 1445-1454.
Glotter E (1991). Withanolides and related ergostane-type steroids. Natural Product Reports 8: 415-440.
Grover A, Samuel G, Bisaria VS, Sundar D (2013). Enhanced withanolide production by overexpression of squalene synthase in Withania somnifera. Journal of Bioscience and Bioengineering 115: 680-685.
Gulen H, Eris A (2004). Effect of heat stress on peroxidase activity and total protein content in strawberry plants. Plant Science 166: 739-744.
Jansen MAK, Hectors K, O’Brien NM, Guisez Y, Potters G (2008). Plant stress and human health: Do human consumers benefit from UV-B acclimated crops? Plant Science 175: 449-458.
Jenkins GI (2009). Signal transduction in responses to UV-B radiation. Annual Review Plant Biology 60: 407-431.
Katoch M, Fazli IS, Suri KA, Ahuja A, Qazi GN (2012). Effect of altitude on picroside content in core collections of Picrorhiza kurrooa from the north western Himalayas. Journal of Natural Medicines 65: 578-582.
Kim O, Bang K, Kim Y, Hyun D, Kim M et al. (2009). Upregulation of ginsenoside and gene expression related to triterpene biosynthesis in ginseng hairy root cultures elicited by methyl jasmonate. Plant Cell Tissue Organ Culture 98: 25-33.
Kim YS, Cho JH, Park S, Han JY, Back K et al. (2011). Gene regulation patterns in triterpene biosynthetic pathway driven by overexpression of squalene synthase and methyl jasmonate elicitation in Bupleurum falcatum. Planta 233:343-355.
Kitchlu S, Ram G, Koul S, Koul K, Gupta KK et al. (2011). Podophyllum lignans array of Podophyllum hexandrum Royle populations from semi-desert alpine region of Zanskar valley in Himalayas. Industrial Crops and Products 33: 584-587.
Kumar A, Kaul MK, Bhan MK, Punit K, Suri KA (2007). Morphological and chemical variations in 25 collections of the Indian medicinal plant, Withania somnifera (L.) Genetic Resources and Crop Evolution 54: 655-660.
Kumar A, Mir BA, Sehgal D, Dar TH, Koul S et al. (2011). Utility of multidisciplinary approach for genome diagnostics of cultivated and wild germplasm resources of medicinal Withania somnifera, and status of new species, W. ashwagandha, in the cultivated taxon. Plant Systematics and Evolution 291: 141-151.
Kumari R, Agrawal SB (2011). Comparative analysis of essential oil composition and oil containing glands in Ocimum sanctum L (Holy basil) under ambient and supplemental level of UV-B through Gas chromatography–mass spectrometry (GC-MS) and Scanning electron microscopy (SEM). Acta Physiologiae Plantarum 33: 1093-1101.
Kumari R, Prasad MNV (2013). Medicinal plant active compounds produced by UV-B exposure. Sustainable Agriculture Reviews 12: 225-254.
Lancaster JE, Reay PF, Norris J, Butler RC (2000). Induction of flavonoids and phenolic acids in apple by UV-B and temperature. Journal of Horticultural Science and Biotechnology 75: 142- 148.
Lee MH, Jeong JH, Seo JW, Shin CG, Kim YS et al. (2004). Enhanced triterpene and phytosterol biosynthesis in Panax ginseng overexpressing squalene synthase gene. Plant Cell Physiology 45: 976-984.
Livak JK, Schmittgen T (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25 (4): 402-408.
Mir BA, Koul S, Soodan AS (2013). Reproductive biology of Withania ashwagandha sp. novo. Industrial Crops and Products 45: 442- 446.
Mirjalili MH, Moyano E, Bonfill M, Cusido RM, Palazon J (2009). Streoidal lactones from Withania somnifera, an ancient plant for novel medicine. Molecules 14: 2373-2393.
Mirjalili MH, Moyano E, Cusido RM, Palazón L (2011). Overexpression of the Arabidopsis thaliana squalene synthase gene in Withania coagulans hairy root cultures. Biologia Plantarum 55: 357-360.
Mishra L, Singh B, Dagenias S, (2000). Scientific basis for the therapeutic use of Withania somnifera (ashwagandha): a review Alternative Medicine Review 5: 334-336.
Morales LO, Tegelberg R, Brosche M, Keinanen M, Lindfors A et al. (2010). Effects of solar UV-A and UV-B radiation on gene expression and phenolic accumulation in Betula pendula leaves. Tree Physiology 30: 923-934.
Neugart S, Fiol M, Schreiner M, Rohn S., Zrenner R et al. (2014). Interaction of moderate UV-B exposure and temperature on the formation of structurally different flavonol glycosides and hydroxycinnamic acid derivatives in kale (Brassica oleracea var. sabellica). Journal of Agricultural Food Chemistry 62: 4054- 4062.
Obata T, Fernie AR (2012). The use of metabolomics to dissect plant responses to abiotic stresses. Cellular and Molecular Life Sciences 69: 3225-3243.
Patel N, Patel P, Kendurkar SV, Thulasiram HV, Khan BM (2015). Overexpression of squalene synthase in Withania somnifera leads to enhanced withanolide biosynthesis. Plant Cell, Tissue and Organ Culture 122: 409-420.
Radman R, Saez T, Bucke C, Keshavarz T (2003). Elicitation of plants and microbial cell systems. Biotechnology Applied Biochemistry 3: 91-102.
Schreiner M, Krumbein A, Mewis I, Ulrichs C, Huyskens-Keil S (2009). Short-term and moderate UV-B radiation effects on secondary plant metabolism in different organs of nasturtium (Tropaeolummajus L.). Innovative Food Science and Emerging Technologies 10: 93-96.
Schreiner M, Mewis I, Huyskens-Keil S, Jansen MAK, Zrenner R et al. (2012). UV -B-induced secondary plant metabolitespotential benefits for plant and human health. Critical Reviews in Plant Sciences 31: 229-240.
Selmar D, Kleinwächter M (2013). Stress enhances the synthesis of secondary plant products: the impact of stress-related overreduction on the accumulation of natural products. Plant Cell Physiology 54 (6): 817-826.
Seo JW, Jeong JH, Shin CH, Lo SC, Han SS et al. (2005). Overexpression of squalene synthase in Eleutherococcus senticosus increases phytosterol and triterpene accumulation. Phytochemistry 66: 869-877.
Sung DY, Kaplan F, Lee KJ, Guy CL (2003). Acquired tolerance to temperature extremes. Trends in Plant Science 8: 179-187.
Takshak S, Agrawal SB (2014). Effect of ultraviolet-B radiation on biomass production, lipid peroxidation, reactive oxygen species, and antioxidants in Withania somnifera. Biology Plantarum 58: 328-334.
Thimmaraju R, Bhagyalakshmi N, Narayan MS, Ravishankar GA (2003). Kinetics of pigment release from hairy root cultures of Beta vulgaris under the influence of pH, sonication, temperature and oxygen stress. Process Biochemistry 38: 1069- 1076.
Tozawa R, Ishibashi S, Osuga J, Yagyu H, Oka T et al. (1999). Embryonic lethality and defective neural tube closure in mice lacking squalene synthase. Journal of Biological Chemistry 274: 30843-30848.
Uchida H, Yamashita H, Kajikawa M, Ohyama K, Nakayachi O et al. (2009). Cloning and characterization of a squalene synthase gene from a petroleum plant, Euphorbia tirucalli L. Planta 229: 1243-1252.
Vogeli U, Chappell J (1988). Induction of sesquiterpene cyclase and suppression of squalene synthetase activities in plant cell cultures treated with fungal elicitor. Plant Physiology 88: 1291- 1296.
Wang Y, Xu WJ, Yan XF, Wang Y (2011). Glucosinolate content and related gene expression in response to enhanced UV-B radiation in Arabidopsis. African Journal of Biotechnology 10 (34): 6481-6491.
Weathers PJ, Diiorio A, Cheetham R, O’Leary M (1990). Recovery of secondary metabolites with minimal loss of cell viability. Progress in Plant Cellular and Molecular Biology 9: 582-586.
Wentzinger LF, Bach TJ, Hartmann MA (2002). Inhibition of squalene synthase and squalene epoxidase in tobacco cells triggers an up-regulation of 3-hydroxy-3-methylglutaryl coenzyme a reductase. Plant Physiology 130: 334-346.
Yücesan B, Mohammed A, Arslan M, Gürel E (2015). Clonal propagation and synthetic seed production from nodal segments of Cape gooseberry (Physalis peruviana L.), a tropical fruit plant. Turkish Journal of Agriculture and Forestry 39: 797-806.
Zhang WJ, Björn LO (2009). The effect of ultraviolet radiation on the accumulation of medicinal compounds in plants. Fitoterapia 80: 207-218.
Zhao J, Davis LC, Verpoorte R (2005). Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnology Advances 23: 283-333.