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Metabolomics and physiological analyses reveal β-sitosterol as an important plant growth regulator inducing tolerance to water stress in white clover

Abstract

Main conclusion

β-sitosterol influences amino acids, carbohydrates, organic acids, and other metabolite metabolism and homeostasis largely contributing to better tolerance to water stress in white clover.

Abstract

β-sitosterol (BS) could act as an important plant growth regulator when plants are subjected to harsh environmental conditions. Objective of this study was to examine effects of BS on growth and water stress tolerance in white clover based on physiological responses and metabolomics. White clover was pretreated with or without BS and then subjected to water stress for 7 days in controlled growth chambers. Physiological analysis demonstrated that exogenous application of BS (120 μM) could significantly improve stress tolerance associated with better growth performance and photosynthesis, higher leaf relative water content, and less oxidative damage in white clover in response to water stress. Metabolic profiling identified 78 core metabolites involved in amino acids, organic acids, sugars, sugar alcohols, and other metabolites in leaves of white clover. For sugars and sugar alcohol metabolism, the BS treatment enhanced the accumulation of fructose, glucose, maltose, and myo-inositol contributing to better antioxidant capacity, growth maintenance, and osmotic adjustment in white clover under water stress. The application of BS was inclined to convert glutamic acid into proline, 5-oxoproline, and chlorophyll instead of going to pyruvate and alanine; the BS treatment did not significantly affect intermediates of tricarboxylic acid cycle (citrate, aconitate, and malate), but promoted the accumulation of other organic acids including lactic acid, glycolic acid, glyceric acid, shikimic acid, galacturonic acid, and quinic acid in white clover subjected to water stress. In addition, cysteine, an important antioxidant metabolite, was also significantly improved by BS in white clover under water stress. These altered amino acids and organic acids metabolism could play important roles in growth maintenance and modulation of osmotic and redox balance against water stress in white clover. Current findings provide a new insight into BS-induced metabolic homeostasis related to growth and water stress tolerance in plants.

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Abbreviations

BS:

β-Sitosterol

OA:

Osmotic adjustment

OP:

Osmotic potential

WUE:

Water use efficiency

References

  1. Abu-Muriefah SS (2015) Effect of sitosterol on growth, metabolism and protein pattern of pepper (Capsicum annuum L.) plants grown under salt stress conditions. Int J Agric Crop Sci 8:94–106

  2. Ambavade SD, Misar AV, Ambavade PD (2014) Pharmacological, nutritional, and analytical aspects of β-sitosterol: a review. Orient Pharm Exp Med 14:193–211

  3. Anderson RC (2006) Evolution and origin of the central grassland of north America: climate, fire, and mammalian grazers. J Torrey Bot Soc 133:626–647

  4. Arnon DI (1949) Copper enzymes in isoloted chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24:1–18

  5. Ashraf M, Foolad MR (2007) Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environ Exp Bot 59:206–216

  6. Barrs HD, Weatherley PE (1968) A re-examination of the relative turgidity technique for estimating water deficits in leaves. Aust J Biol Sci 15:413–428

  7. Baskar AA, Al Numair KS, Gabriel PM, Alsaif MA, Muamar MA, Ignacimuthu S (2012) β-sitosterol prevents lipid peroxidation and improves antioxidant status and histoarchitecture in rats with 1,2-dimethylhydrazine-induced colon cancer. J Med Food 15:335–343

  8. Blum A, Ebercon A (1981) Cell membrane stability as a measure of drought and heat tolerance in wheat. Crop Sci 21:43–47

  9. Bouche N, Fromm H (2004) GABA in plants: just a metabolite? Trends Plant Sci 9:110–115

  10. Chen TH, Murata N (2002) Enhancement of tolerance of abiotic stress by metabolic engineering of betaines and other compatible solutes. Curr Opin Plant Biol 5:250–257

  11. Dhindsa RS, Plumb-Dhindsa P, Thorpe TA (1981) Leaf senescence: correlated with increased levels of membrane permeability and lipid peroxidation, and decreased levels of superoxide dismutase and catalase. J Exp Bot 32:93–101

  12. Elstner EF, Heupel A (1976) Inhibition of nitrite formation from hydroxylammoniumchloride: a simple assay for superoxide dismutase. Anal Biochem 70:616–620

  13. Farooq M, Wahid A, Kobayashi N, Fujita D, Basra SMA (2009) Plant drought stress: effects, mechanisms and management. Agron Sustain Dev 29:185–212

  14. Fawzia AE, Ashraf AE, Samia AH, Hend AE, Nora FS (2016) β-sitosterol ameliorates the chemical constituents of sunflower (Helianthus annuus L.) plants, grown under saline condition. IOSR J Pharm Biol Sci 11:36–45

  15. Gamel RE, Elsayed A, Bashasha J, Haroun S (2006) Priming tomato cultivars in β-sitosterol or gibberellic acid improves tolerance for temperature stress. Int J Bot 13:1–14

  16. Gupta R, Sharma AK, Dobhal MP, Sharma MC, Gupta RS (2011) Antidiabetic and antioxidant potential of β-sitosterol in streptozotocin-induced experimental hyperglycemia. J Diabetes 3:29–37

  17. Ho S, Chao Y, Tong W, Yu S (2001) Sugar coordinately and differentially regulates growth- and stress-related gene expression via a complex signal transduction network and multiple control mechanisms. Plant Physiol 125:877–890

  18. Hoagland CR, Arnon DI (1950) The water culture method for growing plants without soil. Soil Calif Agric Exp Circ 347:1–32

  19. Hu L, Zhang P, Jiang Y, Fu J (2015) Metabolomic analysis revealed differential adaptation to salinity and alkalinity stress in Kentucky bluegrass (Poa pratensis). Plant Mol Biol Rep 33:56–68

  20. Jespersen D, Yu J, Huang B (2017) Metabolic effects of acibenzolar-S-methyl for improving heat or drought stress in creeping bentgrass. Front Plant Sci 8:1224

  21. Jiang Q, Zhang JY, Guo X, Mohamed B, Lloyd S, Joseph B, Wang ZY (2010) Improvement of drought tolerance in white clover (Trifolium repens) by transgenic expression of a transcription factor gene WXP1. Funct Plant Biol 37:157–165

  22. Kinnersley AM, Turano FJ (2000) Gamma aminobutyric acid (GABA) and plant responses to stress. Crit Rev Plant Sci 19:479–509

  23. Kosolapov VM, Chesnokov YV (2015) Possible environmental risks at commercial growing transgenic forage crops. Russ J Plant Physiol 62:143–152

  24. Kumar MS, Ali K, Dahuja A, Tyagi A (2015) Role of phytosterols in drought stress tolerance in rice. Plant Physiol Biochem 96:83–89

  25. Li Z, Yu J, Peng Y, Huang B (2016a) Metabolic pathways regulated by abscisic acid, salicylic acid and γ-aminobutyric acid in association with improved drought tolerance in creeping bentgrass (Agrostis stolonifera). Physiol Plant 159:42–58

  26. Li Z, Yu J, Peng Y, Huang B (2016b) Metabolic pathways regulated by γ-aminobutyric acid (GABA) contributing to heat tolerance in creeping bentgrass (Agrostis stolonifera). Sci Rep 6:30338

  27. Li Z, Zhang Y, Zhang X, Peng Y, Merewitz E, Ma X, Huang L, Yan Y (2016c) The alterations of endogenous polyamines and phytohormones induced by exogenous application of spermidine regulate antioxidant metabolism, metallothionein and relevant genes conferring drought tolerance in white clover. Environ Exp Bot 124:22–38

  28. Li Z, Zhang Y, Zhang XQ, Merewitz E, Peng Y, Ma X, Huang LK, Yan YH (2017) Metabolic pathways regulated by chitosan contributing to drought resistance in white clover. J Proteome Res 16:3039–3052

  29. Lomenick B, Shi H, Huang J, Chen C (2015) Identification and characterization of β-sitosterol target proteins. Bioorg Med Chem Lett 25:4976–4979

  30. López-Martin MC, Gotor C (2008) Knocking out cytosolic cysteine synthesis compromises the antioxidant capacity of the cytosol to maintain discrete concentrations of hydrogen peroxide in Arabidopsis. Plant Physiol 147:562–572

  31. Lytovchenko A, Fernie AR (2002) Carbon assimilation and metabolism in potato leaves deficient in plastidial phosphoglucomutase. Planta 215:802–811

  32. Ma Q, Yue LJ, Zhang JL, Wu GQ, Bao AK, Wang SM (2012) Sodium chloride improves photosynthesis and water status in the succulent xerophyte Zygophyllum xanthoxylum. Tree Physiol 32(1):4–13

  33. Merewitz EB, Du H, Yu W, Liu Y, Gianfagna T, Huang B (2012) Elevated cytokinin content in ipt transgenic creeping bentgrass promotes drought tolerance through regulating metabolite accumulation. J Exp Bot 63:1315–1328

  34. Nakamoto M, Schmit AC, Heintz D, Schaller H, Ohta D (2015) Diversification of sterol methyltransferase enzymes in plants and a role for β-sitosterol in oriented cell plate formation and polarized growth. Plant J 84:860–874

  35. Nam KH, Shin HJ, Pack IS, Park JH, Kim HB, Kim CG (2016) Metabolomic changes in grains of well-watered and drought-stressed transgenic rice. J Sci Food Agr 96:807–814

  36. Nichols SN, Hofmann RW, Williams WM (2015) Physiological drought resistance and accumulation of leaf phenolics in white clover interspecific hybrids. Environ Exp Bot 73:40–47

  37. Nichols SN, Hofmann RW, Williams WM (2017) Drought resistance of Trifolium repens × Trifolium uniflorum interspecific hybrids. Crop Pasture Sci 65:911–921

  38. Qiu Y, Cai G, Su M, Chen T, Zheng X, Xu Y, Ni Y, Zhao A, Xu L, Cai S (2009) Serum metabolite profiling of human colorectal cancer using GC-TOFMS and UPLC–QTOFMS. J Proteome Res 8:4844–4850

  39. Rademacher W (2015) Plant growth regulators: backgrounds and uses in plant production. J Plant Growth Regul 34:845–872

  40. Ramu R, Shirahatti PS, Nayakavadi S, Vadivelan R, Zameer F, Dhananjaya BL, Prasad MN (2016) The effect of a plant extract enriched in stigmasterol and β-sitosterol on glycaemic status and glucose metabolism in alloxan-induced diabetic rats. Food Funct 7:3999–4011

  41. Rosa M, Prado C, Podazza G, Interdonato R, González JA, Hilal M, Prado FE (2009) Soluble sugars-metabolism, sensing and abiotic stress: a complex network in the life of plants. Plant Signal Behav 4:388–393

  42. Sánchezmartín J, Heald J, Kingstonsmith A, Winters A, Rubiales D, Sanz M, Mur LA, Prats E (2015) A metabolomic study in oats (Avena sativa) highlights a drought tolerance mechanism based on salicylate signalling pathways and the modulation of carbon, antioxidant and photo-oxidative metabolism. Plant Cell Environ 38:1434–1452

  43. Shi H, Jiang C, Ye T, Tan D, Reiter RJ, Zhang H, Liu R, Chan Z (2015) Comparative physiological, metabolomic, and transcriptomic analyses reveal mechanisms of improved abiotic stress resistance in bermudagrass [Cynodon dactylon (L). Pers.] by exogenous melatonin. J Integr Plant Biol 66:681–694

  44. Signorelli S, Coitiño EL, Borsani O, Monza J (2014) Molecular mechanisms for the reaction between •OH radicals and proline: insights on the role as reactive oxygen species scavenger in plant stress. J Phys Chem B 118:37–47

  45. Smirnoff N, Cumbes QJ (1989) Hydroxyl radical scavenging activity of compatible solutes. Phytochemistry 28(4):1057–1060

  46. Trenberth KE, Dai A, Schrier GVD, Jones PD, Barichivich J, Briffa KR, Sheffield J (2014) Global warming and changes in drought. Nat Clim Change 4:17–22

  47. Velikova V, Yordanov I, Edreva A (2000) Oxidative stress and some antioxidant systems in acid rain-treated bean plants. Protective role of exogenous polyamines. Plant Sci 151:59–66

  48. Wingler A, Roitsch T (2010) Metabolic regulation of leaf senescence: interactions of sugar signalling with biotic and abiotic stress responses. Plant Biol 10:50–62

  49. Xiong L, Schumaker KS, Zhu JK (2002) Cell signaling during cold, drought, and salt stress. Plant Cell 14:S165–S183

  50. Xu Q, Fan N, Zhuang L, Yu J, Huang B (2018) Enhanced stolon growth and metabolic adjustment in creeping bentgrass with elevated CO2 concentration. Environ Exp Bot 155:87–97

  51. Yildizli A, Çevik S, Ünyayar S (2018) Effects of exogenous myo-inositol on leaf water status and oxidative stress of Capsicum annuum under drought stress. Acta Physiol Plant 40:122

  52. Yong B, Xie H, Li Z, Li YP, Zhang Y, Nie G, Zhang XQ, Ma X, Huang LK, Yan YH (2017) Exogenous application of GABA improves PEG-induced drought tolerance positively associated with GABA-shunt, polyamines, and proline metabolism in white clover. Front Physiol 8:1107

  53. Yongpil H, Yun HJ, Hyokon C, Youngchul C, Hyungkeun K, Myungho J, Taekrim Y, Hyegwang J (2009) Protective mechanisms of 3-caffeoyl, 4-dihydrocaffeoyl quinic acid from Salicornia herbacea against tert-butyl hydroperoxide-induced oxidative damage. Chem-Biol Inter 181:366–376

  54. Youssefian S, Nakamura M, Orudgev E, Kondo N (2001) Increased cysteine biosynthesis capacity of transgenic tobacco overexpressing an O-acetylserine(thiol) lyase modifies plant responses to oxidative stress. Plant Physiol 126:1001–1011

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Acknowledgements

This research was supported by the International Cooperation Project of Sichuan Province (Grant No. 2018HH0067) and the Chunhui Program of the Ministry of Education (Z2017095).

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Correspondence to Yan Peng.

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Li, Z., Cheng, B., Yong, B. et al. Metabolomics and physiological analyses reveal β-sitosterol as an important plant growth regulator inducing tolerance to water stress in white clover. Planta 250, 2033–2046 (2019). https://doi.org/10.1007/s00425-019-03277-1

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Keywords

  • Antioxidant
  • Growth
  • Metabolome
  • Metabolic pathway
  • Oxidative damage
  • Osmotic adjustment
  • Photosynthesis
  • Tricarboxylic acid cycle