Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
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Label-free quantitative proteomic analysis of Panax ginseng leaves upon exposure to heat stress

  • Kim, So Wun (Department of Plant Bioscience, Pusan National University) ;
  • Gupta, Ravi (Department of Plant Bioscience, Pusan National University) ;
  • Min, Cheol Woo (Department of Plant Bioscience, Pusan National University) ;
  • Lee, Seo Hyun (Department of Plant Bioscience, Pusan National University) ;
  • Cheon, Ye Eun (Department of Plant Bioscience, Pusan National University) ;
  • Meng, Qing Feng (Department of Plant Bioscience, Pusan National University) ;
  • Jang, Jeong Woo (Department of Plant Bioscience, Pusan National University) ;
  • Hong, Chi Eun (Department of Herbal Crop Research, Rural Development Administration) ;
  • Lee, Ji Yoon (National Instrumentation Center for Environmental Management, Seoul National University) ;
  • Jo, Ick Hyun (Department of Herbal Crop Research, Rural Development Administration) ;
  • Kim, Sun Tae (Department of Plant Bioscience, Pusan National University)
  • Received : 2018.05.17
  • Accepted : 2018.09.27
  • Published : 2019.01.15
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Abstract

Background: Ginseng is one of the well-known medicinal plants, exhibiting diverse medicinal effects. Its roots possess anticancer and antiaging properties and are being used in the medical systems of East Asian countries. It is grown in low-light and low-temperature conditions, and its growth is strongly inhibited at temperatures above $25^{\circ}C$. However, the molecular responses of ginseng to heat stress are currently poorly understood, especially at the protein level. Methods: We used a shotgun proteomics approach to investigate the effect of heat stress on ginseng leaves. We monitored their photosynthetic efficiency to confirm physiological responses to a high-temperature stress. Results: The results showed a reduction in photosynthetic efficiency on heat treatment ($35^{\circ}C$) starting at 48 h. Label-free quantitative proteome analysis led to the identification of 3,332 proteins, of which 847 were differentially modulated in response to heat stress. The MapMan analysis showed that the proteins with increased abundance were mainly associated with antioxidant and translation-regulating activities, whereas the proteins related to the receptor and structural-binding activities exhibited decreased abundance. Several other proteins including chaperones, G-proteins, calcium-signaling proteins, transcription factors, and transfer/carrier proteins were specifically downregulated. Conclusion: These results increase our understanding of heat stress responses in the leaves of ginseng at the protein level, for the first time providing a resource for the scientific community.

Keywords

References

  1. Choi KT. Botanical characteristics, pharmacological effects and medicinal components of Korean Panax ginseng C. A. Meyer. Acta Pharmacol Sin 2008;29:1109-18. https://doi.org/10.1111/j.1745-7254.2008.00869.x
  2. Lee J-S, Lee J-H, Ahn I-O. Characteristics of resistant lines to high-temperature injury in ginseng (Panax ginseng C. A. Meyer). J Ginseng Res 2010;34:274-81. https://doi.org/10.5142/jgr.2010.34.4.274
  3. Lee KM, Kim HR, Lim H, You YH. Effect of elevated $CO_{2}$ concentration and temperature on the growth and ecophysiological responses of ginseng (Panax ginseng C. A. Meyer). Korean J Crop Sci 2012;57:106-12. https://doi.org/10.7740/kjcs.2012.57.2.106
  4. Mittler R, Finka A, Goloubinoff P. How do plants feel the heat? Trends Biochem Sci 2012;37:118-25. https://doi.org/10.1016/j.tibs.2011.11.007
  5. Shi W, Wang Y, Li J, Zhang H, Ding L. Investigation of ginsenosides in different parts and ages of Panax ginseng. Food Chem 2007;102:664-8. https://doi.org/10.1016/j.foodchem.2006.05.053
  6. Jang IB, Lee DY, Yu J, Park HW, Mo HS, Park KC, Hyun DY, Lee EH, Kim KH, Oh CS. Photosynthesis rates, growth, and ginsenoside contents of 2-yr-old Panax ginseng grown at different light transmission rates in a greenhouse. J Ginseng Res 2015;39:345-53. https://doi.org/10.1016/j.jgr.2015.03.007
  7. Jang IB, Yu J, Kweon KB, Suh SJ. Effect of controlled light environment on the growth and ginsenoside content of Panax ginseng C. A. Meyer. Korean J Med Crop Sci 2016;24:277-83. https://doi.org/10.7783/KJMCS.2016.24.4.277
  8. Pandey A, Mann M. Proteomics to study genes and genomes. Nature 2000;405:837-46. https://doi.org/10.1038/35015709
  9. Ghatak A, Chaturvedi PA, Weckwerth W. Cereal crop proteomics: systemic analysis of crop drought stress responses towards marker-assisted selection breeding. Front Plant Sci 2017;8:757. https://doi.org/10.3389/fpls.2017.00757
  10. Kim SW, Lee SH, Min CW, Jo IH, Bang KH, Hyun DY, Agrawal GK, Rakwal R, Zargar SM, Gupta R, et al. Ginseng (Panax sp.) proteomics: an update. Appl Biol Chem 2017;60:311-20. https://doi.org/10.1007/s13765-017-0283-y
  11. Kim SW, Min CW, Gupta R, Jo IH, Bang KH, Kim YC, Kim KH, Kim ST. Proteomics analysis of early salt-responsive proteins in ginseng (Panax ginseng C. A. Meyer) leaves. Korean J Med Crop Sci 2014;22:398-404. https://doi.org/10.7783/KJMCS.2014.22.5.398
  12. Nam MH, Heo EJ, Kim JY, Kim S, Kwon KH, Seo JB, Kwon O, Yoo JS, Park YM. Proteome analysis of the responses of Panax ginseng C. A. Meyer leaves to high light: use of electrospray ionization quadrupole-time of flight mass spectrometry and expressed sequence tag data. Proteomics 2003;3:2351-67. https://doi.org/10.1002/pmic.200300509
  13. Zhao Q, Chen W, Bian J, Xie H, Li Y, Xu C, Ma J, Guo S, Chen J, Cai X, et al. Proteomics and phosphoproteomics of heat stress-responsive mechanisms in Spinach. Front Plant Sci 2018;9.
  14. Lee DG, Ahsan N, Lee SH, Kang KY, Bahk JD, Lee IJ, Lee BH. A proteomic approach in analyzing heat-responsive proteins in rice leaves. Proteomics 2007;7:3369-83. https://doi.org/10.1002/pmic.200700266
  15. Zou J, Liu C, Chen X. Proteomics of rice in response to heat stress and advances in genetic engineering for heat tolerance in rice. Plant Cell Rep 2011;30:2155-65. https://doi.org/10.1007/s00299-011-1122-y
  16. Valdes-Lopez O, Batek J, Gomez-Hernandez N, Nguyen CT, Isidra-Arellano MC, Zhang N, Joshi T, Xu D, Hixson KK, Weitz KK, et al. Soybean roots grown under heat stress show global changes in their transcriptional and proteomic profiles. Front Plant Sci 2016;7.
  17. Echevarria-Zomeno S, Fernandez-Calvino L, Castro-Sanz AB, Lopez JA, Vazquez J, Castellano MM. Dissecting the proteome dynamics of the early heat stress response leading to plant survival or death in Arabidopsis. Plant Cell Environ 2016;39:1264-78. https://doi.org/10.1111/pce.12664
  18. Li W, Wei Z, Qiao Z, Wu Z, Cheng L, Wang Y. Proteomics analysis of alfalfa response to heat stress. PLoS One 2013;8.
  19. Zhou S, Sauve RJ, Liu Z, Reddy S, Bhatti SA. Heat-induced proteome changes in tomato leaves. J Am Soc Hortic Sci 2012;136:2012.
  20. Wang X, Xu C, Cai X, Wang Q, Dai S. Heat-responsive photosynthetic and signaling pathways in plants: insight from proteomics. Int J Mol Sci 2017;18.
  21. Ghatak A, Chaturvedi P, Paul P, Agrawal GK, Rakwal R, Kim ST, Weckwerth W, Gupta R. Proteomics survey of Solanaceae family: current status and challenges ahead. J Proteomics 2017;169:41-57. https://doi.org/10.1016/j.jprot.2017.05.016
  22. Vilumbrales DM, Skacelova K, Bartak M. Sensitivity of Antarctic freshwater algae to salt stress assessed by fast chlorophyll fluorescence transient. Czech Polar Rep 2013;3:163-72. https://doi.org/10.5817/CPR2013-2-17
  23. Thwe AA, Kasemsap P. Quantification of OJIP fluorescence transient in tomato plants under acute ozone stress. Kasetsart J Nat Sci 2014;48:665-75.
  24. Kim SW, Gupta R, Lee SH, Min CW, Agrawal GK, Rakwal R, Kim JB, Jo IK, Park SY, Kim JK, et al. An Integrated biochemical, proteomics, and metabolomics approach for supporting medicinal value of Panax ginseng fruits. Front Plant Sci 2016;7:1-14.
  25. Kim UG, Jung H-J, Lee S-J, Kim S-T. Crop proteomics: practical method for high resolution of two-dimensional electrophoresis. J Plant Biotechnol 2012;39:81-92. https://doi.org/10.5010/JPB.2012.39.1.081
  26. Min CW, Lee SH, Cheon YE, Han WY, Ko JM, Kang HW, Kim YC, Agrawal GK, Rakwal R, Gupta R, et al. In-depth proteomic analysis of Glycine max seeds during controlled deterioration treatment reveals a shift in seed metabolism. J Proteomics 2017;169:125-35. https://doi.org/10.1016/j.jprot.2017.06.022
  27. Gupta R, Min CW, Kramer K, Agrawal GK, Rakwal R, Park KH, Wang Y, Finkemeier I, Kim ST. A multi-omics analysis of Glycine max leaves reveals alteration in flavonoid and isoflavonoid metabolism upon ethylene and abscisic acid treatment. Proteomics Pract Proteomics 2018;18.
  28. Wisniewski JR, Zougman A, Nagaraj N, Mann M. Universal sample preparation method for proteome analysis. Nat Methods 2009;6:359-62. https://doi.org/10.1038/nmeth.1322
  29. Pajarillo EAB, Kim SH, Lee J-Y, Valeriano VDV, Kang D-K. Quantitative proteogenomics and the reconstruction of the metabolic pathway in Lactobacillus mucosae LM1. Korean J Food Sci Anim Resour 2015;35:692-702. https://doi.org/10.5851/kosfa.2015.35.5.692
  30. Vizcaino JA, Csordas A, Del-Toro N, Dianes JA, Griss J, Lavidas I, Mayer G, Perez-Riverol Y, Reisinger F, Ternent T, et al. 2016 update of the PRIDE database and its related tools. Nucleic Acids Res 2016;44:11033. https://doi.org/10.1093/nar/gkw880
  31. Jo IH, Lee J, Hong CE, Lee DJ, Bae W, Park SG, Ahn YJ, Kim YC, Kim JU, Lee JW, et al. Isoform sequencing provides a more comprehensive view of the Panax ginseng transcriptome. Genes (Basel) 2017;8:228. https://doi.org/10.3390/genes8090228
  32. Cox J, Hein MY, Luber CA, Paron I, Nagaraj N, Mann M. Accurate proteomewide label-free quantification by delayed normalization and maximal peptide ratio extraction, termed MaxLFQ. Mol Cell Proteomics 2014;13:2513-26. https://doi.org/10.1074/mcp.M113.031591
  33. Tyanova S, Temu T, Cox J. The MaxQuant computational platform for mass spectrometry-based shotgun proteomics. Nat Protoc 2016;11:2301-19. https://doi.org/10.1038/nprot.2016.136
  34. Tyanova S, Temu T, Sinitcyn P, Carlson A, Hein MY, Geiger T, Mann M, Cox J. The Perseus computational platform for comprehensive analysis of (prote) omics data. Nat Methods 2016;13:731-40. https://doi.org/10.1038/nmeth.3901
  35. Oh SJ, Koh SC. Analysis of O-J-I-P transients from four subtropical plant species for screening of stress indicators under low temperature. J Environ Sci Int 2005;14:389-95. https://doi.org/10.5322/JES.2005.14.4.389
  36. Kim JH, Moon YR, Lee MH, Kim TH, Lee JW, Chung BY. Application of chlorophyll fluorescence as a useful tool for evaluating the influence of a gamma ray on some horticultural crops. Korean J Hortic Sci Technol 2010;28:126-31.
  37. Stefanov D, Petkova V, Denev ID. Screening for heat tolerance in common bean (Phaseolus vulgaris L.) lines and cultivars using JIP-test. Sci Hortic (Amsterdam) 2011;128:1-6. https://doi.org/10.1016/j.scienta.2010.12.003
  38. Chen LS, Cheng L. Photosystem 2 is more tolerant to high temperature in apple (Malus domestica Borkh.) leaves than in fruit peel. Photosynthetica 2009;47:112-20. https://doi.org/10.1007/s11099-009-0017-4
  39. Zushi K, Kajiwara S, Matsuzoe N. Chlorophyll A fluorescence OJIP transient as a tool to characterize and evaluate response to heat and chilling stress in tomato leaf and fruit. Sci Hortic (Amsterdam) 2012;148:39-46. https://doi.org/10.1016/j.scienta.2012.09.022
  40. Kim NH, Jayakodi M, Lee SC, Choi BS, Jang W, Lee J, Kim HH, Waminal NE, Lakshmanan M, Nguyen BV, et al. Genome and evolution of the shaderequiring medicinal herb Panax ginseng. Plant Biotechnol J 2018.
  41. Jayakodi M, Choi BS, Lee SC, Kim NH, Park JY, Jang W, Lakshmanan M, Mohan SVG, Lee DY, Yang TJ. Ginseng genome database: an open-access platform for genomics of Panax ginseng. BMC Plant Biol 2018.
  42. Donahue JL, Alford SR, Torabinejad J, Kerwin RE, Nourbakhsh A, Ray WK, Hernick M, Huang X, Lyons BM, Hein PP, et al. The Arabidopsis thaliana myoinositol 1-phosphate synthase1 gene is required for myo-inositol synthesis and suppression of cell death. Plant Cell 2010;22:888-903. https://doi.org/10.1105/tpc.109.071779
  43. Gammulla CG, Pascovici D, Atwell BJ, Haynes PA. Differential metabolic response of cultured rice (Oryza sativa) cells exposed to high- and lowtemperature stress. Proteomics 2010;10:3001-19. https://doi.org/10.1002/pmic.201000054
  44. Narusaka Y, Narusaka M, Seki M, Umezawa T, Ishida J, Nakajima M, Enju A, Shinozaki K. Crosstalk in the responses to abiotic and biotic stresses in Arabidopsis: analysis of gene expression in cytochrome P450 gene superfamily by cDNA microarray. Plant Mol Biol 2004;55:327-42. https://doi.org/10.1007/s11103-004-0685-1
  45. Salvucci ME, Crafts-Brandner SJ. Inhibition of photosynthesis by heat stress: the activation state of Rubisco as a limiting factor in photosynthesis. Physiol Plant 2004;120:179-86. https://doi.org/10.1111/j.0031-9317.2004.0173.x
  46. Hutin C, Nussaume L, Moise N, Moya I, Kloppstech K, Havaux M. Early lightinduced proteins protect Arabidopsis from photooxidative stress. Proc Natl Acad Sci 2003;100:4921-6. https://doi.org/10.1073/pnas.0736939100
  47. Han F, Chen H, Li XJ, Yang MF, Liu GS, Shen SH. A comparative proteomic analysis of rice seedlings under various high-temperature stresses. Biochim Biophys Acta Proteins Proteomics 2009;1794:1625-34. https://doi.org/10.1016/j.bbapap.2009.07.013
  48. Wang W, Vinocur B, Shoseyov O, Altman A. Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. Trends Plant Sci 2004;9:244-52. https://doi.org/10.1016/j.tplants.2004.03.006
  49. Jia XY, He LH, Jing RL, Li RZ. Calreticulin: conserved protein and diverse functions in plants. Physiol Plant 2009;136:127-38. https://doi.org/10.1111/j.1399-3054.2009.01223.x
  50. Crofts AJ, Denecke J. Calreticulin and calnexin in plants. Trends Plant Sci 1998;3:396-9. https://doi.org/10.1016/S1360-1385(98)01312-0
  51. Rajan VBV, D'Silva P. Arabidopsis thaliana J-class heat shock proteins: cellular stress sensors. Funct Integr Genom 2009;9:433-46. https://doi.org/10.1007/s10142-009-0132-0
  52. Charng YY. Arabidopsis Hsa32, a novel heat shock protein, is essential for acquired thermotolerance during long recovery after acclimation. Plant Physiol 2006;140:1297-305. https://doi.org/10.1104/pp.105.074898
  53. Jacob P, Hirt H, Bendahmane A. The heat-shock protein/chaperone network and multiple stress resistance. Plant Biotechnol J 2017;15:405-14. https://doi.org/10.1111/pbi.12659
  54. Cao D, Froehlich JE, Zhang H, Cheng CL. The chlorate-resistant and photomorphogenesis-defective mutant CR88 encodes a chloroplast-targeted HSP90. Plant J 2003;33:107-18. https://doi.org/10.1046/j.1365-313X.2003.016011.x
  55. Jaiswal DK, Werth EG, McConnell EW, Hicks LM, Jones AM. Time-dependent, glucose-regulated Arabidopsis regulator of G-protein signaling 1 network. Curr Plant Biol 2016;5:25-35. https://doi.org/10.1016/j.cpb.2015.11.002
  56. Zarsky V, Cvrckova F, Bischoff F, Palme K. At-GDI1 from Arabidopsis thaliana encodes a rab-specific GDP dissociation inhibitor that complements the sec19 mutation of Saccharomyces cerevisiae. FEBS Lett 1997;403:303-8. https://doi.org/10.1016/S0014-5793(97)00072-0
  57. Oh CS. Characteristics of 14-3-3 proteins and their role in plant immunity. Plant Pathol J 2010;26:1-7. https://doi.org/10.5423/PPJ.2010.26.1.001
  58. Larkindale J, Knight MR. Protection against heat stress-induced oxidative damage in Arabidopsis involves calcium, abscisic acid, ethylene, and salicylic acid. Plant Physiol 2002;128:682-95. https://doi.org/10.1104/pp.010320
  59. Kato M, Nagasaki-Takeuchi N, Ide Y, Tomioka R, Maeshima M. PCaPs, possible regulators of PtdInsP signals on plasma membrane. Plant Signal Behav 2010;5:848-50. https://doi.org/10.4161/psb.5.7.11825
  60. Price J, Laxmi A, Martin SK, Jang JC. Global transcription profiling reveals multiple sugar signal transduction mechanisms in Arabidopsis. Plant Cell 2004;16:2128-50. https://doi.org/10.1105/tpc.104.022616
  61. Sang Y. Phospholipase D and phosphatidic acid-mediated generation of superoxide in Arabidopsis. Plant Physiol 2001;126:1449-58. https://doi.org/10.1104/pp.126.4.1449
  62. De Jong CF, Laxalt AM, Bargmann BOR, De Wit PJGM, Joosten MHAJ, Munnik T. Phosphatidic acid accumulation is an early response in the Cf-4/Avr4 interaction. Plant J 2004;39:1-12. https://doi.org/10.1111/j.1365-313X.2004.02110.x
  63. Testerink C, Munnik T. Phosphatidic acid: a multifunctional stress signaling lipid in plants. Trends Plant Sci 2005;10:368-75. https://doi.org/10.1016/j.tplants.2005.06.002
  64. Idanheimo N, Gauthier A, Salojarvi J, Siligato R, Brosche M, Kollist H, Mahonen AP, Kangasjarvi J, Wrzaczek M. The Arabidopsis thaliana cysteine-rich receptor-like kinases CRK6 and CRK7 protect against apoplastic oxidative stress. Biochem Biophys Res Commun 2014;445:457-62. https://doi.org/10.1016/j.bbrc.2014.02.013
  65. Gong M, Li Y-J, Chen S-Z. Abscisic acid-induced thermotolerance in maize seedlings is mediated by calcium and associated with antioxidant systems. J Plant Physiol 1998;153:488-96. https://doi.org/10.1016/S0176-1617(98)80179-X
  66. Robertson AJ, Ishikawa M, Gusta LV, MacKenzie SL. Abscisic acid-induced heat tolerance in Bromus inermis Leyss cell-suspension cultures (heat-stable, abscisic acid-responsive polypeptides in combination with sucrose confer enhanced thermostability). Plant Physiol 1994;105:181-90. https://doi.org/10.1104/pp.105.1.181
  67. Schwechheimer C, Serino G, Callis J, Crosby WL, Lyapina S, Deshaies RJ, Gray WM, Estelle M, Deng XW. Interactions of the COP9 signalosome with the E3 ubiquitin ligase SCFTIR1 in mediating auxin response. Science (80-) 2001;292:1379-82. https://doi.org/10.1126/science.1059776
  68. Lorenzo O, Solano R. Molecular players regulating the jasmonate signalling network. Curr Opin Plant Biol 2005;8:532-40. https://doi.org/10.1016/j.pbi.2005.07.003
  69. Wang J, Gao WY, Zhang J, Zuo BM, Zhang LM, Huang LQ. Advances in study of ginsenoside biosynthesis pathway in Panax ginseng C. A. Meyer. Acta Physiol Plant 2012;34:397-403. https://doi.org/10.1007/s11738-011-0844-3
  70. Schaller A. A cut above the rest: the regulatory function of plant proteases. Planta 2004;220:183-97. https://doi.org/10.1007/s00425-004-1407-2

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