Abstract
Plants adjust their complex molecular, biochemical, and metabolic processes to overcome salt stress. Here, we investigated the proteomic and epigenetic alterations involved in the morphophysiological responses of Pfaffia glomerata, a medicinal plant, to salt stress and the demethylating agent 5-azacytidine (5-azaC). Moreover, we investigated how these changes affected the biosynthesis of 20-hydroxyecdysone (20-E), a pharmacologically important specialized metabolite. Plants were cultivated in vitro for 40 days in Murashige and Skoog medium supplemented with NaCl (50 mM), 5-azaC (25 μM), and NaCl + 5-azaC. Compared with the control (medium only), the treatments reduced growth, photosynthetic rates, and photosynthetic pigment content, with increase in sucrose, total amino acids, and proline contents, but a reduction in starch and protein. Comparative proteomic analysis revealed 282 common differentially accumulated proteins involved in 87 metabolic pathways, most of them related to amino acid and carbohydrate metabolism, and specialized metabolism. 5-azaC and NaCl + 5-azaC lowered global DNA methylation levels and 20-E content, suggesting that 20-E biosynthesis may be regulated by epigenetic mechanisms. Moreover, downregulation of a key protein in jasmonate biosynthesis indicates the fundamental role of this hormone in the 20-E biosynthesis. Taken together, our results highlight possible regulatory proteins and epigenetic changes related to salt stress tolerance and 20-E biosynthesis in P. glomerata, paving the way for future studies of the mechanisms involved in this regulation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- 5-azaC:
-
5-Azacytidine
- 20-E:
-
20-Hydroxyecdysone
- FC:
-
Fold-change
- LRR-RLK:
-
Leucine-rich repeat receptor-like protein kinase
- MS:
-
Murashige & Skoog medium
- ROS:
-
Reactive oxygen species
References
Acosta-Motos JR, Ortuño MF, Bernal-Vicente A, Diaz-Vivancos P, Sanchez-Blanco MJ, Hernandez JA (2017) Plant responses to salt stress: adaptive mechanisms. Agronomy 71:18. https://doi.org/10.3390/agronomy7010018
Agarwal G, Kudapa H, Ramalingam A, Choudhary D, Sinha P, Garg V, Singh VK, Patil GB, Pandey MK, Nguyen HT, Guo B, Sunkar R, Niederhuth CE, Varshney RK (2020) Epigenetics and epigenomics: underlying mechanisms, relevance, and implications in crop improvement. Funct Integr Genomics 20:739–761. https://doi.org/10.1007/s10142-020-00756-7
Arif Y, Singh P, Siddiqui H, Bajguz A, Hayat S (2020) Salinity induced physiological and biochemical changes in plants: an omic approach towards salt stress tolerance. Plant Physiol Biochem 156:64–77. https://doi.org/10.1016/j.plaphy.2020.08.042
Balasubramanian V, Vashisht D, Cletus J, Sakthivel N (2012) Plant β-1,3-glucanases: Their biological functions and transgenic expression against phytopathogenic fungi. Biotechnol Lett 34:1983–1990. https://doi.org/10.1007/s10529-012-1012-6
Bates BC, Kundzewicz ZW, Wu S, Palutikof JP (2008) Climate change and water. Technical paper of the Intergovernmental Panel on Climate Change, IPCC Secretariat, Geneva, pp 210 pp
Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207. https://doi.org/10.1007/BF00018060
Batista DS, Dias LLC, Rêgo MM, Saldanha CW, Otoni WC (2017) Flask sealing on in vitro seed germination and morphogenesis of two types of ornamental pepper explants. Cien Rural 47(3):e20150245. https://doi.org/10.1590/0103-8478cr20150245
Batista DS, Koehler AD, Romanel E, Souza VC, Silva TD, Almeida MC, Maciel TEF, Ferreira PRB, Felipe SHS, Saldanha CW, Maldaner J, Dias LLC, Festucci-Buselli RA, Otoni WC (2019) De novo assembly and transcriptome of Pfaffia glomerata uncovers the role of photoautotrophy and the P450 family genes in 20-hydroxyecdysone production. Protoplasma 256:601–614. https://doi.org/10.1007/s00709-018-1322-1
Baxter CJ, Foyer CH, Turner J, Rolfe SA, Quick WP (2003) Elevated sucrose-phosphate synthase activity in transgenic tobacco sustains photosynthesis in older leaves and alters development. J Exp Bot 54:1813–1820. https://doi.org/10.1093/jxb/erg196
Block AK, Tang HV, Hopkins D, Mendoza J, Solemslie RK, du Toit LJ, Christensen SA (2021) A maize leucine-rich repeat receptor-like protein kinase mediates responses to fungal attack. Planta 254:1–9. https://doi.org/10.1007/S00425-021-03730-0/FIGURES/4
Bolouri-Moghaddam MR, Le Roy K, Xiang L, Rolland F, Van Den Ende W (2010) Sugar signalling and antioxidant network connections in plant cells. FEBS J 277:2022–2037. https://doi.org/10.1111/j.1742-4658.2010.07633.x
Chen L, Huang Y, Xu M, Cheng Z, Zheng J (2017) Proteomic analysis reveals coordinated regulation of anthocyanin biosynthesis through signal transduction and sugar metabolism in black rice leaf. Int J Mol Sci 18:2722. https://doi.org/10.3390/ijms18122722
Chen Q, Wang B, Ding H, Zhang J, Li S (2019) Review: the role of NADP-malic enzyme in plants under stress. Plant Sci 281:206–212. https://doi.org/10.1016/j.plantsci.2019.01.010
Cheng W, Wang Z, Xu F, Ahmad W, Lu G, Su Y, Xu L (2021) Genome-wide identification of LRR-RLK family in saccharum and expression analysis in response to biotic and abiotic stress. Curr Issues Mol Biol 43:1632–1651. https://doi.org/10.3390/CIMB43030116/S1
Corrêa JPO, Vital CE, Pinheiro MVM, Batista DS, Azevedo JFL, Saldanha CW, Cruz ACF, DaMatta FM, Otoni WC (2015) In vitro photoautotrophic potential and ex vitro photosynthetic competence of Pfaffia glomerata (Spreng.) Pedersen accessions. Plant Cell Tiss Organ Cult 121:289–300. https://doi.org/10.1007/s11240-014-0700-4
Damaris RN, Li M, Liu Y, Chen X, Murage H, Yang P (2016) A proteomic analysis of salt stress response in seedlings of two African rice cultivars. Biochim Biophys Acta - Proteins Proteomics 1864:1570–1578. https://doi.org/10.1016/j.bbapap.2016.08.011
Damerval C, De Vienne D, Zivy M, Thiellement H (1986) Technical improvements in two-dimensional electrophoresis increase the level of genetic variation detected in wheat-seedling proteins. Electrophoresis 7:52–54. https://doi.org/10.1002/elps.1150070108
De-la-Peña C, Nic-Can G, Ojeda G, Herrera-Herrera JL, López-Torres A, Wrobel K, Robert-Díaz ML (2012) KNOX1 is expressed and epigenetically regulated during in vitro conditions in Agave spp. BMC Plant Biol 12:1–11. https://doi.org/10.1186/1471-2229-12-203/FIGURES/6
Deng W, Wang Y, Liu Z, Cheng H, Xue Y (2014) HemI: a toolkit for illustrating heatmaps. PLoS ONE 9:e111988. https://doi.org/10.1371/journal.pone.0111988
Dias FCR, Martins ALP, Melo FCSA, Cupertino MC, Gomes MLM, Oliveira JM, Damasceno EM, Silva J, Otoni WC, DaMatta SLP (2019) Hydroalcoholic extract of Pfaffia glomerata alters the organization of the seminiferous tubules by modulating the oxidative state and the microstructural reorganization of the mice testes. J Ethnopharmacol 233:179–189. https://doi.org/10.1016/j.jep.2018.12.047
Dinan L, Dioh W, Veillet S, Lafont R (2021) 20-Hydroxyecdysone, from plant extracts to clinical use: therapeutic potential for the treatment of neuromuscular, cardio-metabolic and respiratory diseases. Biomedicines 9:492. https://doi.org/10.3390/BIOMEDICINES9050492
Diniz AL, Silva DIR, Lembke CG, Costa MDBL, Ten-Caten F, Li F, Vilela RD, Menossi M, Ware D, Endres L, Souza GM (2020) Amino acid and carbohydrate metabolism are coordinated to maintain energetic balance during drought in sugarcane. Int J Mol Sci 21:1–27. https://doi.org/10.3390/ijms21239124
Distler U, Kuharev J, Navarro P, Levin Y, Schild H, Tenzer S (2014) Drift time-specific collision energies enable deep-coverage data-independent acquisition proteomics. Nat Methods 11:167–170. https://doi.org/10.1038/nmeth.2767
Doyle J, Doyle J (1987) Genomic plant DNA preparation from fresh tissue-CTAB method. Phytochem Bull 19:11–15
Erst AA, Zibareva LN, Filonenko ES, Zheleznichenko TV (2019) Influence of Methyl Jasmonate on production of ecdysteroids from hairy roots of Silene linicola C.C. Gmelin Russ J Bioorganic Chem 45:920–926. https://doi.org/10.1134/S1068162019070033
Faria DV, Correia LNF, Batista DS, Vital CE, Heringer AS, De-la-Peña C, Costa MGC, Guerra MP, Otoni WC (2020) 5-Azacytidine downregulates the SABATH methyltransferase genes and augments bixin content in Bixa orellana L. leaves. Plant Cell Tiss Organ Cult 142:425–434. https://doi.org/10.1007/s11240-020-01857-8
Felipe SHS, Batista DS, Vital CE, Chagas K, Silva PO, Silva TD, Fortini EA, Correia LNF, Ávila RT, Maldaner J, Festucci-Buselli RA, DaMatta FM, Otoni WC (2019) Salinity-induced modifications on growth, physiology and 20-hydroxyecdysone levels in Brazilian-ginseng [Pfaffia glomerata (Spreng.) Pedersen]. Plant Physiol Biochem 140:43–54. https://doi.org/10.1016/j.plaphy.2019.05.002
Festucci-Buselli RA, Contim LAS, Barbosa LCA, Stuart J, Otoni WC (2008) Biosynthesis and potential functions of the ecdysteroid 20-hydroxyecdysone — a review. Botany 86:978–987. https://doi.org/10.1139/B08-049
Flores-Ortiz C, Alvarez LM, Undurraga A, Arias D, Durán F, Wegener G, Stange C (2020) Differential role of the two ζ-carotene desaturase paralogs in carrot (Daucus carota): ZDS1 is a functional gene essential for plant development and carotenoid synthesis. Plant Sci 291:110327. https://doi.org/10.1016/j.plantsci.2019.110327
Franco RR, Takata LA, Chagas K, Justino AB, Saraiva AL, Goulart LR, Ávila VMR, Otoni WC, Espíndola FS, Silva CR (2021) A 20-hydroxyecdysone-enriched fraction from Pfaffia glomerata (Spreng.) Pedersen roots alleviates stress, anxiety, and depression in mice. J Ethnopharmacol 267:113599. https://doi.org/10.1016/j.jep.2020.113599
Gao Y, Cui Y, Zhao R, Chen X, Zhang J, Zhao J, Kong L (2022) Cryo-treatment enhances the embryogenicity of mature somatic embryos via the lncRNA–miRNA–mRNA network in White Spruce. Int J Mol Sci 23:1111. https://doi.org/10.3390/IJMS23031111/S1
Genitoni J, Vassaux D, Delaunay A, Citerne S, Portillo Lemus L, Etienne MP, Renault D, Stoeckel S, Barloy D, Maury S (2020) Hypomethylation of the aquatic invasive plant, Ludwigia grandiflora subsp. hexapetala mimics the adaptive transition into the terrestrial morphotype. Physiol Plant 170:280–298. https://doi.org/10.1111/ppl.13162
Goto T, Aoki R, Minamizaki K, Fujita Y (2010) Functional differentiation of two analogous coproporphyrinogen III oxidases for heme and chlorophyll biosynthesis pathways in the Cyanobacterium Synechocystis sp. PCC 6803. Plant Cell Physiol 51:650–663. https://doi.org/10.1093/pcp/pcq023
Griffin PT, Niederhuth CE, Schmitz RE (2016) A comparative analysis of 5-azacytidine- and zebularine-induced DNA demethylation. G3: GenesGenom Genet 6:2773–2780. https://doi.org/10.1534/g3.116.030262
Guan C, Cui X, Liu H, Li X, Li M, Zhang Y (2020) Proline biosynthesis enzyme genes confer salt tolerance to switchgrass (Panicum virgatum L.) in cooperation with polyamines metabolism. Front Plant Sci 11:46. https://doi.org/10.3389/fpls.2020.00046
Gupta B, Huang B (2014) Mechanism of salinity tolerance in plants: physiological, biochemical, and molecular characterization. Int J Genomics. https://doi.org/10.1155/2014/701596
Han B, Xu W, Ahmed N, Yu A, Wang Z, Liu A (2020) Changes and associations of genomic transcription and histone methylation with salt stress in Castor Bean. Plant Cell Physiol 61:1120–1133. https://doi.org/10.1093/pcp/pcaa037
Hildebrandt TM (2018) Synthesis versus degradation: directions of amino acid metabolism during Arabidopsis abiotic stress response. Plant Mol Biol 98:121–135. https://doi.org/10.1007/s11103-018-0767-0
Huang T, Jander G (2017) Abscisic acid-regulated protein degradation causes osmotic stress-induced accumulation of branched-chain amino acids in Arabidopsis thaliana. Planta 246:737–747. https://doi.org/10.1007/s00425-017-2727-3
Isayenkov SV, Maathuis FJM (2019) Plant salinity stress: many unanswered questions remain. Front Plant Sci 10:80. https://doi.org/10.3389/fpls.2019.00080
John R, Shajitha PP, Devassy A, Mathew L (2018) Effect of elicitation and precursor feeding on accumulation of 20-hydroxyecdysone in Achyranthes aspera Linn. cell suspension cultures. Physiol Mol Biol Plants 24:275–284. https://doi.org/10.1007/s12298-018-0506-7
Kosová K, Vítámvás P, Urban MO, Prášil IT, Renaut J (2018) Plant abiotic stress proteomics: the major factors determining alterations in cellular proteome. Front Plant Sci 9:122. https://doi.org/10.3389/fpls.2018.00122
Latzel V, Münzbergová Z, Skuhrovec J, Novák O, Strnad M (2020) Effect of experimental DNA demethylation on phytohormones production and palatability of a clonal plant after induction via jasmonic acid. Oikos 129:1867–1876. https://doi.org/10.1111/oik.07302
Li XL, Wang CR, Li XY, Yao YX, Hao YJ (2013) Modifications of Kyoho grape berry quality under long-term NaCl treatment. Food Chem 139:931–937. https://doi.org/10.1016/j.foodchem.2013.02.038
Li Y, Mukherjee I, Thum KE, Tanurdzic M, Katari MS, Obertello M, Edwards MB, McCombie WR, Martienssen RA, Coruzzi GM (2015) The histone methyltransferase SDG8 mediates the epigenetic modification of light and carbon responsive genes in plants. Genome Biol 16:79. https://doi.org/10.1186/s13059-015-0640-2
Li Z, Hu Y, Chang M, Kashif MH, Tang M, Luo D, Cao S, Lu H, Zhang W, Huang Z, Yue J, Chen P (2021) 5-azacytidine pre-treatment alters DNA methylation levels and induces genes responsive to salt stress in kenaf (Hibiscus cannabinus L.). Chemosphere 271:129562. https://doi.org/10.1016/j.chemosphere.2021.129562
Liao Y, Cui R, Yuan T, Xie Y, Gao Y (2019) Cysteine and methionine contribute differentially to regulate alternative oxidase in leaves of poplar (Populus deltoides x Populus euramericana ‘Nanlin 895’) seedlings exposed to different salinity. J Plant Physiol 240:153017. https://doi.org/10.1016/j.jplph.2019.153017
Lisec J, Schauer N, Kopka J, Willmitzer L, Fernie AR (2006) Gas chromatography mass spectrometry–based metabolite profiling in plants. Nat Protoc 1:387–396. https://doi.org/10.1038/nprot.2006.59
Liu Y, Lu S, Liu K, Wang S, Huang L, Guo L (2019) Proteomics: a powerful tool to study plant responses to biotic stress. Plant Meth. https://doi.org/10.1186/s13007-019-0515-8
López Sánchez A, Stassen JHM, Furci L, Smith LM, Ton J (2016) The role of DNA (de)methylation in immune responsiveness of Arabidopsis. Plant J 88:361–374. https://doi.org/10.1111/tpj.13252
Lu LM, Yang SY, Liu L, Lu YF, Yang SM, Liu F, Ni S, Zeng FC, Ren B, Wang XY, Li LQ (2020) Physiological and quantitative proteomic analysis of NtPRX63-overexpressing tobacco plants revealed that NtPRX63 functions in plant salt resistance. Plant Physiol Biochem 154:30–42. https://doi.org/10.1016/j.plaphy.2020.04.022
Lv GY, Guo XG, Xie LP, Xie CG, Zhang XH, Yang Y, Xiao L, Tang YY, Pan XL, Guo AG, Xu H (2017) Molecular characterization, gene evolution, and expression analysis of the fructose-1, 6-bisphosphate Aldolase (FBA) gene family in wheat (Triticum aestivum L.). Front Plant Sci 8:1030. https://doi.org/10.3389/fpls.2017.01030
Mageroy MH, Wilkinson SW, Tengs T, Cross H, Almvik M, Pétriacq P, Vivian-Smith A, Zhao T, Fossda CG, Krokene P (2020) Molecular underpinnings of methyl jasmonate-induced resistance in Norway spruce. Plant Cell Environ 43:1827–1843. https://doi.org/10.1111/pce.13774
Mozafari A, Ghadakchi A, Ghaderi N (2018) Grape response to salinity stress and role of iron nanoparticle and potassium silicate to mitigate salt induced damage under in vitro conditions. Physiol Mol Biol Plants 24:25–35. https://doi.org/10.1007/s12298-017-0488-x
Munns R, Gilliham M (2015) Salinity tolerance of crops - what is the cost? New Phytol 208:668–673. https://doi.org/10.1111/nph.13519
Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol. https://doi.org/10.1146/annurev.arplant.59.032607.092911
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Neff MM, Chory J (1998) Genetic interactions between phytochrome A, phytochrome B, and cryptochrome 1 during Arabidopsis development. Plant Physiol 118:27–36. https://doi.org/10.1104/pp.118.1.27
Neto AG, Costa JMLC, Belati CC, Vinhólis AHC, Possebom LS, Silva Filho AA, Cunha WR, Carvalho JCT, Bastos JK, Silva MLAE (2005) Analgesic and anti-inflammatory activity of a crude root extract of Pfaffia glomerata (Spreng.) Pedersen. J Ethnopharmacol 96:87–91. https://doi.org/10.1016/j.jep.2004.08.035
Pan T, Liu M, Kreslavski VD, Zharmukhamedov SK, Nie C, Yu M, Kuznetsov VV, Allakhverdiev SI, Shabala S (2020) Non-stomatal limitation of photosynthesis by soil salinity. Crit Rev Environ Sci Technol 54:1–35. https://doi.org/10.1080/10643389.2020.1735231
Panariello BHD, Klein MI, Pavarina AC, Duarte S (2017) Inactivation of genes TEC1 and EFG1 in Candida albicans influences extracellular matrix composition and biofilm morphology. J Oral Microbiol 9:1385372. https://doi.org/10.1080/20002297.2017.1385372
Parashar NC, Parashar G, Nayyar H, Sandhir R (2020) Differential DNA methylation in regulation of deacetylvindoline-4-O-acetyl transferase (DAT) gene in Catharanthus roseus. J Plant Biochem Biotechnol 30:326–335. https://doi.org/10.1007/s13562-020-00592-7
Passamani LZ, Reis RS, Vale EM, Sousa KR, Aragão VPM, Santa-Catarina C, Silveira V (2020) Long-term culture with 2,4-dichlorophenoxyacetic acid affects embryogenic competence in sugarcane callus via changes in starch, polyamine and protein profiles. Plant Cell Tiss Organ Cult 140:415–429. https://doi.org/10.1007/s11240-019-01737-w
Per TS, Khan NA, Reddy PS, Masood A, Hasanuzzaman M, Khan MIR, Anjum NA (2017) Approaches in modulating proline metabolism in plants for salt and drought stress tolerance: phytohormones, mineral nutrients and transgenics. Plant Physiol Biochem 115:125–140. https://doi.org/10.1016/j.plaphy.2017.03.018
Przydacz M, Jones R, Pennington HG, Belmans G, Bruderer M, Greenhill R, Salter T, Wellham PAD, Cota E, Spanu PD (2020) Mode of action of the catalytic site in the N-terminal ribosome-inactivating domain of JIP60. Plant Physiol 183:385–398. https://doi.org/10.1104/pp.19.01029
Reis RS, Vale EM, Sousa KR, Santa-Catarina C, Silveira V (2021) Pretreatment free of 2,4-dichlorophenoxyacetic acid improves the differentiation of sugarcane somatic embryos by affecting the hormonal balance and the accumulation of reserves. Plant Cell Tiss Organ Cult 145:101–115. https://doi.org/10.1007/s11240-020-01995-z
Saddhe AA, Malvankar MR, Karle SB, Kumar K (2019) Reactive nitrogen species: paradigms of cellular signaling and regulation of salt stress in plants. Environ Exp Bot 161:86–97. https://doi.org/10.1016/j.envexpbot.2018.11.010
Samo N, Ebert A, Kopka J, Mozgová I (2021) Plant chromatin, metabolism and development – an intricate crosstalk. Curr Opin Plant Biol 61:102002. https://doi.org/10.1016/j.pbi.2021.102002
Santos AP, Ferreira LJ, Oliveira MM (2017) Concerted flexibility of chromatin structure, methylome, and histone modifications along with plant stress responses. Biology 6:3. https://doi.org/10.3390/biology6010003
Shahid MA, Sarkhosh A, Khan N, Balal RM, Ali S, Rossi L, Gómez C, Mattson N, Nasim W, Garcia-Sanchez F (2020) Insights into the physiological and biochemical impacts of salt stress on plant growth and development. Agronomy 10:938. https://doi.org/10.3390/agronomy10070938
Shen Y, Issakidis-Bourguet E, Zhou DX (2016) Perspectives on the interactions between metabolism, redox, and epigenetics in plants. J Exp Bot 67:5291–5300. https://doi.org/10.1093/jxb/erw310
Silva TD, Batista DS, Castro KM, Fortini EA, Felipe SHS, Fernandes AM, Sousa RMJ, Chagas K, Silva JVS, Correia LNF, Torres-Silva G, Farias LM, Otoni WC (2021) Irradiance-driven 20-hydroxyecdysone production and morphophysiological changes in Pfaffia glomerata plants grown in vitro. Protoplasma 258:151–167. https://doi.org/10.1007/s00709-020-01558-1
Silva TD, Batista DS, Fortini EA, Castro KM, Felipe SHS, Fernandes AM, Sousa RMJ, Chagas K, Silva JVS, Correia LNF, Farias LM, Leite JPV, Rocha DI, Otoni WC (2020) Blue and red light affects morphogenesis and 20-hydroxyecdisone content of in vitro Pfaffia glomerata accessions. J Photochem Photobiol b: Biol 203:111761. https://doi.org/10.1016/j.jphotobiol.2019.111761
Solís MT, El-Tantawy AA, Cano V, Risueño MC, Testillano PS (2015) 5-azacytidine promotes microspore embryogenesis initiation by decreasing global DNA methylation, but prevents subsequent embryo development in rapeseed and barley. Front Plant Sci 6:472. https://doi.org/10.3389/fpls.2015.00472
Sun L, Song G, Guo W, Wang W, Zhao H, Gao T, Lv Q, Yang X, Xu F, Dong Y, Pu L (2019) Dynamic changes in genome-wide histone3 lysine27 trimethylation and gene expression of soybean roots in response to salt stress. Front Plant Sci 10:1031. https://doi.org/10.3389/fpls.2019.01031
Sun SW, Lin YC, Weng YM, Chen MJ (2006) Efficiency improvements on ninhydrin method for amino acid quantification. J Food Compos Anal 19:112–117. https://doi.org/10.1016/j.jfca.2005.04.006
Sun X, Wang Y, Xu L, Li C, Zhang W, Luo X, Jiang H, Liu L (2017) Unraveling the root proteome changes and its relationship to molecular mechanism underlying salt stress response in radish (Raphanus sativus L.). Front Plant Sci 8:1192. https://doi.org/10.3389/fpls.2017.01192
Tóth N, Szabó A, Kacsala P, Héger J, Zádor E (2008) 20-Hydroxyecdysone increases fiber size in a muscle-specific fashion in rat. Phytomedicine 15:691–698. https://doi.org/10.1016/j.phymed.2008.04.015
Trivellini A, Gordillo B, Rodríguez-Pulido FJ, Borghesi E, Ferrante A, Vernieri P, Quijada-Morín N, González-Miret ML, Heredia FJ (2014) Effect of salt stress in the regulation of anthocyanins and color of Hibiscus flowers by digital image analysis. J Agric Food Chem 62:6966–6974. https://doi.org/10.1021/jf502444u
Van der Does D, Boutrot F, Engelsdorf T, Rhodes J, McKenna JF, Vernhettes S, Koevoets I, Tintor N, Veerabagu M, Miedes E, Segonzac C, Roux M, Breda AS, Hardtke CS, Molina A, Rep M, Testerink C, Mouille G, Höfte H, Hamann T, Zipfel C (2017) The Arabidopsis leucine-rich repeat receptor kinase MIK2/LRR-KISS connects cell wall integrity sensing, root growth and response to abiotic and biotic stresses. PLoS Genet 13:e1006832. https://doi.org/10.1371/journal.pgen.1006832
Van Zelm E, Zhang Y, Testerink C (2020) Salt tolerance mechanisms of plants. Annu Rev Plant Biol 71:403–433. https://doi.org/10.1146/annurev-arplant-050718-100005
Wang P, Li C, Wang Y, Huang R, Sun C, Xu Z, Zhu J, Gao X, Deng X, Wang P (2014a) Identification of a geranylgeranyl reductase gene for chlorophyll synthesis in rice. Springerplus 3:201. https://doi.org/10.1186/2193-1801-3-201
Wang QJ, Zheng LP, Zhao PF, Zhao YL, Wang JW (2014b) Cloning and characterization of an elicitor-responsive gene encoding 3-hydroxy-3-methylglutaryl coenzyme a reductase involved in 20-hydroxyecdysone production in cell cultures of Cyanotis arachnoidea. Plant Physiol Biochem 84:1–9. https://doi.org/10.1016/j.plaphy.2014.08.021
Wellburn AR (1994) The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. J Plant Physiol 144:307–313. https://doi.org/10.1016/S0176-1617(11)81192-2
Wu H, Zhang X, Giraldo JP, Shabala S (2018) It is not all about sodium: revealing tissue specificity and signalling roles of potassium in plant responses to salt stress. Plant Soil 431:1–17. https://doi.org/10.1007/s11104-018-3770-y
Xiong Y, Liu X, You Q, Han L, Shi J, Yang J, Cui W, Zhang H, Chao Q, Zhu Y, Duan Y, Xue T, Xue J (2022) Analysis of DNA methylation in potato tuber in response to light exposure during storage. Plant Physiol Biochem 170:218–224. https://doi.org/10.1016/j.plaphy.2021.12.007
Xu J, Wang X, Cao H, Xu H, Xu Q, Deng X (2017) Dynamic changes in methylome and transcriptome patterns in response to methyltransferase inhibitor 5-azacytidine treatment in citrus. DNA Res 24:509–522. https://doi.org/10.1093/dnares/dsx021
Yang D, Huang Z, Jin W, Xia P, Jia Q, Yang Z, Hou Z, Zhang H, Ji W, Han R (2018) DNA methylation: a new regulator of phenolic acids biosynthesis in Salvia miltiorrhiza. Ind Crops Prod 124:402–411. https://doi.org/10.1016/j.indcrop.2018.07.046
Yang Y, Guo Y (2018a) Unraveling salt stress signaling in plants. J Integr Plant Biol 60:796–804. https://doi.org/10.1111/jipb.12689
Yang Y, Guo Y (2018b) Elucidating the molecular mechanisms mediating plant salt-stress responses. New Phytol 217:523–539. https://doi.org/10.1111/nph.14920
Ye M, Kuai P, Hu L, Ye M, Sun H, Erb M, Lou Y (2020) Suppression of a leucine-rich repeat receptor-like kinase enhances host plant resistance to a specialist herbivore. Plant Cell Environ 43:2571–2585. https://doi.org/10.1111/pce.13834
Yuan L, Wang D, Cao L, Yu N, Liu K, Guo Y, Gan S, Chen L (2020) Regulation of leaf longevity by DML3-mediated DNA demethylation. Mol Plant 13:1149–1161. https://doi.org/10.1016/j.molp.2020.06.006
Yuan Y, Zhong M, Shu S, Du N, Sun J, Guo S (2016) Proteomic and physiological analyses reveal putrescine responses in roots of cucumber stressed by NaCl. Front Plant Sci 7:1035. https://doi.org/10.3389/fpls.2016.01035
Zarza X, Atanasov KE, Marco F, Arbona V, Carrasco P, Kopka J, Fotopoulos V, Munnik T, Gómez-Cadenas A, Tiburcio AF, Alcázar R (2017) Polyamine oxidase 5 loss-of-function mutations in Arabidopsis thaliana trigger metabolic and transcriptional reprogramming and promote salt stress tolerance. Plant Cell Environm 40:527–542. https://doi.org/10.1111/pce.12714
Zeng F, Li X, Qie R, Li L, Ma M, Zhan Y (2020) Triterpenoid content and expression of triterpenoid biosynthetic genes in birch (Betula platyphylla Suk) treated with 5-azacytidine. J for Res 31:1843–1850. https://doi.org/10.1007/s11676-019-00966-1
Zhang Z, Mao C, Shi Z, Kou X (2017) The amino acid metabolic and carbohydrate metabolic pathway play important roles during salt-stress response in tomato. Front Plant Sci 8:1231. https://doi.org/10.3389/fpls.2017.01231
Zhu N, Cheng S, Liu X, Du H, Dai M, Zhou DX, Yang W, Zhao Y (2015) The R2R3-type MYB gene OsMYB91 has a function in coordinating plant growth and salt stress tolerance in rice. Plant Sci 236:146–156. https://doi.org/10.1016/j.plantsci.2015.03.023
Zhu Y, Zhang B, Allan AC, Lin-Wang K, Zhao Y, Wang K, Chen K, Xu C (2020) DNA demethylation is involved in the regulation of temperature-dependent anthocyanin accumulation in peach. Plant J 102:965–976. https://doi.org/10.1111/tpj.14680
Acknowledgements
We thank Prof. Takeshi Kamada (Universidade de Rio Verde, Rio Verde, GO, Brazil), Dr. Roberto F. Vieira, and Dr. Rosa Belém Alves Neves (National Center for Genetic Resources and Biotechnology—Embrapa/Cenargen, Brasília, DF, Brazil) for providing the P. glomerata accessions. The Departments of Plant Biology and Biochemistry and Molecular Biology, Universidade Federal de Viçosa, are gratefully acknowledged for providing the facilities for structural, molecular, and biochemical analyses. The Unidad de Biotecnología (CICY, Mexico) is acknowledged for enabling DNA methylation analysis. We would like to thank Editage (www.editage.com) for English language editing.
Funding
This work was supported by the Brazilian agencies Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG, Belo Horizonte, MG, Brazil; Grants no. PRONEX-CAG-APQ-01036–09, CRA-APQ-01651–13, CRA-BPD-00046–14, CBB-APQ-02372–17, APQ-00772–19, CBB-BPD-00020–16 and CRA–RED-00053–16/REDE MINEIRA Estresse em Plantas), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brasília, DF, Brazil; Grant Finance Code 001), and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brasília, DF, Brazil: Grants no. MCT/CNPq 480675/2009–0; PQ 459.529/2014–5; and PQ 313901/2018–0 to WCO).
Author information
Authors and Affiliations
Contributions
EAF, DSB, and WCO designed the study; EAF performed most of the experiments; EAF, TDS, SHSF, LNFC, and LMF performed physiological and biochemical analyses; EAF and TDS evaluated photosynthetic performance; EAF, DSB, DVF, and LNFC performed molecular analyses; EAF and DSB analyzed the data; CDLP and ECC performed DNA methylation analysis; VBP, CSC, and VS performed mass spectrometry and proteomic data analysis; EAF, DSB, and WCO wrote the article with input from all other authors. All authors read and approved the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing interests.
Additional information
Handling Editor: Peter Nick
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
709_2022_1789_MOESM2_ESM.xlsx
Supplementary file2 (XLSX 710 KB) Table S1. Complete list of identified proteins, functional protein annotations, and configuration parameters. https://1drv.ms/x/s!AhsdNhfp1NBbhZZUyUOKBDmDenNemQ?e=DvZqU7
709_2022_1789_MOESM3_ESM.xlsx
Supplementary file3 (XLSX 236 KB) Table S2. List of metabolic pathways identified in the Kyoto Encyclopedia of Genes and Genomes created from the analysis of differentially accumulated proteins. https://1drv.ms/x/s!AhsdNhfp1NBbhZZVGjO3bfjfkWrMZQ?e=T6F5kR
Rights and permissions
About this article
Cite this article
Fortini, E.A., Batista, D.S., Felipe, S.H.S. et al. Physiological, epigenetic, and proteomic responses in Pfaffia glomerata growth in vitro under salt stress and 5-azacytidine. Protoplasma 260, 467–482 (2023). https://doi.org/10.1007/s00709-022-01789-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00709-022-01789-4