Abstract
Key message
Mitigation of deleterious effects of salinity promoted by exogenous proline can be partially explained by changes in proline enzymatic metabolism and expression of specific proline-related genes.
Abstract
Proline accumulation is a usual response to salinity. We studied the ability of exogenous proline to mitigate the salt harmful effects in sorghum (Sorghum bicolor) leaves. Ten-day-old plants were cultivated in Hoagland’s nutrient solution in either the absence or presence of salinity (NaCl at 75 mM) and sprayed with distilled water or 30 mM proline solution. Salinity deleterious effects were alleviated by exogenous proline 14 days after treatment, with a return in growth and recovery of leaf area and photosynthetic parameters. Part of the salinity response reflected an improvement in ionic homeostasis, provided by reduction in Na+ and Cl− ions and increases in K+ and Ca2+ ions as well as increases of compatible solutes. In addition, the application of proline decreased membrane damage and did not increase relative water content. Proline-treated salt-stressed plants displayed increase in proline content, a response counterbalanced by punctual modulation in proline synthesis (down-regulation of Δ1-pyrroline-5-carboxylate synthetase activity) and degradation (up-regulation of proline dehydrogenase activity) enzymes. These responses were correlated with expression of specific proline-related genes (p5cs1 and prodh). Our findings clearly show that proline treatment results in favorable changes, reducing salt-induced damage and improving salt acclimation in sorghum plants.
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Abbreviations
- Ctrl:
-
Control treatment
- DAT:
-
Days after treatments
- GSA:
-
Glutamic-γ-semialdehyde
- HKT:
-
High-affinity K+ transporter
- NHX:
-
Na+/H+ antiporter exchanger
- OAT:
-
Ornithine-δ-aminotransferase
- P:
-
Proline treatment
- P5C:
-
Δ1-pyrroline-5-carboxylate
- P5CDH:
-
Pyrroline-5-carboxylate dehydrogenase
- P5CR:
-
Δ1-pyrroline-5-carboxylate reductase
- P5CS:
-
P5C synthetase
- ProDH:
-
Proline dehydrogenase/proline oxidase
- S:
-
Saline treatment
- SOS1:
-
Salt overly sensitive 1 Na+/H+ antiporter
- SP:
-
Saline/proline treatment
References
Agami RA (2014) Applications of ascorbic acid or proline increase resistance to salt stress in barley seedlings. Biol Plant 58:341–347. https://doi.org/10.1007/s10535-014-0392-y
Ahanger MA, Akram NA, Ashraf M et al (2017) Plant responses to environmental stresses-from gene to biotechnology. AoB Plants 9:1–17. https://doi.org/10.1093/aobpla/plx025
Alyemeni MN, Hayat Q, Hayat S et al (2016) Exogenous proline application enhances the efficiency of nitrogen fixation and assimilation in chickpea plants exposed to cadmium. Legum Res 39:221–227. https://doi.org/10.18805/lr.v0iOF.9291
Ashraf M, Harris PJC (2013) Photosynthesis under stressful environments: an overview. Photosynthetica 51:163–190. https://doi.org/10.1007/s11099-013-0021-6
Bagdi DL, Shaw BP, Sahu BB, Purohit GK, Info P (2015) Real time PCR expression analysis of gene encoding p5cs enzyme and proline metabolism under NaCl salinity in rice. J Environ Biol 36:955–961
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
Ben Rejeb K, Abdelly C, Savouré A (2014) How reactive oxygen species and proline face stress together. Plant Physiol Biochem 80:278–284. https://doi.org/10.1016/j.plaphy.2014.04.007
Benitez LC, Vighi IL, Auler PA et al (2016) Correlation of proline content and gene expression involved in the metabolism of this amino acid under abiotic stress. Acta Physiol Plant 38:267. https://doi.org/10.1007/s11738-016-2291-7
Bieleski RL, Turner NA (1966) Separation and estimation of amino acids in crude plant extracts by thin-layer electrophoresis and chromatography. Anal Biochem 293:278–293. https://doi.org/10.1016/0003-2697(66)90206-5
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Canales J, Rueda-López M, Craven-Bartle B et al (2012) Novel insights into regulation of asparagine synthetase in conifers. Front Plant Sci 3:1–15. https://doi.org/10.3389/fpls.2012.00100
Čatský J (1960) Determination of water deficit in disks cut out from leaf blades. Biol Plant 2:76–78. https://doi.org/10.1007/BF02920701
Deinlein U, Stephan AB, Horie T et al (2014) Plant salt-tolerance mechanisms. Trends Plant Sci 19:371–379. https://doi.org/10.1016/j.tplants.2014.02.001
Dubois M, Gilles KA, Hamilton JK et al (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356. https://doi.org/10.1021/ac60110a016
Fichman Y, Gerdes SY, Kov H (2015) Evolution of proline biosynthesis: enzymology, bioinformatics, genetics, and transcriptional regulation. Biol Rev 1069:1065–1099. https://doi.org/10.1111/brv.12146
Forlani G, Bertazzini M, Zarattini M, Funck D (2015) Functional characterization and expression analysis of rice δ1-pyrroline-5-carboxylate dehydrogenase provide new insight into the regulation of proline and arginine catabolism. Front Plant Sci 6:1–17. https://doi.org/10.3389/fpls.2015.00591
Freitas PAF, Miranda RS, Marques EC et al (2018) Salt tolerance induced by exogenous proline in maize is related to low oxidative damage and favorable ionic homeostasis. J Plant Growth Regul 37:911–924. https://doi.org/10.1007/s00344-018-9787-x
Funck D, Eckard S, Müller G (2010) Non-redundant functions of two proline dehydrogenase isoforms in Arabidopsis. BMC Plant Biol 10:13p. https://doi.org/10.1186/1471-2229-10-70
Gaines MTP, Parker MB, Gascho GJ (1984) Automated determination of chlorides in soil and plant tissue by sodium nitrate. Agron J 76:371–374. https://doi.org/10.2134/agronj1984.00021962007600030005x
Gaufichon L, Rothstein SJ, Suzuki A (2016) Asparagine metabolic pathways in arabidopsis. Plant Cell Physiol 57:675–689. https://doi.org/10.1093/pcp/pcv184
Ginguay A, Cynober L, Curis E et al (2017) Ornithine aminotransferase, an important glutamate-metabolizing enzyme at the crossroads of multiple metabolic pathways. Biology (Basel) 7:E18. https://doi.org/10.3390/biology6010018
Golldack D, Li C, Mohan H, Probst N (2014) Tolerance to drought and salt stress in plants: unraveling the signaling networks. Front Plant Sci 5:1–10. https://doi.org/10.3389/fpls.2014.00151
Gupta B, Huang B (2014) Mechanism of salinity tolerance in plants: physiological, biochemical, and molecular characterization. Int J Genomics 2014:1–18. https://doi.org/10.1155/2014/701596
Gururani MA, Venkatesh J, Tran LSP (2015) Regulation of photosynthesis during abiotic stress-induced photoinhibition. Mol Plant 8:1304–1320. https://doi.org/10.1016/j.molp.2015.05.005
Hayat S, Hayat Q, Alyemeni MN et al (2012) Role of proline under changing environments a review. Plant Signal Behav 7:1456–1466. https://doi.org/10.4161/psb.21949
Hellemans J, Mortier G, De Paepe A et al (2007) qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol 8:R19. https://doi.org/10.1186/gb-2007-8-2-r19
Hildebrandt TM, Nesi AN, Araújo WL et al (2015) Amino acid catabolism in plants. Mol Plant 8:1563–1579. https://doi.org/10.1016/j.molp.2015.09.005
Hoagland DR, Arnon DI (1950) The water culture method for growing plants without soil. Calif Agric Exp Sta Circ 347:1–39
Hu R, Fan C, Li H et al (2009) Evaluation of putative reference genes for gene expression normalization in soybean by quantitative real-time RT-PCR. BMC Mol Biol 10:93. https://doi.org/10.1186/1471-2199-10-93
Kader A, Lindberg S, Kader A, Lindberg S (2017) Cytosolic calcium and pH signaling in plants under salinity stress cytosolic calcium and pH signaling in plants under salinity stress. Plant Signal Behav 2324:233–238. https://doi.org/10.4161/psb.5.3.10740
Kahlaoui B, Hachicha M, Misle E et al (2018) Physiological and biochemical responses to the exogenous application of proline of tomato plants irrigated with saline water. J Saudi Soc Agric Sci 17:17–23. https://doi.org/10.1016/j.jssas.2015.12.002
Kalaji HM, Bosa K, Ko J, Zuk-gołaszewska K (2011) Effects of salt stress on photosystem II efficiency and CO2 assimilation of two Syrian barley landraces. Environ Exp Bot 73:64–72. https://doi.org/10.1016/j.envexpbot.2010.10.009
Kaur G, Asthir B (2015) Proline: a key player in plant abiotic stress tolerance. Biol Plant 59:609–619. https://doi.org/10.1007/s10535-015-0549-3
Kavi-Kishor PB, Sreenivasulu N (2014) Is proline accumulation per se correlated with stress tolerance or is proline homeostasis a more critical issue? Plant Cell Environ 37:300–311. https://doi.org/10.1111/pce.12157
Kim HR, Rho HW, Park JW et al (1994) Assay of ornithine aminotransferase with ninhydrin. Anallytical Biochem 223:205–207. https://doi.org/10.1006/abio.1994.1574
Kudla J, Batistic O, Hashimoto K (2010) Calcium signals: the lead currency of plant information processing. Plant Cell 22:541–563. https://doi.org/10.1105/tpc.109.072686
Lehmann S, Funck D, Szabados L, Rentsch D (2010) Proline metabolism and transport in plant development. Amino Acids 39:949–962. https://doi.org/10.1007/s00726-010-0525-3
Li L, Li L, Wang X et al (2017) Plant growth-promoting endophyte Piriformospora indica alleviates salinity stress in Medicago truncatula. Plant Physiol Biochem 119:211–223. https://doi.org/10.1016/j.plaphy.2017.08.029
Lutts S, Kinet JM, Bouharmout J (1996) NaCl-induced senescence in leaves of rice (Oryza sativa L.) cultivars differing in salinity resistance. Ann Bot 78:389–398. https://doi.org/10.1006/anbo.1996.0134
Malavolta E, Vitti GC, Oliveira SA (1989) Avaliação do estudo nutricional das plantas, princípios e aplicações. Potafos, Piracicaba, p 97
Masclaux-Daubresse C, Daniel-Vedele F, Dechorgnat J et al (2010) Nitrogen uptake, assimilation and remobilization in plants: Challenges for sustainable and productive agriculture. Ann Bot 105:1141–1157. https://doi.org/10.1093/aob/mcq028
Miranda RS, Mesquita RO, Costa JH et al (2017) Integrative control between proton pumps and SOS1 antiporters in roots is crucial for maintaining low Na+ accumulation and salt tolerance in ammonium-supplied Sorghum bicolor. Plant Cell Physiol 58:522–536. https://doi.org/10.1093/pcp/pcw231
Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59:651–681. https://doi.org/10.1146/annurev.arplant.59.032607.092911
Nanjo T, Fujita M, Seki M et al (2003) Toxicity of free proline revealed in an Arabidopsis T-DNA-tagged mutant deficient in proline dehydrogenase. Plant Cell Physiol 44:541–548. https://doi.org/10.1093/pcp/pcg066
Negrão S, Schmöckel SM, Tester M (2017) Evaluating physiological responses of plants to salinity stress. Ann Bot 119:1–11. https://doi.org/10.1093/aob/mcw191
Parihar P, Singh S, Singh R et al (2015) Effect of salinity stress on plants and its tolerance strategies: a review. Environ Sci Pollut Res 22:4056–4075. https://doi.org/10.1007/s11356-014-3739-1
Park HJ, Kim W-Y, Yun D-J (2016) A new insight of salt stress signaling in plant. Mol Cells 39:447–459. https://doi.org/10.14348/molcells.2016.0083
Qamar A, Mysore KS, Senthil-Kumar M (2015) Role of proline and pyrroline-5-carboxylate metabolism in plant defense against invading pathogens. Front Plant Sci 6:1–9. https://doi.org/10.3389/fpls.2015.00503
Reddy PS, Jogeswar G, Rasineni GK et al (2015) Proline over-accumulation alleviates salt stress and protects photosynthetic and antioxidant enzyme activities in transgenic sorghum [Sorghum bicolor (L.) Moench]. Plant Physiol Biochem 94:104–113. https://doi.org/10.1016/j.plaphy.2015.05.014
Roy SJ, Negrão S, Tester M (2014) Salt resistant crop plants. Curr Opin Biotechnol 26:115–124. https://doi.org/10.1016/j.copbio.2013.12.004
Sakuraba H, Takamatsu Y, Satomura T, Kawakami R (2001) Purification, characterization, and application of a novel dye-linked l-proline dehydrogenase from a hyperthermophilic archaeon, Thermococcus profundus. Appl Environ Microb 67:1470–1475. https://doi.org/10.1128/AEM.67.4.1470
Shabala S, Pottosin I (2014) Regulation of potassium transport in plants under hostile conditions: implications for abiotic and biotic stress. Physiol Plant 151:257–279. https://doi.org/10.1111/ppl.12165
Singh M, Kumar J, Singh S et al (2015) Roles of osmoprotectants in improving salinity and drought tolerance in plants: a review. Rev Environ Sci Biotechnol 14:407–426. https://doi.org/10.1007/s11157-015-9372-8
Szabados L, Savoure A (2009) Proline: a multifunctional amino acid. Cell Press 15:89–97. https://doi.org/10.1016/j.tplants.2009.11.009
Székely G, Ábrahám E, Cséplö Á et al (2008) Duplicated P5CS genes of Arabidopsis play distinct roles in stress regulation and developmental control of proline biosynthesis. Plant J 53:11–28. https://doi.org/10.1111/j.1365-313X.2007.03318.x
Teh C-Y, Shaharrudin NA, Ho C-L, Mahmood M (2016) Exogenous proline significantly affects the plant growth and nitrogen assimilation enzymes activities in rice (Oryza sativa) under salt stress. Acta Physiol Plant 38:151. https://doi.org/10.1007/s11738-016-2163-1
Wang K, Liu Y, Dong K, Dong J, Kang J, Yang Q (2011) The effect of NaCl on proline metabolism in Saussurea amara seedlings. Afr J Biotechnol 10:2886–2893. https://doi.org/10.5897/AJB10.2269
Wang H, Tang X, Wang H, Shao H-B (2015) Proline accumulation and metabolism-related genes expression profiles in Kosteletzkya virginica seedlings under salt stress. Front Plant Sci 6:1–9. https://doi.org/10.3389/fpls.2015.00792
Winter G, Todd CD, Trovato M, Forlani G (2015) Physiological implications of arginine. Front Plant Sci 6:1–14. https://doi.org/10.3389/fpls.2015.00534
Wu G-Q, Feng R-J, Li S-J, Du Y-Y (2017) Exogenous application of proline alleviates salt-induced toxicity in sainfoin seedlings. J Anim Plant Sci 27:246–251
Yemm EW, Cocking EC, Ricketts RE (1955) The determination of amino-acids with ninhydrin. Analyst 80:209–214. https://doi.org/10.1039/an9558000209
Yu SM, Lo SF, Ho THD (2015) Source-sink communication: regulated by hormone, nutrient, and stress cross-signaling. Trends Plant Sci 20:844–857. https://doi.org/10.1016/j.tplants.2015.10.009
Zhang L, Becker DF (2015) Connecting proline metabolism and signaling pathways in plant senescence. Front Plant Sci 6:1–8. https://doi.org/10.3389/fpls.2015.00552
Zheng J-L, Zhao L, Bin CW, Zhu SA (2015) Exogenous proline reduces NaCl-induced damage by mediating ionic and osmotic adjustment and enhancing antioxidant defense in Eurya emarginata. Acta Physiol Plant 37:1–10. https://doi.org/10.1007/s11738-015-1921-9
Zörb C, Mühling KH, Kutschera U, Geilfus CM (2015) Salinity stiffens the epidermal cell walls of salt-stressed maize leaves: is the epidermis growth-restricting? PLoS One 10:1–15. https://doi.org/10.1371/journal.pone.0118406
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We thank Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for financial support and scholarships.
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de Freitas, P.A.F., de Carvalho, H.H., Costa, J.H. et al. Salt acclimation in sorghum plants by exogenous proline: physiological and biochemical changes and regulation of proline metabolism. Plant Cell Rep 38, 403–416 (2019). https://doi.org/10.1007/s00299-019-02382-5
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DOI: https://doi.org/10.1007/s00299-019-02382-5