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Characterization and expression analysis of P5CS (Δ1-pyrroline-5-carboxylate synthase) gene in two distinct populations of the Atlantic Forest native species Eugenia uniflora L.

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Abstract

Eugenia uniflora is an Atlantic Forest native species, occurring in contrasting edaphoclimatic environments. The identification of genes involved in response to abiotic factors is very relevant to help in understanding the processes of local adaptation. 1-Pyrroline-5-carboxylate synthetase (P5CS) is one interesting gene to study in this species since it encodes a key enzyme of proline biosynthesis, which is an osmoprotectant during abiotic stress. Applying in silico analysis, we identified one P5CS gene sequence of E. uniflora (EuniP5CS). Phylogenetic analysis, as well as, gene and protein structure investigation, revealed that EuniP5CS is a member of P5CS gene family. Plants of E. uniflora from two distinct environments (restinga and riparian forest) presented differences in the proline accumulation and P5CS expression levels under growth-controlled conditions. Both proline accumulation and gene expression level of EuniP5CS were higher in the genotypes from riparian forest than those from restinga. When these plants were submitted to drought stress, EuniP5CS gene was up-regulated in the plants from restinga, but not in those from riparian forest. These results demonstrated that EuniP5CS is involved in proline biosynthesis in this species and suggest that P5CS gene may be an interesting candidate gene in future studies to understand the processes of local adaptation in E. uniflora.

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References

  1. Wang WX, Vinocur B, Shoseyov O, Altman A (2001) Biotechnology of plant osmotic stress tolerance: physiological and molecular considerattions. Acta Hort 560:285–292

    Article  CAS  Google Scholar 

  2. Mickelbart MV, Hasegawa PM, Bailey-Serres J (2015) Genetic mechanisms of abiotic stress tolerance that translate to crop yield stability. Nat Rev Genet 16:237–251. https://doi.org/10.1038/nrg3901

    Article  CAS  PubMed  Google Scholar 

  3. Lesk C, Rowhani P, Ramankutty N (2016) Influence of extreme weather disasters on global crop production. Nature 529:84–87

    Article  CAS  PubMed  Google Scholar 

  4. Szabados L, Savouré A (2010) Proline: a multifunctional amino acid. Trends Plant Sci 15:89–97. https://doi.org/10.1016/j.tplants.2009.11.009

    Article  CAS  PubMed  Google Scholar 

  5. Smirnoff N, Cumbes QJ (1989) Hydroxyl radical scavening activity of compatible solutes. Phytochimistry 28:1057–1060. https://doi.org/10.1016/0031-9422(89)80182-7

    Article  CAS  Google Scholar 

  6. Matysik J, Alia Bhalu B, Mohanty P (2002) Molecular mechanisms of quenching of reactive oxygen species by proline under stress in plants. Curr Sci 82:525–532

    CAS  Google Scholar 

  7. Chinnusamy V, Jagendorf A, Zhu JK (2005) Understanding and improving salt tolerance in plants. Crop Sci 45:437–448. https://doi.org/10.2135/cropsci2005.0437

    Article  CAS  Google Scholar 

  8. Verbruggen N, Hermans C (2008) Proline accumulation in plants: a review. Amino Acids 35:753–759. https://doi.org/10.1007/s00726-008-0061-6

    Article  CAS  PubMed  Google Scholar 

  9. Delauney AJ, Verma DPS (1993) Proline biosynthesis and osmoregulation in plants. Plant J 4:215–223

    Article  CAS  Google Scholar 

  10. Molinari HBC, Marur CJ, Filho JCB et al (2004) Osmotic adjustment in transgenic citrus rootstock Carrizo citrange (Citrus sinensis Osb. x Poncirus trifoliata L. Raf.) overproducing proline. Plant Sci 167:1375–1381. https://doi.org/10.1016/j.plantsci.2004.07.007

    Article  CAS  Google Scholar 

  11. Kishor PBK, Hong ZL, Miao GH et al (1995) Overexpression of delta-pyrroline-5-carboxylate synthetase increases proline production and confers osmotolerance in transgenic plants. Plant Physiol 108:1387–1394. https://doi.org/10.1104/pp.108.4.1387

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Choudhary NL, Sairam RK, Tyagi A (2005) Expression of delta1-pyrroline-5-carboxylate synthetase gene during drought in rice (Oryza sativa L.). Indian J Biochem Biophys 42:366–370

    CAS  PubMed  Google Scholar 

  13. Guan C, Ji J, Guan W et al (2014) A Lycium chinense-derived P5CS-like gene is regulated by water deficit-induced endogenous abscisic acid and overexpression of this gene enhances tolerance to water deficit stress in Arabidopsis. Mol Breed 34:1109–1124. https://doi.org/10.1007/s11032-014-0103-6

    Article  CAS  Google Scholar 

  14. Turchetto-Zolet AC, Margis-Pinheiro M, Margis R (2009) The evolution of pyrroline-5-carboxylate synthase in plants: a key enzyme in proline synthesis. Mol Genet Genom 281:87–97. https://doi.org/10.1007/s00438-008-0396-4

    Article  CAS  Google Scholar 

  15. Ginzberg I, Stein H, Kapulnik Y et al (1998) Isolation and characterization of two different cDNAs of delta1-pyrroline-5-carboxylate synthase in alfalfa, transcriptionally induced upon salt stress. Plant Mol Biol 38:755–764

    Article  CAS  PubMed  Google Scholar 

  16. Farzaneh M, Jazi FR, Motamed N (2005) Application of dot blotting for detecting the expression of p5cs gene in transgenic olive plant-lets. FEBS J 272:547

    Google Scholar 

  17. Silva-Ortega CO, Ochoa-Alfaro AE, Reyes-Agüero JA et al (2008) Salt stress increases the expression of p5cs gene and induces proline accumulation in cactus pear. Plant Physiol Biochem 46:82–92

    Article  CAS  PubMed  Google Scholar 

  18. Zhuang G-Q, Li B, Guo H-Y et al (2011) Molecular cloning and characterization of P5CS gene from Jatropha curcas L. African J Biotechnol 10:14803–14811. https://doi.org/10.5897/AJB11.2072

    Article  CAS  Google Scholar 

  19. Ramadan AM, Hassanein SE (2014) Characterization of P5CS gene in Calotropis procera plant from the de novo assembled transcriptome contigs of the high-throughput sequencing dataset. Comptes Rendus—Biol 337:683–690. https://doi.org/10.1016/j.crvi.2014.09.002

    Article  Google Scholar 

  20. Cao L, Han L, Zhang H et al (2015) Isolation and characterization of pyrroline-5-carboxylate synthetase gene from perennial ryegrass (Lolium perenne L.). Acta Physiol Plant 37:62. https://doi.org/10.1007/s11738-015-1808-9

    Article  CAS  Google Scholar 

  21. Wang L, Guo Z, Zhang Y et al (2017) Characterization of LhSorP5CS, a gene catalyzing proline synthesis in Oriental hybrid lily Sorbonne: molecular modelling and expression analysis. Bot Stud 58:10. https://doi.org/10.1186/s40529-017-0163-0

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Hmida-Sayari A, Gargouri-Bouzid R, Bidani A et al (2005) Overexpression of Δ1-pyrroline-5-carboxylate synthetase increases proline production and confers salt tolerance in transgenic potato plants. Plant Sci 169:746–752. https://doi.org/10.1016/j.plantsci.2005.05.025

    Article  CAS  Google Scholar 

  23. Xue X, Liu A, Hua X (2009) Proline accumulation and transcriptional regulation of proline biothesynthesis and degradation in Brassica napus. BMB Rep 42:28–34. https://doi.org/10.5483/BMBRep.2009.42.1.028

    Article  CAS  PubMed  Google Scholar 

  24. Peng Z, Lu Q, Verma DPS (1996) Reciprocal regulation of Δ1-pyrroline-5-carboxylate synthetase and proline dehydrogenase genes controls proline levels during and after osmotic stress in plants. Mol Gen Genet 253:334–341. https://doi.org/10.1007/s004380050329

    Article  CAS  PubMed  Google Scholar 

  25. Margis R, Felix D, Caldas JF et al (2002) Genetic differentiation among three neighboring Brazil-cherry (Eugenia uniflora L.) populations within the Brazilian Atlantic rain forest. Biodivers Conserv 11:149–163. https://doi.org/10.1023/A:1014028026273

    Article  Google Scholar 

  26. Salgueiro F, Felix D, Caldas JF et al (2004) Even population differentiation for maternal and biparental gene markers in Eugenia uniflora, a widely distributed species from the Brazilian Coastal Atlantic Rain Forest. Divers Distrib 10:201–210

    Article  Google Scholar 

  27. Rotman AD (1995) Las especies argentinas del género Myrcianthes (Myrtaceae). Bol la Soc Argentina Botánica 31:69–93

    Google Scholar 

  28. Dillenburg LR, Waechter JL, Porto ML (1992) Species composition and structure of a sandy coastal plain forest in Northern Rio Grande do Sul, Brazil. In: Seeliger U (ed) Coastal plant communities of Latin America. Elsevier, Amsterdam, pp 349–366

    Chapter  Google Scholar 

  29. Araújo DSD, Scarano FR, Sá CFC et al (1998) Comunidades vegetais do Parque Nacional da Restinga de Jurujuba. Esteves FA Ecol das lagoas costeiras do Parq Nac da Restin Jurujuba e do Município Macae (RJ) NUPEM/UFRJ, Rio Janeiro 39–62

  30. Barroso GM, Marques MCM (1997) Myrtaceae. In: Marques MCM, Vaz ASF, Marquete R (eds) Flórula da APA Cairuçu, Parati, RJ: espécies vasculares, 14th edn. Jardim Botânico do Rio de Janeiro, Rio de Janeiro, pp 314–382

    Google Scholar 

  31. Scarano FR (2009) Plant communities at the periphery of the Atlantic rain forest: rare-species bias and its risks for conservation. Biol Conserv 142:1201–1208. https://doi.org/10.1016/j.biocon.2009.02.027

    Article  Google Scholar 

  32. Rodrigues RR, Nave AG (2000) Heterogeneidade Florística das Matas Ciliares. In: Rodrigues RR, Leitão Filho HF (eds) Matas Ciliares: conservação e recuperação, 14th edn. São Paulo, EDUSP. EDUSP/FAPESP, São Paulo, pp 45–71

    Google Scholar 

  33. Turchetto-Zolet AC, Salgueiro F, Turchetto C et al (2016) Phylogeography and ecological niche modelling in Eugenia uniflora (Myrtaceae) suggest distinct vegetational responses to climate change between the southern and the northern Atlantic Forest. Bot J Linn Soc 182:670–688

    Article  Google Scholar 

  34. Guzman F, Kulcheski FR, Turchetto-Zolet AC, Margis R (2014) De novo assembly of Eugenia uniflora L. transcriptome and identification of genes from the terpenoid biosynthesis pathway. Plant Sci 229:238–246. https://doi.org/10.1016/j.plantsci.2014.10.003

    Article  CAS  PubMed  Google Scholar 

  35. Langmead B, Trapnell C, Pop M, Salzberg SL (2009) Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol 10:R25. https://doi.org/10.1186/gb-2009-10-3-r25

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Milne I, Bayer M, Cardle L et al (2009) Tablet-next generation sequence assembly visualization. Bioinformatics 26:401–402. https://doi.org/10.1093/bioinformatics/btp666

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32:1792–1797. https://doi.org/10.1093/nar/gkh340

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Stamatakis A (2014) RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 30:1312–1313. https://doi.org/10.1093/bioinformatics/btu033

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Lanfear R, Frandsen PB, Wright AM et al (2016) PartitionFinder 2: new methods for selecting partitioned models of evolution for molecular and morphological phylogenetic analyses. Mol Biol Evol 34:772–773. https://doi.org/10.1093/molbev/msw260

    Article  CAS  Google Scholar 

  40. Gambino G, Perrone I, Gribaudo A (2008) Rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants. Phytochem Anal PCA 19:520–525

    Article  CAS  PubMed  Google Scholar 

  41. Vandesompele J, De Preter K, Pattyn F et al (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. https://doi.org/10.1186/gb-2002-3-7-research0034

    Article  PubMed  PubMed Central  Google Scholar 

  42. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262

    Article  CAS  PubMed  Google Scholar 

  43. Lee MR, Kim CS, Park T et al (2018) Optimization of the ninhydrin reaction and development of a multiwell plate-based high-throughput proline detection assay. Anal Biochem 556:57–62. https://doi.org/10.1016/j.ab.2018.06.022

    Article  CAS  PubMed  Google Scholar 

  44. Zhou H, Qian J, Zhao M-D et al (2016) Cloning and sequence analysis of the Δ1-pyrroline-5-carboxylate synthase gene (MP5CS) from mulberry (Morus alba) and patterns of MP5CS gene expression under abiotic stress conditions. J Hortic Sci Biotechnol 91:100–108. https://doi.org/10.1080/14620316.2015.1110999

    Article  CAS  Google Scholar 

  45. Székely G, Ábrahám E, Cséplo 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

    Article  CAS  PubMed  Google Scholar 

  46. Su M, Li X-F, Ma X-Y et al (2011) Cloning two P5CS genes from bioenergy sorghum and their expression profiles under abiotic stresses and MeJA treatment. Plant Sci 181:652–659. https://doi.org/10.1016/J.PLANTSCI.2011.03.002

    Article  CAS  PubMed  Google Scholar 

  47. Kubala S, Wojtyla L, Quinet M et al (2015) Enhanced expression of the proline synthesis gene P5CSA in relation to seed osmopriming improvement of Brassica napus germination under salinity stress. J Plant Physiol 183:1–12. https://doi.org/10.1016/j.jplph.2015.04.009

    Article  CAS  PubMed  Google Scholar 

  48. Toscano S, Farieri E, Ferrante A, Romano D (2016) Physiological and biochemical responses in two ornamental shrubs to drought stress. Front Plant Sci 7:645. https://doi.org/10.3389/fpls.2016.00645

    Article  PubMed  PubMed Central  Google Scholar 

  49. Chen JB, Wang SM, Jing RL, Mao XG (2009) Cloning the PvP5CS gene from common bean (Phaseolus vulgaris) and its expression patterns under abiotic stresses. J Plant Physiol 166:12–19. https://doi.org/10.1016/j.jplph.2008.02.010

    Article  CAS  PubMed  Google Scholar 

  50. Maghsoudia K, Emama Y, Niazib A, Pessaraklic M, Arvind MJ (2008) P5CS expression level and proline accumulation in the sensitive and tolerant wheat cultivars under control and drought stress conditions in the presence/absence of silicon and salicylic acid. J Plant Interact 13(1):461–471

    Article  Google Scholar 

  51. Fu J, Sun P, Luo Y, Zhou H, Gao J, Zhao D, Pubu Z, Liu J, Hu T (2019) Brassinosteroids enhance cold tolerance in Elymus nutans via mediating redox homeostasis and proline biosynthesis. Environ Exp Bot 167:103831

    Article  CAS  Google Scholar 

  52. Saghfi S, Eivazi AR (2014) Effects of cold stress on proline and soluble carbohydrates in two chickpea cultivars. Int J Curr Microbiol Appl Sci 3(2):591–595

    Google Scholar 

  53. Le Gall H, Fontaine JX, Molinié R, Pelloux J, Mesnard F, Gillet F, Fliniaux O (2017) NMR-based metabolomics to study the cold-acclimation strategy of two miscanthus genotypes. Phytochem Anal 28(1):58–67

    Article  PubMed  Google Scholar 

  54. Taji T, Seki M, Satou M, Sakurai T, Kobayashi M, Ishiyama K, Narusaka Y, Narusaka M, Zhu JK, Shinozaki K (2004) Comparative genomics in salt tolerance between Arabidopsis and Arabidopsis-related halophyte salt cress using Arabidopsis microarray. Plant Physiol 135(3):1697–1709

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Pang Q, Chen S, Dai S, Chen Y, Wang Y, Yan X (2010) Comparative proteomics of salt tolerance in Arabidopsis thaliana and Thellungiella halophila. J Proteome Res 9:2584–2599

    Article  CAS  PubMed  Google Scholar 

  56. De Oliveira AD, Fernandes EJ, De T, Rodrigues JD (2004) Condutância estomática como indicador de estresse hídrico em feijão. Eng Agríc Jaboticabal 25:86–95

    Article  Google Scholar 

  57. Farooq M, Hussain M, Wahid A, Siddique KHM (2012) Drought stress in plants: an overview. In: Aroca R (ed) Plant responses to drought stress. Springer, Singapore, pp 1–33

    Google Scholar 

  58. Chaves MM, Maroco JP, Pereira JS (2003) Understanding plant responses to drought—from genes to the whole plant. Funct Plant Biol 30:239–264. https://doi.org/10.1071/FP02076

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was financially supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; Grant Number: 306202/2016-6) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES; 2806/2014), and Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul [grant number 16/491-9]. We thank Dr. Arthur Germano Fett Neto, who supported us with the experiment of proline determination.

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Authors and Affiliations

  1. Programa de Pós-Graduação em Genética e Biologia Molecular (PPGBM) Departamento de Genética, Instituto de Biociências, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Bento Gonçalves 9500, Prédio 43312, Porto Alegre, 91501-970, Brazil

    Débora Bublitz Anton, Nicole Moreira Vetö, Rogério Margis & Andreia Carina Turchetto-Zolet

  2. Graduação em Biotecnologia, Departamento de Biologia Molecular e Biotecnologia, Instituto de Biociências, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil

    Débora Bublitz Anton

  3. Centro de Biotecnologia e Programa de Pós-Graduação em Biologia Celular e Molecular, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil

    Frank Lino Guzman, Ana Paula Durand Coelho, Guilherme Leitão Duarte & Rogério Margis

  4. Graduação em Agronomia, Faculdade de Agronomia, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil

    Felipe Augusto Krause

  5. Departamento de Biofísica, Instituto de Biociências, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil

    Rogério Margis

  6. Programa de Pós-Graduação em Biologia Celular e do Desenvolvimento (PPGBCD) Departamento de Biologia Celular, Embriologia e Genética, Universidade Federal de Santa Catarina (UFSC), Porto Alegre, Brazil

    Franceli Rodrigues Kulcheski

  7. Laboratório de Ecofisiologia Vegetal, Departamento de Botânica, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil

    Lúcia Rebello Dillenburg

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Anton, D.B., Guzman, F.L., Vetö, N.M. et al. Characterization and expression analysis of P5CS (Δ1-pyrroline-5-carboxylate synthase) gene in two distinct populations of the Atlantic Forest native species Eugenia uniflora L.. Mol Biol Rep 47, 1033–1043 (2020). https://doi.org/10.1007/s11033-019-05195-7

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