Advertisement

Plant Cell Reports

, Volume 32, Issue 11, pp 1729–1741 | Cite as

Molecular and functional characterization of mulberry EST encoding remorin (MiREM) involved in abiotic stress

  • Vibha G. Checker
  • Paramjit KhuranaEmail author
Original Paper

Abstract

Key message

Group1 remorins may help the plants to optimize their growth under adverse conditions by their involvement in mediating osmotic stress responses in plants.

Abstract

Mulberry (Morus indica), a deciduous woody tree, serves as the cardinal component of the sericulture industry. Genomic endeavors in sequencing of mulberry ESTs provided clues to stress-specific clones, but their functional relevance remains fragmentary. Therefore in this study, we assessed the functional significance of a remorin gene family member that was identified in leaf ESTs. Remorins represent a large, plant-specific multigene family gaining importance in recent times with respect to their role in plant–microbe interactions, although their role in response to environmental stresses remains speculative as in vivo functions of remorin genes are limited. Mulberry remorin (MiREM) localizes to plasma membrane and is ubiquitously present in all plant organs. Expression analysis of MiREM by northern analysis reveals that its transcript increases under different abiotic stress conditions especially during dehydration and salt stress, implicating it in regulation of stress signaling pathways. Concomitantly, transgenic Arabidopsis plants overexpressing heterologous remorin show tolerance to dehydration and salinity at the germination and seedling stages as revealed by percentage germination, root inhibition assays, fresh weight and activity of photosystem II. This study predicts the possible function of group 1 remorin gene in mediating osmotic stress thus bringing novel perspectives in understanding the function of remorins in plant abiotic stress responses.

Keywords

Abiotic stresses Arabidopsis Group1 remorin Mulberry 

Notes

Acknowledgments

This work was financially supported by grants received from the Department of Biotechnology (DBT), Government of India, New Delhi. V.G.C. acknowledges University Grants Commission (UGC) for the award of research fellowships.

Supplementary material

299_2013_1483_MOESM1_ESM.ppt (583 kb)
Supplementary material 1 (PPT 583 kb)

References

  1. Bariola PA, Retelska D, Stasiak A, Kammerer RA, Fleming A, Hijri M, Frank S, Farmer EE (2004) Remorins form a novel family of coiled coil-forming oligomeric and filamentous proteins associated with apical, vascular and embryonic tissues in plants. Plant Mol Biol 55:579–594PubMedCrossRefGoogle Scholar
  2. Bray EA (2002) Classification of genes differentially expressed during water-deficit stress in Arabidopsis thaliana: an analysis using microarray and differential expression data. Ann Bot 89:803–811PubMedCrossRefGoogle Scholar
  3. Burkhard P, Stetefeld J, Strelkov SV (2001) Coiled coils: a highly versatile protein folding motif. Trends Cell Biol 11:82–88PubMedCrossRefGoogle Scholar
  4. Checker VG, Chhibbar AK, Khurana P (2012a) Stress-inducible expression of barley Hva1 gene in transgenic mulberry displays enhanced tolerance against drought, salinity and cold stress. Transgenic Res 21:939–957PubMedCrossRefGoogle Scholar
  5. Checker VG, Saeed B, Khurana P (2012b) Analysis of expressed sequence tags from mulberry (Morus indica) roots and implications for comparative transcriptomics and marker identification. Tree Genet Genomes 8(6):1437–1450Google Scholar
  6. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Anal Biochem 162:156–159PubMedCrossRefGoogle Scholar
  7. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743PubMedCrossRefGoogle Scholar
  8. Coaker GL, Willard B, Kinter M, Stockinger EJ, Francis DM (2004) Proteomic analysis of resistance mediated by Rcm 2.0 and Rcm 5.1, two loci controlling resistance to bacterial canker of tomato. Mol Plant Microbe Interact 17:1019–1028PubMedCrossRefGoogle Scholar
  9. Cushman JC, Bohnert HJ (2000) Genomic approaches to plant stress tolerance. Curr Opin Plant Biol 3:117–124PubMedCrossRefGoogle Scholar
  10. Dai X, Xu Y, Ma Q, Xu W, Wang T, Xue Y, Chong K (2007) Overexpression of a R1R2R3 MYB Gene, OsMYB3R-2, increases tolerance to freezing, drought and salt stress in transgenic Arabidopsis. Plant Physiol 143:1739–1751PubMedCrossRefGoogle Scholar
  11. Das M, Chauhan H, Chhibbar A, Rizwanul Haq QM, Khurana P (2011) High-efficiency transformation and selective tolerance against biotic and abiotic stress in mulberry, Morus indica cv. K2, by constitutive and inducible expression of tobacco osmotin. Transgenic Res 20:231–246PubMedCrossRefGoogle Scholar
  12. El Yahyaoui F, Kuster H, Ben Amor B, Hohnjec N, Puhler A, Becker A, Gouzy J, Vernie T, Gough C, Niebel A, Godiard L, Gamas P (2004) Expression profiling in Medicago truncatula identifies more than 750 genes differentially expressed during nodulation, including many potential regulators of the symbiotic program. Plant Physiol 136:3159–3176PubMedCrossRefGoogle Scholar
  13. Farmer EE, Pearce G, Ryan CA (1989) In vitro phosphorylation of plant plasma membrane proteins in response to the proteinase inhibitor inducing factor. Proc Natl Acad Sci USA 86:1539–1542PubMedCrossRefGoogle Scholar
  14. Fedorova M, van de Mortel J, Matsumoto PA, Cho J, Town CD, Vanden Bosch KA, Gantt JS, Vance CP (2002) Genome-wide identification of nodule-specific transcripts in the model legume Medicago truncatula. Plant Physiol 130:519–537PubMedCrossRefGoogle Scholar
  15. Gulyani V, Khurana P (2011) Identification and expression profiling of drought regulated genes in mulberry (Morus sp.) by suppression subtractive hybridization of susceptible and tolerant cultivars. Tree Genet Genomes 7:725–738CrossRefGoogle Scholar
  16. Hema R, Senthil-Kumar M, Shivakumar S, Reddy PC, Udayakumar M (2007) Chlamydomonas reinhardtii, a model system for functional validation of abiotic stress responsive genes. Planta 226:655–670PubMedCrossRefGoogle Scholar
  17. Ivashikina N, Deeken R, Ache P, Kranz E, Pommerrenig B, Sauer N, Hedrich R (2003) Isolation of AtSUC2 promoter-GFP-marked companion cells for patch-clamp studies and expression profiling. Plant J 36:931–945PubMedCrossRefGoogle Scholar
  18. Jacinto T, Farmer EE, Ryan CA (1993) Purification of potato leaf plasma membrane protein pp 34, a protein phosphorylated in response to oligogalacturonide signals for defense and development. Plant Physiol 103:1393–1397PubMedGoogle Scholar
  19. Jain M, Tyagi AK, Khurana JP (2008) Constitutive expression of a meiotic recombination protein gene homolog, OsTOP6A1, from rice confers abiotic stress tolerance in transgenic Arabidopsis plants. Plant Cell Rep 27:767–778PubMedCrossRefGoogle Scholar
  20. Jarsch IK, Ott T (2011) Perspectives on remorin proteins, membrane rafts, and their role during plant–microbe interactions. Mol Plant Microbe Interact 24:7–12PubMedCrossRefGoogle Scholar
  21. Jewell MC, Campbell CB, Godwin ID (2010) Transgenic plants for abiotic stress resistance. In: Kole C et al (eds) Transgenic crop plants. Springer, BerlinGoogle Scholar
  22. Kaplan B, Davydov O, Knight H, Galon Y, Knight MR, Fluhr R, Fromm H (2006) Rapid transcriptome changes induced by cytosolic Ca2+ transients reveal ABRE-related sequences as Ca2+-responsive cis elements in Arabidopsis. Plant Cell 18:2733–2748PubMedCrossRefGoogle Scholar
  23. Khurana P, Checker VG (2011) The advent of genomics in mulberry and perspectives for productivity enhancement. Plant Cell Rep 30:825–838PubMedCrossRefGoogle Scholar
  24. Kim CS, Lee CH, Shin JS, Chung YS, Hyung NI (1997) A simple and rapid method for isolation of high quality genomic DNA from fruit trees and conifers using PVP. Nucleic Acids Res 25:1085–1086PubMedCrossRefGoogle Scholar
  25. Kim DY, Jin JY, Alejandro S, Martinoia E, Lee Y (2010) Overexpression of AtABCG36 improves drought and salt stress resistance in Arabidopsis. Physiol Plant 139:170–180PubMedCrossRefGoogle Scholar
  26. Kistner C, Winzer T, Pitzschke A, Mulder L, Sato S, Kaneko T, Tabata S, Sandal N, Stougaard J, Webb KJ, Szczyglowski K, Parniske M (2005) Seven Lotus japonicus genes required for transcriptional reprogramming of the root during fungal and bacterial symbiosis. Plant Cell 17:2217–2229PubMedCrossRefGoogle Scholar
  27. Knight H, Knight MR (2001) Abiotic stress signalling pathways: specificity and cross-talk. Trends Plant Sci 6:262–267PubMedCrossRefGoogle Scholar
  28. Krause GH, Weiss E (1991) Chlorophyll fluorescence and photosynthesis: the basics. Annu Rev Plant Physiol Plant Mol Biol 42:313–349CrossRefGoogle Scholar
  29. Kreps JA, Wu Y, Chang HS, Zhu T, Wang X, Harper JF (2002) Transcriptome changes for Arabidopsis in response to salt, osmotic, and cold stress. Plant Physiol 130:2129–2141PubMedCrossRefGoogle Scholar
  30. Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132PubMedCrossRefGoogle Scholar
  31. Lal S, Gulyani V, Khurana P (2008) Overexpression of HVA1 gene from barley generates tolerance to salinity and water stress in transgenic mulberry (Morus indica). Transgenic Res 17:651–663PubMedCrossRefGoogle Scholar
  32. Lal S, Ravi V, Khurana JP, Khurana P (2009) Repertoire of leaf expressed sequence tags (ESTs) and partial characterization of stress-related and membrane transporter genes from mulberry (Morus indica L.). Tree Genet Genomes 5:359–374CrossRefGoogle Scholar
  33. Lefebvre B, Timmers T, Mbengue M, Moreau S, Herve C, Toth K, Bittencourt-Silvestre J, Klaus D, Deslandes L, Godiard L, Murray JD, Udvardi MK, Raffaele S, Mongrand S, Cullimore J, Gamas P, Niebel A, Ott T (2010) A remorin protein interacts with symbiotic receptors and regulates bacterial infection. Proc Natl Acad Sci USA 107:2343–2348PubMedCrossRefGoogle Scholar
  34. Lin F, Xu SL, Ni WM, Chu ZQ, Xu ZH, Xue HW (2003) Identification of ABA-responsive genes in rice shoots via cDNA macroarray. Cell Res 13:59–68PubMedCrossRefGoogle Scholar
  35. Liu N, Zhong NQ, Wang GL, Li LJ, Liu XL, He YK, Xia GX (2006) Cloning and functional characterization of PpDBF1 gene encoding a DRE-binding transcription factor from Physcomitrella patens. Planta 226:827–838CrossRefGoogle Scholar
  36. Liu J, Elmore JM, Fuglsang AT, Palmgren MG, Staskawicz BJ, Coaker G (2009) RIN4 functions with plasma membrane H +-ATPases to regulate stomatal apertures during pathogen attack. PLoS Biol 7:e1000139PubMedCrossRefGoogle Scholar
  37. Malakshah SSN, Rezaei MMH, Heidari MM, Salekdeh GHGH (2007) Proteomics reveals new salt responsive proteins associated with rice plasma membrane. Biosci Biotechnol Biochem 71:2144–2154CrossRefGoogle Scholar
  38. Marin M, Ott T (2012) Phosphorylation of intrinsically disordered regions in remorin preteins. Front Plant Sci 3:86PubMedCrossRefGoogle Scholar
  39. Marmagne A, Rouet MA, Ferro M, Rolland N, Alcon C, Joyard J, Garin J, Barbier-Brygoo H, Ephritikhine G (2004) Identification of new intrinsic proteins in Arabidopsis plasma membrane proteome. Mol Cell Proteomics 3:675–691PubMedCrossRefGoogle Scholar
  40. Mason JM, Arndt KM (2004) Coiled coil domains: stability, specificity, and biological implications. Chem Bio Chem 5:170–176PubMedCrossRefGoogle Scholar
  41. Mittler R, Blumwald E (2010) Genetic engineering for modern agriculture: challenge and perspectives. Annu Rev Plant Biol 61:443–462PubMedCrossRefGoogle Scholar
  42. Mukhopadhyay A, Vij S, Tyagi AK (2004) Overexpression of a zinc-finger protein gene from rice confers tolerance to cold, dehydration, and salt stress in transgenic tobacco. Proc Natl Acad Sci USA 101:6309–6314PubMedCrossRefGoogle Scholar
  43. Nelson CJ, Hegeman AD, Harms AC, Sussman MR (2006) A quantitative analysis of Arabidopsis plasma membrane using trypsin-catalyzed 18O labeling. Mol Cell Proteomics 5:1382–1395PubMedCrossRefGoogle Scholar
  44. Ofosu-Anim J, Offei SK, Yamaki S (2006) Pistil receptivity, pollen tube growth and gene expression during early fruit development in sweet pepper (Capsicum annum). Int J Agric Biol 8:576–579Google Scholar
  45. Raffaele S, Mongrand S, Gamas P, Niebel A, Ott T (2007) Genome-wide annotation of remorins, a plant-specific protein family: evolutionary and functional perspectives. Plant Physiol 145:593–600PubMedCrossRefGoogle Scholar
  46. Raffaele S, Bayer E, Lafarge D, Cluzet S, German Retana S, Boubekeur T, Leborgne-Castel N, Carde JP, Lherminier J, Noirot E, Satiat-Jeunemaitre B, Laroche-Traineau J, Moreau P, Ott T, Maule AJ, Reymond P, Simon-Plas F, Farmer EE, Bessoule JJ, Mongrand S (2009a) Remorin, a solanaceae protein resident in membrane rafts and plasmodesmata, impairs potato virus X movement. Plant Cell 21:1541–1555PubMedCrossRefGoogle Scholar
  47. Raffaele S, Bayer E, Mongrand S (2009b) Upregulation of the plant protein remorin correlates with dehiscence and cell maturation: a link with the maturation of plasmodesmata? Plant Signal Behav 4:915–919PubMedCrossRefGoogle Scholar
  48. Reymond P, Kunz B, Paul-Pletzer K, Grimm R, Eckerskorn C, Farmer EE (1996) Cloning of a cDNA encoding a plasma membrane-associated, uronide binding phosphoprotein with physical properties similar to viral movement proteins. Plant Cell 8:2265–2276PubMedGoogle Scholar
  49. Rose A, Schraegle SJ, Stahlberg EA, Meier I (2005) Coiled-coil protein composition of 22 proteomes—differences and common themes in subcellular infrastructure and traffic control. BMC Evol Biol 5:66PubMedCrossRefGoogle Scholar
  50. Sambrook J, Russel DW (2001) Molecular cloning. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  51. Sanchez-Moran E, Mercier R, Higgins JD, Armstrong SJ, Jones GH, Franklin FC (2005) A strategy to investigate the plant meiotic proteome. Cytogenet Genome Res 109:181–189PubMedCrossRefGoogle Scholar
  52. Sazuka T, Keta S, Shiratake K, Yamaki S, Shibata D (2004) A proteomic approach to identification of transmembrane proteins and membrane-anchored proteins of Arabidopsis thaliana by peptide sequencing. DNA Res 11:101–113PubMedCrossRefGoogle Scholar
  53. Seki M, Narusaka M, Ishida J, Nanjo T, Fujita M, Oono Y, Kamiya A, Nakajima M, Enju A, Sakurai T, Satou M, Akiyama K, Taji T, Yamaguchi-Shinozaki K, Carninci P, Kawai J, Hayashizaki Y, Shinozaki K (2002) Monitoring the expression profiles of 7000 Arabidopsis genes under drought, cold and high-salinity stresses using a full-length cDNA microarray. Plant J 31:279–292PubMedCrossRefGoogle Scholar
  54. Seong ES, Baek KH, Oh SK, Sung Hwan Jo SH, Yi SY, Park JM, Joung YH, Lee S, Cho SH, Choi D (2007) Induction of enhanced tolerance to cold stress and disease by overexpression of the pepper CaPIF1 gene in tomato. Physiol Plant 129:555–566CrossRefGoogle Scholar
  55. Sreenivasulu N, Sopory SK, Kavi Kishor PB (2007) Deciphering the regulatory mechanisms of abiotic stress tolerance in plants by genomic approaches. Gene 388:1–13PubMedCrossRefGoogle Scholar
  56. Tóth K, StratilTF MadsenEB, YeJ Popp C, Antolín-Llovera M, GrossmannC JensenOL, SchüßlerA ParniskeM, Ott T (2012) Functional domain analysis of the Remorin protein LjSYMREM1 in Lotus japonicas. PloS One 7:e30817PubMedCrossRefGoogle Scholar
  57. Valot B, Negroni L, Zivy M, Gianinazzi S, Dumas-Gaudot E (2006) A mass spectrometric approach to identify arbuscular mycorrhiza-related proteins in root plasma membrane fractions. Proteomics 6:145–155CrossRefGoogle Scholar
  58. Vicente MRS, Plasencia J (2011) Salicyclic acid beyond defence: its role in plant growth and development. J Exp Bot 62:3321–3338CrossRefGoogle Scholar
  59. Vlot AC, Dempsey DMA, Klessig DF (2009) Salicylic acid, a multifaceted hormone to combat disease. Annu Rev Phytopathol 47:177–206PubMedCrossRefGoogle Scholar
  60. Watson BS, Asirvatham VS, Wang L, Sumner LW (2003) Mapping the proteome of barrel medic (Medicago truncatula). Plant Physiol 131:1104–1123PubMedCrossRefGoogle Scholar
  61. Widjaja I, Naumann K, Roth U, Wolf N, Mackey D, Jl Dangl, Scheel D, Lee J (2009) Combining sub proteome enrichment and rubisco depletion enables identification of low abundance proteins differentially regulated during plant defense. Proteomics 9:138–147PubMedCrossRefGoogle Scholar
  62. Wienkoop S, Saalbach G (2003) Proteome analysis. Novel proteins identified at the peribacteroid membrane from Lotus japonicus root nodules. Plant Physiol 131:1080–1090PubMedCrossRefGoogle Scholar
  63. Yang CJ, Zhang C, Lu YN, Jin JQ, Wang XL (2011) The mechanisms of brassinosteroids’ action: from signal transduction to plant development. Mol Plant 4(4):588–600Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  1. 1.Department of Plant Molecular BiologyUniversity of Delhi South CampusNew DelhiIndia

Personalised recommendations