3 Biotech

, 8:157 | Cite as

Differential expression of leaf proteins in four cultivars of peanut (Arachis hypogaea L.) under water stress

  • Padmavathi A. V. Thangella
  • Srinivas N. B. S. Pasumarti
  • Raghu Pullakhandam
  • Bhanuprakash Reddy Geereddy
  • Manohar Rao Daggu
Original Article


Drought is a major constraint to the productivity of many crops affecting various physiological and biochemical processes. Seventy percent of the peanuts are grown in semiarid tropics that are frequently prone to drought stress. So, we analyzed its effect in 4 cultivars of peanut, with different degrees of drought tolerance, under 10 and 20 days of water stress using two-dimensional gel electrophoresis and mass spectrometry. A total of 189 differentially expressed protein spots were identified in the leaf proteome of all the 4 cultivars using PD Quest Basic software; 74 in ICGV 91114, 41 in ICGS 76, 44 in J 11 and 30 in JL 24. Of these, 30 protein spots were subjected to in-gel trypsin digestion followed by MALDI-TOF that are functionally categorized into 5 groups: molecular chaperones, signal transducers, photosynthetic proteins, defense proteins and detoxification proteins. Of these, 12 proteins were sequenced. Late embryogenesis abundant protein, calcium ion binding protein, sucrose synthase isoform-1, 17.3 kDa heat shock protein and structural maintenance of chromosome proteins were overexpressed only in the 15 and 20 days stressed plants of ICGV 91114 cultivar while cytosolic ascorbate peroxidase was expressed with varying levels in the 10 and 20 days stressed plants of all the 4 cultivars. Signaling protein like 14-3-3 and defense proteins like alpha-methyl-mannoside-specific lectin and mannose/glucose-binding lectins were differentially expressed in the 4 cultivars. Photosynthetic protein like Rubisco was down-regulated in the stressed plants of all 4 cultivars while Photosystem-I reaction center subunit-II of chloroplast precursor protein was overexpressed in only 20 days stressed plants of ICGV 91114, ICGS 76 and J11 cultivars. These differentially expressed proteins could potentially be used as protein markers for screening the peanut germplasm and further crop improvement.


Peanut Water stress 2-DE PMF MALDI-TOF Differential expression 



Late embryogenesis abundant protein-1


Calcium ion binding protein


Sucrose synthase isoform-1


Structural maintenance of chromosome-1


Ascorbate peroxidase-1


Photosystem-I reaction center subunit-II of chloroplast precursor protein

10 DS

10 days water-stressed plants

20 DS

20 Days water-stressed plants


Matrix-assisted laser desorption/ionization-time-of-flight



This work was supported by grants from Council of Scientific and Industrial Research (CSIR), New Delhi, India in the form of Fellowship and also University Grants Commission by providing financial assistance through the Major Research Project.

Author contribution

PT, MRD and GBR conceived of this study and designed the experiments. PT isolated proteins from samples, done all proteomics experiments and analyzed the data and PNBS supported experiments. PT and MRD drafted the manuscript and GBR critically evaluated and proof-read the manuscript. RP supported manuscript corrections.

Complaince with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

13205_2018_1180_MOESM1_ESM.doc (168 kb)
Supplementary material 1 (DOC 167 kb)


  1. Asada K (2006) Production and Scavenging of reactive oxygen species in chloroplasts and their functions. Plant Physiol 141:391–396CrossRefGoogle Scholar
  2. Basha SM, Roberts RM (1981) The glycoproteins of plant seeds: analysis by two dimensional polyacrylamide gel electrophoresis and by their lectin-binding properties. Plant Physiol 67:936–939CrossRefGoogle Scholar
  3. Basha SM, Katam R, Naik KSS (2007) Differential response of peanut genotypes to water stress. Peanut Sci 34:96–104CrossRefGoogle Scholar
  4. Bhushan D, Pandey A, Choudhary MK, Datta A, Chakraborty S, Chakraborty N (2007) Comparative proteomics analysis of differentially expressed proteins in chickpea extracellular matrix during dehydration stress. Mol Cell Proteom 6:1868–1884CrossRefGoogle Scholar
  5. Boston RS, Viitanen PV, Vierling E (1996) Molecular chaperones and protein folding in plants. Plant Mol Biol 32:191–222CrossRefGoogle Scholar
  6. 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 72:248–254CrossRefGoogle Scholar
  7. Bray EA, Bailey-Serres J, Weretilnyk E (2000) Responses to abiotic stress. Biochemistry & molecular biology of plants. In: Gruissem W, Jones R (eds) American society of plant physiologists. Rockville, pp 1158–1203Google Scholar
  8. Britt AB (1999) Molecular genetics of DNA repair in higher plants. Trends Plant Sci 4:20–25CrossRefGoogle Scholar
  9. Castillejo MA, Maldonado AM, Ogueta S, Jorrin JV (2008) Proteomic analysis of responses to drought stress in sunflower (Helianthus annuus) leaves by 2DE gel electrophoresis and mass spectrometry. Open Proteom J 1:59–71CrossRefGoogle Scholar
  10. Chaves MM, Maroco JP, Pereira JS (2003) Understanding plant responses to drought from genes to the whole plant. Funct Plant Biol 30:239–264CrossRefGoogle Scholar
  11. Close TJ (1996) Dehydrins: emergence of a biochemical role of a family of plant dehydration proteins. Physiol Plantarum 4:795–803CrossRefGoogle Scholar
  12. Cooke MS, Evans MD, Dizdaroglu M, Lunec J (2003) Oxidative DNA damage: mechanisms, mutation, and disease. FASEB J 17:1195–1214CrossRefGoogle Scholar
  13. Davletova S, Rizhsky L, Liang H, Shenggiang Z, Oliver DJ, Coutu J, Shulaev V, Schlauch K, Mittler R (2005) Cytolosic ascorbate peroxidase 1 is a central component of the reactive oxygen gene network of Arabidopsis. Plant Cell 17:268–281CrossRefGoogle Scholar
  14. De Vetten NC, Ferl RJ (1994) Two genes encoding GF14 (14-3-3) proteins in Zea mays. Plant Physiol 106:1593–1604CrossRefGoogle Scholar
  15. Dejardin A, Sokolov LN, Kleczkowski LA (1999) Sugar/osmoticum levels modulate differential abscisic acid-independent expression of two stress-responsive sucrose synthase genes in Arabidopsis. Biochem J 344:503–509Google Scholar
  16. Devaiah KM, Bali Geetha, Athmaram TN, Basha MS (2007) Identification of two new genes from drought tolerant peanut up-regulated in response to drought. Plant Growth Regul 52(3):249–258CrossRefGoogle Scholar
  17. Drame KN, Clavel D, Repellin A, Passaquet C, Zuily-Fodil Y (2007) Water deficit induces variation in expression of stress responsive genes in two peanut (Arachis hypogaea L) cultivars with different tolerance to drought. Plant Physiol Biochem 45:236–243CrossRefGoogle Scholar
  18. Emanuelsson O, Soren B, Gunnar VH, Henrik N (2007) Locating proteins in the cell using Target P, Signal P, and related tools. Nat Protoc 2:953–971CrossRefGoogle Scholar
  19. Eriksson J, Chait BT, Fenyo D (2000) A statistical basis for testing the significance of mass spectrometric protein identification results. Anal Chem 72:999–1005CrossRefGoogle Scholar
  20. Faghani E, Gharechahi J, Komatsu S, Mirzaei M, Ramzan et al (2015) Comparative physiology and proteomic analysis of two wheat genotypes contrasting in drought tolerance. J Proteom 114:1–15CrossRefGoogle Scholar
  21. Farooq M, Basra SMA, Wahid A, Cheema ZA, Cheema MA, Khaliq A (2009) Physiological role of exogenously applied glycinebetaine in improving drought tolerance of fine grain aromatic rice (Oryza sativa L). J Agron Crop Sci 194:325–333CrossRefGoogle Scholar
  22. Feller U, Anders I, Mae T (2008) Rubiscolytics: fate of Rubisco after its enzymatic function in a cell is terminated. J Exp Bot 59:1615–1624CrossRefGoogle Scholar
  23. Ferl RJ (1996) 14-3-3 proteins and signal transduction. Annu Rev Plant Physiol Plant Mol Biol 47:49–73CrossRefGoogle Scholar
  24. Govind G, Harshavardhan VT, Patricia JK, Dhanalakshmi R, Senthil KM, Sreenivasulu N, Udayakumar M (2009) Identification and functional validation of a unique set of drought induced genes preferentially expressed in response to gradual water stress in peanut. Mol Genet Genom 281:591–605CrossRefGoogle Scholar
  25. Graan T, Boyer JS (1990) Very high CO2 partially restores photosynthesis in sunflower at low water potentials. Planta 181:378–384CrossRefGoogle Scholar
  26. Granier F (1988) Extraction of plant proteins for two-dimensional electrophoresis. Electrophoresis 9:712–718CrossRefGoogle Scholar
  27. Grimplet J, Wheatley MD, Jouira HB, Deluc LG, Cramer GR, Cushman JC (2009) Proteomic and selected metabolite analysis of grape berry tissues under well watered and water-deficit stress conditions. Proteomics 9(9):2503–2528CrossRefGoogle Scholar
  28. Guo BZ, Xu G, Cao YG, Holbrook CC, Lynch RE (2006) Identification and characterization of phospholipase D and its association with drought susceptibilities in peanut (Arachis hypogaea). Planta 223:512–520CrossRefGoogle Scholar
  29. Ishida T, Kurata T, Okada K, Wada T (2008) A genetic regulatory network in the development of trichomes and root hairs. Annu Rev Plant Biol 59:365–386CrossRefGoogle Scholar
  30. Jain AK, Basha SM, Holbrook CC (2001) Identification of drought responsive transcripts in peanut (Arachis hypogaea L.). Electron J Biotechnol 4(2):59–67CrossRefGoogle Scholar
  31. Jaleel CA, Gopi R, Sankar B, Gomathinayagam M, Panneerselvam R (2008) Differential responses in water use efficiency in two varieties of Catharanthus roseus under drought stress. Comp Rend Biol 331:42–47CrossRefGoogle Scholar
  32. Jensen RG, Bahr JT (1977) Ribulose 1,5-bisphosphate carboxylase-oxygenase. Ann Rev Plant Physiol 28:379–400CrossRefGoogle Scholar
  33. Katam R, Basha SM, Hemanth KN, Vasanthaiah, Naik KSS (2007) Identification of drought tolerant groundnut genotypes employing proteomics approach. J SAT Agric Res 5(1):1–4Google Scholar
  34. Katam R, Basha M, Suravajhala P, Pechan T (2010) Analysis of peanut leaf proteome. J Proteome Res 9:2236–2254CrossRefGoogle Scholar
  35. Katam R, Sakata K, Prashanth S, Pechan Tibor, Devaiah M et al (2016) Comparative leaf proteomics of drought-tolerant and -susceptible peanut in response to water stress. J Proteom 143:209–226CrossRefGoogle Scholar
  36. Ke Y, Han G, He H, Li J (2009) Differential regulation of proteins and phosphoproteins in rice under drought stress. Biochem Biophys Res Commun 379(1):133–138CrossRefGoogle Scholar
  37. Komatsu S, Hossain Z (2013) Organ-specific proteome analysis for identification of abiotic stress response mechanism in crop. Front Plant Sci 4:1–9Google Scholar
  38. Kottapalli KR, Rakwal R, Shibato J, Burow G, Tissue D, Burke J, Puppala N, Burow M, Payton P (2009) Physiology and proteomics of the water-deficit stress response in three contrasting peanut genotypes. Plant Cell Environ 32:380–407CrossRefGoogle Scholar
  39. Kunert KJ, Vorster BJ, Fenta BA, Kibido T, Dionisio G, Foyer CH (2016) Drought stress responses in soybean roots and nodules. Front Plant Sci 7:1015. CrossRefGoogle Scholar
  40. Lannoo N, Van Damme EJM (2014) Lectin domains at the frontiers of plant defense. Front Plant Sci 5:397Google Scholar
  41. Lauer MJ, Boyer JS (1992) Internal CO2 measures directly in leaves: abscisic acid and low leaf water potential cause opposing effects. Plant Physiol 98:1010–1016CrossRefGoogle Scholar
  42. Lehmann AR (2005) The role of SMC proteins in the responses to DNA damage. DNA Rep 4:309–314CrossRefGoogle Scholar
  43. Lis H, Sharon N (1998) Lectins: carbohydrate-specific proteins that mediate cellular recognition. Chem Rev 98:637–674CrossRefGoogle Scholar
  44. Luo M, Liang XQ, Dang P, Holbrook CC, Bausher MG, Lee RD, Guo BZ (2005) Microarray-based screening of differentially expressed genes in peanut in response to Aspergillus parasiticus infection and drought stress. Plant Sci 169:695–703CrossRefGoogle Scholar
  45. Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498CrossRefGoogle Scholar
  46. Morris JS, Clark BN, Gutstein HB (2007) Pinnacle: a fast, automatic and accurate method for detecting and quantifying protein spots in 2-dimensional gel electrophoresis data. Bioinformatics 24:529–536CrossRefGoogle Scholar
  47. Nasmyth K, Haering CH (2005) The structure and function of SMC and kleisin complexes. Annu Rev Biochem 74:595–648CrossRefGoogle Scholar
  48. Nautiyal PC, Nageswara Rao RC, Joshi YC (2002) Moisture-deficit-induced changes in leaf-water content, leaf carbon exchange rate and biomass production in groundnut cultivars differing in specific leaf area. Field Crops Res 74:67–79CrossRefGoogle Scholar
  49. Padmavathi TAV, Rao DM (2013) Differential accumulation of osmolytes in 4 cultivars of Peanut (Arachis hypogaea L.) under drought stress. J Crop Sci Biotechnol 16(2):151–159CrossRefGoogle Scholar
  50. Park CJ, Seo YS (2015) Heat shock proteins: a review of the molecular chaperones for plant immunity. Plant Pathol J 31(4):323–333. CrossRefGoogle Scholar
  51. Parry MAJ, Andralojc PJ, Khan V, Lea PJ, Keys AJ (2002) RuBisCO activity: effects of drought stress. Ann Bot 89:833–839CrossRefGoogle Scholar
  52. Payton P, Kottapalli KR, Rowland D, Faircloth W, Guo B, Burrow M, Puppala N, Gallo M (2009) Gene expression profiling in peanut using high density oligonucleotide microarrays. BMC Genom 10:265CrossRefGoogle Scholar
  53. Perkins DN, Pappin DJC, Creasy DM (1999) Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 20:3551–3567CrossRefGoogle Scholar
  54. Pruthvi V, Rama N, Govind G, Nataraja KN (2013) Expression analysis of drought stress specific genes in Peanut (Arachis hypogaea L). Physiol Mol Biol Plants 19(2):277–281CrossRefGoogle Scholar
  55. Ramanjulu S, Bartels D (2002) Drought and desiccation-induced modulation of gene expression in plants. Plant Cell Environ 25:141–151CrossRefGoogle Scholar
  56. Rampino P, Pataleo S, Gerardi C, Mita G, Perrotta C (2006) Drought stress response in wheat: physiological and molecular analysis of resistant and sensitive genotypes. Plant Cell Environ 29:2143–2152CrossRefGoogle Scholar
  57. Ritchie SW, Nguyan HT, Holaday AS (1990) Leaf water content and gas exchange parameters of two wheat genotypes differing in drought resistance. Crop Sci 30:105–111CrossRefGoogle Scholar
  58. Salekdeh GH, Siopongco J, Wade LJ, Ghareyazie B, Bennett J (2002) Proteomic analysis of rice leaves during drought stress and recovery. Proteomics 2:1131–1145CrossRefGoogle Scholar
  59. Sanchez-Rodriguez E, Rubio-Wilhelmi M, Cervilla LM, Blasco B, Rios JJ, Rosales MA, Romer L, Ruiz JM (2010) Genotypic differences in some physiological parameters symptomatic for oxidative stress under moderate drought in tomato plants. Plant Sci 178:30–40CrossRefGoogle Scholar
  60. Schellmann S, Hulskamp M (2005) Epidermal differentiation: trichomes in Arabidopsis as a model system. Int J Dev Biol 49:579–584CrossRefGoogle Scholar
  61. Sebkova V, Unger C, Hardegger M, Sturm A (1995) Biochemical, physiological, and molecular characterization of sucrose synthase from Daucus carota. Plant Physiol 108:75–83CrossRefGoogle Scholar
  62. Sharp RE, Hsiao TC, Silk WK (1990) Growth of the maize primary root at low water potentials II role of growth and deposition of hexose and potassium in osmotic adjustment. Plant Physiol 93:1337–1346CrossRefGoogle Scholar
  63. Strum A, Lienhard S, Schatte S, Hardegger M (1999) Tissue-specific expression of two genes for sucrose synthase in carrot (Daucus carota L). Plant Mol Biol 39:349–360CrossRefGoogle Scholar
  64. Timperio AM, Eqidi MG, Zolla L (2008) Proteomics applied on plant abiotic stresses: role of heat shock proteins (HSP). J Proteom 71:391CrossRefGoogle Scholar
  65. Tunnacliffe A, Wise MJ (2007) The continuing conundrum of the LEA proteins. Naturwissenschaften 94:791–812CrossRefGoogle Scholar
  66. Valentovic P, Luxova M, Kolarovic L, Gasparikova O (2006) Effect of osmotic stress on compatible solutes content, membrane stability and water relations in two maize cultivars. Plant Soil Environ 52:186–191CrossRefGoogle Scholar
  67. Van Breusegem F, Dat J (2006) Reactive oxygen species in plant cell death. Plant Physiol 141:384–390CrossRefGoogle Scholar
  68. Van Damme EJM, Barre A, Rouge P, Peumans WJ (2004) Cytoplasmic/nuclear plant lectins: a new story. Trends Plant Sci 9:484–489CrossRefGoogle Scholar
  69. Vierling E (1991) The roles of heat shock proteins in plants. Annu Rev Plant Physiol Plant Mol Biol 42:579–620CrossRefGoogle Scholar
  70. Vijayan M, Chandra N (1999) Lectins. Curr Opin Struct Biol 9:707–714CrossRefGoogle Scholar
  71. Wang W, Vinocur B, Shoseyov O, Altman A (2004) Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. Trends Plant Sci 9:244–252CrossRefGoogle Scholar
  72. Winter H, Huber SC (2000) Regulation of sucrose metabolism in higher plants: localization and regulation of activity of key enzymes. Crit Rev Biochem Mol Biol 35:253–289CrossRefGoogle Scholar
  73. Wise MJ, Tunnacliffe A (2004) POPP the question: what do LEA proteins do? Trends Plant Sci 9:13–17CrossRefGoogle Scholar
  74. Xiong L, Zhu JK (2002) Molecular and genetic aspects of plant responses to osmotic stress. Plant Cell Environ 25:131–140CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Padmavathi A. V. Thangella
    • 1
    • 3
  • Srinivas N. B. S. Pasumarti
    • 2
  • Raghu Pullakhandam
    • 2
  • Bhanuprakash Reddy Geereddy
    • 2
  • Manohar Rao Daggu
    • 1
  1. 1.Department of GeneticsOsmania UniversityHyderabadIndia
  2. 2.National Institute of NutritionHyderabadIndia
  3. 3.Department of Microbiology and Plant BiologyUniversity of OklahomaNormanUSA

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