Molecular Biology Reports

, Volume 39, Issue 10, pp 9563–9572 | Cite as

Halopriming mediated salt and iso-osmotic PEG stress tolerance and, gene expression profiling in sugarcane (Saccharum officinarum L.)

  • Vikas Yadav Patade
  • Sujata Bhargava
  • Penna Suprasanna


Seed priming is a well known pre-germination strategy that improves seed performance. However, biochemical and molecular mechanisms underlying priming mediated stress tolerance are little understood. Here, we report results of the study on growth, physiological characteristics and expression of stress responsive genes in salt primed sugarcane cv. Co 86032 plants in response to salt (NaCl, 150 mM) or iso-osmotic (−0.7 MPa) polyethylene glycol-PEG 8000 (20 % w/v) stress exposure for 15 days. Variable growth, osmolyte accumulation and antioxidant capacity was revealed among the primed and non-primed plants. The primed plants showed better tolerance to the salt or PEG stress, as revealed by better growth and lower membrane damage, through better antioxidant capacity as compared to the respective non-primed controls. Further, steady state transcript expression analysis revealed up regulation of sodium proton antiporter (NHX) while, down regulation of sucrose transporter (SUT1), delta 1 -pyrolline-5-carboxylate synthetase (P5CS) and proline dehydrogenase (PDH) in primed plants on exposure to the stress as compared to the non-primed plants. Transcript abundance of catalase (CAT2) decreased by about 25 % in leaves of non-primed stressed plants, however, the expression was maintained in leaves of the stressed primed plants to that of non-stressed controls. Thus, the results indicated priming mediated salt and PEG stress tolerance through altered gene expression leading to improved antioxidant capacity in sugarcane.


Salt priming Iso-osmotic stress Osmotic adjustment Antioxidant defence Gene expression 



Thanks are extended to University Grant Commission, New Delhi, India and Department of Botany, University of Pune, Pune, India for financial support in terms of Research Fellowship to senior author.

Supplementary material

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  1. 1.
    Iqbal M, Ashraf M (2007) Seed treatment with auxins modulates growth and ion partitioning in salt-stressed wheat plants. J Integr Plant Biol 49:1003–1015CrossRefGoogle Scholar
  2. 2.
    Patade VY, Kumari M, Ahmed Z (2011) Chemical seed priming as a simple technique to impart cold and salt stress tolerance in capsicum. J Crop Improv 25:497–503CrossRefGoogle Scholar
  3. 3.
    Patade VY, Kumari M, Ahmed Z (2011) seed priming mediated germination improvement and tolerance to subsequent exposure to cold and salt stress in capsicum. Res J Seed Sci 4:125–136CrossRefGoogle Scholar
  4. 4.
    Casenave EC, Toselli ME (2007) Hydropriming as a pre-treatment for cotton germination under thermal and water stress conditions. Seed Sci Technol 35:88–98Google Scholar
  5. 5.
    Patade VY, Bhargava S, Suprasanna P (2009) Halopriming imparts tolerance in sensitive sugarcane cultivar to salt and PEG induced drought stress. Agric Ecosyst Environ 134:24–28CrossRefGoogle Scholar
  6. 6.
    Patade VY, Kumari M, Ahmed Z (2012) Chemical seed priming as an efficient approach for developing cold tolerance in jatropha. J Crop Improv 26:140–149Google Scholar
  7. 7.
    Beckers GJM, Conrath U (2007) Priming for stress resistance: from the lab to the field. Curr Opin Plant Biol 10:425–431PubMedCrossRefGoogle Scholar
  8. 8.
    Rozbeh F, Farzad S (2006) The effects of NaCl priming on salt tolerance in canola Brassica napus L. seedlings grown under saline conditions. Indian J Crop Sci 11:74–78Google Scholar
  9. 9.
    Afzal I, Basra SMA, Hameed A, Farooq M (2006) Physiological enhancements for alleviation of salt stress in wheat. Pak J Bot 385:1649–1659Google Scholar
  10. 10.
    Conrath U, Beckers GJM, Flors V, Garcia-Agustin P, Jakab G, Mauch F, Newman MA, Pieterse CMJ, Poinssot B, Pozo MJ, Pugin A, Schaffrath U, Ton J, Wendehenne D, Zimmerli L, Mauch-Mani B (2006) Priming: getting ready for battle. Mol Plant-Microbe Interact 19:1062–1071PubMedCrossRefGoogle Scholar
  11. 11.
    Bruce TJA, Matthes MC, Napier JA, Pickett JA (2007) Stressful ‘‘memories’’ of plants: evidence and possible mechanisms. Plant Sci 173:603–608CrossRefGoogle Scholar
  12. 12.
    Chinnusamy V, Schumaker K, Zhu JK (2004) Molecular genetic perspectives on cross-talk and specificity in abiotic stress signaling in plants. J Exp Bot 55:225–236PubMedCrossRefGoogle Scholar
  13. 13.
    Patade VY, Bhargava S, Suprasanna P (2012) Transcript expression profiling of stress responsive genes in response to short-term salt or PEG stress in sugarcane leaves. Mol Biol Rep 39:3311–3318PubMedCrossRefGoogle Scholar
  14. 14.
    Patade VY, Bhargava S, Suprasanna P (2011e) Salt and drought tolerance of sugarcane under iso-osmotic salt and water stress: growth, osmolytes accumulation and antioxidant defense. J Plant Inter (in press)Google Scholar
  15. 15.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Ann Biochem 72:248–254CrossRefGoogle Scholar
  16. 16.
    Patade VY, Rai AN, Suprasanna P (2011). Expression analysis of sugarcane shaggy-like kinase (SuSK) gene identified through cDNA subtractive hybridization in sugarcane (Saccharum officinarum L.). Protoplasma 248:613–621Google Scholar
  17. 17.
    Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402PubMedCrossRefGoogle Scholar
  18. 18.
    Ghiyasi M, Seyahjani AA, Tajbakhsh M, Amirnia R, Salehzadeh H (2008) Effect of osmopriming with polyethylene glycol 8000 on germination and seedling growth of wheat Triticum aestivum L. seeds under salt stress. Res J Biol Sci 310:1249–1251Google Scholar
  19. 19.
    Ghassemi-Golezani K, Aliloo AA, Valizadeh M, Moghaddam M (2008a) Effects of hydro and osmo-priming on seed germination and field emergence of lentil Lens culinaris Medik. Not Bot Hortic Agrobot Cluj 36:29–33Google Scholar
  20. 20.
    Foti R, Aburenia K, Tigerea A, Gotosab J, Gerec J (2008) The efficacy of different seed priming osmotica on the establishment of maize Zea mays L. caryopses. J Arid Environ 72:1127–1130CrossRefGoogle Scholar
  21. 21.
    Ghassemi-Golezani K, Esmaeilpour B (2008) The effect of salt priming on the performance of differentially matured cucumber Cucumis sativus seeds. Not Bot Hortic Agrobot Cluj 36:67–70Google Scholar
  22. 22.
    Sivritepe N, Sivritepe HO, Eris A (2003) The effects of NaCl priming on salt tolerance in melon seedlings grown under saline conditions. Sci Hortic 97:229–237CrossRefGoogle Scholar
  23. 23.
    Neto ADA, Prisco JT, Enéas-Filho J, Medeiros JVR, Gomes-Filho E (2005) Hydrogen peroxide pre-treatment induces salt stress acclimation in maize plants. J Plant Physiol 162:1114–1122CrossRefGoogle Scholar
  24. 24.
    Parra M, Albacete A, Martínez-Andújar C, Pérez-Alfocea F (2007) Increasing plant vigour and tomato fruit yield under salinity by inducing plant adaptation at the earliest seedling stage. Environ Exp Bot 60:77–85CrossRefGoogle Scholar
  25. 25.
    Patanèa C, Valeria C, Salvatore LC (2009) Germination and radicle growth in unprimed and primed seeds of sweet sorghum as affected by reduced water potential in NaCl at different temperatures. Ind Crops Prod 30:1–8CrossRefGoogle Scholar
  26. 26.
    Wahid A, Noreen A, Basra SMA, Gelani S, Farooq M (2008) Priming-induced metabolic changes in sunflower Helianthus annuus achenes improve germination and seedling growth. Bot Stud 49:343–350Google Scholar
  27. 27.
    Shao HB, Liang ZS, Shao MA, Wang BC (2005) Changes of anti-oxidative enzymes and membrane peroxidation for soil water deficits among 10 wheat genotypes at seedling stage. Colloids Surf B 42:107–113CrossRefGoogle Scholar
  28. 28.
    Chen W, Provart NJ, Glazebrook J, Katagiri F, Chang HS, Eulgem T, Mauch F, Luan S, Zou G, Whitham SA, Budworth PR, Tao Y, Xie Z, Chen X, Lam S, Kreps JA, Harper JF, Si-Ammour A, Mauch-Mani B, Heinlein M, Kobayashi K, Hohn T, Dangl JL, Wang X, Zhu T (2002) Expression profile matrix of Arabidopsis transcription factor genes suggests their putative functions in response to environmental stresses. Plant Cell 14:559–574PubMedCrossRefGoogle Scholar
  29. 29.
    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, Ayashizaki 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
  30. 30.
    Knight H, Knight MR (2001) Abiotic stress signaling pathways, specificity and crosstalk. Trends Plant Sci 6:262–267PubMedCrossRefGoogle Scholar
  31. 31.
    Casu RE, Grof CPL, Rae AL, McIntyre CL, Dimmock CM, Manners JM (2003) Identification of a novel sugar transporter homologue strongly expressed in maturing stem vascular tissues of sugarcane by expressed sequence tag and microarray analysis. Plant Mol Biol 52:371–386PubMedCrossRefGoogle Scholar
  32. 32.
    Raven JA (1985) Regulation of pH and generation of osmolarity in vascular plants: a cost benefit analysis in relation to efficiency of use of energy, nitrogen and water. New Phytol 101:25–77CrossRefGoogle Scholar
  33. 33.
    Parks GE, Dietrich MA, Schumaker KS (2002) Increased vacuolar Na+/H+ exchange activity in Salicornia bigelovii Torr. in response to NaCl. J Exp Bot 53:1055–1065PubMedCrossRefGoogle Scholar
  34. 34.
    Weber H, Borisjuk L, Heim U, Sauer N, Wobus U (1997) A role for sucrose transporters during seed development: molecular characterization of a hexose and a sucrose carrier in fava bean seeds. Plant Cell 9:895–908PubMedCrossRefGoogle Scholar
  35. 35.
    Xing Y, Jia W, Zhang J (2007) AtMEK1 mediates stress-induced gene expression of CAT1 catalase by triggering H2O2 production in Arabidopsis. J Exp Bot 58:2969–2981PubMedCrossRefGoogle Scholar
  36. 36.
    Gaxiola RA, Li J, Unurraga S, Dang LM, Allen GJ, Alper SL, Fink GR (2001) Drought- and salt-tolerant plants result from over expression of the AVP1 H+-pump. Proc Natl Acad Sci USA 98:11444–11449PubMedCrossRefGoogle Scholar
  37. 37.
    Silva-Ortega CO, Ochoa-Alfaro AE, Reyes-Agüero JA, Aguado-Santacruz GA, Jiménez-Bremont JF (2008) Salt stress increases the expression of P5CS gene and induces proline accumulation in cactus pear. Plant Physiol Biochem 461:82–92CrossRefGoogle Scholar
  38. 38.
    Nakashima K, Satoh R, Kiyosue T, Yamaguchi-Shinozaki K, Shinozaki K (1998) A gene encoding proline dehydrogenase is not only induced by proline and hypoosmolarity, but is also developmentally regulated in the reproductive organs of Arabidopsis. Plant Physiol 118:1233–1241PubMedCrossRefGoogle Scholar
  39. 39.
    Hong Z, Lakkineni K, Zhang Z, Verma DPS (2000) Removal of feedback inhibition of delta-pyrroline-5-carboxylate synthetase results in increased proline accumulation and protection of plants from osmotic stress. Plant Physiol 122:1129–1136PubMedCrossRefGoogle Scholar
  40. 40.
    Molinari HBC, Marur CJ, Daros E, de Campos MKP, de Carvalho JFRP, Filho JCB, Pereira LFP, Vieira LGE (2008) Evaluation of the stress-inducible production of proline in transgenic sugarcane Saccharum spp.: osmotic adjustment, chlorophyll fluorescence and oxidative stress. Physiol Plant 130:218–229CrossRefGoogle Scholar
  41. 41.
    Frugoli JA, Zhong HH, Nuccio ML, McCourt P, McPeek MA, Thomas TL, McClung CR (1996) Catalase is encoded by a multigene family in Arabidopsis thaliana L. Heynh. Plant Physiol 112:327–336PubMedCrossRefGoogle Scholar
  42. 42.
    Yokoi S, Quintero FJ, Cubero B, Ruiz MT, Bressan RA, Hasegawa PM, Pardo JM (2002) Differential expression and function of Arabidopsis thaliana NHX Na+/H+ antiporters in the salt stress response. Plant J 305:529–539CrossRefGoogle Scholar
  43. 43.
    Polidoros AN, Mylona PV, Scandalios JG (2001) Transgenic tobacco plants expressing the maize CAT2 gene have altered catalase levels that affect plant–pathogen interactions and resistance to oxidative stress. Transgenic Res 10:555–569PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Vikas Yadav Patade
    • 1
    • 2
    • 3
  • Sujata Bhargava
    • 2
  • Penna Suprasanna
    • 1
  1. 1.Functional Plant Biology Section, Nuclear Agriculture and Biotechnology DivisionBhabha Atomic Research Centre, TrombayMumbaiIndia
  2. 2.Botany DepartmentUniversity of PunePuneIndia
  3. 3.Molecular Biology and Genetic Engineering DivisionDefence Institute of Bio-Energy ResearchHaldwani, NainitalIndia

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