Abstract
The reproductive stage of wheat is highly sensitive to the heat stress, especially pollen viability. Here, we report the alterations in the expression of heat shock proteins and antioxidant enzymes in wheat pollen of thermotolerant (C-306) and susceptible (HD2329) cultivars under the heat shock (42 °C, 2 h) with or without ascorbic acid (400 mM) treatment. A significant fold change in expression of catalase (CAT) and ascorbate peroxidase (APX) was observed in the pollens of HD2329 in response to 400 mM ascorbic acid (AsA) before heat stress (T1). The SOD transcript was observed significantly high in C-306 pollens compared to HD2329 in response to T1. The transcripts of high and low molecular weight HSPs (HSP70, HSP90, HSP17 and HSP26) were very high in pollens of C-306 as compared to HD2329. Semi-quantitative RT-PCR showed marked variations in the transcript of HSPs and antioxidant enzymes in pollens of C-306 and HD2329 under T0 and T1 treatments. 1D and 2D polyacrylamide gel electrophoresis of pollens showed more differentially expressed proteins in C-306 than in HD2329 in response to T1. A significant increase in the accumulation of osmolyte (proline), H2O2 and endogenous AsA levels were observed in the pollens of C-306 than in HD2329, in response to T0 and T1 treatments. Immunoblot analysis using monoclonal antibodies of HSP70 and HSP90 showed abundance of HSP70 proteins in the pollens of C-306 and HSP90 proteins in pollens of HD2329. Scanning electron microscopy of pollen under the heat stress showed disintegrated and dehydrated exine layer and alteration in pollen structure from spheroid to ovoid in both the cultivars. The pollen viability was found to be less in HD2329 than in C-306 in response to T1 treatment. A significant increase in the total antioxidant capacity was observed in pollens of C-306 as compared to HD2329 in response to the treatments. Findings indicate that pre-anthesis treatment of 400 mM AsA before HS (42 °C, 2 h) enhances the thermotolerance capacity of wheat pollens, as assessed through biochemical markers; further investigations on similar elicitors may yield useful information on mitigating the effect of the terminal heat on reproductive system of wheat.
Similar content being viewed by others
Abbreviations
- HSP:
-
Heat shock protein
- HS:
-
Heat stress
- qRT-PCR:
-
Quantitative real time PCR
- SEM:
-
Scanning electron microscopy
- AsA:
-
Ascorbic Acid
- ROS:
-
Reactive Oxygen Species
- 2-DE:
-
Two Dimensional Electrophoresis
- 1-DE:
-
One Dimensional Electrophoresis
- PAGE:
-
Polyacrylamide Gel Electrophoresis
- FRAP:
-
Ferric Reducing Antioxidant Power
References
Ashraf M, Foolad MR (2007) Roles of glycinebetaine and proline in improving plant abiotic stress tolerance. Environ Exp Bot 59:206–216
Barnaba’s B, Jager K, Feher A (2008) The effect of drought and heat stress on reproductive processes in cereals. Plant Cell Environ 31:11–38
Barth C, De Tullio M, Conklin PL (2006) The role of ascorbic acid in the control of flowering time and the onset of senescence. J Exp Bot 57:1657–1665
Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207
Benzie IFF, Strain JJ (1999) Ferric reducing/antioxidant power assay: direct measure of the total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. Meth Enzymol 299:15–27
Bradford M (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
Cao F, Cheng H, Cheng S, Li L, Xu F, Yu W, Yuan H (2012) Expression of selected Ginkgo biloba heat shock protein genes after cold treatment could be induced by other abiotic stress. Int J Mol Sci 13(5):5768–5788
Challinor AJ, Wheeler TR, Slingo JM (2005) Simulation of the impact of high temperature stress on the yield of an annual crop. Agric For Meteorol 135:180–189
Chen Y, Liu P, Hoehenwarter W, Lin J (2012) Proteomic and phosphoproteomic analysis of Picea wilsonii pollen development under nutrient limitation. J Proteome Res 11(8):4180–4190
Dolferus R, Ji X, Richards RA (2011) Abiotic stress and control of grain number in cereals. Plant Sci 181:331–341
Firon N, Pressman E, Meir S, Khoury R, Altahan L (2012) Ethylene is involved in maintaining tomato (Solanum lycopersicum) pollen quality under heat-stress conditions. AoB Plants. pls024 doi: 10.1093/aobpla/pls024
Foyer CH, Lopez-Delgardo H, Dat JF, Scott IM (1997) Hydrogen peroxide and glutathione associated mechanisms of acclimatory stress tolerance and signaling. Physiol Plant 100:241–254
Frank G, Pressman E, Ophir R, Althan L, Shaked R, Freedman M, Shen S, Firon N (2009) Transcriptional profiling of maturing tomato (Solanum lycopersicum L.) microspores reveals the involvement of heat shock proteins, ROS scavengers, hormones, and sugars in the heat stress response. J Exp Bot 60:3891–3908
Gechev TS, Hille J (2005) Hydrogen peroxide as a signal controlling plant programmed cell death. JCB 168(1):17–20
Giorno F, Wolters-Arts M, Grillo S, Scharf KD, Vriezen WH, Mariani C (2010) Developmental and heat stress-regulated expression of HsfA2 and small heat shock proteins in tomato anthers. J Exp Bot 61(2):453–462
Grobei MA, Qeli E, Brunner E, Rehrauer H, Zhang R, Roschitzki B, Basler K, Ahrens CH, Grossniklaus U (2009) Deterministic protein inference for shotgun proteomics data provides new insights into Arabidopsis pollen development and function. Gen Res 19:1786–1800
Harper JF, Zinn KE, Tunc-Ozdemir M (2010) Temperature stress and plant sexual reproduction: uncovering the weakest links. J Exp Bot 61(7):1959–1968
Hossain A, Teixeira da Silva JA (2013) Wheat production in Bangladesh: its future in the light of global warming. AoB Plants 5:pls042. doi:10.1093/aobpla/pls042
Jagadish SVK, Muthurajan R, Oane R, Wheeler TR, Heuer S, Bennett J, Craufurd PQ (2010) Physiological and proteomic approaches to address heat tolerance during anthesis in rice (Oryza sativa L). J Exp Bot 61:143–156
Joshi AK, Mishra B, Chatrath R, Ferrara GO, Singh RP (2007) Wheat improvement in India: present status, emerging challenges and future prospects. Euphytica 157:431–446
Kumar RR, Goswami S, Kumar N, Pandey SK, Pandey VC, Sharma SK, Pathak H, Rai RD (2011) Expression of novel ascorbate peroxidase isoenzymes of wheat (Triticum aestivum L) in response to heat stress. Int J Plant Physiol Biochem 3(11):188–194
Kumar RR, Goswami S, Sharma SK, Singh K, Gadpayle KA, Singh SD, Pathak H, Rai RD (2012) Differential expression of heat shock protein and alteration in osmolyte accumulation under heat stress in wheat. J Plant Biochem Biotechnol. doi:10.1007/s13562-012-0106-5
Kurek I, Chang TK, Bertain SM, Madrigal A, Liu L, Lassner MW, Zhu G (2007) Enhanced thermostability of Arabidopsis Rubisco activase improves photosynthesis and growth rates under moderate heat stress. Plant Cell 19:3230–3241
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage. Nat 227:680–685
Lansac AR, Sullivan CY, Johnson BE (1996) Accumulation of free proline in sorghum (Sorghum bicolor) pollen. Can J Bot 74:40–45
Leja M, Wyzgolik G, Kaminska I (2007) Some parameters of antioxidant capacity of red cabbage as related to different forms of nutritive nitrogen. Folia Hort 19(1):15–23
Lobell DB, Burke MB, Tebaldi C, Mastrandrea MD, Falcon WP, Naylor RL (2008) Supporting online materials for: prioritizing climate change adaptation needs for food security in 2030. Sci 319:607–610
Loreto F, Velikova V (2001) Isoprene produced by leaves protects the photosynthetic apparatus against ozone damage, quenches ozone products and reduces lipid peroxidation of cellular membranes. Plant Physiol 127:1781–1787
Miller G, Suzuki N, Ciftci-Yilmaz S, Mittler R (2010) Reactive oxygen species homeostasis and signaling during drought and salinity stresses. Plant Cell Environ 33:453–467
Moot DJ, Henderson AL, Porter JR, Semenov MA (1996) Temperature, CO2 and the growth and development of wheat: changes in the mean and variability of growing conditions. Clim Change 33:351–368
Muller-Moule P, Golan T, Niyogi KK (2004) Ascorbate-deficient mutants of Arabidopsis grows in high light despite chronic photoxidative stress. Plant Physiol 134:1163–1172
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29(9):45
Pressman E, Peet MM, Pharr DM (2002) The effect of heat stress on tomato pollen characteristics is associated with changes in carbohydrate concentration in the developing anthers. Ann Bot 90:631–636
Qin Y, Leydon AR, Manziello A, Pandey R, Mount D, Denic S, Vasic B, Johnson MA, Palanivelu R (2009) Penetration of the stigma and style elicits a novel transcriptome in pollen tubes, pointing to genes critical for growth in a pistil. PLoS Genetics 5:e1000621
Sadasivam S, Manickam A (1996) Biochemical methods. New age international (P) Limited, Publishers, II (ed), New Delhi, 152–160
Sakata T, Takahashi H, Nishiyama I, Higashitani A (2000) Effects of high temperature on the development of pollen mother cells and microspores in barley Hordeum vulgare L. J Plant Res 113:395–402
Sarkar NK, Kim YK, Grover A (2009) Rice sHsp genes: genomic organization and expression profiling under stress and development. BMC Genom 10:393
Sato S, Peet MM, Thomas JF (2002) Determining critical pre- and post-anthesis periods and physiological processes in Lycopersicon esculentum Mill. exposed to moderately elevated temperatures. J Exp Bot 53:1187–1195
Schoffl F, Prandl R, Reindl A (1998) Regulation of the heat shock response. Plant Physiol 117:1135–1141
Schwacke R, Grallath S, Breitkreuz KE, Stransky E, Stransky H, Frommer WB, Rentsch D (1999) LeProT1, a transporter for proline, glycine betaine, and c-amino butyric acid in tomato pollen. Plant Cell 11:377–392
Sheoran IS, Sproule KA, Olson DJH, Ross ARS, Sawhney VK (2006) Proteome profile and functional classification of proteins in Arabidopsis thaliana (Landsberg erecta) mature pollen. Sex Plant Reprod 19:185–196
Smirnoff N (2000) Ascorbic acid: metabolism and functions of a multifacetted molecule. Curr Opin Plant Biol 3:229–235
Stone P (2001) The effects of heat stress on cereal yield and quality. In: AS Basra (Ed) Crop responses and adaptations to temperature stress, food products press, Binghamton, NY, pp 243–291
Thomas JMG, Prasad PVV (2003) Plants and the environment / global warming effects. University of Florida, Gainesville
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 9:244–252
Wu J, Shang Z, Wu J, Jiang X, Moschou PN, Sun W, Roubelakis-Angelakis KA, Zhang S (2010) Spermidine oxidase-derived H2O2 regulates pollen plasma membrane hyperpolarization-activated Ca2+-permeable channels and pollen tube growth. Plant J 63:1042–1053
Acknowledgment
The author’s sincerely thanks Indian Agricultural Research Institute (IARI) and Indian Council of Agriculture Research (ICAR) for providing the financial assistance under the National Initiative for Climate Resilient Agriculture (NICRA) project.
Author information
Authors and Affiliations
Corresponding author
Additional information
Ranjeet R. Kumar and Suneha Goswami equal contribution made by both the authors
Rights and permissions
About this article
Cite this article
Kumar, R.R., Goswami, S., Gadpayle, K.A. et al. Ascorbic acid at pre-anthesis modulate the thermotolerance level of wheat (Triticum aestivum) pollen under heat stress. J. Plant Biochem. Biotechnol. 23, 293–306 (2014). https://doi.org/10.1007/s13562-013-0214-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13562-013-0214-x