Abstract
In this paper, two transgenic wheat lines, PC27 and PC51, containing the maize PEPC gene and its wild-type (WT) were used as experimental material to study the effects of high temperature on their photosynthetic physiological characteristics and metabolome. The results showed that transgenic wheat lines had higher photosynthetic rate (P n) than WT under non-stress treatment (NT) and high temperature stress treatment (HT), and more significantly under HT. The change trends of F v/F m, Ф PSII, and q P were similar to P n, whereas that of non-photochemical quenching (NPQ) was the opposite. Compared with WT, no differences in chlorophyll content between the transgenic wheat and WT were observed under NT, but two transgenic lines had relatively higher contents than WT under HT. The change trends of Chlorophyll a/b radio, the decreased values of F m, Wk, and Vj, and the activity of the antioxidant enzyme were consistent with the chlorophyll content. Compared with WT, transgenic wheat lines exhibited lower rate of superoxide anion production, H2O2 and malondialdehyde content under HT, and no significant differences were observed under NT. The expression pattern of the ZmPEPC gene and wheat endogenous photosynthesis-related genes were in agreement with that of P n. Compared with WT, about 13 different metabolites including one organic acid, six amino acids, four sugars, and two polyols were identified under NT; 25 different metabolites including six organic acids, 12 amino acids, four sugars, and three polyols were identified under HT. Collectively, our results indicate that ZmPEPC gene can enhance photochemical and antioxidant enzyme activity, upregulate the expression of photosynthesis-related genes, delay degradation of chlorophyll, change contents of proline and other metabolites in wheat, and ultimately improves its heat tolerance.
Similar content being viewed by others
References
Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 4(1):1–1
Ashraf M, Harris PJC (2013) Photosynthesis under stressful environments: an overview. Photosynthetica 51(2):163–190
Bandyopadhyay A, Datta K, et al. (2007) Enhanced photosynthesis rate in genetically engineered indica rice expressing pepc gene cloned from maize. Plant Sci 72(6):1204–1209
Black CC Jr (1973) Photosynthetic carbon fixation in relation to net CO2 uptake. Annual Review Plant Physiol 24(1):253–286
Boyer JS (1982) Plant productivity and environment. Science 218(4571):443–448
Brennan T, Frenkel C (1977) Involvement of hydrogen peroxide in the regulation of senescence in pear. Plant Physiol 59(3):411–416
Chen HX, Li WJ, et al. (2004a) Dissipation of excess energy in Mehler-peroxidase reaction in Rumex leaves during salt shock. Photosynthetica 42:117–122
Du H, Wang Z, et al. (2011) Differential metabolic responses of perennial grass Cynodon transvaalensis × Cynodon dactylon (C4) and Poa pratensis (C3) to heat stress. Physiol Plantarum 141(3):251–264
Edgerton MD (2009) Increasing crop productivity to meet global needs for feed, food, and fuel. Plant Physiol 149(1):7–13
Elstner EF, Heupel A (1976) Inhibition of nitrite formation from hydroxylammoni-umchloride: a simple assay for superoxide dismutase. Anal Biochem 70:616–620
Fahnenstich H, Saigo M, et al. (2007) Alteration of organic acid metabolism in Arabidopsis overexpressing the maize C4 NADP-malic enzyme causes accelerated senescence during extended darkness. Plant Physiol 145(3):640–652
Fiehn O (2002) Metabolomics-the link between genotypes and phenotypes. Plant Mol Bio 48(1–2):155–171
Foyer CH, Maud L, et al. (1994) Photooxidative stress in plants. Physiol Plantarum 92(4):696–717
Genty B, Briantais J, et al. (1989) The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochim Biophys Acta 990:87–92
Guy C, Kaplan F, et al. (2008) Metabolomics of temperature stress. Physiol Plantarum 132(2):220–235
Haldimann P, Strasser RJ (1999) Effects of anaerobiosis as probed by the polyphasic chlorophylla fluorescence rise kinetic in pea (Pisum sativum L.). Photosynth Res 62:67–83
Han LL, Xu WG, et al. (2013) Preliminary study on the physiological characteristics of transgenic wheat with maize C4-pepc gene in field conditions. Cereal Res Commun 37:1–11
Häusler RE, Hirsch HJ, et al. (2002) Overexpression of C4-cycle enzymes in transgenic C3 plants: a biotechnological approach to improve C3-photosynthesis. J Exp Bot 53(369):591–607
Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Archives Biochem Biophy 125(1):189–198
Hu L, Li Y, et al. (2012) Improvement of the photosynthetic characteristics of transgenic wheat plants by transformation with the maize C4 phosphoenolpyruvate carboxylase gene. Plant Breed 131:231–349
Hudspeth RL, Grula JW, et al. (1992) Expression of maize phosphoenolpyruvate carboxylase in transgenic tobacco effects on biochemistry and physiology. Plant Physiol 98(2):458–464
Ishimaru K, Ichikawa H, et al. (1997) Analysis of a C4 maize pyruvate, orthophosphate dikinase expressed in C3 transgenic Arabidopsis plants. Plant Sci 129(1):57–64
Ishimaru K, Ohkawa Y, et al. (1998) Elevated pyruvate, orthophosphate dikinase (PPDK) activity alters carbon metabolism in C3 transgenic potatoes with a C4 maize PPDK gene. Physiol Plantarum 103(3):340–346
Kogami H, Shono M, et al. (1994) Molecular and physiological evaluation of transgenic tobacco plants expressing a maize phosphoenolpyruvate carboxylase gene under the control of the cauliflower mosaic virus 35S promoter. Transgenic Res 3(5):287–296
Ku MSB, Agarie S, et al. (1999) High-level expression of maize phosphoenolpyruvate carboxylase in transgenic rice plants. Nat Biotechnol 17(1):76–80
Ku MSB, Cho D, et al. (2000) Photosynthetic performance of transgenic rice plants overexpressing maize C4 photosynthesis enzymes. Studies Plant Sci 7:193–204
Lian L, Wang X, et al. (2014) Physiological and photosynthetic characteristics of indica Hang2 expressing the sugarcane PEPC gene. Mol Biol Rep 41(4):2189–2197
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression date using real-time quantitative PCR and 2−ΔΔCt method. Methods 25:402–408
Long SP, Marshall-Colon A, et al. (2015) Meeting the global food demand of the future by engineering crop photosynthesis and yield potential. Cell 161(1):56–66
Lüttge U (1990) Nocturnal citrate accumulation and its response to environmental stress in the CAM plant Kalanchoe pinnata (Lam.) Pers. Plant Cell Environ 13(9):977–982
Makino A (2011) Photosynthesis, grain yield, and nitrogen utilization in rice and wheat. Plant Physiol 155(1):125–129
Mansour MMF (1998) Protection of plasma membrane of onion epidermal cells by glycinebetaine and proline against NaCl stress. Plant Physiol Bioch 36(10):767–772
Maxwell K, Johnson GN (2000) Chlorophyll fluorescence-a practical guide. J Exp Bot 51:659–668
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8(19):4321–4326
Nayyar H, Gupta D (2006) Differential sensitivity of C3 and C4 plants to water deficit stress: association with oxidative stress and antioxidants. Environ Exp Bot 58(1):106–113
Pospíšil P (2012) Molecular mechanisms of production and scavenging of reactive oxygen species by photosystem II. BBA-Bioenergetics 1817(1):218–231
Qian B, Li X, et al. (2015) Enhanced drought tolerance in transgenic rice over-expressing of maize C4 phosphoenolpyruvate carboxylase gene via NO and Ca2+. J Plant Physiol 175:9–20
Qin N, Xu WG, et al. (2015) Drought tolerance and proteomics studies of transgenic wheat containing the maize C4 phosphoenolpyruvate carboxylase (PEPC) gene. Protoplasma. doi:10.1007/s00709-015-0906-2
Qiu Y, Su M, et al. (2007) Application of ethyl chloroformate derivatization for gas chromatography-mass spectrometry based metabonomic profiling. Anal Chim Acta 583:277–283
Ray DK, Mueller ND, et al. (2013) Yield trends are insufficient to double global crop production by 2050. PLoS One 8(6):e66428
Richards RA (2000) Selectable traits to increase crop photosynthesis and yield of grain crops. J Exp Bot 51(suppl 1):447–458
Sayre KD, Rajaram S, et al. (1997) Yield potential progress in short bread wheats in northwest Mexico. Crop Sci 37:36–42
Shearman VJ, Sylvester-Bradley R, et al. (2005) Physiological processes associated with wheat yield progress in the UK. Crop Sci 45:175–185
Špundová M, Popelková H, et al. (2003) Ultra-structural and functional changes in the chloroplasts of detached barley leaves senescing under dark and light conditions. J Plant Physiol 60(9):1051–1058
Strasser BJ (1997) Donor side capacity of photosystem II probed by chlorophyll a fluorescence transients. Photosynth Res 52:147–155
Strasser RJ, Srivastava A, et al. (2000) The fluorescence transient as a tool to characterize and screen photosynthetic samples [M]. Probing photosynthesis: mechanisms, regulation and adaptation:445–483
Takeuchi K, Akagi H, et al. (2000) Aberrant chloroplasts in transgenic rice plants expressing a high level of maize NADP-dependent malic enzyme. Planta 211:265–274
Tan W, Liu J, et al. (2008) Alterations in photosynthesis and antioxidant enzyme activity in winter wheat subjected to post-anthesis water-logging. Photosynthetica 46:21–27
Wang X, Cai J, et al. (2011) Pre-anthesis high-temperature acclimation alleviates damage to the flag leaf caused by post-anthesis heat stress in wheat. J Plant Physiol 168(6):585–593
Wang YM, Xu WG, et al. (2012) Expression of maize gene encoding C4-pyruvate orthophosphate dikinase (PPDK) and C4-phosphoenolpyruvate carboxylase (PEPC) in transgenic Arabidopsis. Plant Mol Bio Rep 30(6):1367–1374
Wienkoop S, Morgenthal K, et al. (2008) Integration of metabolomic and proteomic phenotypes analysis of data covariance dissects starch and RFO metabolism from low and high temperature compensation response in Arabidopsis thaliana. Mol Cell Proteomic 7(9):1725–1736
Wu Q, Xu WG, et al. (2011) Physiological characteristics of photosynthesis in transgenic wheat with maize-pepc gene under field conditions. Acta Agron Sin 37:2046–2052
Yusuf MA, Kumar D, et al. (2010) Overexpression of γ-tocopherolmethyl transferase gene in transgenic Brassica juncea plants alleviates abiotic stress: physiological and chlorophyll a fluorescence measurements. Biochim Biophys Acta 1797:142–1438
Zhang HF, Xu WG, et al. (2014) Pyramiding expression of maize genes encoding phosphoenolpyruvate carboxylase (PEPC) and pyruvate orthophosphate dikinase (PPDK) synergistically improve the photosynthetic characteristics of transgenic wheat. Protoplasma 251(5):1163–1173
Zhang H, Mao X, et al. (2010) Overexpression of a common wheat gene TaSnRK2.8 enhances tolerance to drought, salt and low temperature in Arabidopsis. PLoS One 5(12):e16041
Zhang X, Wollenweber B, et al. (2008) Water deficits and heat shock effects on photosynthesis of a transgenic Arabidopsis thaliana constitutively expressing ABP9, a bZIP transcription factor. J Exp Bot 59(4):839–848
Acknowledgments
The National Natural Science Foundation of China (no. 31371707), Genetically Modified Organisms Breeding Major Projects of China (no. 2011ZX08002-003), China Agriculture Research System (no.CARS-3-1-9) and Fundamental Research and Advanced Technology Project of Henan Province (no.162300410163) supported this work.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
The authors declare that the submitted manuscript does not contain previously published material and is not under consideration for publication elsewhere. All authors listed have read the complete manuscript and have approved the submission of the paper. The manuscript is a truthful original work without fabrication, fraud, or plagiarism.
Conflict of interest
They authors declare that they have no conflict of interest.
Additional information
Handling Editor: Bhumi Nath Tripathi
Electronic supplementary material
Supplemental Table 1
(DOC 29 kb)
Rights and permissions
About this article
Cite this article
Qi, X., Xu, W., Zhang, J. et al. Physiological characteristics and metabolomics of transgenic wheat containing the maize C4 phosphoenolpyruvate carboxylase (PEPC) gene under high temperature stress. Protoplasma 254, 1017–1030 (2017). https://doi.org/10.1007/s00709-016-1010-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00709-016-1010-y