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Biochar supplementation altered the expression of antioxidant proteins in rice leaf chloroplasts under high-temperature stress
Applied Biological Chemistry volume 67, Article number: 57 (2024)
Abstract
In order to identify the key antioxidant defense systems used to cope with high-temperature stress in rice leaf chloroplasts following biochar supplementation, the present study compared the expression levels of chloroplast proteins related to antioxidant defense in high-temperature stressed rice leaves between without (C0) and with biochar supplementation (C40; 40 g biochar kg–1 soil). A total of sixteen differentially expressed antioxidant chloroplastic proteins were identified. Among them, three antioxidant enzyme proteins and eight thioredoxin proteins were 62–123% and 37–225% higher under the C40 treatment compared to C0, respectively. These results suggest that both antioxidant enzymes and the thioredoxin system are central to the biochar-mediated protection of rice leaves exposed to high-temperature stress.
Introduction
Rice is one of the most important cereals in the world and serves as a staple food for more than half of the world’s population [2]. Global rice demand is projected to increase by 30% by 2050 [19]. Meanwhile, increasing temperature due to climate change poses a threat to rice production. Global temperature is projected to increase by 2 °C by 2050 [1], while global rice yield is projected to be reduced by 3% with a global increase in temperature of 1 °C [22]. Therefore, it is necessary to develop proactive strategies to alleviate the negative effects of global warming on rice production.
Biochar is a carbon (C)-rich solid product formed by the pyrolysis of organic matter in the absence or limited supply of oxygen [8]. The supplementation of biochar to croplands has received wide attention because it offers an option to sequester C in soils and consequently mitigate global warming [9, 18], while also affecting soil properties and crop performance [4, 7, 12]. Our recent study suggests that biochar supplementation can mitigate the negative effects of high-temperature stress on the growth of rice plants [5], but the mechanism by which biochar improves tolerance to high temperature remains little known. Thus, there is a need to elucidate the fundamental understanding of the physiological mechanisms governing this mitigative effect of biochar supplementation.
High-temperature stress induces excessive reactive oxygen species (ROS) production in plant cells and consequently damages cellular and molecular components (e.g., cellular proteins, lipids, and DNA), leading to cell injury or even death [16]. ROS is largely produced in the chloroplast upon high-temperature stress and is mitigated by antioxidant pathways [3, 13, 20]. Antioxidant enzymes such as superoxide dismutase, peroxidase, and glutathione reductase are the most important components for defense against oxidative stress [15]. The thioredoxin system, comprised of thioredoxin, nicotinamide adenine dinucleotide phosphate, and thioredoxin reductase, provides electrons to thiol-dependent antioxidant enzymes (e.g., peroxidase and peroxiredoxin) to remove ROS at high reaction rates and hence is crucial for alleviating oxidative stress-induced cell damage [11]. In the present study, we determined the effect of biochar supplementation on the expression levels of antioxidant proteins (antioxidant enzyme and thioredoxin system proteins) in the chloroplasts of rice leaves under high-temperature stress. Our objective was to identify the key antioxidant defense systems used to cope with high-temperature stress in rice leaf chloroplasts following biochar supplementation.
Materials and methods
The data were obtained from an experiment conducted in 2020. Briefly, a high-yielding hybrid rice variety, Longliangyouhuazhan, was grown in pots (length × width × height = 20 cm × 15 cm × 25 cm) without (C0) and with biochar supplementation (C40, 40 g biochar kg–1 soil). Six consecutive days (6–11 days after transplanting) of daily mean temperatures (35–36 °C) beyond the critical high temperature for rice tillering (33 °C; [6]) were monitored during the experimental period. The experimental details can be found in [5].
The uppermost fully-expanded leaves were sampled from three pots (replicates) for each treatment. The samples were placed in plastic bags and kept cool on ice during transport to the laboratory, after which they were hulled and stored at − 80 °C until proteins were extracted. Total proteins were extracted using the cold acetone method, and then measured via protein DC assay (Bio-Rad) and calculated according to a bovine serum albumin protein standard curve. A proteomic analysis was carried out using iTRAQ (isobaric tags for relative and absolute quantitation) coupled LC-MS/MS (liquid chromatography-mass spectrometry/mass spectrometry). The details of protein extraction and proteomic analysis are provided in Supplementary File S1.
The functions of the identified proteins were defined according to the Gene Ontology (GO) database (http://www.geneontology.org). The differentially expressed proteins related to antioxidant defense (antioxidant enzyme and thioredoxin system proteins) in chloroplasts were selected for further evaluation. The differentially expressed proteins were defined as those that were significantly different (p < 0.05) in the expression level between C40 and C0. The details of bioinformatics analysis are provided in Supplementary File S1.
Results and discussion
A total of sixteen differentially expressed antioxidant proteins in chloroplasts were identified, including five antioxidant enzyme proteins and eleven thioredoxin system proteins (Table 1). These differentially expressed antioxidant proteins had molecular weights of 18.4–60.3 kDa. The sequence coverage of these differentially expressed antioxidant proteins varied between 10.6 and 45.8% and the number of peptides was 2–19.
Among the five antioxidant enzyme proteins, expression levels of two glutathione reductase proteins (A2XAX5 and A2XCU8) were 25–72% lower under C40 conditions compared to C0, whereas those of another three proteins of peroxidase (B8AJE7), superoxide dismutase (B8B8G3), and peroxiredoxin (P0C5D4) were 62–123% higher in C40 leaves compared to C0 (Fig. 1A; Table 1). This result demonstrates that peroxidase, superoxide dismutase, and peroxiredoxin are important components of the chloroplastic antioxidant defense system against high-temperature stress in rice leaves upon biochar supplementation.
Expression levels of antioxidant enzyme proteins (A) and thioredoxin system proteins (B) in chloroplasts of rice leaves exposed to high temperature without (C0) and with biochar supplementation (C40, 40 g biochar kg–1 soil). Columns and bars represent the means and standard deviations for three replicates, respectively. Protein descriptions are provided in Table 1
Among the eleven thioredoxin system proteins, eight thioredoxins (A2WY84, A2XVT3, A2XYX7, A2YL83, A2YUQ6, B8AEW4, B8AG42, and B8BNI2) were 37–225% higher in the expression level under C40 than under C0, while only two thioredoxins (B8ALJ3 and B8B186) and one thioredoxin reductase (B8B1A9) were 23–29% lower under C40 conditions compared to C0 conditions (Fig. 1B; Table 1). These results highlight the prominence of thioredoxin proteins in high temperature-responsive antioxidant activity in the chloroplasts of rice leaves supplemented with biochar.
It has been well documented that nitric oxide (NO) can boost the antioxidant activity in plants under abiotic stress conditions [17]. The synthesis of NO in plants includes two pathways: (1) the enzymatic pathway (nitrate reductase, nitric oxide-like synthase, nitrite-NO reductase, and xanthine oxidase); and (2) the non-enzymatic pathway (nitrification and de-nitrification) [14]. As both these two pathways of NO synthesis are related to nitrogen (N) supply, several studies have shown that N application can increase leaf antioxidant enzyme activities and reduce yield losses in rice under high temperature conditions [10, 21]. In this study, we observed that biochar supplementation increased shoot N uptake and up-regulated leaf N assimilation and transport proteins in rice (see [5] for details), indicating that biochar supplementation may alter the synthesis of NO and in turn regulate the antioxidant defense system in the chloroplasts of rice leaves exposed to high temperature (Fig. 2). However, further investigations are required to confirm this possibility.
The findings of this study suggest that the combined expression of antioxidant enzymes (peroxidase, superoxide dismutase, and peroxiredoxin) and the thioredoxin system are central to the biochar-mediated protection of rice leaves exposed to high-temperature stress. This study provides insight into the physiological mechanisms by which biochar supplementation mitigates the negative effect of high-temperature stress on rice plants.
Data availability
The data used and/or analysed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- C:
-
Carbon
- C0:
-
The treatment without biochar supplementation
- C40:
-
The treatment with supplementation of 40 g biochar kg–1 soil
- N:
-
Nitrogen
- NO:
-
Nitric oxide
- ROS:
-
Reactive oxygen species
References
Diffenbaugh NS, Barnes EA (2023) Data-driven predictions of the time remaining until critical global warming thresholds are reached. Proc Natl Acad Sci USA 120:e2207183120
Fukagawa NK, Ziska LH (2019) Rice: importance for global nutrition. J Nutr Sci Vitaminol 65:S2–S3
Hu S, Ding Y, Zhu C (2020) Sensitivity and responses of chloroplasts to heat stress in plants. Front Plant Sci 11:375
Huang M, Yang L, Qin H, Jiang L, Zou Y (2013) Quantifying the effect of biochar amendment on soil quality and crop productivity in Chinese rice paddies. Field Crops Res 154:172–177
Huang M, Yin X, Chen J, Cao F (2021) Biochar application mitigates the effect of heat stress on rice (Oryza sativa L.) by regulating the root-zone environment. Front Plant Sci 12:711725
Krishnan P, Ramakrishnan B, Reddy KJ, Reddy VR (2011) High-temperature effects on rice growth, yield, and grain quality. Adv Agron 111:87–206
Kuppusamy S, Thavamani P, Megharaj M, Venkateswarlu K, Naidu R (2016) Agronomic and remedial benefits and risks of applying biochar to soil: current knowledge and future research directions. Environ Int 87:1–12
Lehmann J, Joseph S (2009) Biochar for environmental management: an introduction. In: Lehmann J, Joseph S (eds) Biochar for environmental management, science and technology. Earthscan, London, pp 1–12
Lehmann J, Cowie A, Masiello CA, Kammann C, Woolf D, Amonette JE, Cayuela ML, Camps-Arbestain M, Whitman T (2021) Biochar in climate change mitigation. Nat Geosci 14:883–892
Liu K, Deng J, Lu J, Wang X, Lu B, Tian X, Zhang Y (2019) High nitrogen levels alleviate yield loss of super hybrid rice caused by high temperatures during the flowering stage. Front Plant Sci 10:357
Lu J, Holmgren A (2014) The thioredoxin antioxidant system. Free Radical Bio Med 66:75–87
Martínez-Gómez Á, Poveda J, Escobar C (2022) Overview of the use of biochar from main cereals to stimulate plant growth. Front Plant Sci 13:912264
Moustakas M, Sperdouli I, Adamakis IS (2023) Reactive oxygen species in chloroplasts and chloroplast antioxidants under abiotic stress. Front Plant Sci 14:1208247
Niu L, Liao W (2016) Hydrogen peroxide signaling in plant development and abiotic responses: crosstalk with nitric oxide and calcium. Front Plant Sci 7:230
Rajput VD, Harish, Singh RK, Verma KK, Sharma L, Quiroz-Figueroa FR, Meena M, Gour VS, Minkina T, Sushkova S, Mandzhieva S (2021) Recent developments in enzymatic antioxidant defense mechanism in plants with special reference to abiotic stress. Biology 10:267
Sachdev S, Ansari SA, Ansari MI, Fujita M, Hasanuzzaman M (2021) Abiotic stress and reactive oxygen species: Generation, signaling, and defense mechanisms. Antioxidants 10:277
Simontacchi M, Galatro A, Ramos-Artuso F, Santa-Maria GE (2015) Plant survival in a changing environment: the role of nitric oxide in plant responses to abiotic stress. Front Plant Sci 6:977
Woolf D, Amonette JE, Street-Perrott FA, Lehmann J, Joseph S (2010) Sustainable biochar to mitigate global climate change. Nat Commun 1:56
Yuan S, Stuart AM, Laborte AG, Rattalino Edreira JI, Dobermann A, Kien LVN, Thúy LT, Paothong K, Traesang P, Tint KM, San SS, Villafuerte MQ, Quicho ED, Pame ARP, Then R, Flor RJ, Thon N, Agus F, Agustiani N, Deng N, Li T, Grassini P (2022) Southeast Asia must narrow down the yield gap to continue to be a major rice bowl. Nat Food 3:217–226
Zahara N, Hafeez MB, Ghaffar A, Kausar A, Al Zeidi M, Siddique KHM, Farooq M (2023) Plant photosynthesis under heat stress: effects and management. Environ Exp Bot 206:105178
Zhang N, Ren Y, Deng HZ, Dai L, Wang XY, Mo ZW (2023) Regulating effect of nitrogen on grain yield and antioxidant defense of super and non-super fragrant rice under high soil temperature. Appl Ecol Environ Res 21:2521–2538
Zhao C, Liu B, Piao S, Wang X, Lobell DB, Huang Y, Huang M, Yao Y, Bassu S, Ciais P, Durand JL, Elliott J, Ewert F, Janssens IA, Li T, Lin E, Liu Q, Martre P, Müller C, Peng S, Peñuelas J, Ruane AC, Wallach D, Wang T, Wu D, Liu Z, Zhu Y, Zhu Z, Asseng S (2017) Temperature increase reduces global yields of major crops in four independent estimates. Proc Natl Acad Sci USA 114:9326–9331
Acknowledgements
The authors thank other members of the Rice and Product Physiology for their help with this study.
Funding
This study was supported by the Natural Science Foundation of Hunan Province of China (2019JJ50241), the Scientific Research Fund of Hunan Provincial Education Department of China (18C0158), the National Natural Science Foundation of China (31460332), and the Earmarked Fund for China Agriculture Research System (CARS-01).
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MH conceived the study, analysed the data, and wrote the paper; XY, JC and FC performed the experiment.
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Huang, M., Yin, X., Chen, J. et al. Biochar supplementation altered the expression of antioxidant proteins in rice leaf chloroplasts under high-temperature stress. Appl Biol Chem 67, 57 (2024). https://doi.org/10.1186/s13765-024-00911-9
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DOI: https://doi.org/10.1186/s13765-024-00911-9