Research articleEffects of ethylenediurea (EDU) on regulatory proteins in two maize (Zea mays L.) varieties under high tropospheric ozone phytotoxicity
Graphical abstract
Introduction
Ground-level ozone (O3) is increasing at the rate of approximately 0.5–2% per year over the mid-latitudes of the Northern Hemisphere due to rapid industrialization and urbanization in the last three decades (IPCC, 2013; Simpson et al., 2014). Global tropospheric O3 levels were around 50 ppb in the year 2000, already 25% above the AOT40 threshold proven for damage to sensitive plants (Bhatia et al., 2012). Due to its phytotoxicity, tropospheric O3 has been recognized as one of the most hazardous and toxic air pollutants with a higher degree of negative impacts on global agriculture (Ashmore, 2005; Emberson et al., 2009; Singh et al., 2015). Various studies conducted on the Indian crops suggest their high vulnerability to ozone-induced damage, but unfortunately genetic variation among cultivars in response to O3 has hardly been addressed (Oksanen et al., 2013; Peng et al., 2020). Global yield reduction's, due to ambient O3, for maize, rice, wheat, and soybean have been estimated to be 6.1%, 4.4%, 7.1%, and 12.4% (mean of 2010–2012) annually, respectively (Mills et al., 2018). Economic losses for Europe based on ozone assessment studies on 23 crops, were estimated to be US$7.5 billion (Holland et al., 2006) and global crop production losses were estimated to have been 79–121 Mt worth US$11–18 billion (Avnery et al., 2011). Estimating the loss of crop production from ground-level O3 is valuable for understanding the potential benefits of reducing O3 concentration and for projecting future food supply (Burney and Ramanathan, 2014).
Among the different effects of ozone on vegetation, visible injury in leaves is considered a valuable tool for the assessment of ozone impacts in the field and the detection of areas of high risk due to O3 (Schaub et al., 2010). Ozone causes damage by entering the leaf intercellular air spaces via stomata, where it reacts with compounds in the exposed wet cell-wall surfaces, causing the production of damaging radicals and signaling that accelerates senescence (Long and Naidu, 2002; Fiscus et al., 2005). Photosynthetic efficiency and mesophyll conductance are also affected by ozone in the crops (Xu et al., 2019; Peng et al., 2020). This has led to the expectation that O3 damage will be less in C4 plants (maize and sugarcane), given their intrinsically lower stomatal conductance, as well as for plants under drought stress, and in response to rising (CO2) (McKee et al., 2000; Long and Naidu, 2002; Leitao et al., 2007; Yi et al., 2020). Different studies indicate that O3 damages the photosynthetic machinery leading to a progressive loss in the amount as well as activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) (Agrawal et al., 2002; Cho et al., 2008). Light and dark reactions of chloroplast also get affected either directly or indirectly due to high ozone concentration (Fiscus et al., 2005).
Ethylenediurea (EDU) has been widely used as a research tool to reveal and evaluate the ozone-sensitivity in several crops and tree species (Paoletti et al., 2009; Feng et al., 2010; Manning et al., 2011; Oksanen et al., 2013). Protective capability of EDU was observed on reactive oxygen species (ROS) mechanism in wheat (Agrawal et al., 2005; Singh and Agrawal, 2009; Pandey et al., 2019), European Ash (Paoletti et al., 2008), mung bean (Singh et al., 2010a), carrot (Tiwari and Agrawal, 2010), maize (Singh et al., 2018) and in palak (Spinach) (Tiwari and Agrawal, 2009).
The present study comprises the evaluation of regulatory proteins together with morpho-physiological, biochemical, and yield in two maize varieties under EDU treatment. This is the first proteomic study under EDU treatment in the C4 crop. As we know, the morpho-physiological approach which reveals the changes in comparison with the given EDU treatments, whereas proteomic and biochemical response analyses the insight of the plant's metabolism under any prevailing conditions. Proteomic evaluation includes, identifying differential expression of proteins in response to EDU treatment in two maize varieties. The other study parameters include pigments estimation, lipid peroxidation (MDA equivalent content), antioxidants (Ascorbate and Glutathione), and the antioxidative enzymes: Ascorbate peroxidase (APX), Glutathione reductase (GR), catalase (CAT) and superoxide dismutase (SOD). Maize is considered as the third most important crop at the global context of which, two varieties SHM3031 (stress sensitive), and PEHM5 (stress tolerant) were selected for the present study.
Section snippets
Study site, climatic condition and plant material
The study was conducted at CSIR-National Botanical Research Institute garden in Lucknow, city of Uttar Pradesh, India. It is situated along the southern bank of river Gomati at 26055′ N latitude, 80059′ E longitude, and an altitude of 113 m in subtropical climates. Lucknow is characterized by a dry, tropical monsoon climate. Maximum average temperatures varied from 25 to 32 °C, and minimum average temperature varied from 14 to 27 °C, and a minimum of 60% and a maximum of 78% humidity was
Average ozone and AOT40
The average ambient ozone during the study period was 53.21 ppb. It had been recorded that precipitation and cloudy weather were responsible for comparatively low ambient ozone during the study period rather than dry seasons. Precipitation leads to washout of the precursor responsible for ozone formation. The ozone levels since October were higher than until September because of higher precipitation rate in later months (Fig. 1). High ambient O3 concentrations (hourly average) (>40 ppb)
Discussion
We demonstrated harmful effects of tropospheric ozone on maize using EDU as anti-ozonant supplement in the middle IGP region, of India. The higher concentration of ozone in troposphere resulted from high temperature, longer sunshine hours, and less relative humidity. In our study, the maximum O3 concentration (71.38 ppb) was found during October and November, and minimum during peak rainfall months (July, August, and September). The lower O3 concentration during rainy season was appeared due to
Conclusion
In Summary, under conditions with high tropospheric ozone as those prevailing in Lucknow area. EDU conferred an important protection to sensitive maize variety, increasing morpho-physiological performance, maintaining an enhanced antioxidant capacity, and finally leading to higher biomass and/or grain yields. The magnitude of variations in biochemical parameters varied with stages and varieties. SHM3031 showed more induction in non-enzymatic antioxidants with respect to PEHM5 at both the
Author's contribution
VP, and SKG designed the experiment. SKG, MS, and VKM did yield and physiological work. BM did the enzymatic work. SKG, MS,VKM, and FD did proteomic work. SKG, and VP analyzed the data and wrote the paper. SKG, ZJL, and VP revised the manuscript. All the authors approved the paper.
Declaration of competing interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgments
Authors are grateful to Director, CSIR-NBRI for providing necessary facilities. SKG and MS are grateful to CSIR and UGC, respectively for senior research fellowship. Funding for this work was provided by Council of Scientific & Industrial Research (CSIR), New Delhi, India [Project no. PSC112].
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