Exogenous H2O2 alleviates maize seed germination and seedling physiology property under drought stress

Exogenous H2O2 alleviates maize seed germination and seedling physiology property under drought stress | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Exogenous H 2 O 2 alleviates maize seed germination and seedling physiology property under drought stress Zhike Wang This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6202193/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Hydrogen peroxide (H 2 O 2 ) is an important signaling molecule in plant body, which is involved in the regulation of multiple abiotic stresses.This study investigated the ability of exogenous H 2 O 2 to alleviate drought stress in maize ( Zea mays L.) seedlings. With maize seeds as the test material, used the filter paper germination method. The seeds were treated with different concentrations of exogenous H 2 O 2 under 10% polyethylene glycol (PEG-6000) stress to measure seed germination, seedling growth and physiological indicators. The results showed that 10% PEG treatment alone obviously inhibited the seed germination and seedling growth; Treatment with 0.02% H 2 O 2 significantly improved the germination potential and vitality index of maize seeds under PEG stress, promoted the growth of seedling buds and roots, enhanced the activity of APX, SOD and CAT in buds and roots, reduced the MDA content, and promoted the increment of proline and soluble sugars. The above results indicated that exogenous 0.02% H 2 O 2 treatment could activate antioxidant enzyme activity and reduce oxidative damage under drought stress, thus alleviating maize drought tolerance at the seed germination stage. drought stress hydrogen peroxide maize seed germination physiological characteristics Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Drought is one of the most common stress factors in agricultural production in the world, which seriously affects the growth and development of crops and leads to crop production decrease (Terzi et al., 2014 ). Drought can hinder the growth and development of plants, leaf wilting, photosynthesis rate decline, serious can cause plant water shortage death, which has become one of the main factors restricting agricultural production (Sun et al., 2016 ). Drought causes not only changes in individual plant size, but also changes in plant biomass allocation (Iqbal et al., 2018 ). Drought can also lead to the large production of free radicals in plants, causing membrane lipid oxidation, membrane damage and reduced cell membrane stability (Bian and Jiang, 2009 ). Using exogenous substances to stimulate the plant produce a stress resistance response, it is one of the effective ways to improve the plant resistance to external stress (Ozaki et al., 2009 ; Liu et al., 2010 ; Ishibashi et al., 2011 ). Hydrogen peroxide (H 2 O 2 ) is an important signaling molecule in plant body, which is involved in the regulation of plant defense system to stress, and enhance plant tolerance to environmental stress (Gao et al., 2010 ). It has been shown that exogenous H 2 O 2 can enhance plant tolerance to abiotic stress such as heat (Gao et al., 2010 ), salt (Li et al., 2011 ; Liu et al., 2020 ), waterlogging (Andrade et al., 2018 ), drought (He et al., 2009 ) and heavy metal stresses (Hu et al., 2009 ). Corn ( zea mays L.) is an important grain, feed and cash crop in Qingyang city, but it often encounters drought during the sowing period, which leads to low seedling emergence rate, slow seedling growth, production reduction and quality reduction. Seed germination is the initial stage of plant development (Ishibashi et al., 2010 ; Leymarie et al., 2012 ), and seed drought tolerance during sowing can more reflect the plant drought resistance (Ishibashi et al., 2015 ). Reducing the damage of drought stress in the emergence process is beneficial to improve the emergence rate and survival rate (Khan et al., 2017 ). There are few studies on H 2 O 2 in drought tolerance in maize. Therefore, We studied the mitigation effect of exogenous H 2 O 2 on maize under drought stress, and provided practical guidance for the use of exogenous substances in agricultural production to improve crop tolerance to drought stress. Materials and methods Plant materials The corn seed “Longdan 10” used in the experiment was provided by the Physiology Laboratory of Plant, School of Agriculture and Bioengineering, Longdong University. Polyethylene glycol (PEG-6000) and 3% hydrogen peroxide were all analytical reagent. Select the PEG treatment concentration After selection, the corn seeds were sterilized with 0.1% KMnO 4 for 10 min and washed three times in distilled water. Place 50 sterilized seeds in a Petri dish covered with filter paper. They were treated with different concentrations of PEG (0 (distilled water), 1%, 2%, 4%, 6%, 8%, 10%, 15%), a total of 8 treatments.The plates were incubated in an incubator with 25℃ temperature and 85% humidity. Count the number of seeds germinated on time every day. Exogenous HO immersion treatment According to the above screening results, 10% PEG was selected as the semi-lethal concentration of maize seeds. After selection of corn seeds, disinfected by the above method. Place 50 sterilized seeds in a petri dish covered with filter paper. Treatment with 0 (distilled water), 0.01%, 0.02%, 0.04%, 0.08%, 0.2% and 0.3% H 2 O 2 for 12 h, and then 10% PEG stress. Distilled water immersion culture, set as blank control (CK); 10% PEG immersion culture, set as conditioned control (PEG); The test groups were respectively as follows: 0.01% H 2 O 2 + 10% PEG (0.01H + PEG); 0.02% H 2 O 2 + 10% PEG (0.02H + PEG); 0.04% H 2 O 2 + 10% PEG (0.04H + PEG); 0.08% H 2 O 2 + 10% PEG (0.08H + PEG); 0.2% H 2 O 2 + 10% PEG (0.2H + PEG); 0.3% H 2 O 2 + 10% PEG (0.3H + PEG). A total of 8 treatments. The following steps are the same as above. Determination of seed germination rate When the radicle length is half the seed length, it is used as a germination standard. Germination test was performed for 7d and germination potential(GP) was calculated on the 3d, germination rate (GR), germination index (GI) and viability index (VI) was calculated on the 7d (Chen et al., 2020 ; Bu et al., 2024 ). Determination of the seedling growth indicators After the germination test of seeds, 15 seedlings were randomly picked from each group to determine the bud length, root length, bud fresh weight and root fresh weight. Determination of soluble sugars, proline, and malondialdehyde content The content of soluble sugars (SS), proline (PRO) and malondialdehyde (MDA) was determined according to the kit method, which was purchased from Beijing Solaibao Technology Co., Ltd. Determination of the antioxidant enzyme activity The activity of peroxidase (POD), hydrogen peroxide (CAT) and superoxide dismutase (SOD) was determined by the kit, which was purchased from Beijing Solaibao Technology Co., Ltd. Statistical analysis Data collation was performed with Microsoft Excel 2016. Data analysis was performed by one-way ANOVA with SPSS 26.0, and multiple comparisons and differential significance analysis (α = 0.05) were performed by the Duncan method (Wang et al., 2020 ). Data in the graphs are the mean ± SD. Results Effect of different concentrations of PEG stress on seeds germination in maize According to (Fig. 1 ) , the germination index of maize seeds all decreased significantly with increasing PEG concentration compared with the control CK. Seed germination was slightly suppressed when PEG was 2%; when PEG was 10%, germination rate, germination potential, germination index, and root length were nearly semi-lethal, decreasing by 50.08%, 51.83%, 51.09%, 50.24%, respectively, and when PEG was 15%, germination was 28.34%, indicating severely suppressed germination. Therefore, 10% PEG was selected as the semi-lethal drought stress concentration to treat maize seeds with drought stress. Effect of exogenous H 2 O 2 treatment on maize seeds germination under PEG stress According to (Fig. 2 ), PEG treatment significantly suppressed the germination index of maize seeds compared with control CK. Under PEG treatment, with exogenous H 2 O 2 of low concentration (0.01%, 0.02%, 0.04%, and 0.08%) submersion treatment significantly improved the germination index of maize seeds, among them 0.02% H 2 O 2 had the best mitigation effect, and the germination rate, germination potential, germination index and vigor index increased by 76.16%, 66.35%, 87.86% and 243.20% compared with the PEG treatment, respectively. However, the effect of 0.2% H 2 O 2 treatment on the germination index was not significant. This showed that low concentration of exogenous H 2 O 2 treatment alleviated germination of maize seeds under drought stress. Effect of exogenous H 2 O 2 treatment on the growth of maize seedlings under PEG stress As shown from (Fig. 3 ) , compared with the control CK, PEG treatment obviously inhibited the growth of maize seedlings, with root length, bud length, root fresh weight and bud fresh weight decreased by 50.24%, 49.02%, 50.71%, and 57.19%, respectively. Under PEG treatment, the growth of maize seedlings was obviously improved with exogenous H 2 O 2 of low concentration (0.01% and 0.02%) submersion treatment, in which 0.02% H 2 O 2 had the best mitigation effect, and root length, bud length, root fresh weight and bud fresh weight increased by 83.41%, 86.76%, 78.08% and 91.04%, respectively, compared with the PEG treatment. However, the effect of 0.2% H 2 O 2 treatment on seedling growth was not significant. Effect of exogenous H 2 O 2 treatment on osmoregulator in maize seedlings under PEG stress As shown from (Fig. 4 A ) , PEG treatment significantly promoted the increment of proline content in the root and bud of maize seedlings, increated by 20.91% and 21.20% compared with the control CK, respectively. Under PEG treatment, with exogenous H 2 O 2 of low concentration (0.01%, 0.02% and 0.04%) submersion treatment significantly improved the increment of proline content in maize seedlings, of which 0.02% H 2 O 2 had the best mitigation effect, proline content 40.57% and 28.61% higher than PEG treatment, respectively. As shown from (Fig. 4 B ) , PEG treatment significantly inhibited the increment in soluble sugar content in the root and shoot of maize seedlings, reduced by 24.38% and 18.15% compared with the control CK, respectively. Under PEG treatment, with exogenous H 2 O 2 of low concentration (0.01%, 0.02%, and 0.04% and 0.08%) submersion treatment significantly improved the increment of soluble sugar content in maize seedlings, of which 0.02% H 2 O 2 had the best mitigation effect, and the soluble sugar content increased by 95.33% and 66.15% compared with PEG treatment, respectively. Effect of exogenous H 2 O 2 treatment on MDA in maize seedlings under PEG stress As shown from (Fig. 5 ) , PEG treatment obviously promoted the increment of MDA content in the roots and shoots of maize seedlings, increased by 29.06% and 24.60% compared with the control CK, respectively. Under PEG treatment, with exogenous H 2 O 2 of low concentration (0.01%, 0.02%, 0.04%, 0.08% and 0.2%) submersion treatment significantly reduced the increment of MDA content, of which 0.02% H 2 O 2 was the best mitigation effect, and MDA content decreased by 32.15% and 26.49% compared with PEG treatment, respectively. Effect of exogenous H 2 O 2 treatment on antioxidant enzyme activity in maize seedlings under PEG stress As shown in (Fig. 6 ), PEG treatment obviously promoted the activity of POD, CAT, and SOD in maize seedlings, increased by 6.87% and 4.37%, 4.76% and 4.42%, 0.78% and 0.89% compared with the control CK, respectively. Under PEG treatment, the activity of POD, CAT, and SOD was significantly improved with exogenous H 2 O 2 of low concentrations (0.01%, 0.02% and 0.04%) submersion treatment, of which 0.02% H 2 O 2 was the best mitigation effect, increasing by 14.58% and 7.94%, 10.07% and 7.32%, 1.63% and 1.37% over the PEG treatment, respectively. Discussion Drought stress is one of the most common abiotic stresses and has a great impact on both the ecological environment and agricultural production (Guler et al., 2016). Soil moisture is a key environmental factor affecting crop seed germination, seedling growth and yield (Wang et al., 2024 ). This experiment showed that the germination rate of maize seeds decreased significantly with the increasing PEG concentration. It has been shown that H 2 O 2 is involved in the plant abiotic stress response (Černý et al., 2018 ), and can effectively alleviate the damage of drought, salinity, cold damage, and heavy metals (Sachdev et al., 2021 ). This experiment showed that under PEG treatment, with exogenous H 2 O 2 of low concentration (0.01%, 0.02%, 0.04%, and 0.08%) submersion treatment significantly improved the germination index of maize seeds, among them 0.02% H 2 O 2 had the best mitigation effect (Fig. 2 ). However, the effect of 0.2% H 2 O 2 treatment on the germination index was not significant (Fig. 2 ). This showed that low concentration of exogenous H 2 O 2 treatment alleviated germination of maize seeds under drought stress. The growth status of plants can show their own response to the external environment, which can be used as an effective indicator to evaluate whether the plant is drought-resistant (Zhang et al., 2018 ). This experiment showed that PEG treatment obviously suppressed the germination index of maize seeds compared with control CK (Fig. 1 ). Under PEG treatment, with exogenous H 2 O 2 of low concentration (0.01%, 0.02%, 0.04%, and 0.08%) submersion treatment significantly improved the germination index of maize seeds, among them 0.02% H 2 O 2 had the best mitigation effect (Fig. 2 ). However, the effect of 0.2% H 2 O 2 treatment on the germination index was not significant (Fig. 2 ). This showed that low concentration of exogenous H 2 O 2 treatment alleviated germination of maize seeds under drought stress. Under drought stress, plants adapt to water-deficient environments through osmotic regulation (Seleiman et al., 2021 ). Osmotic regulation is a physiological response of plants to environmental water stress, which increases cellular osmotic pressure, reduces water loss rate and improves drought tolerance (Blum., 2016). In drought conditions, soluble sugars and proline are the two main substances that regulate cell osmotic potential (Dghim et al., 2018 ). This experiment showed that PEG treatment obviously promoted the increment of proline content in the root and bud of maize seedlings (Fig. 4 A). Under PEG treatment, with exogenous H 2 O 2 of low concentration (0.01%, 0.02% and 0.04%) submersion treatment significantly improved the increment of proline content in maize seedlings, of which 0.02% H 2 O 2 had the best mitigation effect (Fig. 4 A). When plants are subjected to drought, ROS accumulation disrupts the cell membrane integrity and cellular function, leading to membrane lipid peroxidation (Hussain et al., 2018 ). The MDA content can directly reflect the degree of membrane damage and the ability of plants to adapt to drought (Lee et al., 2009 ). This experiment showed that PEG treatment obviously promoted the increment of MDA content in the roots and shoots of maize seedlings (Fig. 5 ). Under PEG treatment, with exogenous H 2 O 2 of low concentration (0.01%, 0.02%, 0.04%, 0.08% and 0.2%) submersion treatment significantly reduced the increment of MDA content, of which 0.02% H 2 O 2 was the best mitigation effect (Fig. 5 ). In the face of drought-induced oxidative damage, the plant body has enzymatic and non-enzymatic antioxidant systems to maintain redox homeostasis, neutralize free radicals or reactive oxygen species, prevent acute cell damage and maintain cell membrane integrity (Dias et al., 2014 ). This experiment showed that PEG treatment obviously promoted the activity of POD, CAT, and SOD in maize seedlings (Fig. 6 ). Under PEG treatment, the activity of POD, CAT, and SOD was significantly improved with exogenous H 2 O 2 of low concentrations (0.01%, 0.02% and 0.04%) submersion treatment, of which 0.02% H 2 O 2 was the best mitigation effect (Fig. 6 ). Conclusions Corn seed germination and seedling growth decreased significantly with increasing PEG concentration. Under drought stress, with exogenous H 2 O 2 of low concentration (0.02%) submersion treatment significantly improved seed germination and seedling growth, increased the proline content, promoted the activity of POD, CAT and SOD in seedling roots and shoots, and reduced the MDA content of seedlings to improve their drought resistance. Declarations Disclosure Statement No potential conflict of interest was reported by the author. Acknowledgements This work was supported by the Natural Science Foundation of Gansu Province (No. 21JR11RM042) and Longdong University Doctoral Fund (XYBY202007). Contributions Zhike Wang conceived and designed the experiments. Zhike Wang performed laboratory experiments. Zhike Wang performed data analysis and interpretation. Zhike Wang wrote the paper. The author read and approved the final manuscript. References Andrade, C. A., de Souza, K. R. D., de Oliveira Santos, M., da Silva, D. M., & Alves, J. D. (2018). Hydrogen peroxide promotes the tolerance of soybeans to waterlogging. Scientia Horticulturae 232, 40–45. https://doi.org/10.1016/j.scienta.2017.12.048 Bian, S. M., & Jiang, Y. W. (2009). Reactive oxygen species, antioxidant enzyme activities and gene expression patterns in leaves and roots of Kentucky bluegrass in response to drought stress and recovery. Scientia Horticulturae 120(2), 264–270. https://doi.org/10.1016/j.scienta.2008.10.014 Blum, A. (2017). Osmotic adjustment is a prime drought stress adaptive engine in support of plant production. Plant, Cell & Environment 40(1), 4–10. https://doi.org/10.1111/pce.12800 Bu, T., Yang, J. X., Liu, J. X., & Fan, X. F. (2024). NaHS immersion alleviates the stress effect of chromium (III) on alfalfa seeds by affecting active oxygen metabolism. Plant Signaling & Behavior 19(1), e2375673. https://doi.org/10.1080/15592324.2024.2375673 Černý, M., Habánová, H., Berka, M., Luklová, M., & Brzobohatý, B. (2018). Hydrogen peroxide: its role in plant biology and crosstalk with signalling networks. International journal of molecular sciences 19(9), 2812. https://doi.org/10.3390/ijms19092812 Chen, L., Liu, L. T., Lu, B., Ma, T. T., Jiang, D., Li, J., Zhang, K., Sun, H. C., Zhang, Y. J., Bai, Z. Y., & Li, C. D. (2020). Exogenous melatonin promotes seed germination and osmotic regulation under salt stress in cotton ( Gossypium hirsutum L.). Plos One 15(1), e0228241. https://doi.org/ 10.1371/journal.pone.0228241 Dias, M. C., Oliveira, H., Costa, A., & Santos, C. (2014). Improving elms performance under drought stress: The pretreatment with abscisic acid. Environmental and Experimental Botany 100, 64–73. https://doi.org/10.1016/j.envexpbot.2013.12.013 Dghim, F., Abdellaoui, R., Boukhris, M., Neffati, M., & Chaieb, M. (2018). Physiological and biochemical changes in Periploca angustifolia plants under withholding irrigation and rewatering conditions. South African Journal of Botany 114, 241–249. https://doi.org/10.1016/j.sajb.2017.11.007 Gao, Y., Guo, Y. K., Lin, S. H., Fang, Y. Y., & Bai, J. G. (2010). Hydrogen peroxide pretreatment alters the activity of antioxidant enzymes and protects chloroplast ultrastructure in heat-stressed cucumber leaves. Scientia Horticulturae 126(1), 20–26. https://doi.org/10.1016/j.scienta.2010.06.006 Guler, N. S., & Pehlivan, N. (2016). Exogenous low-dose hydrogen peroxide enhances drought tolerance of soybean ( Glycine max L.) through inducing antioxidant system. Acta Biologica Hungarica 67, 169–183. https://doi.org/10.1556/018.67.2016.2.5 He, L. H., Gao, Z. Q., & Li, R. Z. (2009). Pretreatment of seed with H 2 O 2 enhances drought tolerance of wheat ( Triticum aestivum L.) seedlings. African Journal of Biotechnology 8(22), 6151–6157. https://doi.org/10.5897/AJB09.490 Hu, Y. L., Ge, Y., Zhang, C. H, Ju, T., & Cheng, W. D. (2009). Cadmium toxicity and translocation in rice seedlings are reduced by hydrogen peroxide pretreatment. Plant Growth Regulation 59, 51–61. https://doi.org/10.1007/s10725-009-9387-7 Hussain, H. A., Hussain, S., Khaliq, A., Ashraf, U., Anjum, S. A., Men, S. M., & Wang, L. C. (2018). Chilling and drought stresses in crop plants: Implications, cross talk, and potential management opportunities. Frontiers in Plant Science 9, 393. https://doi.org/10.3389/fpls.2018.00393 Iqbal, H., Yaning, C., Waqas, M., Rehman, H., Shareef, M., & Iqbal, S. (2018). Hydrogen peroxide application improves quinoa performance by affecting physiological and biochemical mechanisms under water‐deficit conditions. Journal of Agronomy and Crop Science 204(6), 541–553. https://doi.org/10.1111/jac.12284 Ishibashi, Y., Tawaratsumida, T., Zheng, S. H., Yuasa, T., & Iwaya-inoue, M. (2010). NADPH oxidases act as key enzyme on germination and seedling growth in barley ( Hordeum vulgare L.). Plant Production Science 13(1), 45–52. https://doi.org/10.1626/pps.13.45 Ishibashi, Y., Yamaguchi, H., Yuasa, T., Iwaya-Inoue, M., Arima, S., & Zheng, S. H. (2011). Hydrogen peroxide spraying alleviates drought stress in soybean plants. Journal of Plant Physiology 168 (13), 1562–1567. https://doi.org/10.1016/j.jplph.2011.02.003 Ishibashi, Y., Kasa, S., Sakamoto, M., Aoki, N., Kai, K., Yuasa, T., Hanada, A., Yamaguchi, S., & Iwaya-Inoue, M. (2015). A role for reactive oxygen species produced by NADPH oxidases in the embryo and aleurone cells in barley seed germination. Plos One 10(11): e0143173. https://doi.org/10.1371/journal.pone.0143173 Khan, A., Anwar, Y., Hasan, M. M., Iqbal, A., Ali, M., Alharby, H. F., Hakeem, K. R., & Hasanuzzaman, M. (2017). Attenuation of drought stress in brassica seedlings with exogenous application of Ca 2 + and H 2 O 2 . Plants 6(2), 20. https://doi.org/10.3390/plants6020020 Lee, B. R., Li, L. S., Jung, W. J., Jin, Y. L., Avice, J. C., Ourry, A., & Kim, T. H. (2009).Water deficit-induced oxidative stress and the activation of antioxidant enzymes in white clover leaves. Biologia Plantarum 53, 505–510. https://doi.org/10.1007/s10535-009-0091-2 Leymarie, J., Vitkauskaité, G., Hoang, H. H., Gendreau, E., Chazoule, V., Meimoun, P., Corbineau, F., El-Maarouf-Bouteau, H., & Bailly, C. (2012) Role of reactive oxygen species in the regulation of Arabidopsis seed dormancy. Plant and Cell Physiology 53(1): 96–106. https://doi.org/10.1093/pcp/pcr129 Li, J. T., Qiu, Z. B., Zhang, X. W., & Wang, L. S. (2011). Exogenous hydrogen peroxide can enhance tolerance of wheat seedlings to salt stress. Acta Physiologiae Plantarum 33, 835–842. https://doi.org/10.1093/10.1007/s11738-010-0608-5 Liu, Z. J., Guo, Y. K., & Bai, J. G. (2010). Exogenous hydrogen peroxide changes antioxidant enzyme activity and protects ultrastructure in leaves of two cucumber ecotypes under osmotic stress. Journal of Plant Growth Regulation 29, 171–183. https://doi.org/10.1093/10.1007/s00344-009-9121-8 Liu, L. J, Huang, L., Lin, X. Y., & Sun, C. L. (2020). Hydrogen peroxide alleviates salinity-induced damage through enhancing proline accumulation in wheat seedlings. Plant Cell Reports 39, 567–575. https://doi.org/10.1007/s00299-020-02513-3 Ozaki, K., Uchida, A., Takabe, T., Shinagawa, F., Tanaka, Y., Takabe, T., Hayashi, T., Hattori, T., Rai, A. K., & Takabe, T. (2009). Enrichment of sugar content in melon fruits by hydrogen peroxide treatment. Journal of Plant Physiology 166 (6), 569–578. https://doi.org/10.1016/j.jplph.2008.08.007 Sachdev, S., Ansari, S. A., Ansari, M. I., Fujita, M., & Hasanuzzaman, M. (2021). Abiotic stress and reactive oxygen species: Generation, signaling, and defense mechanisms. Antioxidants 10(2), 277. https://doi.org/10.3390/antiox10020277 Seleiman, M. F., Al-Suhaibani, N., Ali, N., Akmal, M., Alotaibi, M., Refay, Y., Dindaroglu, T., Abdul-Wajid., H. H., & Battaglia, M. L. (2021). Drought stress impacts on plants and different approaches to alleviate its adverse effects. Plants 10(2), 259. https://doi.org/10.3390/plants 10020259 Sun, Y., Wang, H., Liu, S., & Peng, X. (2016). Exogenous application of hydrogen peroxide alleviates drought stress in cucumber seedlings. South African Journal of Botany 106, 23–28. https://doi.org/10.1016/j.sajb.2016.05.008 Terzi, R., Kadioglu, A., Kalaycioglu, E., & Saglam, A. (2014). Hydrogen peroxide pretreatment induces osmotic stress tolerance by influencing osmolyte and abscisic acid levels in maize leaves. Journal of Plant Interactions 9(1), 559–565. https://doi.org/10.1080/17429145.2013.871077 Wang, X. F., Zhao, J. Q., Yuan, P. G., Ding, S. Y., Jiang, L. G., & Xi, Z. M. (2024). Hydrogen peroxide functioned as a redox signaling molecule in the putrescine-promoted drought tolerance in cabernet sauvignon. Scientia Horticulturae 335, 113325. https://doi.org/10.1016/j.scienta.2024. 113325 Wang, Z. K., Ma, R., Zhao, M. S., Wang, F. F., Zhang, N., & Si, H. J. (2020). NO and ABA interaction regulates tuber dormancy and sprouting in potato. Frontiers in Plant Science 11, 311. https://doi.org/10.3389/fpls.2020.00311 Zhang, C. M., Shi, S. L., Wang, B. W., & Zhao, J. F. (2018). Physiological and biochemical changes in different drought-tolerant alfalfa ( Medicago sativa L.) varieties under PEG-induced drought stress. Acta Physiologiae Plantarum, 40, 25. https://doi.org/10.1007/s11738-017-2597-0 Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6202193","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":428094564,"identity":"2e41aec2-4927-40b1-8a89-828cbbced7eb","order_by":0,"name":"Zhike Wang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzklEQVRIiWNgGAWjYBACefnHBww+/vvHzM/eQKQWw4a0hMIZbAfYJXsOEGvNgRyDzzxsB/gNbiQQqYOx4Vjixhk8d6QZbj7eeIOhxiaaoBZ2xubDBh8knhkzzk4rtmA4lpbbQNCWZrY0wxkGzMnM0jlmEowNhwlrYTjGY/6bJ4G5vk3yDLFazvAYGPMcOMzMI8FDpBbDGWwJhjMb0pgleIB+SSDGL/ISzMCobLBhtj9+eOONDzU2RDgMCRhIJJCiHKKFVB2jYBSMglEwMgAAEsJAhD+uun8AAAAASUVORK5CYII=","orcid":"","institution":"Longdong University","correspondingAuthor":true,"prefix":"","firstName":"Zhike","middleName":"","lastName":"Wang","suffix":""}],"badges":[],"createdAt":"2025-03-11 10:53:10","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6202193/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6202193/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":78654705,"identity":"2c37e9fe-f1ae-4b41-830c-a400f5ef7b12","added_by":"auto","created_at":"2025-03-17 09:11:43","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":149258,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of different concentrations of PEG stress on seeds germination of maize.\u003cstrong\u003e \u0026nbsp;(A) \u003c/strong\u003eGermination rate; \u003cstrong\u003e(B) \u003c/strong\u003eGermination potential; \u003cstrong\u003e(C) \u003c/strong\u003eGermination index; \u003cstrong\u003e(D) \u003c/strong\u003eRoot length; \u003cstrong\u003e(E) \u003c/strong\u003eVigour index.The data are the mean ± standard deviation; the different lower-case letters on the different oblong indicate a significant difference (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05). The following table is the same.\u003c/p\u003e","description":"","filename":"image1.png","url":"https://assets-eu.researchsquare.com/files/rs-6202193/v1/c5cf88f1eb5121ae42011a92.png"},{"id":78653874,"identity":"23373f0d-3297-4845-bd59-7868fbcfbca0","added_by":"auto","created_at":"2025-03-17 09:03:43","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":126047,"visible":true,"origin":"","legend":"\u003cp\u003eEffects of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on seeds germination of maize\u003cem\u003e \u003c/em\u003eunder PEG stress.\u003cstrong\u003e (A) \u003c/strong\u003eGermination rate; \u003cstrong\u003e(B) \u003c/strong\u003eGermination potential; \u003cstrong\u003e(C) \u003c/strong\u003eGermination index; \u003cstrong\u003e(D) \u003c/strong\u003eVigour index. CK: distilled water, blank control; PEG: 10% PEG, condition control; 0.01H+PEG: 0.01% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e + 10% PEG; 0.02H+PEG: 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e + 10% PEG; 0.04H+PEG: 0.04% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e +10% PEG; 0.08H+PEG: 0.08% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e + 10% PEG; 0.2H+PEG: 0.2% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e + 10% PEG; 0.3H+PEG: 0.3% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e + 10% PEG, the same below.\u003c/p\u003e","description":"","filename":"image2.png","url":"https://assets-eu.researchsquare.com/files/rs-6202193/v1/6d0df586dc7988f7ec2b9d41.png"},{"id":78654914,"identity":"241013bf-a3c2-470d-ab19-49c921ef9aab","added_by":"auto","created_at":"2025-03-17 09:19:43","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":69684,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on the growth of maize seedlings under PEG stress. \u003cstrong\u003e(A) \u003c/strong\u003eLength; \u003cstrong\u003e(B) \u003c/strong\u003eFresh weight.\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-6202193/v1/8e823929ce78af528ee23444.png"},{"id":78653875,"identity":"5d904b95-efc7-4366-9f66-3bee884988d7","added_by":"auto","created_at":"2025-03-17 09:03:43","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":73359,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on osmoregulator in maize seedlings under PEG stress. \u003cstrong\u003e(A)\u003c/strong\u003e Proline content; \u003cstrong\u003e(B) \u003c/strong\u003eSoluble sugar content.\u003c/p\u003e","description":"","filename":"image4.png","url":"https://assets-eu.researchsquare.com/files/rs-6202193/v1/1cb2a39d0b46bef27253ca2d.png"},{"id":78654701,"identity":"4b60c0e7-6c1d-4e7e-8427-9a445973c33d","added_by":"auto","created_at":"2025-03-17 09:11:43","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":38283,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on MDA in maize seedlings under PEG stress.\u003c/p\u003e","description":"","filename":"image5.png","url":"https://assets-eu.researchsquare.com/files/rs-6202193/v1/b6bae7645f0f122c630135d1.png"},{"id":78653878,"identity":"0de14d6e-8edc-4635-9518-610777e6d50e","added_by":"auto","created_at":"2025-03-17 09:03:43","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":108531,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on antioxidant enzyme activity in maize seedlings under PEG stress. \u003cstrong\u003e\u0026nbsp;(A) \u003c/strong\u003ePOD activity; \u003cstrong\u003e(B) \u003c/strong\u003eCAT activity\u003cstrong\u003e \u003c/strong\u003e; \u003cstrong\u003e(C) \u003c/strong\u003eSOD activity.\u003c/p\u003e","description":"","filename":"image6.png","url":"https://assets-eu.researchsquare.com/files/rs-6202193/v1/47c69e116590dcdfcb98a483.png"},{"id":79009611,"identity":"8b66dbfa-34df-44f4-9aa5-7d1cc598fd46","added_by":"auto","created_at":"2025-03-22 09:46:42","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1276424,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6202193/v1/ae962668-2124-49a2-8028-a9ca4cda547c.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eExogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e alleviates maize seed germination and seedling physiology property under drought stress\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eDrought is one of the most common stress factors in agricultural production in the world, which seriously affects the growth and development of crops and leads to crop production decrease (Terzi et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Drought can hinder the growth and development of plants, leaf wilting, photosynthesis rate decline, serious can cause plant water shortage death, which has become one of the main factors restricting agricultural production (Sun et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Drought causes not only changes in individual plant size, but also changes in plant biomass allocation (Iqbal et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Drought can also lead to the large production of free radicals in plants, causing membrane lipid oxidation, membrane damage and reduced cell membrane stability (Bian and Jiang, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eUsing exogenous substances to stimulate the plant produce a stress resistance response, it is one of the effective ways to improve the plant resistance to external stress (Ozaki et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Liu et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Ishibashi et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) is an important signaling molecule in plant body, which is involved in the regulation of plant defense system to stress, and enhance plant tolerance to environmental stress (Gao et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). It has been shown that exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e can enhance plant tolerance to abiotic stress such as heat (Gao et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), salt (Li et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Liu et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), waterlogging (Andrade et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), drought (He et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) and heavy metal stresses (Hu et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eCorn (\u003cem\u003ezea mays\u003c/em\u003e L.) is an important grain, feed and cash crop in Qingyang city, but it often encounters drought during the sowing period, which leads to low seedling emergence rate, slow seedling growth, production reduction and quality reduction. Seed germination is the initial stage of plant development (Ishibashi et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Leymarie et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), and seed drought tolerance during sowing can more reflect the plant drought resistance (Ishibashi et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Reducing the damage of drought stress in the emergence process is beneficial to improve the emergence rate and survival rate (Khan et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). There are few studies on H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e in drought tolerance in maize. Therefore, We studied the mitigation effect of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e on maize under drought stress, and provided practical guidance for the use of exogenous substances in agricultural production to improve crop tolerance to drought stress.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePlant materials\u003c/h2\u003e \u003cp\u003eThe corn seed \u0026ldquo;Longdan 10\u0026rdquo; used in the experiment was provided by the Physiology Laboratory of Plant, School of Agriculture and Bioengineering, Longdong University. Polyethylene glycol (PEG-6000) and 3% hydrogen peroxide were all analytical reagent.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eSelect the PEG treatment concentration\u003c/h3\u003e\n\u003cp\u003eAfter selection, the corn seeds were sterilized with 0.1% KMnO\u003csub\u003e4\u003c/sub\u003e for 10 min and washed three times in distilled water. Place 50 sterilized seeds in a Petri dish covered with filter paper. They were treated with different concentrations of PEG (0 (distilled water), 1%, 2%, 4%, 6%, 8%, 10%, 15%), a total of 8 treatments.The plates were incubated in an incubator with 25℃ temperature and 85% humidity. Count the number of seeds germinated on time every day.\u003c/p\u003e\n\u003ch3\u003eExogenous HO immersion treatment\u003c/h3\u003e\n\u003cp\u003e According to the above screening results, 10% PEG was selected as the semi-lethal concentration of maize seeds. After selection of corn seeds, disinfected by the above method. Place 50 sterilized seeds in a petri dish covered with filter paper. Treatment with 0 (distilled water), 0.01%, 0.02%, 0.04%, 0.08%, 0.2% and 0.3% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e for 12 h, and then 10% PEG stress. Distilled water immersion culture, set as blank control (CK); 10% PEG immersion culture, set as conditioned control (PEG); The test groups were respectively as follows: 0.01% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;+\u0026thinsp;10% PEG (0.01H\u0026thinsp;+\u0026thinsp;PEG); 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;+\u0026thinsp;10% PEG (0.02H\u0026thinsp;+\u0026thinsp;PEG); 0.04% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;+\u0026thinsp;10% PEG (0.04H\u0026thinsp;+\u0026thinsp;PEG); 0.08% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;+\u0026thinsp;10% PEG (0.08H\u0026thinsp;+\u0026thinsp;PEG); 0.2% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;+\u0026thinsp;10% PEG (0.2H\u0026thinsp;+\u0026thinsp;PEG); 0.3% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u0026thinsp;+\u0026thinsp;10% PEG (0.3H\u0026thinsp;+\u0026thinsp;PEG). A total of 8 treatments. The following steps are the same as above.\u003c/p\u003e\n\u003ch3\u003eDetermination of seed germination rate\u003c/h3\u003e\n\u003cp\u003eWhen the radicle length is half the seed length, it is used as a germination standard. Germination test was performed for 7d and germination potential(GP) was calculated on the 3d, germination rate (GR), germination index (GI) and viability index (VI) was calculated on the 7d (Chen et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Bu et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003eDetermination of the seedling growth indicators\u003c/h3\u003e\n\u003cp\u003eAfter the germination test of seeds, 15 seedlings were randomly picked from each group to determine the bud length, root length, bud fresh weight and root fresh weight.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eDetermination of soluble sugars, proline, and malondialdehyde content\u003c/h2\u003e \u003cp\u003eThe content of soluble sugars (SS), proline (PRO) and malondialdehyde (MDA) was determined according to the kit method, which was purchased from Beijing Solaibao Technology Co., Ltd.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eDetermination of the antioxidant enzyme activity\u003c/h3\u003e\n\u003cp\u003eThe activity of peroxidase (POD), hydrogen peroxide (CAT) and superoxide dismutase (SOD) was determined by the kit, which was purchased from Beijing Solaibao Technology Co., Ltd.\u003c/p\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eData collation was performed with Microsoft Excel 2016. Data analysis was performed by one-way ANOVA with SPSS 26.0, and multiple comparisons and differential significance analysis (α\u0026thinsp;=\u0026thinsp;0.05) were performed by the Duncan method (Wang et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Data in the graphs are the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eEffect of different concentrations of PEG stress on seeds germination in maize\u003c/h2\u003e \u003cp\u003eAccording to (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e, the germination index of maize seeds all decreased significantly with increasing PEG concentration compared with the control CK. Seed germination was slightly suppressed when PEG was 2%; when PEG was 10%, germination rate, germination potential, germination index, and root length were nearly semi-lethal, decreasing by 50.08%, 51.83%, 51.09%, 50.24%, respectively, and when PEG was 15%, germination was 28.34%, indicating severely suppressed germination. Therefore, 10% PEG was selected as the semi-lethal drought stress concentration to treat maize seeds with drought stress.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eEffect of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on maize seeds germination under PEG stress\u003c/h2\u003e \u003cp\u003eAccording to (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), PEG treatment significantly suppressed the germination index of maize seeds compared with control CK. Under PEG treatment, with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentration (0.01%, 0.02%, 0.04%, and 0.08%) submersion treatment significantly improved the germination index of maize seeds, among them 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e had the best mitigation effect, and the germination rate, germination potential, germination index and vigor index increased by 76.16%, 66.35%, 87.86% and 243.20% compared with the PEG treatment, respectively. However, the effect of 0.2% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on the germination index was not significant. This showed that low concentration of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment alleviated germination of maize seeds under drought stress.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eEffect of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on the growth of maize seedlings under PEG stress\u003c/h2\u003e \u003cp\u003eAs shown from (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e, compared with the control CK, PEG treatment obviously inhibited the growth of maize seedlings, with root length, bud length, root fresh weight and bud fresh weight decreased by 50.24%, 49.02%, 50.71%, and 57.19%, respectively. Under PEG treatment, the growth of maize seedlings was obviously improved with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentration (0.01% and 0.02%) submersion treatment, in which 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e had the best mitigation effect, and root length, bud length, root fresh weight and bud fresh weight increased by 83.41%, 86.76%, 78.08% and 91.04%, respectively, compared with the PEG treatment. However, the effect of 0.2% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on seedling growth was not significant.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eEffect of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on osmoregulator in maize seedlings under PEG stress\u003c/h2\u003e \u003cp\u003eAs shown from (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA\u003cb\u003e)\u003c/b\u003e, PEG treatment significantly promoted the increment of proline content in the root and bud of maize seedlings, increated by 20.91% and 21.20% compared with the control CK, respectively. Under PEG treatment, with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentration (0.01%, 0.02% and 0.04%) submersion treatment significantly improved the increment of proline content in maize seedlings, of which 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e had the best mitigation effect, proline content 40.57% and 28.61% higher than PEG treatment, respectively.\u003c/p\u003e \u003cp\u003eAs shown from (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB\u003cb\u003e)\u003c/b\u003e, PEG treatment significantly inhibited the increment in soluble sugar content in the root and shoot of maize seedlings, reduced by 24.38% and 18.15% compared with the control CK, respectively. Under PEG treatment, with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentration (0.01%, 0.02%, and 0.04% and 0.08%) submersion treatment significantly improved the increment of soluble sugar content in maize seedlings, of which 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e had the best mitigation effect, and the soluble sugar content increased by 95.33% and 66.15% compared with PEG treatment, respectively.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eEffect of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on MDA in maize seedlings under PEG stress\u003c/h2\u003e \u003cp\u003eAs shown from (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e, PEG treatment obviously promoted the increment of MDA content in the roots and shoots of maize seedlings, increased by 29.06% and 24.60% compared with the control CK, respectively. Under PEG treatment, with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentration (0.01%, 0.02%, 0.04%, 0.08% and 0.2%) submersion treatment significantly reduced the increment of MDA content, of which 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e was the best mitigation effect, and MDA content decreased by 32.15% and 26.49% compared with PEG treatment, respectively.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eEffect of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on antioxidant enzyme activity in maize seedlings under PEG stress\u003c/h2\u003e \u003cp\u003eAs shown in (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e), PEG treatment obviously promoted the activity of POD, CAT, and SOD in maize seedlings, increased by 6.87% and 4.37%, 4.76% and 4.42%, 0.78% and 0.89% compared with the control CK, respectively. Under PEG treatment, the activity of POD, CAT, and SOD was significantly improved with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentrations (0.01%, 0.02% and 0.04%) submersion treatment, of which 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e was the best mitigation effect, increasing by 14.58% and 7.94%, 10.07% and 7.32%, 1.63% and 1.37% over the PEG treatment, respectively.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eDrought stress is one of the most common abiotic stresses and has a great impact on both the ecological environment and agricultural production (Guler et al., 2016). Soil moisture is a key environmental factor affecting crop seed germination, seedling growth and yield (Wang et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). This experiment showed that the germination rate of maize seeds decreased significantly with the increasing PEG concentration. It has been shown that H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e is involved in the plant abiotic stress response (Čern\u0026yacute; et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), and can effectively alleviate the damage of drought, salinity, cold damage, and heavy metals (Sachdev et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). This experiment showed that under PEG treatment, with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentration (0.01%, 0.02%, 0.04%, and 0.08%) submersion treatment significantly improved the germination index of maize seeds, among them 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e had the best mitigation effect (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). However, the effect of 0.2% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on the germination index was not significant (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). This showed that low concentration of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment alleviated germination of maize seeds under drought stress.\u003c/p\u003e \u003cp\u003eThe growth status of plants can show their own response to the external environment, which can be used as an effective indicator to evaluate whether the plant is drought-resistant (Zhang et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This experiment showed that PEG treatment obviously suppressed the germination index of maize seeds compared with control CK (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Under PEG treatment, with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentration (0.01%, 0.02%, 0.04%, and 0.08%) submersion treatment significantly improved the germination index of maize seeds, among them 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e had the best mitigation effect (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). However, the effect of 0.2% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment on the germination index was not significant (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). This showed that low concentration of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment alleviated germination of maize seeds under drought stress.\u003c/p\u003e \u003cp\u003eUnder drought stress, plants adapt to water-deficient environments through osmotic regulation (Seleiman et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Osmotic regulation is a physiological response of plants to environmental water stress, which increases cellular osmotic pressure, reduces water loss rate and improves drought tolerance (Blum., 2016). In drought conditions, soluble sugars and proline are the two main substances that regulate cell osmotic potential (Dghim et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This experiment showed that PEG treatment obviously promoted the increment of proline content in the root and bud of maize seedlings (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA). Under PEG treatment, with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentration (0.01%, 0.02% and 0.04%) submersion treatment significantly improved the increment of proline content in maize seedlings, of which 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e had the best mitigation effect (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eWhen plants are subjected to drought, ROS accumulation disrupts the cell membrane integrity and cellular function, leading to membrane lipid peroxidation (Hussain et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The MDA content can directly reflect the degree of membrane damage and the ability of plants to adapt to drought (Lee et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). This experiment showed that PEG treatment obviously promoted the increment of MDA content in the roots and shoots of maize seedlings (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Under PEG treatment, with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentration (0.01%, 0.02%, 0.04%, 0.08% and 0.2%) submersion treatment significantly reduced the increment of MDA content, of which 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e was the best mitigation effect (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the face of drought-induced oxidative damage, the plant body has enzymatic and non-enzymatic antioxidant systems to maintain redox homeostasis, neutralize free radicals or reactive oxygen species, prevent acute cell damage and maintain cell membrane integrity (Dias et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). This experiment showed that PEG treatment obviously promoted the activity of POD, CAT, and SOD in maize seedlings (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). Under PEG treatment, the activity of POD, CAT, and SOD was significantly improved with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentrations (0.01%, 0.02% and 0.04%) submersion treatment, of which 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e was the best mitigation effect (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eCorn seed germination and seedling growth decreased significantly with increasing PEG concentration. Under drought stress, with exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e of low concentration (0.02%) submersion treatment significantly improved seed germination and seedling growth, increased the proline content, promoted the activity of POD, CAT and SOD in seedling roots and shoots, and reduced the MDA content of seedlings to improve their drought resistance.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eDisclosure Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo potential conflict of interest was reported by the author.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the Natural Science Foundation of Gansu Province (No. 21JR11RM042) and Longdong University Doctoral Fund (XYBY202007).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eContributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eZhike Wang conceived and designed the experiments. Zhike Wang performed laboratory experiments. Zhike Wang performed data analysis and interpretation. Zhike Wang wrote the paper. The author read and approved the final manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAndrade, C. A., de Souza, K. R. D., de Oliveira Santos, M., da Silva, D. M., \u0026amp; Alves, J. D. (2018). Hydrogen peroxide promotes the tolerance of soybeans to waterlogging. Scientia Horticulturae 232, 40\u0026ndash;45. https://doi.org/10.1016/j.scienta.2017.12.048\u003c/li\u003e\n\u003cli\u003eBian, S. M., \u0026amp; Jiang, Y. W. (2009). Reactive oxygen species, antioxidant enzyme activities and gene expression patterns in leaves and roots of Kentucky bluegrass in response to drought stress and recovery. Scientia Horticulturae 120(2), 264\u0026ndash;270. https://doi.org/10.1016/j.scienta.2008.10.014\u003c/li\u003e\n\u003cli\u003eBlum, A. (2017). Osmotic adjustment is a prime drought stress adaptive engine in support of plant production. \u003cem\u003ePlant, Cell \u0026amp; Environment\u003c/em\u003e 40(1), 4\u0026ndash;10. https://doi.org/10.1111/pce.12800\u003c/li\u003e\n\u003cli\u003eBu, T., Yang, J. X., Liu, J. X., \u0026amp; Fan, X. F. (2024). NaHS immersion alleviates the stress effect of chromium (III) on alfalfa seeds by affecting active oxygen metabolism. Plant Signaling \u0026amp; Behavior 19(1), e2375673. https://doi.org/10.1080/15592324.2024.2375673\u003c/li\u003e\n\u003cli\u003eČern\u0026yacute;, M., Hab\u0026aacute;nov\u0026aacute;, H., Berka, M., Luklov\u0026aacute;, M., \u0026amp; Brzobohat\u0026yacute;, B. (2018). Hydrogen peroxide: its role in plant biology and crosstalk with signalling networks. International journal of molecular sciences 19(9), 2812. https://doi.org/10.3390/ijms19092812\u003c/li\u003e\n\u003cli\u003eChen, L., Liu, L. T., Lu, B., Ma, T. T., Jiang, D., Li, J., Zhang, K., Sun, H. C., Zhang, Y. J., Bai, Z. Y., \u0026amp; Li, C. D. (2020). Exogenous melatonin promotes seed germination and osmotic regulation under salt stress in cotton (\u003cem\u003eGossypium hirsutum\u003c/em\u003e L.). Plos One 15(1), e0228241. https://doi.org/ 10.1371/journal.pone.0228241\u003c/li\u003e\n\u003cli\u003eDias, M. C., Oliveira, H., Costa, A., \u0026amp; Santos, C. (2014). Improving elms performance under drought stress: The pretreatment with abscisic acid. Environmental and Experimental Botany\u003cem\u003e \u003c/em\u003e100, 64\u0026ndash;73. https://doi.org/10.1016/j.envexpbot.2013.12.013\u003c/li\u003e\n\u003cli\u003eDghim, F., Abdellaoui, R., Boukhris, M., Neffati, M., \u0026amp; Chaieb, M. (2018). Physiological and biochemical changes in \u003cem\u003ePeriploca angustifolia\u003c/em\u003e plants under withholding irrigation and rewatering conditions. South African Journal of Botany 114, 241\u0026ndash;249. https://doi.org/10.1016/j.sajb.2017.11.007\u003c/li\u003e\n\u003cli\u003eGao, Y., Guo, Y. K., Lin, S. H., Fang, Y. Y., \u0026amp; Bai, J. G. (2010). Hydrogen peroxide pretreatment alters the activity of antioxidant enzymes and protects chloroplast ultrastructure in heat-stressed cucumber leaves. Scientia Horticulturae 126(1), 20\u0026ndash;26. https://doi.org/10.1016/j.scienta.2010.06.006\u003c/li\u003e\n\u003cli\u003eGuler, N. S., \u0026amp; Pehlivan, N. (2016). Exogenous low-dose hydrogen peroxide enhances drought tolerance of soybean (\u003cem\u003eGlycine max\u003c/em\u003e L.) through inducing antioxidant system. Acta Biologica Hungarica 67, 169\u0026ndash;183. https://doi.org/10.1556/018.67.2016.2.5\u003c/li\u003e\n\u003cli\u003eHe, L. H., Gao, Z. Q., \u0026amp; Li, R. Z. (2009). Pretreatment of seed with H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2 \u003c/sub\u003eenhances drought tolerance of wheat (\u003cem\u003eTriticum aestivum\u003c/em\u003e L.) seedlings. African Journal of Biotechnology 8(22), 6151\u0026ndash;6157. https://doi.org/10.5897/AJB09.490\u003c/li\u003e\n\u003cli\u003eHu, Y. L., Ge, Y., Zhang, C. H, Ju, T., \u0026amp; Cheng, W. D. (2009). Cadmium toxicity and translocation in rice seedlings are reduced by hydrogen peroxide pretreatment. Plant Growth Regulation 59, 51\u0026ndash;61. https://doi.org/10.1007/s10725-009-9387-7\u003c/li\u003e\n\u003cli\u003eHussain, H. A., Hussain, S., Khaliq, A., Ashraf, U., Anjum, S. A., Men, S. M., \u0026amp; Wang, L. C. (2018). Chilling and drought stresses in crop plants: Implications, cross talk, and potential management opportunities. Frontiers in Plant Science 9, 393. https://doi.org/10.3389/fpls.2018.00393\u003c/li\u003e\n\u003cli\u003eIqbal, H., Yaning, C., Waqas, M., Rehman, H., Shareef, M., \u0026amp; Iqbal, S. (2018). Hydrogen peroxide application improves quinoa performance by affecting physiological and biochemical mechanisms under water‐deficit conditions. Journal of Agronomy and Crop Science\u003cem\u003e \u003c/em\u003e204(6), 541\u0026ndash;553. https://doi.org/10.1111/jac.12284\u003c/li\u003e\n\u003cli\u003eIshibashi, Y., Tawaratsumida, T., Zheng, S. H., Yuasa, T., \u0026amp; Iwaya-inoue, M. (2010). NADPH oxidases act as key enzyme on germination and seedling growth in barley (\u003cem\u003eHordeum vulgare\u003c/em\u003e L.). \u003cem\u003ePlant Production Science\u003c/em\u003e 13(1), 45\u0026ndash;52. https://doi.org/10.1626/pps.13.45 \u003c/li\u003e\n\u003cli\u003eIshibashi, Y., Yamaguchi, H., Yuasa, T., Iwaya-Inoue, M., Arima, S., \u0026amp; Zheng, S. H. (2011). Hydrogen peroxide spraying alleviates drought stress in soybean plants. Journal of Plant Physiology 168 (13), 1562\u0026ndash;1567. https://doi.org/10.1016/j.jplph.2011.02.003\u003c/li\u003e\n\u003cli\u003eIshibashi, Y., Kasa, S., Sakamoto, M., Aoki, N., Kai, K., Yuasa, T., Hanada, A., Yamaguchi, S., \u0026amp; Iwaya-Inoue, M. (2015). A role for reactive oxygen species produced by NADPH oxidases in the embryo and aleurone cells in barley seed germination. Plos One 10(11): e0143173. https://doi.org/10.1371/journal.pone.0143173\u003c/li\u003e\n\u003cli\u003eKhan, A., Anwar, Y., Hasan, M. M., Iqbal, A., Ali, M., Alharby, H. F., Hakeem, K. R., \u0026amp; Hasanuzzaman, M. (2017). Attenuation of drought stress in \u003cem\u003ebrassica\u003c/em\u003e seedlings with exogenous application of Ca\u003csup\u003e2 +\u003c/sup\u003e and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e. Plants 6(2), 20. https://doi.org/10.3390/plants6020020\u003c/li\u003e\n\u003cli\u003eLee, B. R., Li, L. S., Jung, W. J., Jin, Y. L., Avice, J. C., Ourry, A., \u0026amp; Kim, T. H. (2009).Water deficit-induced oxidative stress and the activation of antioxidant enzymes in white clover leaves. Biologia Plantarum 53, 505\u0026ndash;510. https://doi.org/10.1007/s10535-009-0091-2\u003c/li\u003e\n\u003cli\u003eLeymarie, J., Vitkauskait\u0026eacute;, G., Hoang, H. H., Gendreau, E., Chazoule, V., Meimoun, P., Corbineau, F., El-Maarouf-Bouteau, H., \u0026amp; Bailly, C. (2012) Role of reactive oxygen species in the regulation of \u003cem\u003eArabidopsis \u003c/em\u003eseed dormancy.\u003cem\u003e \u003c/em\u003ePlant and Cell Physiology 53(1): 96\u0026ndash;106. https://doi.org/10.1093/pcp/pcr129\u003c/li\u003e\n\u003cli\u003eLi, J. T., Qiu, Z. B., Zhang, X. W., \u0026amp; Wang, L. S. (2011). Exogenous hydrogen peroxide can enhance tolerance of wheat seedlings to salt stress. Acta Physiologiae Plantarum 33, 835\u0026ndash;842. https://doi.org/10.1093/10.1007/s11738-010-0608-5\u003c/li\u003e\n\u003cli\u003eLiu, Z. J., Guo, Y. K., \u0026amp; Bai, J. G. (2010). Exogenous hydrogen peroxide changes antioxidant enzyme activity and protects ultrastructure in leaves of two cucumber ecotypes under osmotic stress. Journal of Plant Growth Regulation\u003cem\u003e \u003c/em\u003e29, 171\u0026ndash;183. https://doi.org/10.1093/10.1007/s00344-009-9121-8\u003c/li\u003e\n\u003cli\u003eLiu, L. J, Huang, L., Lin, X. Y., \u0026amp; Sun, C. L. (2020). Hydrogen peroxide alleviates salinity-induced damage through enhancing proline accumulation in wheat seedlings. Plant Cell Reports 39, 567\u0026ndash;575. https://doi.org/10.1007/s00299-020-02513-3\u003c/li\u003e\n\u003cli\u003eOzaki, K., Uchida, A., Takabe, T., Shinagawa, F., Tanaka, Y., Takabe, T., Hayashi, T., Hattori, T., Rai, A. K., \u0026amp; Takabe, T. (2009). Enrichment of sugar content in melon fruits by hydrogen peroxide treatment. Journal of Plant Physiology 166 (6), 569\u0026ndash;578. https://doi.org/10.1016/j.jplph.2008.08.007\u003c/li\u003e\n\u003cli\u003eSachdev, S., Ansari, S. A., Ansari, M. I., Fujita, M., \u0026amp; Hasanuzzaman, M. (2021). Abiotic stress and reactive oxygen species: Generation, signaling, and defense mechanisms. Antioxidants\u003cem\u003e \u003c/em\u003e10(2), 277. https://doi.org/10.3390/antiox10020277\u003c/li\u003e\n\u003cli\u003eSeleiman, M. F., Al-Suhaibani, N., Ali, N., Akmal, M., Alotaibi, M., Refay, Y., Dindaroglu, T., Abdul-Wajid., H. H., \u0026amp; Battaglia, M. L. (2021). Drought stress impacts on plants and different approaches to alleviate its adverse effects. Plants\u003cem\u003e \u003c/em\u003e10(2), 259. https://doi.org/10.3390/plants 10020259\u003c/li\u003e\n\u003cli\u003eSun, Y., Wang, H., Liu, S., \u0026amp; Peng, X. (2016). Exogenous application of hydrogen peroxide alleviates drought stress in cucumber seedlings. South African Journal of Botany 106, 23\u0026ndash;28. https://doi.org/10.1016/j.sajb.2016.05.008\u003c/li\u003e\n\u003cli\u003eTerzi, R., Kadioglu, A., Kalaycioglu, E., \u0026amp; Saglam, A. (2014). Hydrogen peroxide pretreatment induces osmotic stress tolerance by influencing osmolyte and abscisic acid levels in maize leaves. Journal of Plant Interactions\u003cem\u003e \u003c/em\u003e9(1), 559\u0026ndash;565. https://doi.org/10.1080/17429145.2013.871077\u003c/li\u003e\n\u003cli\u003eWang, X. F., Zhao, J. Q., Yuan, P. G., Ding, S. Y., Jiang, L. G., \u0026amp; Xi, Z. M. (2024). Hydrogen peroxide functioned as a redox signaling molecule in the putrescine-promoted drought tolerance in cabernet sauvignon. Scientia Horticulturae 335, 113325. https://doi.org/10.1016/j.scienta.2024. 113325\u003c/li\u003e\n\u003cli\u003eWang, Z. K., Ma, R., Zhao, M. S., Wang, F. F., Zhang, N., \u0026amp; Si, H. J. (2020). NO and ABA interaction regulates tuber dormancy and sprouting in potato. Frontiers in Plant Science 11, 311. https://doi.org/10.3389/fpls.2020.00311\u003c/li\u003e\n\u003cli\u003eZhang, C. M., Shi, S. L., Wang, B. W., \u0026amp; Zhao, J. F. (2018). Physiological and biochemical changes in different drought-tolerant alfalfa (\u003cem\u003eMedicago sativa\u003c/em\u003e L.) varieties under PEG-induced drought stress. Acta Physiologiae Plantarum, 40, 25. https://doi.org/10.1007/s11738-017-2597-0\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"drought stress, hydrogen peroxide, maize, seed germination, physiological characteristics","lastPublishedDoi":"10.21203/rs.3.rs-6202193/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6202193/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eHydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) is an important signaling molecule in plant body, which is involved in the regulation of multiple abiotic stresses.This study investigated the ability of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e to alleviate drought stress in maize (\u003cem\u003eZea mays\u003c/em\u003e L.) seedlings. With maize seeds as the test material, used the filter paper germination method. The seeds were treated with different concentrations of exogenous H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e under 10% polyethylene glycol (PEG-6000) stress to measure seed germination, seedling growth and physiological indicators. The results showed that 10% PEG treatment alone obviously inhibited the seed germination and seedling growth; Treatment with 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e significantly improved the germination potential and vitality index of maize seeds under PEG stress, promoted the growth of seedling buds and roots, enhanced the activity of APX, SOD and CAT in buds and roots, reduced the MDA content, and promoted the increment of proline and soluble sugars. The above results indicated that exogenous 0.02% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment could activate antioxidant enzyme activity and reduce oxidative damage under drought stress, thus alleviating maize drought tolerance at the seed germination stage.\u003c/p\u003e","manuscriptTitle":"Exogenous H2O2 alleviates maize seed germination and seedling physiology property under drought stress","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-17 09:03:38","doi":"10.21203/rs.3.rs-6202193/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a72903f8-a044-4301-ac68-a8c9887bc0a0","owner":[],"postedDate":"March 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-03-22T09:38:29+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-17 09:03:38","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6202193","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6202193","identity":"rs-6202193","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}