Spermidine synthase genes from mulberry play an important role in drought stress tolerance by altering antioxidant enzymes and osmotic regulation
Dan Liu
A B , Qiang Lin B , Changyu Qiu B , Xiaomei Lu B and Ningjia He A *
A
B
Abstract
Our previous experiments confirmed that two mulberry spermidine synthase (SPDS) genes (MnSPDS1 and MnSPDS2) that encode functional proteins are highly expressed under drought stress. In this study, the functions of MnSPDS1/MnSPDS2 in the drought stress response were further explored by silencing and overexpressing these genes in mulberry and tobacco, respectively. Compared with the wild-type (WT) plants, the MnSPDS1/MnSPDS2-overexpression tobacco plants were more tolerant to drought stress and showed a higher spermidine content (P < 0.05). Moreover, overexpression of MnSPDS1/MnSPDS2 at the physiological level alleviated membrane damage caused by drought and improved osmotic regulation and antioxidant capacity. In addition, correlation analysis showed that the content of spermidine was positively correlated with the expression levels of MnSPDS1 and MnSPDS2, with correlation coefficients of 0.762 and 0.715, respectively. Moreover, drought injury was more serious in the MnSPDS-silenced seedlings than in the WT seedlings after drought treatment. These results suggest that MnSPDS genes play important roles in the drought stress response and are valuable for molecular breeding to enhance the drought tolerance of mulberry.
Keywords: drought stress, drought tolerance, gene expression, gene silencing, molecular breeding, mulberry, spermidine, spermidine synthase.
References
Alcázar R, Marco F, Cuevas JC, Patron M, Ferrando A, Carrasco P, Tiburcio AF, Altabella T (2006) Involvement of polyamines in plant response to abiotic stress. Biotechnology Letters 28(23), 1867-1876.
| Crossref | Google Scholar | PubMed |
Alcázar R, Altabella T, Marco F, Bortolotti C, Reymond M, Koncz C, Carrasco P, Tiburcio AF (2010) Polyamines: molecules with regulatory functions in plant abiotic stress tolerance. Planta 231(6), 1237-1249.
| Crossref | Google Scholar | PubMed |
Benesch R (1954) Methods of biochemical analysis. Journal of the American Chemical Society 76, 5576.
| Crossref | Google Scholar |
Blum A (2017) Osmotic adjustment is a prime drought stress adaptive engine in support of plant production. Plant, Cell & Environment 40, 4-10.
| Crossref | Google Scholar | PubMed |
Chen M, Fu Y, Mou Q, An J, Zhu X, Ahmed T, Zhang S, Basit F, Hu J, Guan Y (2021) Spermidine induces expression of stress associated proteins (SAPs) genes and protects rice seed from heat stress-induced damage during grain-filling. Antioxidants 10, 1544.
| Crossref | Google Scholar | PubMed |
Crabos A, Huang Y, Boursat T, Maurel C, Ruffel S, Krouk G, Boursiac Y (2023) Distinct early transcriptional regulations by turgor and osmotic potential in the roots of Arabidopsis. Journal of Experimental Botany 74(18), 5917-5930.
| Crossref | Google Scholar | PubMed |
Cruz de Carvalho MH (2008) Drought stress and reactive oxygen species: production, scavenging and signaling. Plant Signaling & Behavior 3(3), 156-165.
| Crossref | Google Scholar |
Duan J, Li J, Guo S, Kang Y (2008) Exogenous spermidine affects polyamine metabolism in salinity-stressed Cucumis sativus roots and enhances short-term salinity tolerance. Journal of Plant Physiology 165(15), 1620-1635.
| Crossref | Google Scholar |
Dudley HW, Rosenheim O, Starling WW (1926) The chemical constitution of spermine III. Structure and synthesis. Biochemical Journal 20(5), 1082-1094.
| Crossref | Google Scholar |
Dudley HW, Rosenheim O, Starling WW (1927) The constitution and synthesis of spermidine, a newly discovered base isolated from animal tissues. Biochemical Journal 21(1), 97-103.
| Crossref | Google Scholar |
Fu XZ, Chen CW, Yin W, Liu JH, Moriguchi T (2011) Ectopic expression of MdSPDS1 in sweet orange (Citrus sinensis Osbeck) reduces canker susceptibility: involvement of H2O2 production and transcriptional alteration. BMC Plant Biology 11(1), 55.
| Crossref | Google Scholar |
Goufo P, Moutinho-Pereira JM, Jorge TF, Correia CM, Oliveira MR, Rosa EAS, António C, Henrique T (2017) Cowpea (Vigna unguiculata L. Walp.) metabolomics: osmoprotection as a physiological strategy for drought stress resistance and improved yield. Frontiers in Plant Science 8, 586.
| Crossref | Google Scholar |
Gupta K, Dey A, Gupta B (2013) Plant polyamines in abiotic stress responses. Acta Physiologiae Plantarum 35(7), 2015-2036.
| Crossref | Google Scholar |
Gupta K, Sengupta A, Chakraborty M, Gupta B (2016) Hydrogen peroxide and polyamines act as double edged swords in plant abiotic stress responses. Frontiers in Plant Science 7, 1314.
| Crossref | Google Scholar |
Hu Q, Fu Y, Guan Y, Lin C, Cao D, Hu W, Sheteiwy M, Hu J (2016) Inhibitory effect of chemical combinations on seed germination and pre-harvest sprouting in hybrid rice. Plant Growth Regulation 80, 281-289.
| Crossref | Google Scholar |
Jiang D, Hou J, Gao W, Tong X, Li M, Chu X, Chen G (2021) Exogenous spermidine alleviates the adverse effects of aluminum toxicity on photosystem II through improved antioxidant system and endogenous polyamine contents. Ecotoxicology and Environmental Safety 207, 111265.
| Crossref | Google Scholar |
Knörzer OC, Burner J, Boger P (2010) Alterations in the antioxidative system of suspension-cultured soybean cells (Glycine max) induced by oxidative stress. Physiologia Plantarum 97, 388-396.
| Crossref | Google Scholar |
Kubiś J (2008) Exogenous spermidine differentially alters activities of some scavenging system enzymes, H2O2 and superoxide radical levels in water-stressed cucumber leaves. Journal of Plant Physiology 165(4), 397-406.
| Crossref | Google Scholar |
Li J, Hu L, Zhang L, Pan X, Hu X (2015) Exogenous spermidine is enhancing tomato tolerance to salinity–alkalinity stress by regulating chloroplast antioxidant system and chlorophyll metabolism. BMC Plant Biology 15, 303.
| Crossref | Google Scholar |
Liu D, Meng S, Xiang ZH, Yang G, He N (2019) An R1R2R3 MYB transcription factor, MnMYB3R1, regulates the polyphenol oxidase gene in mulberry (Morus notabilis). International Journal of Molecular Sciences 20, 2602.
| Crossref | Google Scholar | PubMed |
Liu D, Zeng Y, Qiu C, Lin Q (2021) Molecular cloning and adversity stress expression analysis of SPDS genes in mulberry (Morus notabilis). Russian Journal of Plant Physiology 68(6), 1186-1193.
| Crossref | Google Scholar |
Majumdar R, Shao L, Minocha R, Long S, Minocha SC (2013) Ornithine: the overlooked molecule in the regulation of polyamine metabolism. Plant and Cell Physiology 54(6), 990-1004.
| Crossref | Google Scholar |
Mostofa MG, Yoshida N, Fujita M (2014) Spermidine pretreatment enhances heat tolerance in rice seedlings through modulating antioxidative and glyoxalase systems. Plant Growth Regulation 73(1), 31-44.
| Crossref | Google Scholar |
Nahar K, Hasanuzzaman M, Rahman A, Alam MM, Mahmud J-A, Suzuki T, Fujita M (2016) Polyamines confer salt tolerance in mung bean (Vigna radiata L.) by reducing sodium uptake, improving nutrient homeostasis, antioxidant defense, and methylglyoxal detoxification systems. Frontiers in Plant Science 7, 1104.
| Crossref | Google Scholar | PubMed |
Neilya MH, Matsukuraa C, Maucourt M, Bernillonb S, Deborde C, Moing A, Yina YG, Saitoa T, Mori K, Asamizua E, Rolinb D, Moriguchi T, Ezura H (2011) Enhanced polyamine accumulation alters carotenoid metabolism at the transcriptional level in tomato fruit over-expressing spermidine synthase. Journal of Plant Physiology 168, 242-252.
| Crossref | Google Scholar |
Ozturk M, Turkyilmaz Unal B, García-Caparrós P, Khursheed A, Gul A, Hasanuzzaman M (2021) Osmoregulation and its actions during the drought stress in plants. Physiologia Plantarum 172(2), 1321-1335.
| Crossref | Google Scholar |
Pottosin I, Shabala S (2014) Polyamines control of cation transport across plant membranes: implications for ion homeostasis and abiotic stress signaling. Frontiers in Plant Science 5, 154.
| Crossref | Google Scholar |
Ramegowda V, Mysore KS, Senthil-Kumar M (2014) Virus-induced gene silencing is a versatile tool for unraveling the functional relevance of multiple abiotic-stress-responsive genes in crop plants. Frontiers in Plant Science 5, 323.
| Crossref | Google Scholar | PubMed |
Ren J, Li C, Xiu Q, Xu M, Liu H (2024) Overexpression of wheat spermidine synthase gene enhances wheat resistance to Fusarium head blight. Phytopathology Research 6, 24.
| Crossref | Google Scholar |
Richards FJ, Coleman EG (1952) Occurrence of putrescine in potassium-deficient barley. Nature 170(4324), 460.
| Crossref | Google Scholar |
Roychoudhury A, Basu S, Sengupta DN (2011) Amelioration of salinity stress by exogenously applied spermidine or spermine in three varieties of indica rice differing in their level of salt tolerance. Journal of Plant Physiology 168(4), 317-328.
| Crossref | Google Scholar |
Shan X, Zhou H, Sang T, Shu S, Sun J, Guo S (2016) Effects of exogenous spermidine on carbon and nitrogen metabolism in tomato seedlings under high temperature. Journal of the American Society for Horticultural Science 141(4), 381-388.
| Crossref | Google Scholar |
Shehata SA, Omar HS, Elfaidy AGS, El-Sayed SSF, Abuarab ME, Abdeldaym EA (2022) Grafting enhances drought tolerance by regulating stress-responsive gene expression and antioxidant enzyme activities in cucumbers. BMC Plant Biology 22(1), 408.
| Crossref | Google Scholar |
Shi J, Fu XZ, Peng T, Huang X-S, Fan Q-J, Liu J-H (2010) Spermine pretreatment confers dehydration tolerance of citrus in vitro plants via modulation of antioxidative capacity and stomatal response. Tree Physiology 30(7), 914-922.
| Crossref | Google Scholar | PubMed |
Shi H, Ye T, Chan Z (2013) Comparative proteomic and physiological analyses reveal the protective effect of exogenous polyamines in the bermudagrass (Cynodon dactylon) response to salt and drought stresses. Journal of Proteome Research 12(11), 4951-4964.
| Crossref | Google Scholar |
Stewart RRC, Bewley JD (1980) Lipid peroxidation associated with accelerated aging of soybean axes. Plant Physiology 65, 245-248.
| Crossref | Google Scholar |
Tang C, Zhang R, Hu X, Song J, Li B, Ou D, Hu X, Zhao Y (2019) Exogenous spermidine elevating cadmium tolerance in Salix matsudana involves cadmium detoxification and antioxidant defense. International Journal of Phytoremediation 21(4), 305-315.
| Crossref | Google Scholar |
Wang LC (1972) Polyamines in soybeans. Plant Physiology 50(1), 152-156.
| Crossref | Google Scholar | PubMed |
Wang P, Xu Z, Zhang Y, Ma Y, Yang J, Zhou F, Gao Y, Li G, Hu X (2022) Over-expression of spermidine synthase 2 (SlSPDS2) in tomato plants improves saline-alkali stress tolerance by increasing endogenous polyamines content to regulate antioxidant enzyme system and ionic homeostasis. Plant Physiology and Biochemistry 192, 172-185.
| Crossref | Google Scholar |
Wang W, Liu W, Wang B (2023) Identification of CDK gene family and functional analysis of CqCDK15 under drought and salt stress in quinoa. BMC Genomics 24(1), 461.
| Crossref | Google Scholar |
Wen XP, Pang XM, Matsuda N, Kita M, Inoue H, Hao YJ, Honda C, Moriguchi T (2008) Over-expression of the apple spermidine synthase gene in pear confers multiple abiotic stress tolerance by altering polyamine titers. Transgenic Research 17(2), 251-263.
| Crossref | Google Scholar |
Wen XP, Ban Y, Inoue H, Matsuda N, Moriguchi T (2009) Aluminum tolerance in a spermidine synthase-overexpressing transgenic European pear is correlated with the enhanced level of spermidine via alleviating oxidative status. Environmental and Experimental Botany 66, 471-478.
| Crossref | Google Scholar |
Wen XP, Ban Y, Inoue H, Matsuda N, Moriguchi T (2010) Spermidine levels are implicated in heavy metal tolerance in a spermidine synthase overexpressing transgenic European pear by exerting antioxidant activities. Transgenic Research 19(1), 91-103.
| Crossref | Google Scholar |
Wu M, Tian T, Liu H, Zhang H, Qiu Z, Qiao G, Wen X (2020) Shelter covering altered polyamines accumulation of cherry (Prunus pseudocerasus Lindl.) via elevating the expression of spermidine synthase (SPDS) gene. Scientia Horticulturae 270, 109440.
| Crossref | Google Scholar |
Xu Y, Jian C, Li K, Tian Y, Zhu K, Zhang W, Wang W, Wang Z, Yang J (2021) The role of polyamines in regulating amino acid biosynthesis in rice grains. Food and Energy Security 10(4), e306.
| Crossref | Google Scholar |
Yamamoto A, Shim IS, Fujihara S (2012) Chilling-stress responses by rice seedlings grown with different ammonium concentrations and its relationship to leaf spermidine content. Journal of Plant Biology 55(3), 191-197.
| Crossref | Google Scholar |
Yang J, Wang P, Li X, Zhou D, Cai X, Hu X, Hu S (2023) Regulation of cold resistance by the polyamine biosynthetic gene SlSPDS2 via modulating the antioxidant enzyme system and osmotic regulatory substances in Solanum lycopersicum. Environmental and Experimental Botany 216, 105531.
| Crossref | Google Scholar |
Zhang J, Xie M, Yu G, Wang D, Xu Z, Liang L, Xiao J, Xie Y, Tang Y, Sun G, Sun B, Huang Z, Lai Y, Li H (2023) CaSPDS, a spermidine synthase gene from pepper (Capsicum annuum L.), plays an important role in response to cold stress. International Journal of Molecular Sciences 24, 5013.
| Crossref | Google Scholar |
Zhu GL, Deng XW, Zuo WN (1983) Determination of free proline in plants. Plant Physiology Communications 1, 35-37.
| Google Scholar |
Zhu X, Wang B, Liu W, Wei X, Wang X, Du X, Liu H (2023) Genome-wide analysis of AP2/ERF gene and functional analysis of CqERF24 gene in drought stress in quinoa. International Journal of Biological Macromolecules 253(8), 127582.
| Crossref | Google Scholar |
Zou D, Min Y, Liu Y, Wei X, Wang J (2020) Identification of a spermidine synthase gene from soybean by recombinant expression, transcriptional verification, and sequence analysis. Journal of Agricultural and Food Chemistry 68, 2366-2372.
| Crossref | Google Scholar | PubMed |
