Abstract
Key message
Transformation of MruGSTU39 in M. ruthenica and alfalfa enhanced growth and survival of transgenic plants by up-regulating GST and glutathione peroxidase activity to detoxify ROS under drought stress.
Abstract
Glutathione S-transferases (GSTs) are ubiquitous supergene family which play crucial roles in detoxification of reactive oxygen species (ROS). Despite studies on GSTs, few studies have focused on them in perennial, wild plant species with high tolerance to environmental stress. Here, we identified 66 MruGST genes from the genome of Medicago ruthenica, a perennial legume species native to temperate grasslands with high tolerance to environmental stress. These genes were divided into eight classes based on their conserved domains, phylogenetic tree and gene structure, with the tau class being the most numerous. Duplication analysis revealed that GST family in M. ruthenica was expanded by segmental and tandem duplication. Several drought-responsive MruGSTs were identified by transcriptomic analyses. Of them, expression of MruGSTU39 was up-regulated much more in a tolerant accession by drought stress. Transformation of MruGSTU39 in M. ruthenica and alfalfa (Medicago sativa) enhanced growth and survival of transgenic seedlings than their wild-type counterparts under drought. We demonstrated that MruGSTU39 can detoxify ROS to reduce its damage to membrane by up-regulating activities of GST and glutathione peroxidase. Our findings provide full-scale knowledge on GST family in the wild legume M. ruthenica with high tolerance to drought, and highlight improvement tolerance of legume forages to drought using genomic information of M. ruthenica.
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Data availability
The transcriptome data have been deposited in GenBank under accession PRJNA589938.
References
Ahmad MZ, Nasir JA, Ahmed S, Ahmad B, Sana A, Salman S, Shah Z, Yang CY (2020) Genome-wide analysis of glutathione S-transferase gene family in G. max. Biologia 75:1691–1705. https://doi.org/10.2478/s11756-020-00463-5
Alexieva V, Sergiev I, Mapelli S, Karanov E (2001) The effect of drought and ultraviolet radiation on growth and stress markers in pea and wheat. Plant Cell Environ 24:1337–1344. https://doi.org/10.1046/j.1365-3040.2001.00778.x
Anders S, Pyl PT, Huber W (2015) HTSeq-a Python framework to work with high-throughput sequencing data. Bioinformatics 31:166–169. https://doi.org/10.1093/bioinformatics/btu638
Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399. https://doi.org/10.1146/annurev.arplant.55.031903.141701
Brozynska M, Furtado A, Henry RJ (2016) Genomics of crop wild relatives: expanding the gene pool for crop improvement. Plant Biotechnol J 14:1070–1085. https://doi.org/10.1111/pbi.12454
Campbell TA, Bao G, Xia ZL (1997) Agronomic evaluation of Medicago ruthenica collected in inner mongolia. Crop Sci 37:599–604. https://doi.org/10.2135/cropsci1997.0011183X003700020048x
Chen C, Chen H, Zhang Y, Thomas HR, Frank MH, He Y, Xia R (2020a) TBtools: an integrative toolkit developed for interactive analyses of big biological data. Mol Plant 13:1194–1202. https://doi.org/10.1016/j.molp.2020.06.009
Chen H, Zeng Y, Yang Y, Huang L, Tang B, Zhang H, Hao F, Liu W, Li Y, Liu Y, Zhang X, Zhang R, Zhang Y, Wang K, He H, Wang Z, Fan G, Yang H, Bao A, Shang Z, Chen J, Wang W, Qiu Q (2020b) Allele-aware chromosome-level genome assembly and efficient transgene-free genome editing for the autotetraploid cultivated alfalfa. Nat Commun 11:2494. https://doi.org/10.1038/s41467-020-16338-x
Collino DJ, Dardanelli JL, De Luca MJ, Racca RW (2005) Temperature and water availability effects on radiation and water use efficiencies in alfalfa (Medicago sativa L.). Aust J Exp Agr 45:383–390. https://doi.org/10.1071/EA04050
Cummins I, Dixon DP, Freitag-Pohl S, Skipsey M, Edwards R (2011) Multiple roles for plant glutathione transferases in xenobiotic detoxification. Drug Metab Rev 43:266–280. https://doi.org/10.3109/03602532.2011.552910
Dixon DP, Lapthorn A, Edwards R (2002) Plant glutathione transferases. Genome Biol 3:3004. https://doi.org/10.1186/gb-2002-3-3-reviews3004
Dong YT, Li C, Zhang Y, He QL, Daud MK, Chen JH, Zhu SJ (2016) Glutathione S-transferase gene family in Gossypium raimondii and G. arboreum: comparative genomic study and their expression under salt stress. Front Plant Sci 7:139. https://doi.org/10.3389/Fpls.2016.00139
Edwards R, Dixon DP, Walbot V (2000) Plant glutathione S-transferases: enzymes with multiple functions in sickness and in health. Trends Plant Sci 5:193–198. https://doi.org/10.1016/S1360-1385(00)01601-0
Gao J, Chen BW, Lin HJ, Liu Y, Wei Y, Chen FB, Li WB (2020) Identification and characterization of the Glutathione S-Transferase (GST) family in radish reveals a likely role in anthocyanin biosynthesis and heavy metal stress tolerance. Gene 743:144484. https://doi.org/10.1016/J.Gene.2020.144484
Ghangal R, Rajkumar MS, Garg R, Jain M (2020) Genome-wide analysis of glutathione S-transferase gene family in chickpea suggests its role during seed development and abiotic stress. Mol Biol Rep 47:2749–2761. https://doi.org/10.1007/s11033-020-05377-8
Han XM, Yang ZL, Liu YJ, Yang HL, Zeng QY (2018) Genome-wide profiling of expression and biochemical functions of the Medicago glutathione S-transferase gene family. Plant Physiol Bioch 126:126–133. https://doi.org/10.1016/j.plaphy.2018.03.004
Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database: 1999. Nucleic Acids Res 27:297–300. https://doi.org/10.1093/Nar/27.1.297
Hoffmann B, Trinh TH, Leung J, Kondorosi A, Kondorosi E (1997) A new Medicago truncatula line with superior in vitro regeneration, transformation, and symbiotic properties isolated through cell culture selection. Mol Plant Microbe in 10:307–315. https://doi.org/10.1094/Mpmi.1997.10.3.307
Howe EA, Sinha R, Schlauch D, Quackenbush J (2011) RNA-Seq analysis in MeV. Bioinformatics 27:3209–3210. https://doi.org/10.1093/bioinformatics/btr490
Jain M, Ghanashyam C, Bhattacharjee A (2010) Comprehensive expression analysis suggests overlapping and specific roles of rice glutathione S-transferase genes during development and stress responses. BMC Genomics 11:73. https://doi.org/10.1186/1471-2164-11-73
Kissoudis C, Kalloniati C, Flemetakis E, Madesis P, Labrou NE, Tsaftaris A, Nianiou-Obeidat I (2015) Stress-inducible GmGSTU4 shapes transgenic tobacco plants metabolome towards increased salinity tolerance. Acta Physiol Plant 37:102. https://doi.org/10.1007/S11738-015-1852-5
Kramer GF, Norman HA, Krizek DT, Mirecki RM (1991) Influence of UV-B radiation on polyamines, lipid-peroxidation and membrane-lipids in cucumber. Phytochemistry 30:2101–2108. https://doi.org/10.1016/0031-9422(91)83595-C
Krzywinski M, Schein J, Birol I, Connors J, Gascoyne R, Horsman D, Jones SJ, Marra MA (2009) Circos: an information aesthetic for comparative genomics. Genome Res 19:1639–1645. https://doi.org/10.1101/gr.092759.109
Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 35:1547–1549. https://doi.org/10.1093/molbev/msy096
Lallement PA, Meux E, Gualberto JM, Dumarcay S, Favier F, Didierjean C, Saul F, Haouz A, Morel-Rouhier M, Gelhaye E, Rouhier N, Hecker A (2015) Glutathionyl-hydroquinone reductases from poplar are plastidial proteins that deglutathionylate both reduced and oxidized glutathionylated quinones. Febs Lett 589:37–44. https://doi.org/10.1016/j.febslet.2014.11.021
Lan T, Yang ZL, Yang X, Liu YJ, Wang XR, Zeng QY (2009) Extensive functional diversification of the Populus glutathione S-transferase supergene family. Plant Cell 21:3749–3766. https://doi.org/10.1105/tpc.109.070219
Liu YJ, Han XM, Ren LL, Yang HL, Zeng QY (2013) Functional divergence of the glutathione S-transferase supergene family in Physcomitrella patens reveals complex patterns of large gene family evolution in land plants. Plant Physiol 161:773–786. https://doi.org/10.1104/pp.112.205815
Liu M, Li Y, Ma YL, Zhao Q, Stiller J, Feng Q, Tian QL, Liu DC, Han B, Liu CJ (2020) The draft genome of a wild barley genotype reveals its enrichment in genes related to biotic and abiotic stresses compared to cultivated barley. Plant Biotechnol J 18:443–456. https://doi.org/10.1111/pbi.13210
Meng YY, Hou YL, Wang H, Ji RH, Liu B, Wen JQ, Niu LF, Lin H (2017) Targeted mutagenesis by CRISPR/Cas9 system in the model legume Medicago truncatula. Plant Cell Rep 36:371–374. https://doi.org/10.1007/s00299-016-2069-9
Mulder N, Apweiler R (2007) InterPro and InterProScan: tools for protein sequence classification and comparison. Methods Mol Biol 396:59–70. https://doi.org/10.1007/978-1-59745-515-2_5
Nianiou-Obeidat I, Madesis P, Kissoudis C, Voulgari G, Chronopoulou E, Tsaftaris A, Labrou NE (2017) Plant glutathione transferase-mediated stress tolerance: functions and biotechnological applications. Plant Cell Rep 36:791–805. https://doi.org/10.1007/s00299-017-2139-7
Rezaei MK, Shobbar ZS, Shahbazi M, Abedini R, Zare S (2013) Glutathione S-transferase (GST) family in barley: identification of members, enzyme activity, and gene expression pattern. J Plant Physiol 170:1277–1284. https://doi.org/10.1016/j.jplph.2013.04.005
Roxas VP, Smith RK, Allen ER, Allen RD (1997) Overexpression of glutathione S-transferase glutathione peroxidase enhances the growth of transgenic tobacco seedlings during stress. Nat Biotechnol 15:988–991. https://doi.org/10.1038/Nbt1097-988
Sappl PG, Carroll AJ, Clifton R, Lister R, Whelan J, Millar AH, Singh KB (2009) The Arabidopsis glutathione transferase gene family displays complex stress regulation and co-silencing multiple genes results in altered metabolic sensitivity to oxidative stress. Plant J 58:53–68. https://doi.org/10.1111/j.1365-313X.2008.03761.x
Sharma R, Sahoo A, Devendran R, Jain M (2014) Over-expression of a rice tau class glutathione S-transferase gene improves tolerance to salinity and oxidative stresses in Arabidopsis. PLoS ONE 9:e92900. https://doi.org/10.1371/journal.pone.0092900
Shen C, Du H, Chen Z, Lu H, Zhu F, Chen H, Meng X, Liu Q, Liu P, Zheng L, Li X, Dong J, Liang C, Wang T (2020) The chromosome-level genome sequence of the autotetraploid alfalfa and resequencing of core germplasms provide genomic resources for alfalfa research. Mol Plant 13:1250–1261. https://doi.org/10.1016/j.molp.2020.07.003
Small E, Jomphe M (1989) A synopsis of the genus Medicago (Leguminosae). Can J Bot 67:3260–3294. https://doi.org/10.1139/B89-405
Sylvestre-Gonon E, Law SR, Schwartz M, Robe K, Keech O, Didierjean C, Dubos C, Rouhier N, Hecker A (2019) Functional, structural and biochemical features of plant serinyl-glutathione transferases. Front Plant Sci 10:608. https://doi.org/10.3389/Fpls.2019.00608
Szklarczyk D, Gable AL, Lyon D, Junge A, Wyder S, Huerta-Cepas J, Simonovic M, Doncheva NT, Morris JH, Bork P, Jensen LJ, Mering C (2019) STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res 47:D607–D613. https://doi.org/10.1093/nar/gky1131
Tang HB, Bowers JE, Wang XY, Ming R, Alam M, Paterson AH (2008) Perspective - synteny and collinearity in plant genomes. Science 320:486–488. https://doi.org/10.1126/science.1153917
Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680. https://doi.org/10.1093/nar/22.22.4673
Wang T, Zhang WH (2020) Agrobacterium rhizogenes-mediated transformation of hairy roots in Medicago ruthenica. Acta Agrestia Sinica 28:268–272. https://doi.org/10.11733/j.issn.1007-0435.2020.01.032
Wang T, Ren L, Li C, Zhang D, Zhang X, Zhou G, Gao D, Chen R, Chen Y, Wang Z, Shi F, Farmer AD, Li Y, Zhou M, Young ND, Zhang W-H (2021) The genome of a wild Medicago species provides insights into the tolerant mechanisms of legume forage to environmental stress. BMC Biol 19:96. https://doi.org/10.1186/s12915-021-01033-0
Yang Q, Han XM, Gu JK, Liu YJ, Yang MJ, Zeng QY (2019a) Functional and structural profiles of GST gene family from three Populus species reveal the sequence-function decoupling of orthologous genes. New Phytol 221:1060–1073. https://doi.org/10.1111/nph.15430
Yang Q, Liu YJ, Zeng QY (2019b) Overexpression of three orthologous glutathione S-transferases from Populus increased salt and drought resistance in Arabidopsis. Biochem Syst Ecol 83:57–61. https://doi.org/10.1016/j.bse.2019.01.001
Zhang W, Hou L, Yang J, Song S, Mao X, Zhang Q, Bai W, Pan Q, Zhou Q (2018) Establishment and management of alfalfa pasture in cold regions of China. Chin Sci Bull 63:1651–1663. https://doi.org/10.1360/N972017-01181
Acknowledgements
We thank Dr. Hao Lin (Chinese Academy of Agricultural Sciences) for providing pFGC5941-Cas9 vector of Crispr/Cas9 system. We thank Dr. Mengyan Zhou (Novogene Bioinformatics Institute) for helping us perform the bioinformatics analysis. We thank Dr. Xiuzhi Xia (Chinese Academy of Agricultural Sciences) for providing the advice for hairy root transformation.
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The research was supported by the National Natural Science Foundation of China (32070351, 31830011) and the Science and Technology Program of Inner Mongolia, China (2021GG0372).
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TW conceived the research and designed the experiments. TW and DZ performed the experiments. TW, LC and JW analyzed the data. TW and WHZ wrote the manuscript. All authors discussed the results and approved the final manuscript.
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Wang, T., Zhang, D., Chen, L. et al. Genome-wide analysis of the Glutathione S-Transferase family in wild Medicago ruthenica and drought-tolerant breeding application of MruGSTU39 gene in cultivated alfalfa. Theor Appl Genet 135, 853–864 (2022). https://doi.org/10.1007/s00122-021-04002-x
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DOI: https://doi.org/10.1007/s00122-021-04002-x