Abstract
Key message
Overexpression of FvC5SD improves drought tolerance in soybean.
Abstract
Drought stress is one of the most important abiotic stress factors that influence soybean crop quality and yield. Therefore, the creation of drought-tolerant soybean germplasm resources through genetic engineering technology is effective in alleviating drought stress. FvC5SD is a type of C-5 sterol desaturase gene that is obtained from the edible fungus Flammulina velutipes. This gene has good tolerance to the effects of stresses, including drought and low temperature, in yeast cells and tomato. In this study, we introduced the FvC5SD gene into the soybean variety Shennong9 through the Agrobacterium-mediated transformation of soybean to identify drought-tolerant transgenic soybean varieties. PCR, RT-PCR, and Southern blot analysis results showed that T-DNA was inserted into the soybean genome and stably inherited by the progeny. The ectopic expression of FvC5SD under the control of a CaMV 35S promoter in transgenic soybean plants enhanced the plant’s tolerance to dehydration and drought. Under drought conditions, the transgenic plants accumulated lower levels of reactive oxygen species and exhibited higher activities and expression levels of enzymes and cell than wild-type soybean. iTRAQ analysis of the comparative proteomics showed that some exogenous genes coding either functional or regulatory proteins were induced in the transgenic lines under drought stress. FvC5SD overexpression can serve as a direct and efficient target in improving drought tolerance in soybean and may be an important biotechnological strategy for trait improvement in soybean and other crops.
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References
Aharoni A, Dixit S, Jetter R, Thoenes E, van Arkel G, Pereira A (2004) The SHINE clade of AP2 domain transcription factors activates wax biosynthesis, alters cuticle properties, and confers drought tolerance when overexpressed in Arabidopsis. Plant Cell 16:2463–2480. https://doi.org/10.1105/tpc.104.022897
Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in beta vulgaris. Plant Physiol 24:1–15
Attitalla IH (2011) Modified CTAB method for high quality genomic DNA extraction from medicinal plants. Pak J Biol Sci 14:998–999
Berkowitz GA (1987) Chloroplast acclimation to low osmotic potential. Plant Cell Rep 6:208–211. https://doi.org/10.1007/BF00268481
Bourdenx B et al (2011) Overexpression of Arabidopsis ECERIFERUM1 promotes wax very-long-chain alkane biosynthesis and influences plant response to biotic and abiotic stresses. Plant Physiol 156:29–45. https://doi.org/10.1104/pp.111.172320
Darnet S, Rahier A (2004) Plant sterol biosynthesis: identification of two distinct families of sterol 4alpha-methyl oxidases. Biochem J 378:889–898. https://doi.org/10.1042/bj20031572
de Paiva Rolla AA et al (2014) Phenotyping soybean plants transformed with rd29A:AtDREB1A for drought tolerance in the greenhouse and field. Transgenic Res 23:75–87. https://doi.org/10.1007/s11248-013-9723-6
Deshmukh R et al (2014) Integrating omic approaches for abiotic stress tolerance in soybean. Front Plant Sci 5:244. https://doi.org/10.3389/fpls.2014.00244
Edwards K, Johnstone C, Thompson C (1991) A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Res 19:1349
Funke T, Han H, Healy-Fried ML, Fischer M, Schonbrunn E (2006) Molecular basis for the herbicide resistance of Roundup Ready crops. Proc Natl Acad Sci USA 103:13010–13015. https://doi.org/10.1073/pnas.0603638103
Gerhardt R, Stitt M, Heldt HW (1987) Subcellular metabolite levels in spinach leaves: regulation of sucrose synthesis during diurnal alterations in photosynthetic partitioning. Plant Physiol 83:399–407
Guttikonda SK et al (2014) Overexpression of AtDREB1D transcription factor improves drought tolerance in soybean. Mol Biol Rep 41:7995–8008. https://doi.org/10.1007/s11033-014-3695-3
Irigoyen J et al (1992) Water stress induced changes in concentrations of proline and total soluble sugars in nodulated alfalfa (Medicago sativa) plants. Physiol Plant 84:55–60
Islam MA, Du H, Ning J, Ye H, Xiong L (2009) Characterization of Glossy1-homologous genes in rice involved in leaf wax accumulation and drought resistance. Plant Mol Biol 70:443–456. https://doi.org/10.1007/s11103-009-9483-0
Kamthan A, Kamthan M, Azam M, Chakraborty N, Chakraborty S, Datta A (2012) Expression of a fungal sterol desaturase improves tomato drought tolerance, pathogen resistance and nutritional quality. Sci Rep 2:951. https://doi.org/10.1038/srep00951
Kamthan A, Chaudhuri A, Kamthan M, Datta A (2016) Genetically modified (GM) crops: milestones and new advances in crop improvement. Theor Appl Genet 129:1639–1655. https://doi.org/10.1007/s00122-016-2747-6
Kamthan A, Kamthan M, Datta A (2017) Expression of C-5 sterol desaturase from an edible mushroom in fisson yeast enhances its ethanol and thermotolerance. PLoS One 12:e0173381. https://doi.org/10.1371/journal.pone.0173381
Kim HJ et al (2018a) Confirmation of drought tolerance of ectopically expressed AtABF3 gene in Soybean. Mol Cells 41:413–422. https://doi.org/10.14348/molcells.2018.2254
Kim HJ et al (2018b) Erratum to: confirmation of drought tolerance of ectopically expressed AtABF3 gene in Soybean. Mol Cells 41:703. https://doi.org/10.14348/molcells.2018.1254
Le Gall H, Philippe F, Domon JM, Gillet F, Pelloux J, Rayon C (2015) Cell wall metabolism in response to abiotic stress. Plants (Basel) 4:112–166. https://doi.org/10.3390/plants4010112
Lee HJ et al (2013) Overexpression of the glutamine synthetase gene modulates oxidative stress response in rice after exposure to cadmium stress. Plant Cell Rep 32:1521–1529. https://doi.org/10.1007/s00299-013-1464-8
Leite JP et al (2014) Overexpression of the activated form of the AtAREB1 gene (AtAREB1DeltaQT) improves soybean responses to water deficit. Genet Mol Res 13:6272–6286. https://doi.org/10.4238/2014.august.15.10
Li Y et al (2013) Expression of an Arabidopsis molybdenum cofactor sulphurase gene in soybean enhances drought tolerance and increases yield under field conditions. Plant Biotechnol J 11:747–758. https://doi.org/10.1111/pbi.12066
Li Y et al (2017) Overexpression of GmFDL19 enhances tolerance to drought and salt stresses in soybean. PLoS One 12:e0179554. https://doi.org/10.1371/journal.pone.0179554
Manavalan LP, Guttikonda SK, Tran LS, Nguyen HT (2009) Physiological and molecular approaches to improve drought resistance in soybean. Plant Cell Physiol 50:1260–1276. https://doi.org/10.1093/pcp/pcp082
Mansour MMF, Ali EF (2017) Evaluation of proline functions in saline conditions. Phytochemistry 140:52–68. https://doi.org/10.1016/j.phytochem.2017.04.016
Miflin BJ, Habash DZ (2002) The role of glutamine synthetase and glutamate dehydrogenase in nitrogen assimilation and possibilities for improvement in the nitrogen utilization of crops. J Exp Bot 53:979–987
Miyazaki Y et al (1999) Cloning, sequencing, expression and allelic sequence diversity of ERG3 (C-5 sterol desaturase gene) in Candida albicans. Gene 236:43–51
Ogata T, Nagatoshi Y, Yamagishi N, Yoshikawa N, Fujita Y (2017) Virus-induced down-regulation of GmERA1A and GmERA1B genes enhances the stomatal response to abscisic acid and drought resistance in soybean. PLoS One 12:e0175650. https://doi.org/10.1371/journal.pone.0175650
Paz MM, Martinez JC, Kalvig AB, Fonger TM, Wang K (2006) Improved cotyledonary node method using an alternative explant derived from mature seed for efficient Agrobacterium-mediated soybean transformation. Plant Cell Rep 25:206–213. https://doi.org/10.1007/s00299-005-0048-7
Rodrigues NF, Fonseca GC, Kulcheski FR, Margis R (2017) Salt stress affects mRNA editing in soybean chloroplasts. Genet Mol Biol 40:200–208. https://doi.org/10.1590/1678-4685-GMB-2016-0055
Suutari M, Liukkonen K, Laakso S (1990) Temperature adaptation in yeasts: the role of fatty acids. J Gen Microbiol 136:1469–1474. https://doi.org/10.1099/00221287-136-8-1469
Taton M, Rahier A (1996) Plant sterol biosynthesis: identification and characterization of higher plant delta 7-sterol C5(6)-desaturase. Arch Biochem Biophys 325:279–288. https://doi.org/10.1006/abbi.1996.0035
Thao NP, Tran LS (2012) Potentials toward genetic engineering of drought-tolerant soybean. Crit Rev Biotechnol 32:349–362. https://doi.org/10.3109/07388551.2011.643463
Valente MA et al (2009) The ER luminal binding protein (BiP) mediates an increase in drought tolerance in soybean and delays drought-induced leaf senescence in soybean and tobacco. J Exp Bot 60:533–546. https://doi.org/10.1093/jxb/ern296
Valliyodan B, Ye H, Song L, Murphy M, Shannon JG, Nguyen HT (2017) Genetic diversity and genomic strategies for improving drought and waterlogging tolerance in soybeans. J Exp Bot 68:1835–1849. https://doi.org/10.1093/jxb/erw433
van Leeuwen M, Kremens RL, van Aardt J (2015) Tracking diurnal variation in photosynthetic down-regulation using low cost spectroscopic instrumentation. Sensors (Basel) 15:10616–10630. https://doi.org/10.3390/s150510616
Weber RL et al (2014) Expression of an osmotin-like protein from Solanum nigrum confers drought tolerance in transgenic soybean. BMC Plant Biol 14:343. https://doi.org/10.1186/s12870-014-0343-y
Yamada T, Takagi K, Ishimoto M (2012) Recent advances in soybean transformation and their application to molecular breeding and genomic analysis. Breed Sci 61:480–494. https://doi.org/10.1270/jsbbs.61.480
Yang L, Tang R, Zhu J, Liu H, Mueller-Roeber B, Xia H, Zhang H (2008) Enhancement of stress tolerance in transgenic tobacco plants constitutively expressing AtIpk2beta, an inositol polyphosphate 6-/3-kinase from Arabidopsis thaliana. Plant Mol Biol 66:329–343. https://doi.org/10.1007/s11103-007-9267-3
Zhang JY, Broeckling CD, Blancaflor EB, Sledge MK, Sumner LW, Wang ZY (2005) Overexpression of WXP1, a putative Medicago truncatula AP2 domain-containing transcription factor gene, increases cuticular wax accumulation and enhances drought tolerance in transgenic alfalfa (Medicago sativa). Plant J 42:689–707. https://doi.org/10.1111/j.1365-313x.2005.02405.x
Zhang XX et al (2013) OsDREB2A, a rice transcription factor, significantly affects salt tolerance in transgenic soybean. PLoS One 8:e83011. https://doi.org/10.1371/journal.pone.0083011
Zhao C, Haigh AM, Holford P, Chen ZH (2018) Roles of chloroplast retrograde signals and ion transport in plant drought tolerance. Int J Mol Sci. https://doi.org/10.3390/ijms19040963
Acknowledgements
This research work was supported by the Agricultural Science and Technology Innovation Project of Jilin Province (CXGC2017ZY02), the National Natural Science Foundation of China (31501327), and the Ministry of Agriculture of China for Transgenic Research (2016ZX08004-004).
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Zhang, L., Li, T., Wang, Y. et al. FvC5SD overexpression enhances drought tolerance in soybean by reactive oxygen species scavenging and modulating stress-responsive gene expression. Plant Cell Rep 38, 1039–1051 (2019). https://doi.org/10.1007/s00299-019-02424-y
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DOI: https://doi.org/10.1007/s00299-019-02424-y