Ectopic expression of secretory peptide PdEPF3 in Arabidopsis confers drought tolerance with reduced stomatal density

Sha Liu, Fuli Jia, Zhiyin Jiao, Jingjing Wang, Xinli Xia, Weilun Yin

Abstract


Drought is one of the most prevalent environmental stresses that affect plant growth and development. Improvement in drought tolerance is associated with reduced stomatal density and higher water use efficiency (WUE). In this study, an epidermal patterning factor (EPF), PdEPF3, from a fast-growing poplar clone, NE-19 [Populus nigra × (Populus deltoides × Populus nigra)], was characterized. Quantitative reverse transcription polymerase chain reaction showed that the transcription of PdEPF3 was induced by drought. We further found that the transgenic Arabidopsis overexpressing PdEPF3 had an earlier seedling germination and longer primary roots under osmotic stress treatments, compared with the WT and mutant epf1-1. In addition, ectopic overexpression of PdEPF3 in Arabidopsis plants was able to enhance drought tolerance. This observation was associated with the reduced stomatal density of transgenic plants, which would limit transpiration and reduce water loss, consequently improving the WUE of plants. Interestingly, the reduction of stomatal density in transgenic plants overexpressing PdEPF3 did not affect their photosynthetic capacity. These results indicate that PdEPF3 could be used in transgenic breeding to enhance plant drought tolerance.

Keywords


drought; PdEPF3; stomatal density; water use efficiency (WUE)

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References


Gleick PH. The world’s water: the biennial report of fresh water resources. Washington, DC: Island Press; 1998.

Yu H, Chen X, Hong YY, Wang Y, Xu P, Ke SD, et al. Activated expression of an Arabidopsis HD-START protein confers drought tolerance with improved root system and reduced stomatal density. Plant Cell. 2008;20(4):1134–1151. https://doi.org/10.1105/tpc.108.058263

Zhangbin Z, Ping X, Hongbo S, Mengjun L, Zhenyan F, Liye C. Advances and prospects: biotechnologically improving crop water use efficiency. Crit Rev Biotechnol. 2011;31(3):281–293. https://doi.org/10.3109/07388551.2010.531004

Todaka D, Nakashima K, Shinozaki K, Yamaguchi-Shinozaki K. Toward understanding transcriptional regulatory networks in abiotic stress responses and tolerance in rice. Rice. 2012;5(1):6. https://doi.org/10.1186/1939-8433-5-6

Nadeau JA, Sack FD. Stomatal development in Arabidopsis. In: Somerville CR, Meyerowitz EM, editors. The Arabidopsis book. Rockville, MD: American Society of Plant Biologists; 2002. https://doi.org/10.1199/tab.0066

Xiong L, Schumaker KS, Zhu JK. Cell signaling during cold, drought, and salt stress. Plant Cell. 2002;14(suppl 1):S165–S183. https://doi.org/10.1105/tpc.000596

Farooq M, Wahid A, Lee DJ. Exogenously applied polyamines increase drought tolerance of rice by improving leaf water status, photosynthesis and membrane properties. Acta Physiol Plant. 2009;31(5):937–945. https://doi.org/10.1007/s11738-009-0307-2

Masle J, Gilmore SR, Farquhar GD. The ERECTA gene regulates plant transpiration efficiency in Arabidopsis. Nature. 2005;436(7052):866. https://doi.org/10.1038/nature03835

Yoo CY, Pence HE, Jin JB, Miura K, Gosney MJ, Hasegawa PM, et al. The Arabidopsis GTL1 transcription factor regulates water use efficiency and drought tolerance by modulating stomatal density via transrepression of SDD1. Plant Cell. 2010;22(12):4128–4141. https://doi.org/10.1105/tpc.110.078691

Franks PJ, W Doheny‐Adams T, Britton‐Harper ZJ, Gray JE. Increasing water‐use efficiency directly through genetic manipulation of stomatal density. New Phytol. 2015;207(1):188–195. https://doi.org/10.1111/nph.13347

Hepworth C, Doheny‐Adams T, Hunt L, Cameron DD, Gray JE. Manipulating stomatal density enhances drought tolerance without deleterious effect on nutrient uptake. New Phytol. 2015;208(2):336–341. https://doi.org/10.1111/nph.13598

Wang C, Liu S, Dong Y, Zhao Y, Geng A, Xia X, et al. PdEPF1 regulates water‐use efficiency and drought tolerance by modulating stomatal density in poplar. Plant Biotechnol J. 2016;14(3):849–860. https://doi.org/10.1111/pbi.12434

Li Y, Li H, Li Y, Zhang S. Improving water-use efficiency by decreasing stomatal conductance and transpiration rate to maintain higher ear photosynthetic rate in drought-resistant wheat. Crop J. 2017;5(3):231–239. https://doi.org/10.1016/j.cj.2017.01.001

Hughes J, Hepworth C, Dutton C, Dunn JA, Hunt L, Stephens J, et al. Reducing stomatal density in barley improves drought tolerance without impacting on yield. Plant Physiol. 2017;174(2):776–787. https://doi.org/10.1104/pp.16.01844

Sachs T. Pattern formation in plant tissues. New York, NY: Cambridge University Press; 1991. https://doi.org/10.1017/CBO9780511574535

Geisler M, Nadeau J, Sack FD. Oriented asymmetric divisions that generate the stomatal spacing pattern in Arabidopsis are disrupted by the too many mouths mutation. Plant Cell. 2000;12(11):2075–2086. https://doi.org/10.1105/tpc.12.11.2075

Sugano SS, Shimada T, Imai Y, Okawa K, Tamai A, Mori M, et al. Stomagen positively regulates stomatal density in Arabidopsis. Nature. 2010;463(7278):241–244. https://doi.org/10.1038/nature08682

Lee JS, Kuroha T, Hnilova M, Khatayevich D, Kanaoka M, McAbee JM, et al. Direct interaction of ligand–receptor pairs specifying stomatal patterning. Genes Dev. 2012;26(2):126–136. https://doi.org/10.1101/gad.179895.111

Lin G, Zhang L, Han Z, Yang X, Liu W, Li E, et al. A receptor-like protein acts as a specificity switch for the regulation of stomatal development. Genes Dev. 2017;31(9):927–938. https://doi.org/10.1101/gad.297580.117

Liu S, Wang C, Jia F, An Y, Liu C, Xia X, et al. Secretory peptide PdEPF2 enhances drought tolerance by modulating stomatal density and regulates ABA response in transgenic Arabidopsis thaliana. Plant Cell Tissue Organ Cult. 2016;125(3):419–431. https://doi.org/10.1007/s11240-016-0957-x

Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue culture. Physiol Plant. 1962;15(3):473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x

Chaves MM, Maroco JP, Pereira JS. Understanding plant responses to drought – from genes to the whole plant. Funct Plant Biol. 2003;30(3):239–264. https://doi.org/10.1071/FP02076

Zhang X, Henriques R, Lin SS, Niu QW, Chua NH. Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method. Nat Protoc. 2006;1(2):641. https://doi.org/10.1038/nprot.2006.97

Chen J, Xia X, Yin W. Expression profiling and functional characterization of a DREB2-type gene from Populus euphratica. Biochem Biophys Res Commun. 2009;378(3):483–487. https://doi.org/10.1016/j.bbrc.2008.11.071

Han X, Tang S, An Y, Zheng DC, Xia XL, Yin WL. Overexpression of the poplar NF-YB7 transcription factor confers drought tolerance and improves water-use efficiency in Arabidopsis. J Exp Bot. 2013;64(14):4589–4601. https://doi.org/10.1093/jxb/ert262

Hetherington AM, Woodward FI. The role of stomata in sensing and driving environmental change. Nature. 2003;424(6951):901. https://doi.org/10.1038/nature01843

Supratim B, Venkategowda R, Anuj K, Andy P. Plant adaptation to drought stress:. F1000Res. 2016;5:1554. https://doi.org/10.12688/f1000research.7678.1

Woodward FI. Stomatal numbers are sensitive to increases in CO2 from pre-industrial levels. Nature. 1987;327(6123):617–618. https://doi.org/10.1038/327617a0

Tanaka Y, Sugano SS, Shimada T, Hara-Nishimura I. Enhancement of leaf photosynthetic capacity through increased stomatal density in Arabidopsis. New Phytol. 2013;198(3):757–764. https://doi.org/10.1111/nph.12186

Sharp RE, Davies WJ. Root growth and water uptake by maize plants in drying soil. J Exp Bot. 1985;36(9):1441–1456. https://doi.org/10.1093/jxb/36.9.1441

Zhao W, Sun Y, Kjelgren R, Liu X. Response of stomatal density and bound gas exchange in leaves of maize to soil water deficit. Acta Physiol Plant. 2015;37(1):1704. https://doi.org/10.1007/s11738-014-1704-8

Hara K, Kajita R, Torii KU, Bergmann DC, Kakimoto T. The secretory peptide gene EPF1 enforces the stomatal one-cell-spacing rule. Genes Dev. 2007;21(14):1720–1725. https://doi.org/10.1101/gad.1550707

Brodribb TJ, McAdam SAM, Jordan GJ, Field TS. Evolution of stomatal responsiveness to CO2 and optimization of water‐use efficiency among land plants. New Phytol. 2009;183(3):839–847. https://doi.org/10.1111/j.1469-8137.2009.02844.x

Franks PJ, Beerling DJ. Maximum leaf conductance driven by CO2 effects on stomatal size and density over geologic time. Proc Natl Acad Sci USA. 2009;106(25):10343–10347. https://doi.org/10.1073/pnas.0904209106

Yu L, Chen X, Wang Z, Wang Y, Zhu Q, Li S, et al. Arabidopsis Enhanced Drought Tolerance1/HOMEODOMAIN GLABROUS11 confers drought tolerance in transgenic rice without yield penalty. Plant Physiol. 2013;162(3):1378–1391. https://doi.org/10.1104/pp.113.217596