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Overexpression of LOV KELCH PROTEIN 2 confers dehydration tolerance and is associated with enhanced expression of dehydration-inducible genes in Arabidopsis thaliana

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Abstract

Key message

The overexpression of LKP2 confers dehydration tolerance in Arabidopsis thaliana ; this is likely due to enhanced expression of dehydration-inducible genes and reduced stomatal opening.

Abstract

LOV KELCH PROTEIN 2 (LKP2) modulates the circadian rhythm and flowering time in plants. In this study, we observed that LKP2 overexpression enhanced dehydration tolerance in Arabidopsis. Microarray analysis demonstrated that expression of water deprivation-responsive genes was higher in the absence of dehydration stress in transgenic Arabidopsis plants expressing green fluorescent protein–tagged LKP2 (GFP-LKP2) than in control transgenic plants expressing GFP. After dehydration followed by rehydration, GFP-LKP2 plants developed more leaves and roots and exhibited higher survival rates than control plants. In the absence of dehydration stress, four dehydration-inducible genes, namely DREB1A, DREB1B, DREB1C, and RD29A, were expressed in GFP-LKP2 plants, whereas they were not expressed or were expressed at low levels in control plants. Under dehydration stress, the expression of DREB2B and RD29A peaked faster in the GFP-LKP2 plants than in control plants. The stomatal aperture of GFP-LKP2 plants was smaller than that of control plants. These results suggest that the dehydration tolerance of GFP-LKP2 plants is caused by upregulation of DREB1A–C/CBF1–3 and their downstream targets; restricted stomatal opening in the absence of dehydration stress also appears to contribute to the phenotype. The rapid and high expression of DREB2B and its downstream target genes also likely accounts for some features of the GFP-LKP2 phenotype. Our results suggest that LKP2 can be used for biotechnological applications not only to adjust the flowering time control but also to enhance dehydration tolerance.

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References

  • Abe H, Yamaguchi-Shinozaki K, Urao T, Iwasaki T, Hosokawa D, Shinozaki K (1997) Role of arabidopsis MYC and MYB homologs in drought- and abscisic acid-regulated gene expression. Plant Cell 9:1859–1868

    PubMed Central  CAS  PubMed  Google Scholar 

  • Abe H, Urao T, Ito T, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Arabidopsis AtMYC2 (bHLH) and AtMYB2 (MYB) function as transcriptional activators in abscisic acid signaling. Plant Cell 15:63–78

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Abe M, Kobayashi Y, Yamamoto S, Daimon Y, Yamaguchi A, Ikeda Y, Ichinoki H, Notaguchi M, Goto K, Araki T (2005) FD, a bZIP protein mediating signals from the floral pathway integrator FT at the shoot apex. Science 309:1052–1056

    Article  CAS  PubMed  Google Scholar 

  • Ando E, Ohnishi M, Wang Y, Matsushita T, Watanabe A, Hayashi Y, Fujii M, Ma JF, Inoue S, Kinoshita T (2013) TWIN SISTER OF FT, GIGANTEA, and CONSTANS have a positive but indirect effect on blue light-induced stomatal opening in Arabidopsis. Plant Physiol 162:1529–1538

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Baudry A, Ito S, Song YH, Strait AA, Kiba T, Lu S, Henriques R, Pruneda-Paz JL, Chua NH, Tobin EM, Kay SA, Imaizumi T (2010) F-box proteins FKF1 and LKP2 act in concert with ZEITLUPE to control Arabidopsis clock progression. Plant Cell 22:606–622

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Chen M, Chory J (2011) Phytochrome signaling mechanisms and the control of plant development. Trends Cell Biol 21:664–671

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Covington MF, Maloof JN, Straume M, Kay SA, Harmer SL (2008) Global transcriptome analysis reveals circadian regulation of key pathways in plant growth and development. Genome Biol 9:R130

    Article  PubMed Central  PubMed  Google Scholar 

  • Dodd AN, Salathia N, Hall A, Kévei E, Tóth R, Nagy F, Hibberd JM, Millar AJ, Webb AA (2005) Plant circadian clocks increase photosynthesis, growth, survival, and competitive advantage. Science 309:630–633

    Article  CAS  PubMed  Google Scholar 

  • Dong MA, Farré EM, Thomashow MF (2011) Circadian clock-associated 1 and late elongated hypocotyl regulate expression of the C-repeat binding factor (CBF) pathway in Arabidopsis. Proc Natl Acad Sci USA 108:7241–7246

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Fowler S, Thomashow MF (2002) Arabidopsis transcriptome profiling indicates that multiple regulatory pathways are activated during cold acclimation in addition to the CBF cold response pathway. Plant Cell 14:1675–1690

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Franklin K, Quail PH (2010) Phytochrome functions in Arabidopsis development. J Exp Bot 61:11–24

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Fujita M, Fujita Y, Maruyama K, Seki M, Hiratsu K, Ohme-Takagi M, Tran LS, Yamaguchi-Shinozaki K, Shinozaki K (2004) A dehydration-induced NAC protein, RD26, is involved in a novel ABA-dependent stress-signaling pathway. Plant J 39:863–876

    Article  CAS  PubMed  Google Scholar 

  • Fujita Y, Fujita M, Satoh R, Maruyama K, Parvez MM, Seki M, Hiratsu K, Ohme-Takagi M, Shinozaki K, Yamaguchi-Shinozaki K (2005) AREB1 is a transcription activator of novel ABRE-dependent ABA signaling that enhances drought stress tolerance in Arabidopsis. Plant Cell 17:3470–3488

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • González CV, Ibarra SE, Piccoli PN, Botto JF, Boccalandro HE (2012) Phytochrome B increases drought tolerance by enhancing ABA sensitivity in Arabidopsis thaliana. Plant, Cell Environ 35:1958–1968

    Article  Google Scholar 

  • Imaizumi T, Tran HG, Swartz TE, Briggs WR, Kay SA (2003) FKF1 is essential for photoperiodic-specific light signalling in Arabidopsis. Nature 426:302–306

    Article  CAS  PubMed  Google Scholar 

  • Jarillo JA, Capel J, Tang RH, Yang HQ, Alonso JM, Ecker JR, Cashmore AR (2001) An Arabidopsis circadian clock component interacts with both CRY1 and phyB. Nature 410:487–490

    Article  CAS  PubMed  Google Scholar 

  • Kiba T, Henriques R, Sakakibara H, Chua NH (2007) Targeted degradation of PSEUDO-RESPONSE REGULATOR5 by an SCFZTL complex regulates clock function and photomorphogenesis in Arabidopsis thaliana. Plant Cell 19:2516–2530

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Kinoshita T, Ono N, Hayashi Y, Morimoto S, Nakamura S, Soda M, Kato Y, Ohnishi M, Nakano T, Inoue S, Shimazaki K (2011) FLOWERING LOCUS T regulates stomatal opening. Curr Biol 21:1232–1238

    Article  CAS  PubMed  Google Scholar 

  • Kiyosue T, Wada M (2000) LKP1 (LOV kelch protein 1): a factor involved in the regulation of flowering time in Arabidopsis. Plant J 23:807–815

    Article  CAS  PubMed  Google Scholar 

  • Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki K, Shinozaki K (1998) Two transcription factors, DREB1 and DREB2, with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought- and low-temperature-responsive gene expression, respectively, in Arabidopsis. Plant Cell 10:1391–1406

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Más P, Kim WY, Somers DE, Kay SA (2003) Targeted degradation of TOC1 by ZTL modulates circadian function in Arabidopsis thaliana. Nature 426:567–570

    Article  PubMed  Google Scholar 

  • Michael TP, Breton G, Hazen SP, Priest H, Mockler TC, Kay SA, Chory J (2008) A morning-specific phytohormone gene expression program underlying rhythmic plant growth. PLoS Biol 6:e225

    Article  PubMed Central  PubMed  Google Scholar 

  • Miyazaki Y, Yoshizumi T, Takase T, Matsui M, Kiyosue T (2011) Overexpression of LOV KELCH PROTEIN 2 enhances cell elongation and increases cell number and ploidy in the hypocotyl of Arabidopsis thaliana. Plant Biotech 28:267–272

    Article  CAS  Google Scholar 

  • Mockler TC, Michael TP, Priest HD, Shen R, Sullivan CM, Givan SA, McEntee C, Kay SA, Chory J (2007) The DIURNAL project: DIURNAL and circadian expression profiling, model-based pattern matching, and promoter analysis. Cold Spring Harb Symp Quant Biol 72:353–363

    Article  CAS  PubMed  Google Scholar 

  • Nakamichi N, Kusano M, Fukushima A, Kita M, Ito S, Yamashino T, Saito K, Sakakibara H, Mizuno T (2009) Transcript profiling of an Arabidopsis PSEUDO RESPONSE REGULATOR arrhythmic triple mutant reveals a role for the circadian clock in cold stress response. Plant Cell Physiol 50:447–462

    Article  CAS  PubMed  Google Scholar 

  • Nakamichi N, Kiba T, Kamioka M, Suzuki T, Yamashino T, Higashiyama T, Sakakibara H, Mizuno T (2012) Transcriptional repressor PRR5 directly regulates clock-output pathways. Proc Natl Acad Sci USA 109:17123–17128

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Nelson DC, Lasswell J, Rogg LE, Cohen MA, Bartel B (2000) FKF1, a clock-controlled gene that regulates the transition to flowering in Arabidopsis. Cell 101:331–340

    Article  CAS  PubMed  Google Scholar 

  • Oh S, Warnasooriya SN, Montgomery BL (2014) Mesophyll-localized phytochromes gate stress- and light-inducible anthocyanin accumulation in Arabidopsis thaliana. Plant Signal Behav 9:e28013

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Qin F, Shinozaki K, Yamaguchi-Shinozaki K (2011) Achievements and challenges in understanding plant abiotic stress responses and tolerance. Plant Cell Physiol 52:1569–1582

    Article  CAS  PubMed  Google Scholar 

  • Saibo NJ, Lourenço T, Oliveira MM (2009) Transcription factors and regulation of photosynthetic and related metabolism under environmental stresses. Ann Bot 103:609–623

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Sakuma Y, Maruyama K, Osakabe Y, Qin F, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2006) Functional analysis of an Arabidopsis transcription factor, DREB2A, involved in drought-responsive gene expression. Plant Cell 18:1292–1309

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Schultz TF, Kiyosue T, Yanovsky M, Wada M, Kay SA (2001) A role for LKP2 in the circadian clock of Arabidopsis. Plant Cell 13:2659–2670

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Somers DE, Schultz TF, Milnamow M, Kay SA (2000) ZEITLUPE encodes a novel clock-associated PAS protein from Arabidopsis. Cell 101:319–329

    Article  CAS  PubMed  Google Scholar 

  • Somers DE, Kim WY, Geng R (2004) The F-box protein ZEITLUPE confers dosage-dependent control on the circadian clock, photomorphogenesis, and flowering time. Plant Cell 16:769–782

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Suetsugu N, Wasda M (2013) Evolution of three LOV blue light receptor families in green plants and photosynthetic stramenopiles: Phototropin, ZTL/FKF1/LKP2 and aureochrome. Plant Cell Physiol 54:8–23

    Article  CAS  PubMed  Google Scholar 

  • Takase T, Nishiyama Y, Tanihigashi H, Ogura Y, Miyazaki Y, Yamada Y, Kiyosue T (2011) LOV KELCH PROTEIN2 and ZEITLUPE repress Arabidopsis photoperiodic flowering under non-inductive conditions, dependent on FLAVIN-BINDING KELCH REPEAT F-BOX1. Plant J 67:608–621

    Article  CAS  PubMed  Google Scholar 

  • Tran LS, Nakashima K, Sakuma Y, Simpson SD, Fujita Y, Maruyama K, Fujita M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2004) Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cis-element in the early responsive to dehydration stress 1 promoter. Plant Cell 16:2481–2498

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Uno Y, Furihata T, Abe H, Yoshida R, Shinozaki K, Yamaguchi-Shinozaki K (2000) Arabidopsis basic leucine zipper transcription factors involved in an abscisic acid-dependent signal transduction pathway under drought and high-salinity conditions. Proc Natl Acad Sci USA 97:11632–11637

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Urao T, Yamaguchi-Shinozaki K, Urao S, Shinozaki K (1993) An Arabidopsis myb homolog is induced by dehydration stress and its gene product binds to the conserved MYB recognition sequence. Plant Cell 5:1529–1539

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Wang L, Fujiwara S, Somers DE (2010) PRR5 regulates phosphorylation, nuclear import and subnuclear localization of TOC1 in the Arabidopsis circadian clock. EMBO J 29:1903–1915

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Xoconostle-Cázares B, Ramírez-Ortega FA, Flores-Elenes L, Ruiz-Medrano R (2010) Drought tolerance in crop plants. American J Plant Physiol 5:241–256

    Article  Google Scholar 

  • Yamaguchi-Shinozaki K, Shinozaki K (2006) Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses. Annu Rev Plant Biol 57:781–803

    Article  CAS  PubMed  Google Scholar 

  • Yasuhara M, Mitsui S, Hirano H, Takanabe R, Tokioka Y, Ihara N, Komatsu A, Seki M, Shinozaki K, Kiyosue T (2004) Identification of ASK and clock-associated proteins as molecular partners of LKP2 (LOV kelch protein 2) in Arabidopsis. J Exp Bot 55:2015–2027

    Article  CAS  PubMed  Google Scholar 

  • Yoshida T, Fujita Y, Sayama H, Kidokoro S, Maruyama K, Mizoi J, Shinozaki K, Yamaguchi-Shinozaki K (2010) AREB1, AREB2, and ABF3 are master transcription factors that cooperatively regulate ABRE-dependent ABA signaling involved in drought stress tolerance and require ABA for full activation. Plant J 61:672–685

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We thank Dr. K. Shimazaki (Kyushu University) and Dr. T. Kinoshita (Nagoya University) for their suggestions on the measurement of stomatal aperture. This research was partially supported by a fund from Ministry of Education, Culture, Sports, Science and Technology of Japan.

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Life Science, Faculty of Science, Gakushuin University, 1-5-1 Mejiro, Toshima-ku, Tokyo, 171-8588, Japan

    Yuji Miyazaki, Tomoyuki Takase & Tomohiro Kiyosue

  2. Experimental Plant Division, Department of Biological Systems, RIKEN BioResource Center, 3-1-1 Koyadai, Tsukuba, 305-0074, Ibaraki, Japan

    Hiroshi Abe & Masatomo Kobayashi

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  1. Yuji Miyazaki
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Correspondence to Tomohiro Kiyosue.

Additional information

Communicated by P. Lakshmanan.

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299_2015_1746_MOESM1_ESM.tif

Supplemental Fig. S1 Sixteen-day-old Arabidopsis plants (GFP-LKP2, lines 1 and 2; control, Col and GFP) grown in the absence of dehydration stress. (A) Plants grown on GM agar plates. (B) Leaf and root numbers of these plants (n = 10; mean ± SE). Different letters indicate statistical difference (p < 0.05, Student–Newman–Keuls test) (TIFF 2097 kb)

299_2015_1746_MOESM2_ESM.tif

Supplemental Fig. S2 Stomatal aperture of GFP-LKP2 (lines 1 and 2) and control (Col and GFP) plants at ZT20 under long-day (16-h light/8-h dark) conditions (n = 20; mean ± SE; p < 0.05, Student–Newman–Keuls test) (TIFF 101 kb)

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Miyazaki, Y., Abe, H., Takase, T. et al. Overexpression of LOV KELCH PROTEIN 2 confers dehydration tolerance and is associated with enhanced expression of dehydration-inducible genes in Arabidopsis thaliana . Plant Cell Rep 34, 843–852 (2015). https://doi.org/10.1007/s00299-015-1746-4

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