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Molecular cloning and functional analysis of a blue light receptor gene MdCRY2 from apple (Malus domestica)

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MdCRY2 was isolated from apple fruit skin, and its function was analyzed in MdCRY2 transgenic Arabidopsis. The interaction between MdCRY2 and AtCOP1 was found by yeast two-hybrid and BiFC assays.

Abstract

Cryptochromes are blue/ultraviolet-A (UV-A) light receptors involved in regulating various aspects of plant growth and development. Investigations of the structure and functions of cryptochromes in plants have largely focused on Arabidopsis (Arabidopsis thaliana), tomato (Solanum lycopersicum), pea (Pisum sativum), and rice (Oryza sativa). However, no data on the function of CRY2 are available in woody plants. In this study, we isolated a cryptochrome gene, MdCRY2, from apple (Malus domestica). The deduced amino acid sequences of MdCRY2 contain the conserved N-terminal photolyase-related domain and the flavin adenine dinucleotide (FAD) binding domain, as well as the C-terminal DQXVP-acidic-STAES (DAS) domain. Relationship analysis indicates that MdCRY2 shows the highest similarity to the strawberry FvCRY protein. The expression of MdCRY2 is induced by blue/UV-A light, which represents a 48-h circadian rhythm. To investigate the function of MdCRY2, we overexpressed the MdCRY2 gene in a cry2 mutant and wild type (WT) Arabidopsis, assessed the phenotypes of the resulting transgenic plants, and found that MdCRY2 functions to regulate hypocotyl elongation, root growth, flower initiation, and anthocyanin accumulation. Furthermore, we examined the interaction between MdCRY2 and AtCOP1 using a yeast two-hybrid assay and a bimolecular fluorescence complementation assay. These data provide functional evidence for a role of blue/UV-A light-induced MdCRY2 in controlling photomorphogenesis in apple.

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References

  • Ahmad M, Cashmore AR (1993) HY4 gene of A. thaliana encodes a protein with characteristics of a blue-light photoreceptor. Nature 366:162–166

    Article  CAS  PubMed  Google Scholar 

  • Allan AC, Hellens RP, Laing WA (2008) MYB transcription factors that colour our fruit. Trends Plant Sci 13:99–102

    Article  CAS  PubMed  Google Scholar 

  • Briggs WR, Christie JM (2002) Phototropins 1 and 2: versatile plant blue-light receptors. Trends Plant Sci 7(5):204–210

    Article  CAS  PubMed  Google Scholar 

  • Brudler R, Hitomi K, Daiyasu H, Toh H, Kucho K, Ishiura M, Kanehisa M, Roberts VA, Todo T, Tainer JA, Getzoff ED (2003) Identification of a new cryptochrome class. Structure, function, and evolution. Mol Cell 11:59–67

    Article  CAS  PubMed  Google Scholar 

  • Canamero RC, Bakrim N, Bouly JP, Garay A, Dudkin EE, Habricot Y, Ahmad M (2006) Cryptochrome photoreceptors cry1 and cry2 antagonistically regulate primary root elongation in Arabidopsis thaliana. Planta 224:995–1003

    Article  CAS  PubMed  Google Scholar 

  • Cashmore AR (2003) Cryptochromes: enabling plants and animals to determine circadian time. Cell 114:537–543

    Article  CAS  PubMed  Google Scholar 

  • Chatterjee M, Sharma P, Khurana JP (2006) Cryptochrome1 from Brassica napus is up-regulated by blue light and controls hypocotyl/stem growth and anthocyanin accumulation. Plant Physiol 141:61–74

    Article  CAS  PubMed  Google Scholar 

  • Christie JM, Arvai AS, Baxter KJ, Heilmann M, Pratt AJ, O’Hara A, Kelly SM, Hothorn M, Smith BO, Hitomi K, Jenkins KI, Getzoff ED (2012) Plant UVR8 photo-receptor senses UV-B by tryptophan-mediated disruption of cross-dimer salt bridges. Science 335:1492–1496

    Article  CAS  PubMed  Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    Article  CAS  PubMed  Google Scholar 

  • Correll MJ, Kiss JZ (2005) The roles of phytochromes in elongation and gravitropism of roots. Plant Cell Physiol 46(2):317–323

    Article  CAS  PubMed  Google Scholar 

  • Espley RV, Hellens RP, Putterill J, Stevenson DE, Kutty-Amma S, Allan AC (2007) Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10. Plant J 49:414–427

    Article  CAS  PubMed  Google Scholar 

  • Franklin KA, Larner VS, Whitelam GC (2005) The signal transducing photoreceptors of plants. Int J Dev Biol 49:653–664

    Article  CAS  PubMed  Google Scholar 

  • Guo H, Yang H, Mockler TC, Lin C (1998) Regulation of flowering time by Arabidopsis photoreceptors. Science 279:1360–1363

    Article  CAS  PubMed  Google Scholar 

  • Gyula P, Schafer E, Nagy F (2003) Light perception and signaling in higher plants. Curr Opin Plant Biol 6:446–452

    Article  CAS  PubMed  Google Scholar 

  • Hong SH, Kim HJ, Ryu JS, Choi H, Jeong S, Shin J, Choi G, Nam HG (2008) CRY1 inhibits COP1-mediated degradation of BIT1, a MYB transcription factor, to activate blue light-dependent gene expression in Arabidopsis. Plant J 55:361–371

    Article  CAS  PubMed  Google Scholar 

  • Jang S, Marchal V, Panigrahi KC, Wenkel S, Soppe W, Deng XW, Valverde F, Coupland G (2008) Arabidopsis COP1 shapes the temporal pattern of CO accumulation conferring a photoperiodic flowering response. EMBO J 27:1277–1288

    Article  CAS  PubMed  Google Scholar 

  • Kleine T, Lockhart P, Batschauer A (2003) An Arabidopsis protein closely related to Synechocystis cryptochrome is targeted to organelles. Plant J 35(1):93–103

    Article  CAS  PubMed  Google Scholar 

  • Li YY, Mao K, Zhao C, Zhao XY, Zhang HL, Shu HR, Hao YJ (2012) MdCOP1 ubiquitin E3 ligases interact with MdMYB1 to regulate light-induced anthocyanin biosynthesis and red fruit coloration in apple. Plant Physiol 160:1011–1022

    Article  CAS  PubMed  Google Scholar 

  • Lin C (2000) Plant blue-light receptors. Trends Plant Sci 5(8):337–342

    Article  CAS  PubMed  Google Scholar 

  • Lin C (2002) Blue light receptors and signal transduction. Plant Cell 14:207–225

    Google Scholar 

  • Lin C, Shalitin D (2003) Cryptochrome structure and signal transduction. Annu Rev Plant Biol 54:469–496

    Article  CAS  PubMed  Google Scholar 

  • Lin C, Ahmad M, Gordon D, Cashmore A (1995) Expression of an Arabidopsis cryptochrome gene in transgenic tobacco results in hypersensitivity to blue, UV-A, and green light. Proc Natl Acad Sci USA 92:8423–8427

    Article  CAS  PubMed  Google Scholar 

  • Lin C, AhmadM Chan J, Cashmore AR (1996) CRY2, a second member of the Arabidopsis cryptochrome gene family. Plant Physiol 110:1047

    Article  Google Scholar 

  • Lin C, Yang H, Guo H, Mockler T, Chen J, Cashmore AR (1998) Enhancement of blue-light sensitivity of Arabidopsis seedlings by a blue light receptor cryptochrome 2. Proc Natl Acad Sci USA 95:2686–2690

    Article  CAS  PubMed  Google Scholar 

  • Liu LJ, Zhang YC, Li QH, Sang Y, Mao J, Lian HL, Wang L, Yang HQ (2008) COP1-mediated ubiquitination of CONSTANS is implicated in cryptochrome regulation of flowering in Arabidopsis. Plant Cell 20:292–306

    Article  CAS  PubMed  Google Scholar 

  • Liu H, Liu B, Zhao C, Pepper M, Lin C (2011) The action mechanisms of plant cryptochromes. Trends Plant Sci 16(12):684–691

    Article  CAS  PubMed  Google Scholar 

  • Mandoli DF, Ford GA, Waldron LJ, Nemson JA, Briggs WR (1990) Some spectral properties of several soil types: implications for photomorphogenesis. Plant Cell Environ 13:287–294

    Article  Google Scholar 

  • Osterlund MT, Hardtke CS, Wei N, Deng XW (2000) Targeted destabilization of HY5 during light-regulated development of Arabidopsis. Nature 405:462–466

    Article  CAS  PubMed  Google Scholar 

  • Partch CL, Sancar A (2005) Cryptochromes and circadian photoreception in animals. Methods Enzymol 393:726–745

    Article  CAS  PubMed  Google Scholar 

  • Platten JD, Foo E, Elliott RC, Hecht V, Reid JB, Weller JL (2005) Cryptochrome 1 contributes to blue-light sensing in pea. Plant Physiol 139:1472–1482

    Article  CAS  PubMed  Google Scholar 

  • Quattrocchio F, Wing J, Woude K, Souer E, Vetten N, Mol J, Koes R (1999) Molecular analysis of the anthocyanin2 gene of petunia and its role in the evolution of flower color. Plant Cell 11(8):1433–1444

    CAS  PubMed  Google Scholar 

  • Rizzini L, Favory JJ, Cloix C, Faggionato D, O’Hara A, Kaiserli E, Baumeister R, Schäfer E, Nagy F, Jenkins GI, Ulm R (2011) Perception of UV-B by the Arabidopsis UVR8 protein. Science 332:103–106

    Article  CAS  PubMed  Google Scholar 

  • Sakamoto K, Briggs WR (2002) Cellular and subcellular localization of phototropin1. Plant Cell 14(8):1723–1735

    Article  CAS  PubMed  Google Scholar 

  • Seo HS, Yang JY, Ishikawa M, Bolle C, Ballesteros ML, Chua NH (2003) LAF1 ubiquitination by COP1 controls photomorphogenesis and is stimulated by SPA1. Nature 423:995–999

    Article  CAS  PubMed  Google Scholar 

  • Toth R, Kevei E, Hall A, Millar AJ, Nagy F, Kozma-Bognar L (2001) Circadian clock-regulated expression of phytochrome and cryptochrome genes in Arabidopsis. Plant Physiol 127:1607–1616

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Ma LG, Li JM, Zhao HY, Deng XW (2001) Direct interaction of Arabidopsis cryptochromes with COP1 in light control development. Science 294:154–158

    Article  CAS  PubMed  Google Scholar 

  • Wray GA, Hahn MW, Abouheif E, Balhoff JP, Pizer M, Rockman MV, Romano LA (2003) The evolution of transcriptional regulation in eukaryotes. Mol Biol Evol 20(9):1377–1419

    Article  CAS  PubMed  Google Scholar 

  • Wu G, Spalding EP (2007) Separate functions for nuclear and cytoplasmic cryptochrome 1 during photomorphogenesis of Arabidopsis seedlings. Proc Natl Acad Sci USA 104:18813–18818

    Article  CAS  PubMed  Google Scholar 

  • Wu D, Hu Q, Yan Z, Chen W, Yan C, Huang X, Zhang J, Yang P, Deng H, Wang J, Deng XW, Shi Y (2012) Structural basis of ultraviolet-B perception by UVR8. Nature 484:214–219

    Article  PubMed  Google Scholar 

  • Xu P, Xiang Y, Zhu H, Xu H, Zhang Z, Zhang C, Zhang L, Ma Z (2009) Wheat cryptochromes: subcellular localization and involvement in photomorphogenesis and osmotic stress responses. Plant Physiol 149:760–774

    Article  CAS  PubMed  Google Scholar 

  • Yang HQ, Tang RH, Cashmore AR (2001) The signaling mechanism of Arabidopsis CRY1 involves direct interaction with COP1. Plant Cell 13:2573–2587

    CAS  PubMed  Google Scholar 

  • Yang J, Lin R, Sullivan J, Hoecker U, Liu B, Xu L, Deng XW, Wang H (2005) Light regulates COP1-mediated degradation of HFR1, a transcription factor essential for light signaling in Arabidopsis. Plant Cell 17:804–821

    Article  CAS  PubMed  Google Scholar 

  • Yu X, Shalitin D, Liu X, Maymon M, Klejnot J, Yang H, Lopez J, Zhao X, Bendehakkalu KT, Lin C (2007) Derepression of the NC80 motif is critical for the photoactivation of Arabidopsis CRY2. Proc Natl Acad Sci USA 104:7289–7294

    Article  CAS  PubMed  Google Scholar 

  • Yu X, Sayegh R, Maymon M, Warpeha K, Klejnot J, Yang H, Huang J, Lee J, Kaufman L, Lin C (2009) Formation of nuclear bodies of Arabidopsis CRY2 in response to blue light is associated with its blue light-dependent degradation. Plant Cell 21:118–130

    Article  CAS  PubMed  Google Scholar 

  • Yu X, Liu H, Klejnot J, Lin C (2010) The cryptochrome blue-light receptors. The Arabidopsis book 8:e0135. doi:10.1199/tab.0135

    PubMed  Google Scholar 

  • Zhang Q, Li H, Li R, Hu R, Fan C, Chen F, Wang Z, Liu X, Fu Y, Lin C (2008) Association of the circadian rhythmic expression of GmCRY1a with a latitudinal cline in photoperiodic flowering of soybean. Proc Natl Acad Sci USA 105:21028–21033

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Prof. Hongquan Yang of Shanghai Jiao Tong University, China, for providing Arabidopsis cry2 mutant. This work was supported by National Basic Research Program of China (2011CB100600), National Natural Science Foundation of China (31272142), and Program for Changjiang Scholars and Innovative Research Team in University (IRT1155).

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Authors and Affiliations

  1. State Key Laboratory of Crop Biology, National Research Center for Apple Engineering and Technology, College of Horticulture Science and Engineering, Shandong Agricultural University, Tai-An, 271018, Shandong, China

    Yuan-Yuan Li, Cheng Zhao, Xian-Yan Zhao, Rui-Fen Zhang, Hua-Lei Zhang, Huai-Rui Shu & Yu-Jin Hao

  2. College of Biological Science and Technology, Beijing Forestry University, Beijing, 100083, China

    Ke Mao

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  1. Yuan-Yuan Li
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Correspondence to Yu-Jin Hao.

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Communicated by K. Chong.

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299_2013_1387_MOESM1_ESM.tif

Figure S1. Expression of AtPAP1, AtBIT1, AtCO, AtHY5, AtCHS, and AtDFR in WT and MdCRY2 overexpressing Arabidopsis. AtACTIN was used as the external control (TIFF 112 kb)

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Li, YY., Mao, K., Zhao, C. et al. Molecular cloning and functional analysis of a blue light receptor gene MdCRY2 from apple (Malus domestica). Plant Cell Rep 32, 555–566 (2013). https://doi.org/10.1007/s00299-013-1387-4

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