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Identification of the ASR gene family from Brachypodium distachyon and functional characterization of BdASR1 in response to drought stress

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A genome-wide investigation identified five B. distachyon ASR genes. BdASR1 may be a transcription factor that confers drought resistance by activating antioxidant systems involving ROS-scavenging enzymes and non-enzymatic antioxidants.

Abstract

Abscisic acid-, stress-, and ripening-induced (ASR) proteins belong to a family of plant-specific, small, and hydrophilic proteins with important roles in responses to abiotic stresses. Although several ASR genes involved in drought tolerance have been characterized in various plant species, the mechanisms regulating ASR activities are still uncharacterized. Additionally, no research on Brachypodium distachyon ASR proteins have been completed. In this study, five B. distachyon BdASR genes were identified through genome-wide analyses. Phylogenetic analyses revealed that BdASR genes originated from tandem and whole genome duplications. Expression analyses revealed the BdASR genes responded to various abiotic stresses, including cold, drought, and salinity, as well as signaling molecules such as abscisic acid, ethylene, and H2O2. BdASR1, which localizes to the nucleus and is transcriptionally active, was functionally characterized. BdASR1 overexpression considerably enhanced drought tolerance in transgenic tobacco plants, which was accompanied by increased superoxide dismutase, catalase, and peroxidase activities, as well as an increased abundance of antioxidants such as ascorbate, tocopherols, and glutathione. BdASR1 may function as a transcription factor that provides drought stress resistance by inducing the production of reactive oxygen species-scavenging enzymes and non-enzymatic antioxidants.

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References

  • Abogadallah GM (2010) Insights into the significance of antioxidative defense under salt stress. Plant Signaling Behav 5:369–374

    Article  CAS  Google Scholar 

  • Ahmad P, Sarwat M, Sharma S (2008) Reactive oxygen species, antioxidants and signaling in plants. J Plant Biol 51:167–173

    Article  CAS  Google Scholar 

  • Anjum SA, Xie X, Wang L, Saleem MF, Man C, Lei W (2011) Morphological, physiological and biochemical responses of plants to drought stress. Afr J Agric Res 6:2026–2032

    Google Scholar 

  • Bari R, Jones JD (2009) Role of plant hormones in plant defence responses. Plant Mol Biol 69:473–488

    Article  CAS  PubMed  Google Scholar 

  • Blokhina O, Virolainen E, Fagerstedt KV (2003) Antioxidants, oxidative damage and oxygen deprivation stress: a review. Ann Bot NLM 91:179–194

    Article  CAS  Google Scholar 

  • Buchanan BB, Balmer Y (2005) Redox regulation: a broadening horizon. Annu Rev Plant Biol 56:187–220

    Article  CAS  PubMed  Google Scholar 

  • Cakir B, Agasse A, Gaillard C, Saumonneau A, Delrot S, Atanassova R (2003) A grape ASR protein involved in sugar and abscisic acid signaling. Plant Cell 15:2165–2180

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Canel C, Bailey-Serres JN, Roose ML (1995) Pummelo fruit transcript homologous to ripening-induced genes. Plant Physiol 108:1323–1324

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Carrari F, Fernie AR, Iusem ND (2004) Heard it through the grapevine? ABA and sugar cross-talk: the ASR story. Trends Plant Sci 9:57–59

    Article  CAS  PubMed  Google Scholar 

  • Chang S, Puryear JD, Dias M, Funkhouser EA, Newton RJ, Cairney J (1996) Gene expression under water deficit in loblolly pine (Pinus taeda): isolation and characterization of cDNA clones. Physiol Plantarum 97:139–148

    Article  CAS  Google Scholar 

  • Dai JR, Liu B, Feng DR, Liu HY, He YM, Qi KB, Wang HB, Wang JF (2011) MpAsr encodes an intrinsically unstructured protein and enhances osmotic tolerance in transgenic Arabidopsis. Plant Cell Rep 30:1219–1230

    Article  CAS  PubMed  Google Scholar 

  • Dominguez PG, Carrari F (2015) ASR1 transcription factor and its role in metabolism. Plant Signaling Behav 10:e992751

    Article  Google Scholar 

  • Dowdle J, Ishikawa T, Gatzek S, Rolinski S, Smirnoff N (2007) Two genes in Arabidopsis thaliana encoding GDP-l-galactose phosphorylase are required for ascorbate biosynthesis and seedling viability. Plant J 52:673–689

    Article  CAS  PubMed  Google Scholar 

  • Frankel N, Carrari F, Hasson E, Iusem ND (2006) Evolutionary history of the Asr gene family. Gene 378:74–83

    Article  CAS  PubMed  Google Scholar 

  • Frankel N, Nunes-Nesi A, Balbo I, Mazuch J, Centeno D, Iusem ND, Fernie AR, Carrari F (2007) ci21A/Asr1 expression influences glucose accumulation in potato tubers. Plant Mol Biol 63:719–730

    Article  CAS  PubMed  Google Scholar 

  • Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Bioch 48:909–930

    Article  CAS  Google Scholar 

  • Golan I, Dominguez PG, Konrad Z, Shkolnik-Inbar D, Carrari F, Bar-Zvi D (2014) Tomato ABSCISIC ACID STRESS RIPENING (ASR) gene family revisited. PLoS ONE 9:e107117

    Article  PubMed  PubMed Central  Google Scholar 

  • Gu Z, Cavalcanti A, Chen FC, Bouman P, Li WH (2002) Extent of gene duplication in the genomes of Drosophila, nematode, and yeast. Mol Biol Evol 19:256–262

    Article  CAS  PubMed  Google Scholar 

  • Halliwell B, Gutteridge J (1984) Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J 219:1–14

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Henry IM, Carpentier SC, Pampurova S, Van Hoylandt A, Panis B, Swennen R, Remy S (2011) Structure and regulation of the Asr gene family in banana. Planta 234:785–798

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hong SH, Kim IJ, Yang DC, Chung WI (2002) Characterization of an abscisic acid responsive gene homologue from Cucumis melo. J Exp Bot 53:2271–2272

    Article  CAS  PubMed  Google Scholar 

  • Hong SY, Seo PJ, Yang MS, Xiang F, Park CM (2008) Exploring valid reference genes for gene expression studies in Brachypodium distachyon by real-time PCR. BMC Plant Biol 8:112

    Article  PubMed  PubMed Central  Google Scholar 

  • Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SC, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231

    Article  CAS  Google Scholar 

  • Hsu YF, Yu SC, Yang CY, Wang CS (2011) Lily ASR protein-conferred cold and freezing resistance in Arabidopsis. Plant Physiol Bioch 49:937–945

    Article  CAS  Google Scholar 

  • Hu W, Huang C, Deng X, Zhou S, Chen L, Li Y, Wang C, Ma Z, Yuan Q, Wang Y, Cai R, Liang X, Yang G, He G (2013) TaASR1, a transcription factor gene in wheat, confers drought stress tolerance in transgenic tobacco. Plant Cell Environ 36:1449–1464

    Article  CAS  PubMed  Google Scholar 

  • Hundertmark M, Hincha DK (2008) LEA (late embryogenesis abundant) proteins and their encoding genes in Arabidopsis thaliana. BMC Genom 9:118

    Article  Google Scholar 

  • Iusem ND, Bartholomew DM, Hitz WD, Scolnik PA (1993) Tomato (Lycopersicon esculentum) transcript induced by water deficit and ripening. Plant Physiol 102:1353–1354

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jeanneau M, Gerentes D, Foueillassar X, Zivy M, Vidal J, Toppan A, Perez P (2002) Improvement of drought tolerance in maize: towards the functional validation of the Zm-Asr1 gene and increase of water use efficiency by over-expressing C4–PEPC. Biochimie 84:1127–1135

    Article  CAS  PubMed  Google Scholar 

  • Jha B, Agarwal PK, Reddy PS, Lal S, Sopory SK, Reddy MK (2009) Identification of salt-induced genes from Salicornia brachiata, an extreme halophyte through expressed sequence tags analysis. Genes Genet Syst 84:111–120

    Article  CAS  PubMed  Google Scholar 

  • Kalifa Y, Perlson E, Gilad A, Konrad Z, Scolnik P, Bar-Zvi D (2004a) Over-expression of the water and salt stress-regulated Asr1 gene confers an increased salt tolerance. Plant, Cell Environ 27:1459–1468

    Article  CAS  Google Scholar 

  • Kalifa Y, Gilad A, Konrad Z, Zaccai M, Scolnik P, Bar-Zvi D (2004b) The water-and salt-stress-regulated Asr1 (abscisic acid stress ripening) gene encodes a zinc-dependent DNA-binding protein. Biochem J 381:373–378

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kim IS, Kim YS, Yoon HS (2012) Rice ASR1 protein with reactive oxygen species scavenging and chaperone-like activities enhances acquired tolerance to abiotic stresses in Saccharomyces cerevisiae. Mol Cells 33:285–293

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Konrad Z, Bar-Zvi D (2008) Synergism between the chaperone-like activity of the stress regulated ASR1 protein and the osmolyte glycine-betaine. Planta 227:1213–1219

    Article  CAS  PubMed  Google Scholar 

  • Larkin M, Blackshields G, Brown N, Chenna R, Mcgettigan PA, Mcwilliam H, Valentin F, Wallace IM, Wilm A, Lopez R (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948

    Article  CAS  PubMed  Google Scholar 

  • Liu X, Wang Z, Wang L, Wu R, Phillips J, Deng X (2009) LEA 4 group genes from the resurrection plant Boea hygrometrica confer dehydration tolerance in transgenic tobacco. Plant Sci 176:90–98

    Article  CAS  Google Scholar 

  • Liu HY, Dai JR, Feng DR, Liu B, Wang HB, Wang JF (2010) Characterization of a novel plantain Asr gene, MpAsr, that is regulated in response to infection of Fusarium oxysporum f. sp. cubense and abiotic stresses. J Integr Plant Biol 52:315–323

    Article  CAS  PubMed  Google Scholar 

  • Liu J, Jia C, Dong F, Wang J, Zhang J, Xu Y, Xu B, Jin Z (2013) Isolation of an abscisic acid senescence and ripening inducible gene from litchi and functional characterization under water stress. Planta 237:1025–1036

    Article  CAS  PubMed  Google Scholar 

  • Livak K, Schmittgen T (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Maskin L, Frankel N, Gudesblat G, Demergasso MJ, Pietrasanta LI, Iusem ND (2007) Dimerization and DNA-binding of ASR1, a small hydrophilic protein abundant in plant tissues suffering from water loss. Biochem Biophys Res Commun 352:831–835

    Article  CAS  PubMed  Google Scholar 

  • Miller G, Suzuki N, Ciftci-Yilmaz S, Mittler R (2010) Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant Cell Environ 33:453–467

    Article  CAS  PubMed  Google Scholar 

  • Moore K, Roberts LJ (1998) Measurement of lipid peroxidation. Free Radical Res 28:659–671

    Article  CAS  Google Scholar 

  • Nagalakshmi N, Prasad M (2001) Responses of glutathione cycle enzymes and glutathione metabolism to copper stress in Scenedesmus bijugatus. Plant Sci 160:291–299

    Article  CAS  PubMed  Google Scholar 

  • Padmanabhan V, Dias DM, Newton RJ (1997) Expression analysis of a gene family in loblolly pine (Pinus taeda L.) induced by water deficit stress. Plant Mol Biol 35:801–807

    Article  CAS  PubMed  Google Scholar 

  • Philippe R, Courtois B, Mcnally KL, Mournet P, El-Malki R, Le Paslier MC, Fabre D, Billot C, Brunel D, Glaszmann J-C (2010) Structure, allelic diversity and selection of Asr genes, candidate for drought tolerance, in Oryza sativa L. and wild relatives. Theor Appl Genet 121:769–787

    Article  CAS  PubMed  Google Scholar 

  • Pitzschke A, Djamei A, Bitton F, Hirt H (2009) A major role of the MEKK1–MKK1/2–MPK4 pathway in ROS signalling. Mol Plant 2:120–137

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Riccardi F, Gazeau P, De Vienne D, Zivy M (1998) Protein changes in response to progressive water deficit in maize quantitative variation and polypeptide identification. Plant Physiol 117:1253–1263

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rippert P, Scimemi C, Dubald M, Matringe M (2004) Engineering plant shikimate pathway for production of tocotrienol and improving herbicide resistance. Plant Physiol 134:92–100

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Saumonneau A, Agasse A, Bidoyen M-T, Lallemand M, Cantereau A, Medici A, Laloi M, Atanassova R (2008) Interaction of grape ASR proteins with a DREB transcription factor in the nucleus. FEBS Lett 582:3281–3287

    Article  CAS  PubMed  Google Scholar 

  • Shao HB, Chu LY, Lu ZH, Kang CM (2008) Primary antioxidant free radical scavenging and redox signaling pathways in higher plant cells. Int J Biol Sci 4:8

    Article  CAS  PubMed Central  Google Scholar 

  • Shen G, Pang Y, Wu W, Deng Z, Liu X, Lin J, Zhao L, Sun X, Tang K (2005) Molecular cloning, characterization and expression of a novel Asr gene from Ginkgo biloba. Plant Physiol Bioch 43:836–843

    Article  CAS  Google Scholar 

  • Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tiwari V, Chaturvedi AK, Mishra A, Jha B (2015) Introgression of the SbASR-1 gene cloned from a Halophyte Salicornia brachiata enhances salinity and drought endurance in transgenic groundnut (Arachis hypogaea) and acts as a transcription factor. PLoS One 10:e0131567

    Article  PubMed  PubMed Central  Google Scholar 

  • Torres-Schumann S, Godoy JA, Pintor-Toro JA (1992) A probable lipid transfer protein gene is induced by NaCl in stems of tomato plants. Plant Mol Biol 18:749–757

    Article  CAS  PubMed  Google Scholar 

  • Treviño MB, O’connell MA (1998) Three drought-responsive members of the nonspecific lipid-transfer protein gene family in Lycopersicon pennellii show different developmental patterns of expression. Plant Physiol 116:1461–1468

    Article  PubMed  PubMed Central  Google Scholar 

  • Vaidyanathan R, Kuruvilla S, Thomas G (1999) Characterization and expression pattern of an abscisic acid and osmotic stress responsive gene from rice. Plant Sci 140:21–30

    Article  CAS  Google Scholar 

  • Van Berkel J, Salamini F, Gebhardt C (1994) Transcripts accumulating during cold storage of potato (Solanum tuberosum L.) tubers are sequence related to stress-responsive genes. Plant Physiol 104:445–452

    Article  PubMed  PubMed Central  Google Scholar 

  • Van Ha C, Leyva-González MA, Osakabe Y, Tran UT, Nishiyama R, Watanabe Y, Tanaka M, Seki M, Yamaguchi S, Van Dong N (2014) Positive regulatory role of strigolactone in plant responses to drought and salt stress. Pro Natl Acad Sci USA 111:851–856

    Article  Google Scholar 

  • Virlouvet L, Jacquemot M-P, Gerentes D, Corti H, Bouton S, Gilard F, Valot B, Trouverie J, Tcherkez G, Falque M, Damerval C, Rogowsky P, Perez P, Noctor G, Zivy M, Coursol S (2011) The ZmASR1 protein influences branched-chain amino acid biosynthesis and maintains kernel yield in maize under water-limited conditions. Plant Physiol 157:917–936

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vogel JP, Garvin DF, Mockler TC, Schmutz J, Rokhsar D, Bevan MW, Barry K, Lucas S, Harmon-Smith M, Lail K (2010) Genome sequencing and analysis of the model grass Brachypodium distachyon. Nature 463:763–768

    Article  CAS  Google Scholar 

  • Wang HJ, Jauh GY, Hsu YH, Wang CS (2003) The nuclear localization signal of a pollen-specific, desiccation-associated protein of lily is necessary and sufficient for nuclear targeting. Bot Bull Acad Sinica 44:123–128

    CAS  Google Scholar 

  • Wang HJ, Hsu CM, Jauh GY, Wang CS (2005) A lily pollen ASR protein localizes to both cytoplasm and nuclei requiring a nuclear localization signal. Physiol Plantarum 123:314–320

    Article  CAS  Google Scholar 

  • Yang CY, Chen YC, Jauh GY, Wang CS (2005) A lily ASR protein involves abscisic acid signaling and confers drought and salt resistance in Arabidopsis. Plant Physiol 139:836–846

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yang SH, Kim SH, Berberich T, Kusano T (2012) Identification and properties of a small protein that interacts with a tobacco bZIP-type transcription factor TBZF. Plant Biotechnol 29:395–399

    Article  Google Scholar 

  • Zhang L, Hu W, Wang Y, Feng R, Zhang Y, Liu J, Jia C, Miao H, Zhang J, Xu B (2015) The MaASR gene as a crucial component in multiple drought stress response pathways in Arabidopsis. Funct Integr Genomic 15:247–260

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by National Genetically Modified New Varieties of Major Projects of China (Grant no. 2016ZX08010004-004, International S & T Cooperation Key Projects of MoST (Grant no. 2009DFB30340), Research Fund for the Doctoral Program of Higher Education of China (Grant no. 2012014211075) and Open Research Fund of State Key Laboratory of Hybrid Rice in Wuhan University (Grant no. KF201302).

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    1. The Genetic Engineering International Cooperation Base of Chinese Ministry of Science and Technology, The Key Laboratory of Molecular Biophysics of Chinese Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, China

      Lianzhe Wang, Wei Hu, Jialu Feng, Xiaoyue Yang, Quanjun Huang, Jiajing Xiao, Yang Liu, Guangxiao Yang & Guangyuan He

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    1. Lianzhe Wang
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    Correspondence to Guangyuan He.

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    Communicated by H. S. Judelson.

    Lianzhe Wang, Wei Hu, Jialu Feng contributed equally to this work.

    Electronic supplementary material

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    Motif analysis of BdASRs. (TIFF 357 kb)

    299_2016_1954_MOESM2_ESM.jpg

    Organ-specific expression patterns of BdASR genes. Relative expressions of BdASR genes were analyzed in roots, stems, leaves and spikelets by RT-qPCR. BdUBC18 was used as an internal control. Normalized mRNA levels in roots were set to 1, and fold changes were used to indicate the transcript level. In addition, as BdASR5 had almost no detectable expression in roots, the expression in spikelets was set to 1. Data are means ± SD calculated from three replicates. (JPEG 110 kb)

    299_2016_1954_MOESM3_ESM.tif

    Expression of BdASR1 in transgenic tobacco lines. Expression of BdASR1 in WT, VC and three independent transgenic lines (OE8, OE9 and OE12) with RT-PCR. NtActin was used as an internal control. (TIFF 128 kb)

    299_2016_1954_MOESM4_ESM.jpg

    Data of unchanged antioxidant enzyme activity and antioxidants levels under drought and oxidative stresses. (A) CAT activity under drought stress. (B) SOD activity under oxidative stress. (C) V E content under oxidative stress. The indices were measured in controls and transgenic lines under normal, drought and oxidative stresses. Data are means ± SD calculated from three replicates. Significant differences between the OE and control lines are indicated as *P < 0.05; **P < 0.01. (JPEG 266 kb)

    299_2016_1954_MOESM5_ESM.jpg

    Expression levels of NtLEA5, NtGMP and NtGSH2 genes of transgenic lines under normal, hyperosmotic and MV treatments. One-week-old seedlings were subjected to normal, hyperosmotic and MV treatments for one week and then used for RT-qPCR. NtActin was used as an internal control. Fold changes were used to indicate relative expression levels between WT and transgenic lines. Data are means ± SD calculated from three replicates. Significant differences between the OE and control lines are indicated as *P < 0.05; **P < 0.01. (JPEG 172 kb)

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    Wang, L., Hu, W., Feng, J. et al. Identification of the ASR gene family from Brachypodium distachyon and functional characterization of BdASR1 in response to drought stress. Plant Cell Rep 35, 1221–1234 (2016). https://doi.org/10.1007/s00299-016-1954-6

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