Genome-Wide Quantitative Identification of DNA Differentially Methylated Sites in Arabidopsis Seedlings Growing at Different Water Potential

Background In eukaryotes, the combinatorial usage of cis-regulatory elements enables the assembly of composite genetic switches to integrate multifarious, convergent signals within a single promoter. Plants as sessile organisms, incapable of seeking for optimal conditions, rely on the use of this resource to adapt to changing environments. Emerging evidence suggests that the transcriptional responses of plants to stress are associated with epigenetic processes that govern chromatin accessibility. However, the extent at which specific chromatin modifications contribute to gene regulation has not been assessed. Methodology/Principal Findings In the present work, we combined methyl-sensitive-cut counting and RNA-seq to follow the transcriptional and epigenetic response of plants to simulated drought. Comprehensive genome wide evidence supports the notion that the methylome is widely reactive to water potential. The predominant changes in methylomes were loci in the promoters of genes encoding for proteins suited to cope with the environmental challenge. Conclusion/Significance These selective changes in the methylome with corresponding changes in gene transcription suggest drought sets in motion an instructive mechanism guiding epigenetic machinery toward specific effectors genes.

[1]  Israel Steinfeld,et al.  BMC Bioinformatics BioMed Central , 2008 .

[2]  O. Mathieu,et al.  Transgenerational Stability of the Arabidopsis Epigenome Is Coordinated by CG Methylation , 2007, Cell.

[3]  P. Verslues,et al.  LWR 1 and LWR 2 Are Required for Osmoregulation and Osmotic Adjustment in Arabidopsis 1 , 2004 .

[4]  S. Jacobsen,et al.  Genetic analyses of DNA methyltransferases in Arabidopsis thaliana. , 2006, Cold Spring Harbor symposia on quantitative biology.

[5]  Patrick Achard,et al.  Integration of Plant Responses to Environmentally Activated Phytohormonal Signals , 2006, Science.

[6]  K. Shinozaki,et al.  Gene networks involved in drought stress response and tolerance. , 2006, Journal of experimental botany.

[7]  Karsten M. Borgwardt,et al.  Spontaneous epigenetic variation in the Arabidopsis thaliana methylome , 2011, Nature.

[8]  N. Tuteja,et al.  Cold, salinity and drought stresses: an overview. , 2005, Archives of biochemistry and biophysics.

[9]  Christophe Maurel,et al.  Early Effects of Salinity on Water Transport in Arabidopsis Roots. Molecular and Cellular Features of Aquaporin Expression1 , 2005, Plant Physiology.

[10]  Han Yong Lee,et al.  Non-specific phytohormonal induction of AtMYB44 and suppression of jasmonate-responsive gene activation in Arabidopsis thaliana , 2010, Molecules and cells.

[11]  K. Humbeck,et al.  Stress induced and nuclear localized HIPP26 from Arabidopsis thaliana interacts via its heavy metal associated domain with the drought stress related zinc finger transcription factor ATHB29 , 2008, Plant Molecular Biology.

[12]  J. Greally,et al.  Optimized design and data analysis of tag-based cytosine methylation assays , 2010, Genome Biology.

[13]  A. Peeters,et al.  The Arabidopsis aldehyde oxidase 3 (AAO3) gene product catalyzes the final step in abscisic acid biosynthesis in leaves. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Tianyuan Wang,et al.  Expanded methyl-sensitive cut counting reveals hypomethylation as an epigenetic state that highlights functional sequences of the genome , 2011, Proceedings of the National Academy of Sciences.

[15]  Manabu Ishitani,et al.  Regulation of Osmotic Stress-responsive Gene Expression by theLOS6/ABA1 Locus inArabidopsis * , 2002, The Journal of Biological Chemistry.

[16]  Erik Alexandersson,et al.  Transcriptional regulation of aquaporins in accessions of Arabidopsis in response to drought stress. , 2010, The Plant journal : for cell and molecular biology.

[17]  Zhikang Li,et al.  Drought-induced site-specific DNA methylation and its association with drought tolerance in rice (Oryza sativa L.) , 2010, Journal of experimental botany.

[18]  S. Shigeoka,et al.  Arabidopsis heat shock transcription factor A2 as a key regulator in response to several types of environmental stress. , 2006, The Plant journal : for cell and molecular biology.

[19]  R. E. Sharp,et al.  Root growth maintenance during water deficits: physiology to functional genomics. , 2004, Journal of experimental botany.

[20]  S. Kim,et al.  ABF2, an ABRE-binding bZIP factor, is an essential component of glucose signaling and its overexpression affects multiple stress tolerance. , 2004, The Plant journal : for cell and molecular biology.

[21]  I. Henderson,et al.  RNAi, DRD1, and Histone Methylation Actively Target Developmentally Important Non-CG DNA Methylation in Arabidopsis , 2006, PLoS genetics.

[22]  P. Verslues,et al.  LWR1 and LWR2 Are Required for Osmoregulation and Osmotic Adjustment in Arabidopsis1 , 2004, Plant Physiology.

[23]  Jon Reinders,et al.  Unlocking the Arabidopsis epigenome , 2009, Epigenetics.

[24]  Xiaofeng Cao,et al.  ARGONAUTE4 Control of Locus-Specific siRNA Accumulation and DNA and Histone Methylation , 2003, Science.

[25]  K. Shinozaki,et al.  Regulation of drought tolerance by gene manipulation of 9-cis-epoxycarotenoid dioxygenase, a key enzyme in abscisic acid biosynthesis in Arabidopsis. , 2001, The Plant journal : for cell and molecular biology.

[26]  Frank Johannes,et al.  Assessing the Impact of Transgenerational Epigenetic Variation on Complex Traits , 2009, PLoS genetics.

[27]  S. Song,et al.  Overexpression of AtMYB44 Enhances Stomatal Closure to Confer Abiotic Stress Tolerance in Transgenic Arabidopsis1[C][W][OA] , 2007, Plant Physiology.

[28]  W. Reik,et al.  Epigenetic Reprogramming in Plant and Animal Development , 2010, Science.

[29]  Erin T. Hamanishi,et al.  Clone history shapes Populus drought responses , 2011, Proceedings of the National Academy of Sciences.

[30]  J. Paszkowski,et al.  Maintenance of CpG methylation is essential for epigenetic inheritance during plant gametogenesis , 2003, Nature Genetics.

[31]  J. Paszkowski,et al.  Release of epigenetic gene silencing by trans-acting mutations in Arabidopsis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[32]  M. Fromm,et al.  Multiple exposures to drought 'train' transcriptional responses in Arabidopsis , 2012, Nature Communications.

[33]  M. Mirouze,et al.  Epigenetic contribution to stress adaptation in plants. , 2011, Current opinion in plant biology.

[34]  Jianhua Zhu,et al.  Methods and concepts in quantifying resistance to drought, salt and freezing, abiotic stresses that affect plant water status. , 2006, The Plant journal : for cell and molecular biology.

[35]  R. E. Sharp,et al.  Growth of Arabidopsis thaliana seedlings under water deficit studied by control of water potential in nutrient-agar media. , 2000, Journal of experimental botany.

[36]  Madeleine P. Ball,et al.  Corrigendum: Targeted and genome-scale strategies reveal gene-body methylation signatures in human cells , 2009, Nature Biotechnology.

[37]  Masakazu Satou,et al.  Arabidopsis transcriptome analysis under drought, cold, high-salinity and ABA treatment conditions using a tiling array. , 2008, Plant & cell physiology.

[38]  R. Fischer,et al.  DNA Methylation Is Critical for Arabidopsis Embryogenesis and Seed Viability , 2006, The Plant Cell Online.

[39]  C. Ponting,et al.  Regulation of chromatin structure by site-specific histone H3 methyltransferases , 2000, Nature.

[40]  S. Rothstein,et al.  The role of epigenetic processes in controlling flowering time in plants exposed to stress. , 2011, Journal of experimental botany.

[41]  P. Verslues Quantification of water stress-induced osmotic adjustment and proline accumulation for Arabidopsis thaliana molecular genetic studies. , 2010, Methods in molecular biology.

[42]  P. Verslues,et al.  Mechanisms independent of abscisic acid (ABA) or proline feedback have a predominant role in transcriptional regulation of proline metabolism during low water potential and stress recovery. , 2010, Plant, cell & environment.

[43]  H. Takatsuji,et al.  RNA-directed DNA methylation induces transcriptional activation in plants , 2009, Proceedings of the National Academy of Sciences.

[44]  Yuan Gao,et al.  MOM: maximum oligonucleotide mapping , 2009, Bioinform..

[45]  M. Tan Analysis of DNA methylation of maize in response to osmotic and salt stress based on methylation-sensitive amplified polymorphism. , 2010, Plant physiology and biochemistry : PPB.

[46]  Matthew D. Schultz,et al.  Transgenerational Epigenetic Instability Is a Source of Novel Methylation Variants , 2011, Science.

[47]  S. Song,et al.  Overexpression of AtMYB 44 Enhances Stomatal Closure to Confer Abiotic Stress Tolerance in Transgenic Arabidopsis 1 [ C ] [ W ] [ OA ] , 2008 .

[48]  Jian-Kang Zhu,et al.  Epigenetic regulation of stress responses in plants. , 2009, Current opinion in plant biology.