Long-Term Waterlogging as Factor Contributing to Hypoxia Stress Tolerance Enhancement in Cucumber: Comparative Transcriptome Analysis of Waterlogging Sensitive and Tolerant Accessions

Waterlogging (WL), excess water in the soil, is a phenomenon often occurring during plant cultivation causing low oxygen levels (hypoxia) in the soil. The aim of this study was to identify candidate genes involved in long-term waterlogging tolerance in cucumber using RNA sequencing. Here, we also determined how waterlogging pre-treatment (priming) influenced long-term memory in WL tolerant (WL-T) and WL sensitive (WL-S) i.e., DH2 and DH4 accessions, respectively. This work uncovered various differentially expressed genes (DEGs) activated in the long-term recovery in both accessions. De novo assembly generated 36,712 transcripts with an average length of 2236 bp. The results revealed that long-term waterlogging had divergent impacts on gene expression in WL-T DH2 and WL-S DH4 cucumber accessions: after 7 days of waterlogging, more DEGs in comparison to control conditions were identified in WL-S DH4 (8927) than in WL-T DH2 (5957). Additionally, 11,619 and 5007 DEGs were identified after a second waterlogging treatment in the WL-S and WL-T accessions, respectively. We identified genes associated with WL in cucumber that were especially related to enhanced glycolysis, adventitious roots development, and amino acid metabolism. qRT-PCR assay for hypoxia marker genes i.e., alcohol dehydrogenase (adh), 1-aminocyclopropane-1-carboxylate oxidase (aco) and long chain acyl-CoA synthetase 6 (lacs6) confirmed differences in response to waterlogging stress between sensitive and tolerant cucumbers and effectiveness of priming to enhance stress tolerance.

[1]  Anna Kołton,et al.  Selection of Tomato and Cucumber Accessions for Waterlogging Sensitivity through Morpho-Physiological Assessment at an Early Vegetative Stage , 2020, Agronomy.

[2]  Chun-Gen Hu,et al.  Comparative Transcriptome Analysis of Two Contrasting Kiwifruit (Actinidia) Genotypes under Waterlogging Stress , 2020 .

[3]  P. Perata,et al.  The Many Facets of Hypoxia in Plants , 2020, Plants.

[4]  Loc Van Nguyen,et al.  Variation in root growth responses of sweet potato to hypoxia and waterlogging , 2020 .

[5]  E. Braga,et al.  Long-term transcriptional memory in rice plants submitted to salt shock , 2020, Planta.

[6]  I. Feussner,et al.  Wax biosynthesis upon danger: its regulation upon abiotic and biotic stress. , 2020, The New phytologist.

[7]  H. Çelik,et al.  Epigenetic memory and priming in plants , 2020, Genetica.

[8]  S. Xiao,et al.  Long-Chain acyl-CoA Synthetase LACS2 Contributes to Submergence Tolerance by Modulating Cuticle Permeability in Arabidopsis , 2020, Plants.

[9]  J. Tucker,et al.  Genome-Wide Analysis of Gene Expression Provides New Insights into Waterlogging Responses in Barley (Hordeum vulgare L.) , 2020, Plants.

[10]  M. Baier,et al.  Cold priming uncouples light- and cold-regulation of gene expression in Arabidopsis thaliana , 2020, BMC Plant Biology.

[11]  Francesco Licausi,et al.  Editorial: Crop Response to Waterlogging , 2019, Front. Plant Sci..

[12]  Linhai Wang,et al.  Transcriptomic profiling of sesame during waterlogging and recovery , 2019, Scientific Data.

[13]  Wei Li,et al.  Identification of Genes/Proteins Related to Submergence Tolerance by Transcriptome and Proteome Analyses in Soybean , 2019, Scientific Reports.

[14]  P. Somta,et al.  Comparative Transcriptome Analysis of Waterlogging-Sensitive and Tolerant Zombi Pea (Vigna vexillata) Reveals Energy Conservation and Root Plasticity Controlling Waterlogging Tolerance , 2019, Plants.

[15]  C. Sams,et al.  Waterlogging Causes Early Modification in the Physiological Performance, Carotenoids, Chlorophylls, Proline, and Soluble Sugars of Cucumber Plants , 2019, Plants.

[16]  Devanand L. Luthria,et al.  Biochemical and Anatomical Investigation of Sesbania herbacea (Mill.) McVaugh Nodules Grown under Flooded and Non-Flooded Conditions , 2019, International journal of molecular sciences.

[17]  T. Schmülling,et al.  Stress priming, memory, and signalling in plants. , 2019, Plant, cell & environment.

[18]  S. Shabala,et al.  Soil and Crop Management Practices to Minimize the Impact of Waterlogging on Crop Productivity , 2019, Front. Plant Sci..

[19]  J. Zwiazek,et al.  Stable expression of aquaporins and hypoxia-responsive genes in adventitious roots are linked to maintaining hydraulic conductance in tobacco (Nicotiana tabacum) exposed to root hypoxia , 2019, PloS one.

[20]  Chengdao Li,et al.  Proteomic analysis reveals response of differential wheat (Triticum aestivum L.) genotypes to oxygen deficiency stress , 2019, BMC Genomics.

[21]  M. Alves-Ferreira,et al.  Using transcriptomics to assess plant stress memory , 2018, Theoretical and Experimental Plant Physiology.

[22]  Jing Yang,et al.  Expression of long non-coding RNA and mRNA in the hippocampus of mice with type 2 diabetes , 2018, Molecular medicine reports.

[23]  Xiaomin Du,et al.  De novo transcriptomic analysis to identify differentially expressed genes during the process of aerenchyma formation in Typha angustifolia leaves. , 2018, Gene.

[24]  Chao Lv,et al.  Morpho-anatomical and physiological responses to waterlogging stress in different barley (Hordeum vulgare L.) genotypes , 2018, Plant Growth Regulation.

[25]  Ajay Saini,et al.  Heat-stress priming and alternative splicing-linked memory , 2018, Journal of experimental botany.

[26]  S. Komatsu,et al.  An Integrated Approach of Proteomics and Computational Genetic Modification Effectiveness Analysis to Uncover the Mechanisms of Flood Tolerance in Soybeans , 2018, International journal of molecular sciences.

[27]  L. Voesenek,et al.  A stress recovery signaling network for enhanced flooding tolerance in Arabidopsis thaliana , 2018, Proceedings of the National Academy of Sciences.

[28]  K. Arora,et al.  In Silico Characterization and Functional Validation of Cell Wall Modification Genes Imparting Waterlogging Tolerance in Maize , 2017, Bioinformatics and biology insights.

[29]  K. Arora,et al.  RNAseq revealed the important gene pathways controlling adaptive mechanisms under waterlogged stress in maize , 2017, Scientific Reports.

[30]  J. Ji,et al.  Comparative RNA-seq based transcriptome profiling of waterlogging response in cucumber hypocotyls reveals novel insights into the de novo adventitious root primordia initiation , 2017, BMC Plant Biology.

[31]  G. Jeena,et al.  MaRAP2‐4, a waterlogging‐responsive ERF from Mentha, regulates bidirectional sugar transporter AtSWEET10 to modulate stress response in Arabidopsis , 2017, Plant biotechnology journal.

[32]  J. Lämke,et al.  Epigenetic and chromatin-based mechanisms in environmental stress adaptation and stress memory in plants , 2017, Genome Biology.

[33]  A. Zebarjadi,et al.  Effect of waterlogging at different growth stages on some morphological traits of wheat varieties , 2017, International Journal of Biometeorology.

[34]  J. Ji,et al.  Inheritance and quantitative trail loci mapping of adventitious root numbers in cucumber seedlings under waterlogging conditions , 2016, Molecular Genetics and Genomics.

[35]  Joost T. van Dongen,et al.  Community recommendations on terminology and procedures used in flooding and low oxygen stress research. , 2017, The New phytologist.

[36]  Wei Wang,et al.  Ectopic expression of wheat expansin gene TaEXPA2 improved the salt tolerance of transgenic tobacco by regulating Na+ /K+ and antioxidant competence. , 2017, Physiologia plantarum.

[37]  D. Mykles,et al.  A Comparison of Resources for the Annotation of a De Novo Assembled Transcriptome in the Molting Gland (Y-Organ) of the Blackback Land Crab, Gecarcinus lateralis. , 2016, Integrative and comparative biology.

[38]  Xiaotian Ma,et al.  Comparative Proteomic Analysis Provides Insight into the Key Proteins Involved in Cucumber (Cucumis sativus L.) Adventitious Root Emergence under Waterlogging Stress , 2016, Front. Plant Sci..

[39]  L. Voesenek,et al.  Transcriptomes of Eight Arabidopsis thaliana Accessions Reveal Core Conserved, Genotype- and Organ-Specific Responses to Flooding Stress1[OPEN] , 2016, Plant Physiology.

[40]  T. Trewavas Plant Intelligence: An Overview , 2016 .

[41]  A. Ding,et al.  Expansins: roles in plant growth and potential applications in crop improvement , 2016, Plant Cell Reports.

[42]  Diep Ganguly,et al.  Reconsidering plant memory: Intersections between stress recovery, RNA turnover, and epigenetics , 2016, Science Advances.

[43]  Liwei Hu,et al.  Gamma-aminobutyric acid mediates nicotine biosynthesis in tobacco under flooding stress☆ , 2016, Plant diversity.

[44]  P. Mazzafera,et al.  Flooding of the root system in soybean: biochemical and molecular aspects of N metabolism in the nodule during stress and recovery , 2016, Amino Acids.

[45]  G. Lu,et al.  Comparison of transcriptomes undergoing waterlogging at the seedling stage between tolerant and sensitive varieties of Brassica napus L. , 2015 .

[46]  Alexander Dobin,et al.  Mapping RNA‐seq Reads with STAR , 2015, Current protocols in bioinformatics.

[47]  Lizhong He,et al.  The effect of exogenous calcium on mitochondria, respiratory metabolism enzymes and ion transport in cucumber roots under hypoxia , 2015, Scientific Reports.

[48]  U. Conrath,et al.  Priming for enhanced defense. , 2015, Annual review of phytopathology.

[49]  L. Tran,et al.  Hydrogen peroxide priming modulates abiotic oxidative stress tolerance: insights from ROS detoxification and scavenging , 2015, Front. Plant Sci..

[50]  S. Salzberg,et al.  StringTie enables improved reconstruction of a transcriptome from RNA-seq reads , 2015, Nature Biotechnology.

[51]  F. Gillet,et al.  Cell Wall Metabolism in Response to Abiotic Stress , 2015, Plants.

[52]  A. Maass,et al.  Transcriptome sequencing of Prunus sp. rootstocks roots to identify candidate genes involved in the response to root hypoxia , 2015, Tree Genetics & Genomes.

[53]  W. Huber,et al.  Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 , 2014, Genome Biology.

[54]  N. Yao,et al.  Arabidopsis acyl-CoA-binding protein ACBP3 participates in plant response to hypoxia by modulating very-long-chain fatty acid metabolism , 2014, The Plant journal : for cell and molecular biology.

[55]  M. Shih,et al.  Ethylene plays an essential role in the recovery of Arabidopsis during post-anaerobiosis reoxygenation. , 2014, Plant, cell & environment.

[56]  Xiang Li,et al.  Antisense suppression of cucumber (Cucumis sativus L.) sucrose synthase 3 (CsSUS3) reduces hypoxic stress tolerance. , 2014, Plant, cell & environment.

[57]  R. Reiter,et al.  The RNA‐seq approach to discriminate gene expression profiles in response to melatonin on cucumber lateral root formation , 2014, Journal of pineal research.

[58]  Meiling Liu,et al.  Regulation of flavanone 3-hydroxylase gene involved in the flavonoid biosynthesis pathway in response to UV-B radiation and drought stress in the desert plant, Reaumuria soongorica. , 2013, Plant physiology and biochemistry : PPB.

[59]  Damian Szklarczyk,et al.  eggNOG v4.0: nested orthology inference across 3686 organisms , 2013, Nucleic Acids Res..

[60]  Mukesh Jain,et al.  Deep Transcriptome Sequencing of Wild Halophyte Rice, Porteresia coarctata, Provides Novel Insights into the Salinity and Submergence Tolerance Factors , 2013, DNA research : an international journal for rapid publication of reports on genes and genomes.

[61]  A. Ismail,et al.  Tolerance of anaerobic conditions caused by flooding during germination and early growth in rice (Oryza sativa L.) , 2013, Front. Plant Sci..

[62]  Colin N. Dewey,et al.  De novo transcript sequence reconstruction from RNA-seq using the Trinity platform for reference generation and analysis , 2013, Nature Protocols.

[63]  M. Sauter Root responses to flooding. , 2013, Current opinion in plant biology.

[64]  Junping Gao,et al.  RhEXPA4, a rose expansin gene, modulates leaf growth and confers drought and salt tolerance to Arabidopsis , 2013, Planta.

[65]  N. Zhang,et al.  Melatonin promotes water‐stress tolerance, lateral root formation, and seed germination in cucumber (Cucumis sativus L.) , 2013, Journal of pineal research.

[66]  Dongdong Niu,et al.  Induction of Drought Tolerance in Cucumber Plants by a Consortium of Three Plant Growth-Promoting Rhizobacterium Strains , 2012, PloS one.

[67]  S. Réhman,et al.  A comparative proteomics analysis in roots of soybean to compatible symbiotic bacteria under flooding stress , 2012, Amino Acids.

[68]  Guangchuang Yu,et al.  clusterProfiler: an R package for comparing biological themes among gene clusters. , 2012, Omics : a journal of integrative biology.

[69]  Xuehao Chen,et al.  Identification of differentially expressed genes in cucumber (Cucumis sativus L.) root under waterlogging stress by digital gene expression profile. , 2012, Genomics.

[70]  M. Nakazono,et al.  Mechanisms for coping with submergence and waterlogging in rice , 2012, Rice.

[71]  N. Suzuki,et al.  ROS and redox signalling in the response of plants to abiotic stress. , 2012, Plant, cell & environment.

[72]  Jing Li,et al.  Identification of hypoxic-responsive proteins in cucumber roots using a proteomic approach. , 2012, Plant physiology and biochemistry : PPB.

[73]  S. Brunak,et al.  SignalP 4.0: discriminating signal peptides from transmembrane regions , 2011, Nature Methods.

[74]  Colin N. Dewey,et al.  RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome , 2011, BMC Bioinformatics.

[75]  Y. Ha-Lee,et al.  Expression Profile Analysis of Hypoxia Responses in Arabidopsis Roots and Shoots , 2011, Journal of Plant Biology.

[76]  K. Shinozaki,et al.  Transcriptional responses to flooding stress in roots including hypocotyl of soybean seedlings , 2011, Plant Molecular Biology.

[77]  Robert D. Finn,et al.  HMMER web server: interactive sequence similarity searching , 2011, Nucleic Acids Res..

[78]  J. Zhao,et al.  A soybean β-expansin gene GmEXPB2 intrinsically involved in root system architecture responses to abiotic stresses. , 2011, The Plant journal : for cell and molecular biology.

[79]  A. Katiyar,et al.  Comparative analysis of drought-responsive transcriptome in Indica rice genotypes with contrasting drought tolerance. , 2011, Plant biotechnology journal.

[80]  J. Yiu,et al.  Exogenous catechin increases antioxidant enzyme activity and promotes flooding tolerance in tomato (Solanum lycopersicum L.) , 2011, Plant and Soil.

[81]  Xuexia Chen,et al.  Modulation of chlorophyll contents and anti-oxidant systems in two cucumber varieties under waterlogging stress and subsequent drainage , 2011 .

[82]  Yong-lian Zheng,et al.  Identification of transcriptome induced in roots of maize seedlings at the late stage of waterlogging , 2010, BMC Plant Biology.

[83]  P. Perata,et al.  Hormonal interplay during adventitious root formation in flooded tomato plants. , 2010, The Plant journal : for cell and molecular biology.

[84]  Asan,et al.  The genome of the cucumber, Cucumis sativus L. , 2009, Nature Genetics.

[85]  Davis J. McCarthy,et al.  edgeR: a Bioconductor package for differential expression analysis of digital gene expression data , 2009, Bioinform..

[86]  L. Hoffmann,et al.  Gene expression changes related to the production of phenolic compounds in potato tubers grown under drought stress. , 2009, Phytochemistry.

[87]  V. Beneš,et al.  The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. , 2009, Clinical chemistry.

[88]  Adam J. Carroll,et al.  Differential Response of Gray Poplar Leaves and Roots Underpins Stress Adaptation during Hypoxia1[W] , 2008, Plant Physiology.

[89]  E. Birney,et al.  Pfam: the protein families database , 2013, Nucleic Acids Res..

[90]  G. Mortier,et al.  qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data , 2007, Genome Biology.

[91]  J. Dat,et al.  Sensing and signalling during plant flooding. , 2004, Plant physiology and biochemistry : PPB.

[92]  J. Browse,et al.  The Acyl-CoA Synthetase Encoded by LACS2 Is Essential for Normal Cuticle Development in Arabidopsis , 2004, The Plant Cell Online.

[93]  E. Dennis,et al.  Enhanced Low Oxygen Survival in Arabidopsis through Increased Metabolic Flux in the Fermentative Pathway1 , 2003, Plant Physiology.

[94]  O. Blokhina,et al.  Antioxidants, oxidative damage and oxygen deprivation stress: a review. , 2003, Annals of botany.

[95]  Yi Lee,et al.  Expression of α-Expansin and Expansin-Like Genes in Deepwater Rice1 , 2002, Plant Physiology.

[96]  J. Browse,et al.  Arabidopsis Contains Nine Long-Chain Acyl-Coenzyme A Synthetase Genes That Participate in Fatty Acid and Glycerolipid Metabolism1 , 2002, Plant Physiology.

[97]  B. Winkel-Shirley,et al.  Biosynthesis of flavonoids and effects of stress. , 2002, Current opinion in plant biology.

[98]  M. Ashburner,et al.  Gene Ontology: tool for the unification of biology , 2000, Nature Genetics.

[99]  R. Rasheed,et al.  Chemical Priming for Multiple Stress Tolerance , 2019, Priming and Pretreatment of Seeds and Seedlings.

[100]  Biswajit Pradhan,et al.  Alcohol Dehydrogenase (ADh) Enzyme is a Potent Biochemical Marker for Submergence Tolerance in Rice (Oryza sativa L.) During Seedling Stage of Growth , 2019, International Journal of Current Research and Review.

[101]  Sudhir Kumar,et al.  Seed Priming: An Emerging Technology to Impart Abiotic Stress Tolerance in Crop Plants , 2018 .

[102]  M. J. García,et al.  Hypoxia and bicarbonate could limit the expression of iron acquisition genes in Strategy I plants by affecting ethylene synthesis and signaling in different ways. , 2014, Physiologia plantarum.

[103]  S. Shabala,et al.  Comparison of growth and physiological responses to waterlogging and subsequent recovery in six barley genotypes , 2003 .

[104]  Maria Jesus Martin,et al.  The SWISS-PROT protein knowledgebase and its supplement TrEMBL in 2003 , 2003, Nucleic Acids Res..

[105]  Hiroyuki Ogata,et al.  KEGG: Kyoto Encyclopedia of Genes and Genomes , 1999, Nucleic Acids Res..

[106]  N. Hoffman,et al.  Ethylene biosynthesis and its regulation in higher plants , 1984 .