A Transcript and Metabolite Atlas of Blackcurrant Fruit Development Highlights Hormonal Regulation and Reveals the Role of Key Transcription Factors

Blackcurrant fruit collected at six stages of development were assessed for changes in gene expression using custom whole transcriptome microarrays and for variation in metabolite content using a combination of liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry. Principal components analysis demonstrated that fruit development could be clearly defined according to their transcript or metabolite profiles. During early developmental stages, metabolite profiles were dominated by amino acids and tannins, whilst transcript profiles were enriched in functions associated with cell division, anatomical structure morphogenesis and cell wall metabolism. During mid fruit development, fatty acids accumulated and transcript profiles were consistent with seed and embryo development. At the later stages, sugars and anthocyanins accumulated consistent with transcript profiles that were associated with secondary metabolism. Transcript data also indicated active signaling during later stages of fruit development. A targeted analysis of signaling networks revealed a dynamic activation and repression of almost 60 different transcripts encoding transcription factors across the course of fruit development, many of which have been demonstrated as pivotal to controlling such processes in other species. Transcripts associated with cytokinin and gibberellin were highly abundant at early fruit development, whilst those associated with ABA and ethylene tended to be more abundant at later stages. The data presented here provides an insight into fruit development in blackcurrant and provides a foundation for further work in the elucidation of the genetic basis of fruit quality.

[1]  S. Matsuo,et al.  Roles and regulation of cytokinins in tomato fruit development , 2012, Journal of experimental botany.

[2]  Paul T. Tarr,et al.  Control of plant stem cell function by conserved interacting transcriptional regulators , 2014, Nature.

[3]  S. Fabris,et al.  New enzymatic method for the determination of total phenolic content in tea and wine. , 2004, Journal of agricultural and food chemistry.

[4]  C. Chervin,et al.  Ethylene signalling receptors and transcription factors over the grape berry development: gene expression profiling , 2010 .

[5]  T. Sun,et al.  Role of the gibberellin receptors GID1 during fruit-set in Arabidopsis. , 2014, The Plant journal : for cell and molecular biology.

[6]  J. Renaudin,et al.  Endoreduplication and Growth of Fleshy Fruits , 2010 .

[7]  R. Brennan,et al.  Improving Fruit Quality in Rubus and Ribes through Breeding , 2009 .

[8]  Jungmook Kim,et al.  AtC3H14, a plant-specific tandem CCCH zinc-finger protein, binds to its target mRNAs in a sequence-specific manner and affects cell elongation in Arabidopsis thaliana. , 2014, The Plant journal : for cell and molecular biology.

[9]  J. Sheen,et al.  Molecular identification of phenylalanine ammonia‐lyase as a substrate of a specific constitutively active Arabidopsis CDPK expressed in maize protoplasts , 2001, FEBS letters.

[10]  C. Hardtke,et al.  The Arabidopsis transcription factor HY5 integrates light and hormone signaling pathways. , 2004, The Plant journal : for cell and molecular biology.

[11]  L. Mariño-Ramírez,et al.  Development and Characterization of Microsatellite Markers for the Cape Gooseberry Physalis peruviana , 2011, PloS one.

[12]  D. Gonzalez,et al.  TCP15 modulates cytokinin and auxin responses during gynoecium development in Arabidopsis. , 2015, The Plant journal : for cell and molecular biology.

[13]  I. Sussex,et al.  Function of the apetala-1 gene during Arabidopsis floral development. , 1990, The Plant cell.

[14]  Jian Zhao Flavonoid transport mechanisms: how to go, and with whom. , 2015, Trends in plant science.

[15]  Willy Govaerts,et al.  Atypical E2F activity restrains APC/CCCS52A2 function obligatory for endocycle onset , 2008, Proceedings of the National Academy of Sciences.

[16]  T. Laux,et al.  Transcriptional activation of Arabidopsis axis patterning genes WOX8/9 links zygote polarity to embryo development. , 2011, Developmental cell.

[17]  A. Sharma,et al.  Role of plant hormones and their interplay in development and ripening of fleshy fruits. , 2013, Journal of experimental botany.

[18]  P. Mattila,et al.  High variability in flavonoid contents and composition between different North-European currant (Ribes spp.) varieties. , 2016, Food chemistry.

[19]  O. M. Heide,et al.  Influence of Controlled Postflowering Temperature and Daylength on Individual Phenolic Compounds in Four Black Currant Cultivars. , 2016, Journal of agricultural and food chemistry.

[20]  S. Cutler,et al.  Abscisic acid: emergence of a core signaling network. , 2010, Annual review of plant biology.

[21]  J. Cushman,et al.  Transcriptomic and metabolite analyses of Cabernet Sauvignon grape berry development , 2007, BMC Genomics.

[22]  M. L. Ruiz del Castillo,et al.  Genotypic variation in fatty acid content of blackcurrant seeds. , 2002, Journal of agricultural and food chemistry.

[23]  Yuling Jiao,et al.  Regulation of inflorescence architecture by cytokinins , 2014, Front. Plant Sci..

[24]  G. Gheysen,et al.  Tightly controlled WRKY23 expression mediates Arabidopsis embryo development , 2013, EMBO reports.

[25]  K. Malik,et al.  Ectopic expression of an Arabidopsis calmodulin-like domain protein kinase-enhanced NADPH oxidase activity and oxidative burst in tomato protoplasts. , 2001, Molecular plant-microbe interactions : MPMI.

[26]  S. Hayat,et al.  Salicylic Acid: Biosynthesis, Metabolism and Physiological Role in Plants , 2007 .

[27]  Zhangjun Fei,et al.  Gene expression in developing watermelon fruit , 2008, BMC Genomics.

[28]  R. Franks,et al.  Novel functional roles for PERIANTHIA and SEUSS during floral organ identity specification, floral meristem termination, and gynoecial development , 2014, Front. Plant Sci..

[29]  G. Bryan,et al.  The Metabolic and Developmental Roles of Carotenoid Cleavage Dioxygenase4 from Potato1[W] , 2010, Plant Physiology.

[30]  A. Törrönen,et al.  High-performance liquid chromatography (HPLC) analysis of phenolic compounds in berries with diode array and electrospray ionization mass spectrometric (MS) detection: ribes species. , 2003, Journal of agricultural and food chemistry.

[31]  Gregory M Symons,et al.  Grapes on Steroids. Brassinosteroids Are Involved in Grape Berry Ripening1 , 2005, Plant Physiology.

[32]  Reijo Karjalainen,et al.  Lipophilic components in black currant seed and pomace extracts , 2012 .

[33]  W. Gruissem,et al.  Fruits: A Developmental Perspective. , 1993, The Plant cell.

[34]  Jie Ren,et al.  Suppression of 9-cis-Epoxycarotenoid Dioxygenase, Which Encodes a Key Enzyme in Abscisic Acid Biosynthesis, Alters Fruit Texture in Transgenic Tomato1[W][OA] , 2012, Plant Physiology.

[35]  The use of genotyping by sequencing in blackcurrant (Ribes nigrum): developing high-resolution linkage maps in species without reference genome sequences , 2014, Molecular Breeding.

[36]  C. Atkinson,et al.  Linking ascorbic acid production in Ribes nigrum with fruit development and changes in sources and sinks. , 2013, Annals of botany.

[37]  D. Choi,et al.  Non-climacteric fruit ripening in pepper: increased transcription of EIL-like genes normally regulated by ethylene , 2010, Functional & Integrative Genomics.

[38]  F. Speleman,et al.  Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes , 2002, Genome Biology.

[39]  Joost T. van Dongen,et al.  A Trihelix DNA Binding Protein Counterbalances Hypoxia-Responsive Transcriptional Activation in Arabidopsis , 2014, PLoS biology.

[40]  A. Fernie,et al.  Metabolic Profiling during Peach Fruit Development and Ripening Reveals the Metabolic Networks That Underpin Each Developmental Stage1[C][W] , 2011, Plant Physiology.

[41]  Caihong Yu,et al.  CFL1, a WW Domain Protein, Regulates Cuticle Development by Modulating the Function of HDG1, a Class IV Homeodomain Transcription Factor, in Rice and Arabidopsis[W] , 2011, Plant Cell.

[42]  A. Bacic,et al.  SEUSS and SEUSS-LIKE 2 coordinate auxin distribution and KNOXI activity during embryogenesis. , 2014, The Plant journal : for cell and molecular biology.

[43]  Kashif Ali,et al.  Transcript and metabolite analysis in Trincadeira cultivar reveals novel information regarding the dynamics of grape ripening , 2011, BMC Plant Biology.

[44]  C. Ávila,et al.  Deciphering the Role of Aspartate and Prephenate Aminotransferase Activities in Plastid Nitrogen Metabolism1[C][W][OPEN] , 2013, Plant Physiology.

[45]  Yunbiao Jiang,et al.  Comprehensive Analysis of ABA Effects on Ethylene Biosynthesis and Signaling during Tomato Fruit Ripening , 2016, PloS one.

[46]  E. Farmer,et al.  Interplant communication: airborne methyl jasmonate induces synthesis of proteinase inhibitors in plant leaves. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[47]  Zheng Qing Fu,et al.  NPR3 and NPR4 are receptors for the immune signal salicylic acid in plants , 2012, Nature.

[48]  Richard E. Reisinger,et al.  Currants and Gooseberries , 1915, Bulletin of popular information - Arnold Arboretum, Harvard University..

[49]  C. Hackett,et al.  Candidate genes associated with bud dormancy release in blackcurrant (Ribes nigrum L.) , 2010, BMC Plant Biology.

[50]  Leng Xiangpeng,et al.  Comparison and verification of the genes involved in ethylene biosynthesis and signaling in apple, grape, peach, pear and strawberry , 2016, Acta Physiologiae Plantarum.

[51]  K. Ljung,et al.  Arabidopsis gulliver1/SUPERROOT2-7 identifies a metabolic basis for auxin and brassinosteroid synergy. , 2014, The Plant journal : for cell and molecular biology.

[52]  F. Stampar,et al.  Changes in fruit quality parameters of four Ribes species during ripening. , 2015, Food chemistry.

[53]  V. Sundaresan,et al.  Auxin Import and Local Auxin Biosynthesis Are Required for Mitotic Divisions, Cell Expansion and Cell Specification during Female Gametophyte Development in Arabidopsis thaliana , 2015, PloS one.

[54]  A. E. Hall,et al.  Ethylene perception by the ERS1 protein in Arabidopsis. , 2000, Plant physiology.

[55]  G. Casadoro,et al.  Different ethylene receptors show an increased expression during the ripening of strawberries: does such an increment imply a role for ethylene in the ripening of these non-climacteric fruits? , 2005, Journal of experimental botany.

[56]  J. Ozga,et al.  Hormonal Interactions in Fruit Development , 2003, Journal of Plant Growth Regulation.

[57]  R. Viola,et al.  L-Ascorbic acid accumulation in fruit of Ribes nigrum occurs by in situ biosynthesis via the L-galactose pathway. , 2007, Functional plant biology : FPB.

[58]  M. Fujisawa,et al.  Transcriptional Regulation of Fruit Ripening by Tomato FRUITFULL Homologs and Associated MADS Box Proteins[W] , 2014, Plant Cell.

[59]  R. Hancock,et al.  Treatment with fungicides influences phytochemical quality of blackcurrant juice , 2012 .

[60]  A. Vianello,et al.  Plant Flavonoids—Biosynthesis, Transport and Involvement in Stress Responses , 2013, International journal of molecular sciences.

[61]  Yang-Dong Guo,et al.  The role of abscisic acid in fruit ripening and responses to abiotic stress. , 2013, Journal of experimental botany.

[62]  J. Giovannoni,et al.  MOLECULAR BIOLOGY OF FRUIT MATURATION AND RIPENING. , 2001, Annual review of plant physiology and plant molecular biology.

[63]  K. David,et al.  A dynamic interplay between phytohormones is required for fruit development, maturation, and ripening , 2013, Front. Plant Sci..

[64]  M. Bayer,et al.  Identification, utilisation and mapping of novel transcriptome-based markers from blackcurrant (Ribes nigrum) , 2011, BMC Plant Biology.

[65]  Q. Ngo,et al.  Genetic and molecular identification of genes required for female gametophyte development and function in Arabidopsis , 2005, Development.

[66]  A. Aharoni,et al.  The SHINE Clade of AP2 Domain Transcription Factors Activates Wax Biosynthesis, Alters Cuticle Properties, and Confers Drought Tolerance when Overexpressed in Arabidopsis w⃞ , 2004, The Plant Cell Online.

[67]  R. Martienssen,et al.  The FRUITFULL MADS-box gene mediates cell differentiation during Arabidopsis fruit development. , 1998, Development.

[68]  T. Altmann,et al.  Metabolic changes in fruits of the tomato dx mutant. , 2006, Phytochemistry.

[69]  Yiguo Hong,et al.  A tomato HD-Zip homeobox protein, LeHB-1, plays an important role in floral organogenesis and ripening , 2008, The Plant journal : for cell and molecular biology.

[70]  A. Fernie,et al.  Reconfiguration of the Achene and Receptacle Metabolic Networks during Strawberry Fruit Development1[C][W] , 2008, Plant Physiology.

[71]  A. Korycinska,et al.  Genetic diversity within a secondary gene pool for Ribes nigrum L. revealed by RAPD and ISSR markers , 2000 .

[72]  Yuan-Yue Shen,et al.  Abscisic Acid Plays an Important Role in the Regulation of Strawberry Fruit Ripening1[W][OA] , 2011, Plant Physiology.

[73]  R. Hancock,et al.  A high-throughput monolithic HPLC method for rapid vitamin C phenotyping of berry fruit. , 2006, Phytochemical analysis : PCA.

[74]  A. Ochoa‐Leyva,et al.  Mango (Mangifera indica L.) cv. Kent fruit mesocarp de novo transcriptome assembly identifies gene families important for ripening , 2015, Front. Plant Sci..

[75]  R. Brennan Currants and Gooseberries , 2017 .

[76]  G. Galla,et al.  Computational annotation of genes differentially expressed along olive fruit development , 2009, BMC Plant Biology.

[77]  A. Fernie,et al.  Gibberellin biosynthesis and signalling during development of the strawberry receptacle. , 2011, The New phytologist.

[78]  B. Donèche,et al.  Relation between hormonal balance and polygalacturonase activity in grape berry , 2005 .

[79]  V. Sadras,et al.  Metabolic effects of elevated temperature on organic acid degradation in ripening Vitis vinifera fruit , 2014, Journal of experimental botany.

[80]  G. Hu,et al.  Comparative transcriptome analyses of a late-maturing mandarin mutant and its original cultivar reveals gene expression profiling associated with citrus fruit maturation , 2017, PeerJ.

[81]  U. Albrecht,et al.  Comparative transcriptome analysis during early fruit development between three seedy citrus genotypes and their seedless mutants , 2017, Horticulture Research.

[82]  C. Hackett,et al.  RAPD fingerprinting of blackcurrant (Ribes nigrum L.) cultivars , 1995, Theoretical and Applied Genetics.

[83]  M. Bevan,et al.  The Ubiquitin Receptors DA1, DAR1, and DAR2 Redundantly Regulate Endoreduplication by Modulating the Stability of TCP14/15 in Arabidopsis , 2015, Plant Cell.

[84]  M. Stitt,et al.  Genome-Wide Identification and Testing of Superior Reference Genes for Transcript Normalization in Arabidopsis1[w] , 2005, Plant Physiology.

[85]  G. Coupland,et al.  Arabidopsis DOF transcription factors act redundantly to reduce CONSTANS expression and are essential for a photoperiodic flowering response. , 2009, Developmental cell.

[86]  D. Grierson Ethylene and the Control of Fruit Ripening , 2013 .

[87]  V. Chalifa-Caspi,et al.  Metabolite and transcript profiling of berry skin during fruit development elucidates differential regulation between Cabernet Sauvignon and Shiraz cultivars at branching points in the polyphenol pathway , 2014, BMC Plant Biology.

[88]  H. Klee,et al.  Characterization of three members of the Arabidopsis carotenoid cleavage dioxygenase family demonstrates the divergent roles of this multifunctional enzyme family. , 2006, The Plant journal : for cell and molecular biology.

[89]  C. Hackett,et al.  The development of a genetic linkage map of blackcurrant (Ribes nigrum L.) and the identification of regions associated with key fruit quality and agronomic traits , 2008, Euphytica.

[90]  Jyan-chyun Jang,et al.  The Arabidopsis tandem CCCH zinc finger proteins AtTZF4, 5 and 6 are involved in light-, abscisic acid- and gibberellic acid-mediated regulation of seed germination. , 2013, Plant, cell & environment.

[91]  Nathan D. Miller,et al.  Auxin and Ethylene Induce Flavonol Accumulation through Distinct Transcriptional Networks1[C][W][OA] , 2011, Plant Physiology.

[92]  박종대,et al.  Brassinosteroids , 2003, Springer Netherlands.

[93]  K. Engel,et al.  Analysis and Sensory Evaluation of Volatile Constituents of Fresh Blackcurrant (Ribes nigrum L.) Fruits. , 2017, Journal of agricultural and food chemistry.

[94]  G. King,et al.  A SQUAMOSA MADS Box Gene Involved in the Regulation of Anthocyanin Accumulation in Bilberry Fruits1[W][OA] , 2010, Plant Physiology.

[95]  Cornelius S. Barry,et al.  Pleiotropic Phenotypes of the sticky peel Mutant Provide New Insight into the Role of CUTIN DEFICIENT2 in Epidermal Cell Function in Tomato1[W][OA] , 2012, Plant Physiology.

[96]  Yuhai Cui,et al.  Identification and characterization of microRNAs and their targets from expression sequence tags of Ribes nigrum , 2016, Canadian Journal of Plant Science.

[97]  Monika Nenon A Dynamic Interplay , 2019, Gender, Collaboration, and Authorship in German Culture.

[98]  A. Trubuil,et al.  Specialization of Oleosins in Oil Body Dynamics during Seed Development in Arabidopsis Seeds[W][OPEN] , 2014, Plant Physiology.

[99]  Pilar Cubas,et al.  Arabidopsis BRANCHED1 Acts as an Integrator of Branching Signals within Axillary Buds[W] , 2007, The Plant Cell Online.

[100]  Zhou Du,et al.  agriGO: a GO analysis toolkit for the agricultural community , 2010, Nucleic Acids Res..

[101]  A. Jauneau,et al.  In Vivo Interference with AtTCP20 Function Induces Severe Plant Growth Alterations and Deregulates the Expression of Many Genes Important for Development[C][W] , 2008, Plant Physiology.

[102]  A. Handa,et al.  Hormonal Regulation of Tomato Fruit Development: A Molecular Perspective , 2005, Journal of Plant Growth Regulation.

[103]  Lingfei Xu,et al.  Histological, hormonal and transcriptomic reveal the changes upon gibberellin-induced parthenocarpy in pear fruit , 2018, Horticulture Research.

[104]  S. Hayat,et al.  Salicylic acid: a plant hormone. , 2007 .

[105]  A. Fortes,et al.  Complex Interplay of Hormonal Signals during Grape Berry Ripening , 2015, Molecules.

[106]  M. Stefova,et al.  Separation, characterization and quantification of phenolic compounds in blueberries and red and black currants by HPLC-DAD-ESI-MSn. , 2011, Journal of agricultural and food chemistry.

[107]  E. Johansson,et al.  Phenols and ascorbic acid in black currants (Ribes nigrum L.): variation due to genotype, location, and year. , 2013, Journal of agricultural and food chemistry.

[108]  A. Liston,et al.  Ribes (Grossulariaceae) phylogeny as indicated by restriction-site polymorphisms of PCR-amplified chloroplast DNA , 1999, Plant Systematics and Evolution.

[109]  Short-term response in leaf metabolism of perennial ryegrass (Lolium perenne) to alterations in nitrogen supply , 2013, Metabolomics.

[110]  Anupama Singh,et al.  Tissue specific and abiotic stress regulated transcription of histidine kinases in plants is also influenced by diurnal rhythm , 2015, Front. Plant Sci..

[111]  Richard J. Pattison,et al.  Evaluating auxin distribution in tomato (Solanum lycopersicum) through an analysis of the PIN and AUX/LAX gene families. , 2012, The Plant journal : for cell and molecular biology.

[112]  C. Hackett,et al.  The development of a PCR-based marker linked to resistance to the blackcurrant gall mite (Cecidophyopsis ribis Acari: Eriophyidae) , 2008, Theoretical and Applied Genetics.

[113]  A. Dale,et al.  Horticulture of Ribes , 2010 .

[114]  C. Simpson,et al.  Physiological, biochemical and molecular responses of the potato (Solanum tuberosum L.) plant to moderately elevated temperature. , 2014, Plant, cell & environment.

[115]  J. Vrebalov,et al.  A MADS-Box Gene Necessary for Fruit Ripening at the Tomato Ripening-Inhibitor (Rin) Locus , 2002, Science.

[116]  C. Bonghi,et al.  The use of microarray μPEACH1.0 to investigate transcriptome changes during transition from pre-climacteric to climacteric phase in peach fruit , 2006 .

[117]  M. Gore,et al.  CAROTENOID CLEAVAGE DIOXYGENASE4 Is a Negative Regulator of β-Carotene Content in Arabidopsis Seeds[W] , 2013, Plant Cell.

[118]  Chuanyou Li,et al.  PIF4 and PIF5 Transcription Factors Link Blue Light and Auxin to Regulate the Phototropic Response in Arabidopsis[C][W][OPEN] , 2013, Plant Cell.

[119]  Nobutaka Mitsuda,et al.  MIXTA-Like Transcription Factors and WAX INDUCER1/SHINE1 Coordinately Regulate Cuticle Development in Arabidopsis and Torenia fournieri[C][W] , 2013, Plant Cell.

[120]  Riki Kawaguchi,et al.  Genome-wide analysis of transcript abundance and translation in Arabidopsis seedlings subjected to oxygen deprivation. , 2005, Annals of botany.

[121]  K. Yano,et al.  Comparative transcriptome analysis reveals distinct ethylene–independent regulation of ripening in response to low temperature in kiwifruit , 2018, BMC Plant Biology.

[122]  M. Zanor,et al.  Integrated Analysis of Metabolite and Transcript Levels Reveals the Metabolic Shifts That Underlie Tomato Fruit Development and Highlight Regulatory Aspects of Metabolic Network Behavior1[W] , 2006, Plant Physiology.

[123]  Lei Gao,et al.  Comparative transcriptome analysis reveals key genes potentially related to soluble sugar and organic acid accumulation in watermelon , 2018, PloS one.

[124]  R. Viola,et al.  Ascorbic Acid Content of Blackcurrant Fruit is Influenced by Both Genetic and Environmental Factors , 2009 .

[125]  J. Ozga,et al.  Gene Expression and Metabolite Profiling of Developing Highbush Blueberry Fruit Indicates Transcriptional Regulation of Flavonoid Metabolism and Activation of Abscisic Acid Metabolism1[W][OA] , 2011, Plant Physiology.

[126]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[127]  S. E. Perry,et al.  Identification of Direct Targets of FUSCA3, a Key Regulator of Arabidopsis Seed Development1[C][W][OA] , 2013, Plant Physiology.

[128]  P. Carbonero,et al.  Leaf expansion in Arabidopsis is controlled by a TCP-NGA regulatory module likely conserved in distantly related species. , 2015, Physiologia plantarum.

[129]  J. Giovannoni,et al.  Regulatory Networks Controlling Ripening , 2013 .

[130]  Zoran Nikoloski,et al.  Integrative Comparative Analyses of Transcript and Metabolite Profiles from Pepper and Tomato Ripening and Development Stages Uncovers Species-Specific Patterns of Network Regulatory Behavior[W][OA] , 2012, Plant Physiology.

[131]  Peter Hedden,et al.  Gibberellin biosynthesis and its regulation. , 2012, The Biochemical journal.

[132]  J. Goodrich,et al.  ZHOUPI controls embryonic cuticle formation via a signalling pathway involving the subtilisin protease ABNORMAL LEAF-SHAPE1 and the receptor kinases GASSHO1 and GASSHO2 , 2013, Development.

[133]  J. Giovannoni Genetic Regulation of Fruit Development and Ripening , 2004, The Plant Cell Online.

[134]  W. Vriezen,et al.  ABA-deficiency results in reduced plant and fruit size in tomato. , 2012, Journal of plant physiology.

[135]  B. Coombe The Development of Fleshy Fruits , 1976 .