Sedentary plant-parasitic cyst nematodes are biotrophs that cause significant losses in agriculture. Parasitism is based on modifications of host root cells that lead to the formation of a hypermetabolic feeding site (a syncytium) from which nematodes withdraw nutrients. The host cell cycle is activated in an initial cell selected by the nematode for feeding, followed by activation of neighboring cells and subsequent expansion of feeding site through fusion of hundreds of cells. It is generally assumed that nematodes manipulate production and signaling of the plant hormone cytokinin to activate cell division. In fact, nematodes have been shown to produce cytokinin in vitro; however, whether the hormone is secreted into host plants and plays a role in parasitism remained unknown. Here, we analyzed the spatiotemporal activation of cytokinin signaling during interaction between the cyst nematode, Heterodera schachtii, and Arabidopsis using cytokinin-responsive promoter:reporter lines. Our results showed that cytokinin signaling is activated not only in the syncytium but also in neighboring cells to be incorporated into syncytium. An analysis of nematode infection on mutants that are deficient in cytokinin or cytokinin signaling revealed a significant decrease in susceptibility of these plants to nematodes. Further, we identified a cytokinin-synthesizing isopentenyltransferase gene in H. schachtii and show that silencing of this gene in nematodes leads to a significant decrease in virulence due to a reduced expansion of feeding sites. Our findings demonstrate the ability of a plant-parasitic nematode to synthesize a functional plant hormone to manipulate the host system and establish a long-term parasitic interaction.

Department of Biology Graduate School of Science Osaka University Toyonaka Osaka 560 0043 Japan

Department of Botany Warsaw University of Life Sciences PL 02787 Warsaw Poland;

Division of Plant Sciences and Bond Life Sciences Center University of Missouri Columbia MO 65211;

Institute of Biology Applied Genetics Dahlem Centre of Plant Sciences Freie Universität Berlin D 14195 Berlin Germany;

Laboratory of Growth Regulators Centre of the Region Haná for Biotechnological and Agricultural Research Faculty of Science Palacký University and Institute of Experimental Botany Academy of Sciences of the Czech Republic CZ 78371 Olomouc Czech Republic;

Rheinische Friedrich Wilhelms University of Bonn Department of Molecular Phytomedicine D 53115 Bonn Germany;

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Sasser JN, Freckman DW. A world perspective on nematology: The role of the society. J Nematol. 1986;18(4):596–597.

de Almeida-Engler J, et al. Molecular markers and cell cycle inhibitors show the importance of cell cycle progression in nematode-induced galls and syncytia. Plant Cell. 1999;11(5):793–807. PubMed PMC

Vieira P, et al. The cyclin-dependent kinase inhibitor KRP6 induces mitosis and impairs cytokinesis in giant cells induced by plant-parasitic nematodes in Arabidopsis. Plant Cell. 2014;26(6):2633–2647. PubMed PMC

de Almeida-Engler J, Engler G, Gheysen G. Unravelling the plant cell cycle in nematode induced feeding sites. In: Jones J, Gheysen G, Fenoll C, editors. Genomics and Molecular Genetics of Plant-Nematode Interactions. Springer; Dordrecht, The Netherlands: 2011. pp. 349–368.

Goverse A, et al. Cell cycle activation by plant parasitic nematodes. Plant Mol Biol. 2000;43(5-6):747–761. PubMed

Ha S, Vankova R, Yamaguchi-Shinozaki K, Shinozaki K, Tran LS. Cytokinins: Metabolism and function in plant adaptation to environmental stresses. Trends Plant Sci. 2012;17(3):172–179. PubMed

Kieber JJ, Schaller GE. Cytokinins. Arabidopsis Book. 2014;12:e0168. PubMed PMC

Miyawaki K, et al. Roles of Arabidopsis ATP/ADP isopentenyltransferases and tRNA isopentenyltransferases in cytokinin biosynthesis. Proc Natl Acad Sci USA. 2006;103(44):16598–16603. PubMed PMC

Inoue T, et al. Identification of CRE1 as a cytokinin receptor from Arabidopsis. Nature. 2001;409(6823):1060–1063. PubMed

Riefler M, Novak O, Strnad M, Schmülling T. Arabidopsis cytokinin receptor mutants reveal functions in shoot growth, leaf senescence, seed size, germination, root development, and cytokinin metabolism. Plant Cell. 2006;18(1):40–54. PubMed PMC

Higuchi M, et al. In planta functions of the Arabidopsis cytokinin receptor family. Proc Natl Acad Sci USA. 2004;101(23):8821–8826. PubMed PMC

Nishimura C, et al. Histidine kinase homologs that act as cytokinin receptors possess overlapping functions in the regulation of shoot and root growth in Arabidopsis. Plant Cell. 2004;16(6):1365–1377. PubMed PMC

Dimalla GG, Vanstaden J. Cytokinins in root-knot nematode, Meloidogyne incognita. Plant Sci Lett. 1977;10(1):25–29.

Vanstaden J, Dimalla GG. Comparison of endogenous cytokinins in roots and xylem exudate of nematode resistant and susceptible tomato cultivars. J Exp Bot. 1977;28(107):1351–1356.

Lohar DP, et al. Cytokinins play opposite roles in lateral root formation, and nematode and rhizobial symbioses. Plant J. 2004;38(2):203–214. PubMed

Walters DR, McRoberts N. Plants and biotrophs: A pivotal role for cytokinins? Trends Plant Sci. 2006;11(12):581–586. PubMed

Abelenda JA, Prat S. Cytokinins: Determinants of sink storage ability. Curr Biol. 2013;23(13):R561–R563. PubMed

De Meutter J, et al. Identification of cytokinins produced by the plant parasitic nematodes Heterodera schachtii and Meloidogyne incognita. Mol Plant Pathol. 2003;4(4):271–277. PubMed

Zürcher E, et al. A robust and sensitive synthetic sensor to monitor the transcriptional output of the cytokinin signaling network in planta. Plant Physiol. 2013;161(3):1066–1075. PubMed PMC

D’Agostino IB, et al. Characterization of the response of the Arabidopsis response regulator gene family to cytokinin. Plant Physiol. 2000;124(4):1706–1717. PubMed PMC

Stolz A, et al. The specificity of cytokinin signalling in Arabidopsis thaliana is mediated by differing ligand affinities and expression profiles of the receptors. Plant J. 2011;67(1):157–168. PubMed

Werner T, et al. Root-specific reduction of cytokinin causes enhanced root growth, drought tolerance, and leaf mineral enrichment in Arabidopsis and tobacco. Plant Cell. 2010;22(12):3905–3920. PubMed PMC

Werner T, et al. Cytokinin-deficient transgenic Arabidopsis plants show multiple developmental alterations indicating opposite functions of cytokinins in the regulation of shoot and root meristem activity. Plant Cell. 2003;15(11):2532–2550. PubMed PMC

Colon-Carmona A, You R, Haimovitch-Gal T, Doerner P. Spatio-temporal analysis of mitotic activity with a labile cyclin-GUS fusion protein. Plant J. 1999;20(4):503–508. PubMed

Golinowski W, Grundler FMW, Sobczak M. Changes in the structure of Arabidopsis thaliana during female development of the plant-parasitic nematode Heterodera schachtii. Protoplasma. 1996;194(1):103–116.

Elling AA, et al. Divergent evolution of arrested development in the dauer stage of Caenorhabditis elegans and the infective stage of Heterodera glycines. Genome Biol. 2007;8(10):R211. PubMed PMC

Absmanner B, Stadler R, Hammes UZ. Phloem development in nematode-induced feeding sites: The implications of auxin and cytokinin. Front Plant Sci. 2013;4:241. PubMed PMC

von Schwartzenberg K, et al. Cytokinins in the bryophyte Physcomitrella patens: Analyses of activity, distribution, and cytokinin oxidase/dehydrogenase overexpression reveal the role of extracellular cytokinins. Plant Physiol. 2007;145(3):786–800. PubMed PMC

Lindner A, et al. Isopentenyltransferase-1 (IPT1) knockout in Physcomitrella together with phylogenetic analyses of IPTs provide insights into evolution of plant cytokinin biosynthesis. J Exp Bot. 2014;65(9):2533–2543. PubMed PMC

Kakimoto T. Identification of plant cytokinin biosynthetic enzymes as dimethylallyl diphosphate: ATP/ADP isopentenyltransferases. Plant Cell Physiol. 2001;42(7):677–685. PubMed

Giron D, Frago E, Glevarec G, Pieterse CMJ, Dicke M. Cytokinins as key regulators in plant-microbe-insect interactions: Connecting plant growth and defence. Funct Ecol. 2013;27(3):599–609.

Siddique S, et al. Parasitic worms stimulate host NADPH oxidases to produce reactive oxygen species that limit plant cell death and promote infection. Sci Signal. 2014;7(320):ra33. PubMed

Urwin PE, Lilley CJ, Atkinson HJ. Ingestion of double-stranded RNA by preparasitic juvenile cyst nematodes leads to RNA interference. Mol Plant Microbe Interact. 2002;15(8):747–752. PubMed

Sukno SA, et al. Quantitative detection of double-stranded RNA-mediated gene silencing of parasitism genes in Heterodera glycines. J Nematol. 2007;39(2):145–152. PubMed PMC

Pfaffl MW. A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Res. 2001;29(9):e45. PubMed PMC

Lozano-Torres JL, et al. Apoplastic venom allergen-like proteins of cyst nematodes modulate the activation of basal plant innate immunity by cell surface receptors. PLoS Pathog. 2014;10(12):e1004569. PubMed PMC

Svacinova J, et al. A new approach for cytokinin isolation from Arabidopsis tissues using miniaturized purification: Pipette tip solid-phase extraction. Plant Methods. 2012;8(1):17. PubMed PMC

Rittenberg D, Foster GL. A new procedure for quantitative analysis by isotope dilution, with application to the determination of amino acids and fatty acids. J Biol Chem. 1940;133(3):737–774.

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