Rearrangement of Actin Cytoskeleton Mediates Invasion of Lotus japonicus Roots by Mesorhizobium loti[C][W]

Infection thread–dependent invasion of legume roots by rhizobia leads to internalization of bacteria into the plant cells, which is one of the salient features of root nodule symbiosis. We found that two genes, Nap1 (for Nck-associated protein 1) and Pir1 (for 121F-specific p53 inducible RNA), involved in actin rearrangements were essential for infection thread formation and colonization of Lotus japonicus roots by its natural microsymbiont, Mesorhizobium loti. nap1 and pir1 mutants developed an excess of uncolonized nodule primordia, indicating that these two genes were not essential for the initiation of nodule organogenesis per se. However, both the formation and subsequent progression of infection threads into the root cortex were significantly impaired in these mutants. We demonstrate that these infection defects were due to disturbed actin cytoskeleton organization. Short root hairs of the mutants had mostly transverse or web-like actin filaments, while bundles of actin filaments in wild-type root hairs were predominantly longitudinal. Corroborating these observations, temporal and spatial differences in actin filament organization between wild-type and mutant root hairs were also observed after Nod factor treatment, while calcium influx and spiking appeared unperturbed. Together with various effects on plant growth and seed formation, the nap1 and pir1 alleles also conferred a characteristic distorted trichome phenotype, suggesting a more general role for Nap1 and Pir1 in processes establishing cell polarity or polar growth in L. japonicus.

[1]  S. Tabata,et al.  CYCLOPS, a mediator of symbiotic intracellular accommodation , 2008, Proceedings of the National Academy of Sciences.

[2]  H. Kouchi,et al.  Transposition of a 600 thousand-year-old LTR retrotransposon in the model legume Lotus japonicus , 2008, Plant Molecular Biology.

[3]  J. Downie,et al.  Coordinating nodule morphogenesis with rhizobial infection in legumes. , 2008, Annual review of plant biology.

[4]  E. Blancaflor,et al.  Improved imaging of actin filaments in transgenic Arabidopsis plants expressing a green fluorescent protein fusion to the C- and N-termini of the fimbrin actin-binding domain 2. , 2007, The New phytologist.

[5]  S. Thirup,et al.  LysM domains mediate lipochitin–oligosaccharide recognition and Nfr genes extend the symbiotic host range , 2007, The EMBO journal.

[6]  J. F. Marsh,et al.  Medicago truncatula NIN Is Essential for Rhizobial-Independent Nodule Organogenesis Induced by Autoactive Calcium/Calmodulin-Dependent Protein Kinase1 , 2007, Plant Physiology.

[7]  S. Tabata,et al.  A Novel Ankyrin-Repeat Membrane Protein, IGN1, Is Required for Persistence of Nitrogen-Fixing Symbiosis in Root Nodules of Lotus japonicus1[OA] , 2007, Plant Physiology.

[8]  S. Tabata,et al.  NUCLEOPORIN85 Is Required for Calcium Spiking, Fungal and Bacterial Symbioses, and Seed Production in Lotus japonicus , 2007, The Plant Cell Online.

[9]  S. Tabata,et al.  A Gain-of-Function Mutation in a Cytokinin Receptor Triggers Spontaneous Root Nodule Organogenesis , 2007, Science.

[10]  J. Perry,et al.  Identification of symbiotically defective mutants of Lotus japonicus affected in infection thread growth. , 2006, Molecular plant-microbe interactions : MPMI.

[11]  J. Šamaj,et al.  Vesicular trafficking, cytoskeleton and signalling in root hairs and pollen tubes. , 2006, Trends in plant science.

[12]  J. Perry,et al.  Lotus japonicus Nodulation Requires Two GRAS Domain Regulators, One of Which Is Functionally Conserved in a Non-Legume1[C][W] , 2006, Plant Physiology.

[13]  Bogumil J. Karas,et al.  Genetic suppressors of the Lotus japonicus har1-1 hypernodulation phenotype. , 2006, Molecular plant-microbe interactions : MPMI.

[14]  R. Sachidanandam,et al.  Comprehensive splice-site analysis using comparative genomics , 2006, Nucleic acids research.

[15]  J. Downie,et al.  Analysis of Nod-factor-induced calcium signaling in root hairs of symbiotically defective mutants of Lotus japonicus. , 2006, Molecular plant-microbe interactions : MPMI.

[16]  M. Hayashi,et al.  New nodulation mutants responsible for infection thread development in Lotus japonicus. , 2006, Molecular plant-microbe interactions : MPMI.

[17]  Satoshi Tabata,et al.  Deregulation of a Ca2+/calmodulin-dependent kinase leads to spontaneous nodule development , 2006, Nature.

[18]  P. Hussey,et al.  Control of the actin cytoskeleton in plant cell growth. , 2006, Annual review of plant biology.

[19]  S. Davis Faculty Opinions recommendation of Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. , 2006 .

[20]  S. Tabata,et al.  A nucleoporin is required for induction of Ca2+ spiking in legume nodule development and essential for rhizobial and fungal symbiosis. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[21]  H. Kouchi,et al.  Positional cloning identifies Lotus japonicus NSP2, a putative transcription factor of the GRAS family, required for NIN and ENOD40 gene expression in nodule initiation. , 2006, DNA research : an international journal for rapid publication of reports on genes and genomes.

[22]  Bogumil J. Karas,et al.  Genetics of symbiosis in Lotus japonicus: recombinant inbred lines, comparative genetic maps, and map position of 35 symbiotic loci. , 2006, Molecular plant-microbe interactions : MPMI.

[23]  T. Baskin,et al.  Directional cell expansion--turning toward actin. , 2005, Current opinion in plant biology.

[24]  A. K. Grennan Putative Arabidopsis arp2/3 complex controls leaf cell morphogenesis. , 2005, Plant physiology.

[25]  P. Hussey,et al.  Arp2/3 and SCAR: plants move to the fore , 2005, Nature Reviews Molecular Cell Biology.

[26]  L. Schauser,et al.  LORE1, an active low-copy-number TY3-gypsy retrotransposon family in the model legume Lotus japonicus. , 2005, The Plant journal : for cell and molecular biology.

[27]  Laurie G. Smith,et al.  Spatial control of cell expansion by the plant cytoskeleton. , 2005, Annual review of cell and developmental biology.

[28]  T. Winzer,et al.  Seven Lotus japonicus Genes Required for Transcriptional Reprogramming of the Root during Fungal and Bacterial Symbiosisw⃞ , 2005, The Plant Cell Online.

[29]  J. F. Marsh,et al.  Nodulation Signaling in Legumes Requires NSP2, a Member of the GRAS Family of Transcriptional Regulators , 2005, Science.

[30]  T. Bisseling,et al.  NSP1 of the GRAS Protein Family Is Essential for Rhizobial Nod Factor-Induced Transcription , 2005, Science.

[31]  S. Tabata,et al.  The Sulfate Transporter SST1 Is Crucial for Symbiotic Nitrogen Fixation in Lotus japonicus Root Nodules , 2005, The Plant Cell Online.

[32]  H. Kouchi,et al.  Microtubule Dynamics in Living Root Hairs: Transient Slowing by Lipochitin Oligosaccharide Nodulation Signalsw⃞ , 2005, The Plant Cell Online.

[33]  Bogumil J. Karas,et al.  Invasion of Lotus japonicus root hairless 1 by Mesorhizobium loti Involves the Nodulation Factor-Dependent Induction of Root Hairs1[w] , 2005, Plant Physiology.

[34]  S. Tabata,et al.  Characterization of the Lotus japonicus Symbiotic Mutant lot1 That Shows a Reduced Nodule Number and Distorted Trichomes1 , 2005, Plant Physiology.

[35]  D. Bird,et al.  Root-knot nematodes and bacterial Nod factors elicit common signal transduction events in Lotus japonicus. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[36]  Naoya Takeda,et al.  Plastid proteins crucial for symbiotic fungal and bacterial entry into plant roots , 2005, Nature.

[37]  D. Szymanski Breaking the WAVE complex: the point of Arabidopsis trichomes. , 2005, Current opinion in plant biology.

[38]  Adrienne R Hardham,et al.  The Cytoskeleton as a Regulator and Target of Biotic Interactions in Plants , 2004, Plant Physiology.

[39]  M. Bevan,et al.  Arabidopsis NAP and PIR Regulate Actin-Based Cell Morphogenesis and Multiple Developmental Processes1 , 2004, Plant Physiology.

[40]  T. Brembu,et al.  NAPP and PIRP Encode Subunits of a Putative Wave Regulatory Protein Complex Involved in Plant Cell Morphogenesis , 2004, The Plant Cell Online.

[41]  D. Szymanski,et al.  Interchangeable functions of Arabidopsis PIROGI and the human WAVE complex subunit SRA1 during leaf epidermal development , 2004, Development.

[42]  P. Hussey,et al.  Arabidopsis NAP1 Is Essential for Arp2/3-Dependent Trichome Morphogenesis , 2004, Current Biology.

[43]  D. Gage Infection and Invasion of Roots by Symbiotic, Nitrogen-Fixing Rhizobia during Nodulation of Temperate Legumes , 2004, Microbiology and Molecular Biology Reviews.

[44]  D. Szymanski,et al.  DISTORTED2 encodes an ARPC2 subunit of the putative Arabidopsis ARP2/3 complex. , 2004, The Plant journal : for cell and molecular biology.

[45]  S. Gygi,et al.  Purification and architecture of the ubiquitous Wave complex. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[46]  A. Edwards,et al.  A Ca2+/calmodulin-dependent protein kinase required for symbiotic nodule development: Gene identification by transcript-based cloning. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[47]  T. Bisseling,et al.  A Putative Ca2+ and Calmodulin-Dependent Protein Kinase Required for Bacterial and Fungal Symbioses , 2004, Science.

[48]  B. Roe,et al.  Medicago truncatula DMI1 Required for Bacterial and Fungal Symbioses in Legumes , 2004, Science.

[49]  Jonathan D. G. Jones,et al.  The maize transposable element Ac is mobile in the legume Lotus japonicus , 1995, Plant Molecular Biology.

[50]  G. Wasteneys,et al.  Remodeling the cytoskeleton for growth and form: an overview with some new views. , 2003, Annual review of plant biology.

[51]  S. Tabata,et al.  Plant recognition of symbiotic bacteria requires two LysM receptor-like kinases , 2003, Nature.

[52]  S. Tabata,et al.  A receptor kinase gene of the LysM type is involved in legumeperception of rhizobial signals , 2003, Nature.

[53]  E. Johannes,et al.  The distributional changes and role of microtubules in Nod factor-challenged Medicago sativa root hairs , 2003, Planta.

[54]  D. Szymanski,et al.  Requirements for Arabidopsis ATARP2 and ATARP3 during Epidermal Development , 2003, Current Biology.

[55]  J. Esseling,et al.  Nod Factor-Induced Root Hair Curling: Continuous Polar Growth towards the Point of Nod Factor Application1 , 2003, Plant Physiology.

[56]  J. Mathur,et al.  Arabidopsis CROOKED encodes for the smallest subunit of the ARP2/3 complex and controls cell shape by region specific fine F-actin formation , 2003, Development.

[57]  M. Hayashi,et al.  crinkle, a Novel Symbiotic Mutant That Affects the Infection Thread Growth and Alters the Root Hair, Trichome, and Seed Development in Lotus japonicus 1 , 2003, Plant Physiology.

[58]  A. Emons,et al.  Unstable F-Actin Specifies the Area and Microtubule Direction of Cell Expansion in Arabidopsis Root Hairs Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.007039. , 2003, The Plant Cell Online.

[59]  Alexandre V. Podtelejnikov,et al.  Mechanism of regulation of WAVE1-induced actin nucleation by Rac1 and Nck , 2002, Nature.

[60]  A. Kereszt,et al.  A receptor kinase gene regulating symbiotic nodule development , 2002, Nature.

[61]  S. Tabata,et al.  A plant receptor-like kinase required for both bacterial and fungal symbiosis , 2002, Nature.

[62]  S. Akao,et al.  Root, root hair, and symbiotic mutants of the model legume Lotus japonicus. , 2002, Molecular plant-microbe interactions : MPMI.

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

[64]  M. Kawaguchi,et al.  Providing the basis for genomics in Lotus japonicus: the accessions Miyakojima and Gifu are appropriate crossing partners for genetic analyses , 2001, Molecular Genetics and Genomics.

[65]  K Van Gestel,et al.  A comparison of F-actin labeling methods for light microscopy in different plant specimens: multiple techniques supplement each other. , 2001, Micron.

[66]  H. Spaink,et al.  Cell biological changes of outer cortical root cells in early determinate nodulation. , 2001, Molecular plant-microbe interactions : MPMI.

[67]  A. Emons,et al.  Lipochito-oligosaccharide nodulation factors stimulate cytoplasmic polarity with longitudinal endoplasmic reticulum and vesicles at the tip in vetch root hairs. , 2000, Molecular plant-microbe interactions : MPMI.

[68]  Leif Schauser,et al.  A plant regulator controlling development of symbiotic root nodules , 1999, Nature.

[69]  T. Bisseling,et al.  Rhizobium Nod Factors Induce an Increase in Sub-apical Fine Bundles of Actin Filaments in Vicia sativa Root Hairs within Minutes , 1999 .

[70]  Zhenbiao Yang,et al.  Control of Pollen Tube Tip Growth by a Rop GTPase–Dependent Pathway That Leads to Tip-Localized Calcium Influx , 1999, Plant Cell.

[71]  R. Iggo,et al.  Increased apoptosis induction by 121F mutant p53 , 1999, EMBO Journal.

[72]  A. Timmers,et al.  Refined analysis of early symbiotic steps of the Rhizobium-Medicago interaction in relationship with microtubular cytoskeleton rearrangements. , 1999, Development.

[73]  Adrian R. Krainer,et al.  AT-AC Pre-mRNA Splicing Mechanisms and Conservation of Minor Introns in Voltage-Gated Ion Channel Genes , 1999, Molecular and Cellular Biology.

[74]  T. Thykjær,et al.  Symbiotic mutants deficient in nodule establishment identified after T-DNA transformation of Lotus japonicus , 1998, Molecular and General Genetics MGG.

[75]  F. Dazzo,et al.  Nodule Organogenesis and Symbiotic Mutants of the Model Legume Lotus japonicus , 1998 .

[76]  F. Sánchez,et al.  Rearrangement of actin microfilaments in plant root hairs responding to rhizobium etli nodulation signals , 1998, Plant physiology.

[77]  E. Kondorosi,et al.  The role of ion fluxes in Nod factor signalling in Medicago sativa , 1998 .

[78]  S. Akao,et al.  Two Ineffective-Nodulating Mutants of Lotus japonicus—Different Phenotypes Caused by the Blockage of Endocytotic Bacterial Release and Nodule Maturation , 1997 .

[79]  D. Ehrhardt,et al.  Calcium Spiking in Plant Root Hairs Responding to Rhizobium Nodulation Signals , 1996, Cell.

[80]  J. Kijne,et al.  Nod factors produced by Rhizobium leguminosarum biovar viciae induce ethylene-related changes in root cortical cells of Vicia sativa ssp. nigra. , 1995, European journal of cell biology.

[81]  C. Ronson,et al.  Nodulating strains of Rhizobium loti arise through chromosomal symbiotic gene transfer in the environment. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[82]  D. Ehrhardt,et al.  Effects of Nod Factors on Alfalfa Root Hair Ca ++ and H + Currents and on Cytoskeletal Behavior , 1994 .

[83]  A. Osbourn,et al.  Advances in Molecular Genetics of Plant-Microbe Interactions , 1994, Current Plant Science and Biotechnology in Agriculture.

[84]  J. Stougaard,et al.  Lotus japonicus, an autogamous, diploid legume species for classical and molecular genetics , 1992 .

[85]  F. Sánchez,et al.  Control of Nodulin Genes in Root-Nodule Development and Metabolism , 1991 .

[86]  S. Camut,et al.  Rhizobium meliloti Genes Encoding Catabolism of Trigonelline Are Induced under Symbiotic Conditions. , 1990, The Plant cell.

[87]  W. Broughton,et al.  Control of leghaemoglobin synthesis in snake beans. , 1971, The Biochemical journal.

[88]  G. Fåhraeus The infection of clover root hairs by nodule bacteria studied by a simple glass slide technique. , 1957, Journal of general microbiology.