Glutamine synthetase I-deficiency in Mesorhizobium loti differentially affects nodule development and activity in Lotus japonicus

2014 ◽  
Vol 171 (5) ◽  
pp. 104-108 ◽  
Author(s):  
Sirinapa Chungopast ◽  
Pilunthana Thapanapongworakul ◽  
Hiroyuki Matsuura ◽  
Tan Van Dao ◽  
Toshimasa Asahi ◽  
...  
Keyword(s):  
Glutamine Synthetase ◽  
Lotus Japonicus ◽  
Nodule Development ◽  
Mesorhizobium Loti ◽  
Glutamine Synthetase I

scholarly journals Defects in Rhizobial Cyclic Glucan and Lipopolysaccharide Synthesis Alter Legume Gene Expression During Nodule Development

2008 ◽  
Vol 21 (1) ◽  
pp. 50-60 ◽  
Author(s):  
Alejandra L. D'Antuono ◽  
Thomas Ott ◽  
Lene Krusell ◽  
Vera Voroshilova ◽  
Rodolfo A. Ugalde ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mutant Strain ◽  
Early Recognition ◽  
Lotus Japonicus ◽  
Cdna Array ◽  
Nodule Development ◽  
Marker Genes ◽  
Wild Type ◽  
Mesorhizobium Loti ◽  
Expression Of Genes

cDNA array technology was used to compare transcriptome profiles of Lotus japonicus roots inoculated with a Mesorhizobium loti wild-type and two mutant strains affected in cyclic β(1-2) glucan synthesis (cgs) and in lipopolysaccharide synthesis (lpsβ2). Expression of genes associated with the development of a fully functional nodule was significantly affected in plants inoculated with the cgs mutant. Array results also revealed that induction of marker genes for nodule development was delayed when plants were inoculated with the lpsβ2 mutant. Quantitative real-time reverse-transcriptase polymerase chain reaction was used to quantify gene expression of a subset of genes involved in plant defense response, redox metabolism, or genes that encode for nodulins. The majority of the genes analyzed in this study were more highly expressed in roots inoculated with the wild type compared with those inoculated with the cgs mutant strain. Some of the genes exhibited a transient increase in transcript levels during intermediate steps of normal nodule development while others displayed induced expression during the final steps of nodule development. Ineffective nodules induced by the glucan mutant showed higher expression of phenylalanine ammonia lyase than wild-type nodules. Differences in expression pattern of genes involved in early recognition and signaling were observed in plants inoculated with the M. loti mutant strain affected in the synthesis of cyclic glucan.

scholarly journals A Novel Fix¯ Symbiotic Mutant of Lotus japonicus, Ljsym105, Shows Impaired Development and Premature Deterioration of Nodule Infected Cells and Symbiosomes

2006 ◽  
Vol 19 (7) ◽  
pp. 780-788 ◽  
Author(s):  
Md. Shakhawat Hossain ◽  
Yosuke Umehara ◽  
Hiroshi Kouchi
Keyword(s):  
Nitrogen Deficiency ◽  
Lotus Japonicus ◽  
Nodule Development ◽  
Acetylene Reduction Activity ◽  
Nitrogen Fixing ◽  
Wild Type ◽  
Electron Microscopic ◽  
Model Legume ◽  
Mesorhizobium Loti ◽  
Infected Cells

Nitrogen-fixing symbiosis between legume plants and rhizobia is established through complex interactions between two symbiotic partners. To identify the host legume genes that play crucial roles in such interactions, we isolated a novel Fix¯ mutant, Ljsym105, from a model legume Lotus japonicus MG-20. The Ljsym105 plants displayed nitrogen-deficiency symptoms after inoculation with Mesorhizobium loti under nitrogen-free conditions, but their growth recovered when supplied with nitrogen-rich nutrients. Ljsym105 was recessive and monogenic and mapped on the upper portion of chromosome 4. The mutant Ljsym105 formed an increased number of small and pale-pink nodules. Nitrogenase (acetylene reduction) activity per nodule fresh weight was low but retained more than 50% of that of the wild-type nodules. Light and electron microscopic observations revealed that the Ljsym105 nodule infected cells were significantly smaller than those of wild-type plants, contained enlarged symbiosomes with multiple bacteroids, and underwent deterioration of the symbiosomes prematurely as well as disintegration of the whole infected cell cytoplasm. These results indicate that the ineffectiveness of the Ljsym105 nodules is primarily due to impaired growth of infected cells accompanied with the premature senescence induced at relatively early stages of nodule development. These symbiotic phenotypes are discussed in respect to possible functions of the LjSym105 locus in the symbiotic interactions required for establishment of the nitrogen-fixing symbiosis.

scholarly journals Photorespiratory Metabolism and Nodule Function: Behavior of Lotus japonicus Mutants Deficient in Plastid Glutamine Synthetase

2012 ◽  
Vol 25 (2) ◽  
pp. 211-219 ◽  
Author(s):  
Margarita García-Calderón ◽  
Maurizio Chiurazzi ◽  
M. Rosario Espuny ◽  
Antonio J. Márquez
Keyword(s):  
Nitrogen Fixation ◽  
Glutamine Synthetase ◽  
Carbon Metabolism ◽  
Lotus Japonicus ◽  
Acetylene Reduction Activity ◽  
Symbiotic Association ◽  
Atmospheric Nitrogen ◽  
Mesorhizobium Loti ◽  
Reduction Activity ◽  
Air Atmosphere

Two photorespiratory mutants of Lotus japonicus deficient in plastid glutamine synthetase (GS2) were examined for their capacity to establish symbiotic association with Mesorhizobium loti bacteria. Biosynthetic glutamine synthetase (GS) activity was reduced by around 40% in crude nodule extracts from mutant plants as compared with the wild type (WT). Western blot analysis further confirmed the lack of GS2 polypeptide in mutant nodules. The decrease in GS activity affected the nodular carbon metabolism under high CO2 (suppressed photorespiration) conditions, although mutant plants were able to form nodules and fix atmospheric nitrogen. However, when WT and mutant plants were transferred to an ordinary air atmosphere (photorespiratory active conditions) the nodulation process and nitrogen fixation were substantially affected, particularly in mutant plants. The number and fresh weight of mutant nodules as well as acetylene reduction activity showed a strong inhibition compared with WT plants. Optical microscopy studies from mutant plant nodules revealed the anticipated senescence phenotype linked to an important reduction in starch and sucrose levels. These results show that, in Lotus japonicus, photorespiration and, particularly, GS2 deficiency result in profound limitations in carbon metabolism that affect the nodulation process and nitrogen fixation.

scholarly journals Genetic Suppressors of the Lotus japonicus har1-1 Hypernodulation Phenotype

2006 ◽  
Vol 19 (10) ◽  
pp. 1082-1091 ◽  
Author(s):  
Jeremy Murray ◽  
Bogumil Karas ◽  
Loretta Ross ◽  
Andreas Brachmann ◽  
Cameron Wagg ◽  
...  
Keyword(s):  
Glomus Intraradices ◽  
Lotus Japonicus ◽  
Introgression Line ◽  
Nodule Development ◽  
Genetic Dissection ◽  
Kinase Gene ◽  
Mesorhizobium Loti ◽  
Genetic Suppressors ◽  
Nodulation Capacity ◽  
Mutant Lines

Lotus japonicus har1 mutants respond to inoculation with Mesorhizobium loti by forming an excessive number of nodules due to genetic lesions in the HAR1 autoregulatory receptor kinase gene. In order to expand the repertoire of mutants available for the genetic dissection of the root nodule symbiosis (RNS), a screen for suppressors of the L. japonicus har1-1 hypernodulation phenotype was performed. Of 150,000 M2 plants analyzed, 61 stable L. japonicus double-mutant lines were isolated. In the context of the har1-1 mutation, 26 mutant lines were unable to form RNS, whereas the remaining 35 mutant lines carried more subtle symbiotic phenotypes, either forming white ineffective nodules or showing reduced nodulation capacity. When challenged with Glomus intraradices, 18 of the 61 suppressor lines were unable to establish a symbiosis with this arbuscular mycorrhiza fungus. Using a combined approach of genetic mapping, targeting induced local lesions in genomics, and sequencing, all non-nodulating mutant lines were characterized and shown to represent new alleles of at least nine independent symbiotic loci. The class of mutants with reduced nodulation capacity was of particular interest because some of them may specify novel plant functions that regulate nodule development in L. japonicus. To facilitate mapping of the latter class of mutants, an introgression line, in which the har1-1 allele was introduced into a polymorphic background of L. japonicus ecotype MG20, was constructed.

scholarly journals The Mesorhizobium loti purB Gene Is Involved in Infection Thread Formation and Nodule Development in Lotus japonicus

2007 ◽  
Vol 189 (22) ◽  
pp. 8347-8352 ◽  
Author(s):  
Shin Okazaki ◽  
Yoshiyuki Hattori ◽  
Kazuhiko Saeki
Keyword(s):  
Lotus Japonicus ◽  
Infection Thread ◽  
Nodule Development ◽  
Nodule Formation ◽  
Mesorhizobium Loti ◽  
Infection Threads ◽  
Thread Formation ◽  
Infection Thread Formation

ABSTRACT The purB and purH mutants of Mesorhizobium loti exhibited purine auxotrophy and nodulation deficiency on Lotus japonicus. In the presence of adenine, only the purH mutant induced nodule formation and the purB mutant produced few infection threads, suggesting that 5-aminoimidazole-4-carboxamide ribonucleotide biosynthesis catalyzed by PurB is required for the establishment of symbiosis.

scholarly journals Blue Light Perception by Both Roots and Rhizobia Inhibits Nodule Formation in Lotus japonicus

2016 ◽  
Vol 29 (10) ◽  
pp. 786-796 ◽  
Author(s):  
Aya Shimomura ◽  
Ayumi Naka ◽  
Nobuyuki Miyazaki ◽  
Sayaka Moriuchi ◽  
Susumu Arima ◽  
...  
Keyword(s):  
Blue Light ◽  
Red Light ◽  
Lotus Japonicus ◽  
Inhibitory Effect ◽  
Nodule Development ◽  
Nodule Formation ◽  
Mesorhizobium Loti ◽  
Empty Vector ◽  
Legume Nodulation ◽  
Strong Inhibitory Effect

In many legumes, roots that are exposed to light do not form nodules. Here, we report that blue light inhibits nodulation in Lotus japonicus roots inoculated with Mesorhizobium loti. Using RNA interference, we suppressed the expression of the phototropin and cryptochrome genes in L. japonicus hairy roots. Under blue light, plants transformed with an empty vector did not develop nodules, whereas plants exhibiting suppressed expression of cry1 and cry2 genes formed nodules. We also measured rhizobial growth to investigate whether the inhibition of nodulation could be caused by a reduced population of rhizobia in response to light. Although red light had no effect on rhizobial growth, blue light had a strong inhibitory effect. Rhizobial growth under blue light was partially restored in signature-tagged mutagenesis (STM) strains in which LOV-HK/PAS- and photolyase-related genes were disrupted. Moreover, when Ljcry1A and Ljcry2B-silenced plants were inoculated with the STM strains, nodulation was additively increased. Our data show that blue light receptors in both the host plant and the symbiont have a profound effect on nodule development. The exact mechanism by which these photomorphogenetic responses function in the symbiosis needs further study, but they are clearly involved in optimizing legume nodulation.

scholarly journals Lotus japonicus LjKUP Is Induced Late During Nodule Development and Encodes a Potassium Transporter of the Plasma Membrane

2004 ◽  
Vol 17 (7) ◽  
pp. 789-797 ◽  
Author(s):  
Guilhem Desbrosses ◽  
Claudia Kopka ◽  
Thomas Ott ◽  
Michael K. Udvardi
Keyword(s):  
Plasma Membrane ◽  
Symbiotic Nitrogen Fixation ◽  
Ion Homeostasis ◽  
Lotus Japonicus ◽  
Nodule Development ◽  
Mesorhizobium Loti ◽  
Potassium Transporter ◽  
Membrane Location ◽  
High Level ◽  
Arabidopsis Cells

The KUP family of potassium transporters in plants is large but poorly characterized. We isolated and characterized the first KUP transporter from a legume, LjKUP of Lotus japonicus. Although expressed throughout plants, LjKUP transcript levels were highest in nodules. Induction of LjKUP expression occurred late during nodule development, at a time of rapid organ expansion. A high level of LjKUP expression was maintained in mature, full-sized nodules. However, induction of LjKUP expression was independent of symbiotic nitrogen fixation (SNF), and occurred in ineffective nodules resulting from mutations in either the plant or its microsymbiont, Mesorhizobium loti. Heterologous expression of LjKUP in Escherichia coli showed that the protein is able to transport potassium. Transient expression of a GFP-LjKUP fusion protein in Arabidopsis cells indicated a plasma membrane location for the transporter. Taken together, the results indicate that LjKUP is a potassium transporter of the plasma membrane, which may play roles in cell expansion during nodule development and in ion homeostasis during SNF.

scholarly journals ALotus japonicusE3 ligase interacts with the Nod factor receptor 5 and positively regulates nodulation

2018 ◽  
Author(s):  
Daniela Tsikou ◽  
Estrella E. Ramirez ◽  
Ioanna S. Psarrakou ◽  
Jaslyn E. Wong ◽  
Dorthe B. Jensen ◽  
...  
Keyword(s):  
Lotus Japonicus ◽  
E3 Ligase ◽  
Kinase Domain ◽  
Plant Interactions ◽  
Post Translational Modification ◽  
Direct Role ◽  
Nod Factor ◽  
Mesorhizobium Loti ◽  
Signaling Systems ◽  
Nodulation Capacity

SUMMARYPost-translational modification of receptor proteins is involved in activation and de-activation of signaling systems in plants. Both ubiquitination and deubiquitination have been implicated in plant interactions with pathogens and symbionts. Here we presentLjPUB13, a PUB-ARMADILLO repeat E3 ligase that specifically ubiquitinates the kinase domain of the Nod Factor receptor NFR5 and has a direct role in nodule organogenesis events inLotus japonicus. Phenotypic analyses of three LORE1 retroelement insertion plant lines revealed thatpub13plants display delayed and reduced nodulation capacity and retarded growth.LjPUB13expression is spatially regulated during symbiosis withMesorhizobium loti, with increased levels in young developing nodules. Thus,LjPUB13 is an E3 ligase with a positive regulatory role during the initial stages of nodulation inL. japonicus.

scholarly journals Correction: Reassimilation of Photorespiratory Ammonium in Lotus japonicus Plants Deficient in Plastidic Glutamine Synthetase

PLoS ONE ◽  
2016 ◽  
Vol 11 (5) ◽  
pp. e0156568 ◽  
Author(s):  
Carmen M. Pérez-Delgado ◽  
Margarita García-Calderón ◽  
Antonio J. Márquez ◽  
Marco Betti
Keyword(s):  
Glutamine Synthetase ◽  
Lotus Japonicus

scholarly journals Lotus japonicus Contains Two Distinct ENOD40 Genes That Are Expressed in Symbiotic, Nonsymbiotic, and Embryonic Tissues

2000 ◽  
Vol 13 (9) ◽  
pp. 987-994 ◽  
Author(s):  
Emmanouil Flemetakis ◽  
Nektarios Kavroulakis ◽  
Nicolette E. M. Quaedvlieg ◽  
Herman P. Spaink ◽  
Maria Dimou ◽  
...  
Keyword(s):  
Root Nodule ◽  
Vascular System ◽  
Lotus Japonicus ◽  
Mrna Accumulation ◽  
Mesorhizobium Loti ◽  
Embryonic Tissues ◽  
Early Nodulin ◽  
Gene Transcripts ◽  
Young Seed

ENOD40, an early nodulin gene, has been postulated to play a significant role in legume root nodule ontogenesis. We have isolated two distinct ENOD40 genes from Lotus japonicus. The transcribed regions of the two ENOD40 genes share 65% homology, while the two promoters showed no significant homology. Both transcripts encode a putative dodecapeptide similar to that identified in other legumes forming determinate nodules. Both ENOD40 genes are coordinately expressed following inoculation of roots with Mesorhizobium loti or treatment with purified Nod factors. In the former case, mRNA accumulation could be detected up to 10 days following inoculation while in the latter case the accumulation was transient. High levels of both ENOD40 gene transcripts were found in nonsymbiotic tissues such as stems, fully developed flowers, green seed pods, and hypocotyls. A relatively lower level of both transcripts was observed in leaves, roots, and cotyledons. In situ hybridization studies revealed that, in mature nodules, transcripts of both ENOD40 genes accumulate in the nodule vascular system; additionally, in young seed pods strong signal is observed in the ovule, particularly in the phloem and epithelium, as well as in globular stage embryos.

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