scholarly journals Involvement of Diamine Oxidase and Peroxidase in Insolubilization of the Extracellular Matrix: Implications for Pea Nodule Initiation by Rhizobium leguminosarum

2000 ◽  
Vol 13 (4) ◽  
pp. 413-420 ◽  
Author(s):  
J.-P. Wisniewski ◽  
E. A. Rathbun ◽  
J. P. Knox ◽  
N. J. Brewin
Keyword(s):  
Diamine Oxidase ◽  
Rhizobium Leguminosarum ◽  
Plant Cell Wall ◽  
Extracellular Enzymes ◽  
Host Cells ◽  
Soluble Form ◽  
Post Inoculation ◽  
Root Tips ◽  
Pea Seedlings ◽  
Infection Threads

Rhizobium leguminosarum colonizes host cells and tissues through infection threads, which are tubular in-growths of the plant cell wall. Monoclonal antibody MAC265 recognizes a plant matrix glycoprotein (MGP) associated with the lumen of these infection threads. This glycoprotein is also released in soluble form from the root tips of pea seedlings. In the presence of hydrogen peroxide, release of glycoprotein from root tips was not observed. Extractability from root tips was therefore used as the basis for investigating the peroxide-driven insolubilization of MGP and the possible involvement of two extracellular enzymes, peroxidase (POD) and diamine oxidase (DAO), was investigated. Release of MGP from root tips was enhanced by application of POD and DAO inhibitors (salicylhy-droxamic acid and o-phenanthroline, respectively). Furthermore, release of MGP was inhibited by pretreatment of roots with putrescine (the substrate of DAO) and also by application of a partially purified extract of DAO from pea shoots. Following inoculation of pea roots with R. le-guminosarum, elevated levels of DAO transcript were observed by reverse transcriptase-polymerase chain reaction (RT-PCR), but these then dropped to a low level from 4 to 10 days post inoculation, rising again in more mature nodules. In situ hybridization studies indicated that the bulk of the transcription was associated with the infected tissue in the center of the nodule. On the basis of these observations, we postulate that DAO may be involved in the peroxide-driven hardening of MGP in the lumen of infection threads and in the intercellular matrix.

scholarly journals Identification of Symbiotically Defective Mutants of Lotus japonicus Affected in Infection Thread Growth

2006 ◽  
Vol 19 (12) ◽  
pp. 1444-1450 ◽  
Author(s):  
Fabien Lombardo ◽  
Anne B. Heckmann ◽  
Hiroki Miwa ◽  
Jillian A. Perry ◽  
Koji Yano ◽  
...  
Keyword(s):  
Root Hair ◽  
Plant Cell Wall ◽  
Plant Root ◽  
Lotus Japonicus ◽  
Cortical Cell ◽  
Host Cells ◽  
Infection Thread ◽  
Mycorrhizal Symbiosis ◽  
Symbiotic Interaction ◽  
Infection Threads

During the symbiotic interaction between legumes and rhizobia, the host cell plasma membrane and associated plant cell wall invaginate to form a tunnel-like infection thread, a structure in which bacteria divide to reach the plant root cortex. We isolated four Lotus japonicus mutants that make infection pockets in root hairs but form very few infection threads after inoculation with Mesorhizobium loti. The few infection threads that did initiate in the mutants usually did not progress further than the root hair cell. These infection-thread deficient (itd) mutants were unaffected for early symbiotic responses such as calcium spiking, root hair deformation, and curling, as well as for the induction of cortical cell division and the arbuscular mycorrhizal symbiosis. Complementation tests and genetic mapping indicate that itd2 is allelic to Ljsym7, whereas the itd1, itd3, and itd4 mutations identified novel loci. Bacterial release into host cells did occur occasionally in the itd1, itd2, and itd3 mutants suggesting that some infections may succeed after a long period and that infection of nodule cells could occur normally if the few abnormal infection threads that were formed reached the appropriate nodule cells.

scholarly journals Identification of a Family of Extensin-Like Glycoproteins in the Lumen of Rhizobium-Induced Infection Threads in Pea Root Nodules

2002 ◽  
Vol 15 (4) ◽  
pp. 350-359 ◽  
Author(s):  
Elizabeth A. Rathbun ◽  
Michael J. Naldrett ◽  
Nicholas J. Brewin
Keyword(s):  
Posttranscriptional Regulation ◽  
Rhizobium Leguminosarum ◽  
Root Nodules ◽  
Regulation Of Gene Expression ◽  
Pcr Primers ◽  
Amino Acid Sequences ◽  
Sequence Conservation ◽  
Host Cells ◽  
Cdna Clones ◽  
Infection Threads

Rhizobium leguminosarum bv. viciae normally gains access to pea host cells through tubular cell wall ingrowths termed infection threads. Matrix glycoprotein (MGP), a major component of the infection thread lumen, is also secreted from the tips of uninoculated roots and can be released into solution under reducing conditions. Monoclonal antibody MAC265, which recognizes MGP through a carbohydrate epitope, was used for immunoaffinity purification of the glycoprotein from pea roots. Following treatment with chymotrypsin, a peptide fragment was obtained and subjected to N-terminal sequencing. Using PCR primers based on this sequence, cDNA clones were isolated with RNA from inoculated roots and nodules. DNA sequencing of 30 of these clones revealed a family of closely related and repetitive polypeptides with (hydroxy)proline-rich motifs. The cDNA sequences showed over 70% identity with the deduced amino acid sequences of plant extensins, particularly with VfNDS-E from Vicia faba and MtN12 from Medicago truncatula, both of which are strongly upregulated in legume root nodules. Root nodule extensins from pea were of variable length but showed strong sequence conservation of the N-terminus, of the C-terminus, and of a central domain comprising 33 amino acids that were sometimes reiterated. The distribution of tyrosine residues suggested the possible importance of intramolecular and intermolecular cross-linking. There was strong sequence conservation with MtN12 in the 3′-untranslated region, suggesting a possible involvement in posttranscriptional regulation of gene expression.

Early production of rhizopine in nodules induced bySinorhizobium melilotistrain L5-30

2001 ◽  
Vol 47 (2) ◽  
pp. 165-171 ◽  
Author(s):  
K Heinrich ◽  
M H Ryder ◽  
P J Murphy
Keyword(s):  
Sinorhizobium Meliloti ◽  
Rhizobium Leguminosarum ◽  
Fold Increase ◽  
Gas Chromatography Mass Spectrometry ◽  
Post Inoculation ◽  
Infection Threads ◽  
Nodulation Competitiveness ◽  
Meliloti Strain ◽  
Early Production ◽  
Early Effects

The rhizopine L-3-O-methyl-scyllo-inosamine (3-O-MSI) is metabolized by approximately 10% of the strains of Rhizobium leguminosarum bv. viciae and Sinorhizobium meliloti. Rhizopine strains enjoy a substantial competitive advantage in nodulation, which is manifest before 14 days post-inoculation, implying that rhizopine is produced before this time. We were able to detect this compound in the roots of alfalfa (Medicago sativum L. cv. Hunter River) four days after germination (six days post-infection) with S. meliloti strain L5-30 by gas chromatography-mass spectrometry (GC-MS). At four days, nodules were not visible, and the concentration of rhizopine was extremely low, estimated at 67 pg/gfw (picograms/gram fresh weight). The amount increased gradually but remained low until 16 days, when there was a 50-fold increase from day four, by which time nodules were well established. This pattern of synthesis is consistent with previous studies indicating that rhizopine synthesis is regulated by nifA/ntrA regulatory genes, which are maximally expressed in bacteroids at the onset of nitrogen fixation. However, the low level of rhizopine synthesis must be responsible for the early effects on competition for nodulation. Production of rhizopine at this time most likely results from micro-aerobic induction of mos genes in free-living bacteria, either in the infection threads or in the rhizosphere.Key words: Medicago sativum, nodulation competitiveness, Rhizobium, rhizopine, Sinorhizobium meliloti.

scholarly journals Structure and Development of the Legume-Rhizobial Symbiotic Interface in Infection Threads

Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1050
Author(s):  
Anna V. Tsyganova ◽  
Nicholas J. Brewin ◽  
Viktor E. Tsyganov
Keyword(s):  
Cell Wall ◽  
Plant Cell Wall ◽  
Host Cells ◽  
Infection Thread ◽  
Root Hair Cell ◽  
Infection Threads ◽  
Legume Symbiosis ◽  
And Control ◽  
Insight Into

The intracellular infection thread initiated in a root hair cell is a unique structure associated with Rhizobium-legume symbiosis. It is characterized by inverted tip growth of the plant cell wall, resulting in a tunnel that allows invasion of host cells by bacteria during the formation of the nitrogen-fixing root nodule. Regulation of the plant-microbial interface is essential for infection thread growth. This involves targeted deposition of the cell wall and extracellular matrix and tight control of cell wall remodeling. This review describes the potential role of different actors such as transcription factors, receptors, and enzymes in the rearrangement of the plant-microbial interface and control of polar infection thread growth. It also focuses on the composition of the main polymers of the infection thread wall and matrix and the participation of reactive oxygen species (ROS) in the development of the infection thread. Mutant analysis has helped to gain insight into the development of host defense reactions. The available data raise many new questions about the structure, function, and development of infection threads.

scholarly journals Development of Functional Symbiotic White Clover Root Hairs and Nodules Requires Tightly Regulated Production of Rhizobial Cellulase CelC2

2011 ◽  
Vol 24 (7) ◽  
pp. 798-807 ◽  
Author(s):  
Marta Robledo ◽  
José I. Jiménez-Zurdo ◽  
M. José Soto ◽  
Encarnación Velázquez ◽  
Frank Dazzo ◽  
...  
Keyword(s):  
Cell Wall ◽  
White Clover ◽  
Rhizobium Leguminosarum ◽  
Plant Cell Wall ◽  
Root Nodules ◽  
Root Hairs ◽  
Nitrogen Fixing ◽  
Plant Host ◽  
Root Cortex ◽  
Infection Threads

The establishment of rhizobia as nitrogen-fixing endosymbionts within legume root nodules requires the disruption of the plant cell wall to breach the host barrier at strategic infection sites in the root hair tip and at points of bacterial release from infection threads (IT) within the root cortex. We previously found that Rhizobium leguminosarum bv. trifolii uses its chromosomally encoded CelC2 cellulase to erode the noncrystalline wall at the apex of root hairs, thereby creating the primary portal of its entry into white clover roots. Here, we show that a recombinant derivative of R. leguminosarum bv. trifolii ANU843 that constitutively overproduces the CelC2 enzyme has increased competitiveness in occupying aberrant nodule-like root structures on clover that are inefficient in nitrogen fixation. This aberrant symbiotic phenotype involves an extensive uncontrolled degradation of the host cell walls restricted to the expected infection sites at tips of deformed root hairs and significantly enlarged infection droplets at termini of wider IT within the nodule infection zone. Furthermore, signs of elevated plant host defense as indicated by reactive oxygen species production in root tissues were more evident during infection by the recombinant strain than its wild-type parent. Our data further support the role of the rhizobial CelC2 cell wall–degrading enzyme in primary infection, and show evidence of its importance in secondary symbiotic infection and tight regulation of its production to establish an effective nitrogen-fixing root nodule symbiosis.

scholarly journals Extracellular Glycanases of Rhizobium leguminosarum Are Activated on the Cell Surface by an Exopolysaccharide-Related Component

2000 ◽  
Vol 182 (5) ◽  
pp. 1304-1312 ◽  
Author(s):  
Angeles Zorreguieta ◽  
Christine Finnie ◽  
J. Allan Downie
Keyword(s):  
Cell Wall ◽  
Agrobacterium Tumefaciens ◽  
Cell Surface ◽  
Rhizobium Leguminosarum ◽  
Plant Cell Wall ◽  
Agar Medium ◽  
Wild Type ◽  
Close Proximity ◽  
Mutant Strains ◽  
Colony Surface

ABSTRACT Rhizobium leguminosarum secretes two extracellular glycanases, PlyA and PlyB, that can degrade exopolysaccharide (EPS) and carboxymethyl cellulose (CMC), which is used as a model substrate of plant cell wall cellulose polymers. When grown on agar medium, CMC degradation occurred only directly below colonies of R. leguminosarum, suggesting that the enzymes remain attached to the bacteria. Unexpectedly, when a PlyA-PlyB-secreting colony was grown in close proximity to mutants unable to produce or secrete PlyA and PlyB, CMC degradation occurred below that part of the mutant colonies closest to the wild type. There was no CMC degradation in the region between the colonies. By growing PlyB-secreting colonies on a lawn of CMC-nondegrading mutants, we could observe a halo of CMC degradation around the colony. Using various mutant strains, we demonstrate that PlyB diffuses beyond the edge of the colony but does not degrade CMC unless it is in contact with the appropriate colony surface. PlyA appears to remain attached to the cells since no such diffusion of PlyA activity was observed. EPS defective mutants could secrete both PlyA and PlyB, but these enzymes were inactive unless they came into contact with an EPS+ strain, indicating that EPS is required for activation of PlyA and PlyB. However, we were unable to activate CMC degradation with a crude EPS fraction, indicating that activation of CMC degradation may require an intermediate in EPS biosynthesis. Transfer of PlyB to Agrobacterium tumefaciens enabled it to degrade CMC, but this was only observed if it was grown on a lawn ofR. leguminosarum. This indicates that the surface ofA. tumefaciens is inappropriate to activate CMC degradation by PlyB. Analysis of CMC degradation by other rhizobia suggests that activation of secreted glycanases by surface components may occur in other species.

scholarly journals The Role of Arabinogalactan Type II Degradation in Plant-Microbe Interactions

2021 ◽  
Vol 12 ◽  
Author(s):  
Maria Guadalupe Villa-Rivera ◽  
Horacio Cano-Camacho ◽  
Everardo López-Romero ◽  
María Guadalupe Zavala-Páramo
Keyword(s):  
Cell Wall ◽  
Plant Defense ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Degradation Products ◽  
Hydrolytic Enzymes ◽  
Pathogenic Fungi ◽  
Plant Interactions ◽  
Root Tips ◽  
Microbe Interactions

Arabinogalactans (AGs) are structural polysaccharides of the plant cell wall. A small proportion of the AGs are associated with hemicellulose and pectin. Furthermore, AGs are associated with proteins forming the so-called arabinogalactan proteins (AGPs), which can be found in the plant cell wall or attached through a glycosylphosphatidylinositol (GPI) anchor to the plasma membrane. AGPs are a family of highly glycosylated proteins grouped with cell wall proteins rich in hydroxyproline. These glycoproteins have important and diverse functions in plants, such as growth, cellular differentiation, signaling, and microbe-plant interactions, and several reports suggest that carbohydrate components are crucial for AGP functions. In beneficial plant-microbe interactions, AGPs attract symbiotic species of fungi or bacteria, promote the development of infectious structures and the colonization of root tips, and furthermore, these interactions can activate plant defense mechanisms. On the other hand, plants secrete and accumulate AGPs at infection sites, creating cross-links with pectin. As part of the plant cell wall degradation machinery, beneficial and pathogenic fungi and bacteria can produce the enzymes necessary for the complete depolymerization of AGs including endo-β-(1,3), β-(1,4) and β-(1,6)-galactanases, β-(1,3/1,6) galactanases, α-L-arabinofuranosidases, β-L-arabinopyranosidases, and β-D-glucuronidases. These hydrolytic enzymes are secreted during plant-pathogen interactions and could have implications for the function of AGPs. It has been proposed that AGPs could prevent infection by pathogenic microorganisms because their degradation products generated by hydrolytic enzymes of pathogens function as damage-associated molecular patterns (DAMPs) eliciting the plant defense response. In this review, we describe the structure and function of AGs and AGPs as components of the plant cell wall. Additionally, we describe the set of enzymes secreted by microorganisms to degrade AGs from AGPs and its possible implication for plant-microbe interactions.

Effects of Killing by Heat or Desiccation on Membrane Structure in Pea Roots

1975 ◽  
Vol 2 (2) ◽  
pp. 225 ◽  
Author(s):  
MS Buttrose ◽  
JG Swift
Keyword(s):  
Membrane Structure ◽  
Structural Protein ◽  
Root Tips ◽  
Freeze Fracturing ◽  
Lipid Structure ◽  
Pea Seedlings ◽  
Thin Sectioning ◽  
Pea Seeds ◽  
Pea Roots ◽  
Dead Tissue

Radicles of dry pea seeds were killed by heating at temperatures up to 160°C, and root tips excised from pea seedlings were killed by desiccation. The effect of these treatments on the appearance of membranes was studied in the electron microscope after fixation and thin-sectioning and after freeze- fracturing, After fixation, membranes from dead tissue appeared modified in various ways compared with membranes from control tissue, whereas after freeze-fracturing they appeared unchanged. It is argued that heating and desiccation denature or disorganize structural protein in membranes but that they do not affect the lipid structure of the membrane.

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