Ultrastructure of soybean nodules. I: release of rhizobia from the infection thread

1977 ◽  
Vol 23 (5) ◽  
pp. 573-582 ◽  
Author(s):  
B. Bassett ◽  
R. N. Goodman ◽  
A. Novacky
Keyword(s):  
Root Nodules ◽  
Infection Process ◽  
Infection Thread ◽  
Rhizobium Japonicum ◽  
Wall Material ◽  
Cell Infection ◽  
Host Cytoplasm ◽  
Transmission Electron ◽  
Membrane Envelope ◽  
Membrane Bound

Root nodules on soybeans (var. Clark 63) were examined by transmission electron microscopy 10–12 days after seed inoculation and planting. The cell infection process appeared identical in both effective nodules, induced by Rhizobium japonicum strain 138 (USDA) and in ineffective nodules, induced by strain 8-0 (Iowa). Electron micrographs are presented which suggest that rhizobia are freed from the infection thread by disintegration of the thread wall and compartmentalization of the disintegrated wall material in membrane-bound vesicles derived from the membrane surrounding the thread. As the thread wall is removed in this manner, the bacteria are released into the host cytoplasm by a process which encloses each in an envelope also derived from the thread membrane. Any thread wall material remaining around a bacterium after it has dissociated from the thread is removed from the envelope space by vesiculation of the membrane envelope. Thus, it appears that endocytosis of both the bacteria and the material composing the infection thread wall occurs during release of rhizobia into the host cell.

A light and electron microscope study of the fungal endophytes in the sporophyte and gametophyte of Lycopodium cernuum with observations on the gametophyte–sporophyte junction

1992 ◽  
Vol 70 (1) ◽  
pp. 58-72 ◽  
Author(s):  
Jeffrey G. Duckett ◽  
Roberto Ligrone
Keyword(s):  
Root Nodules ◽  
Fungal Endophytes ◽  
Fungal Endophyte ◽  
Peninsular Malaysia ◽  
Epidermal Cells ◽  
Host Cells ◽  
Wall Material ◽  
Intercellular Spaces ◽  
Host Cytoplasm ◽  
Fungal Associations

The ventral epidermal cells of the photosynthetic, surface-living gametophytes of Lycopodium cernuum, collected from moist shaded banks in Peninsular Malaysia, contain an aseptate fungus. In some cells the hyphae are thick walled and form coils encapsulated by a thin layer of host wall material. In others the fungus is thin walled and shows limited differentiation into larger trunk hyphae and arbuscules. The adjacent host cytoplasm, separated from the fungus by a granular interfacial matrix, contains numerous chloroplasts, mitochondria, and microtubules. The hyphae contact the substratum via the ventral walls of the epidermal cells and the rhizoids are free from infection. In the protocorm and root nodules, aseptate hyphae initially colonize mucilage-filled schizogenous intercellular spaces. Subsequent invasion of the host cells is associated with the development of massive overgrowths of host wall material. The fungal associations in L. cernuum share a mixture of attributes otherwise found in different angiosperm mycorrhizae and in mycotrophic relationships in liverworts. Wall ingrowths are present in both the gametophyte and sporophyte cells in the placenta of L. cernuum. The very limited development of the placenta, compared with L. appressum, certain bryophytes and ferns, the diminutive size, and early senescence of the gametophytes of L. cernuum are all linked to the presence of the protocorm. This massive absorptive organ, homologous to a foot, in terms of its position in sporophyte ontogeny, but external to the parent gametophyte, derives its nutrition partly from photosynthesis and partly from its fungal endophyte. Key words: chloroplasts, Lycopodium, mycorrhiza, pteridophytes, root nodules, symbiosis, transfer cells.

Structure and host–actinomycete interactions in developing root nodules of Comptonia peregrina

1978 ◽  
Vol 56 (5) ◽  
pp. 502-531 ◽  
Author(s):  
William Newcomb ◽  
R. L. Peterson ◽  
Dale Callaham ◽  
John G. Torrey
Keyword(s):  
Host Cell ◽  
Root Nodules ◽  
Host Cells ◽  
Electron Microscopic ◽  
Cortical Cells ◽  
Host Cytoplasm ◽  
Transmission Electron ◽  
Microscopic Studies ◽  
Host Plasma Membrane ◽  
Soil Actinomycete

Correlated fluorescence, bright-field, transmission electron, and scanning electron microscopic studies were made on developing root nodules of Comptonia peregrina (L.) Coult. (Myricaceae) produced by a soil actinomycete which invades the root and establishes a symbiosis leading to fixation of atmospheric dinitrogen. After entering the host via a root hair infection, the hyphae of the endophyte perforate root cortical cells by local degradation of host cell walls and penetration of the host cytoplasm. The intracellular hyphae are always surrounded by host plasma membrane and a thick polysaccharide material termed the capsule. (For convenience, term intracellular refers to the endophyte being inside a Comptonia cell as distinguished from being intercellular, i.e.. between host cells, even though the former is actually extracellular as the endophyte is separated from the host cytoplasm by the host plasmalemma.) Numerous profiles of vesiculate rough endoplasmic reticulum (RER) occur near the growing hyphae. Although the capsule shows a positive Thiery reaction indicating its polysaccharide nature, the fibrillar contents of the RER do not, leaving uncertain whether the capsule results from polymers derived from the RER. Amyloplasts of the cortical cells lose their starch deposits during hyphal proliferation. The hyphae branch extensively in specific layers of the cortex, penetrating much of the host cytoplasm. At this stage, hyphal ends become swollen and form septate club-shaped vesicles within the periphery of the host cells. Lipid-like inclusions and Thiery-positive particles, possibly glycogen, are observed in the hyphae at this time. Associated with hyphal development is an increase in average host cell volume, although nuclear volume appears to remain constant. Concomitant with vesicle maturation, the mitochondrial population increases sharply, suggesting a possible relationship to vesicle function. The intimate interactions between host and endophyte during development of the symbiotic relationship are emphasized throughout.

A correlated light and electron microscopic study of symbiotic growth and differentiation in Pisum sativum root nodules

1976 ◽  
Vol 54 (18) ◽  
pp. 2163-2186 ◽  
Author(s):  
William Newcomb
Keyword(s):  
Electron Microscopy ◽  
Pisum Sativum ◽  
Rhizobium Leguminosarum ◽  
Root Nodules ◽  
Light And Electron Microscopy ◽  
Infection Thread ◽  
Electron Microscopic ◽  
Cortical Cells ◽  
Garden Pea ◽  
Host Cytoplasm

Plants of the garden pea Pisum sativum cv. Little Marvel were grown in aeroponic culture to facilitate observations and microscopy and were inoculated with Rhizobium leguminosarum, and nodules were sampled at five weekly intervals for light and electron microscopy. The invasion of the cortical cells by the infection thread, the structure of the infection thread, and the release of bacteria from it into the host cytoplasm and the subsequent symbiotic growth and differentiation of the two organisms are described in detail. The fine structure of the nodule is correlated with light microscopic observations and morphogenesis. A restriction in the use of the term 'vesicle' is proposed because of the current multiple and confusing usage of the term. The loss of the nodule meristem and its morphogenetic significance are discussed.

Early events in the infection of soybean by Rhizobium japonicum. Time course and cytology of the initial infection process

1982 ◽  
Vol 60 (2) ◽  
pp. 152-161 ◽  
Author(s):  
B. Gillian Turgeon ◽  
Wolfgang D. Bauer
Keyword(s):  
Root Hair ◽  
Time Course ◽  
Cortical Cell ◽  
Root Hairs ◽  
Infection Process ◽  
Infection Thread ◽  
Rhizobium Japonicum ◽  
Outer Cortex ◽  
Light Microscope Level ◽  
Infection Threads

The time course of early infection events in Glycine max following inoculation with Rhizobium japonicum is described. Bacteria became attached to epidermal cells and root hairs within minutes of inoculation. Marked root hair curling occurred within 12 h. Infection thread formation was visible at the light microscope level of resolution about 24 h after inoculation. Infections were observed in short, tightly curled root hairs. These root hairs had not yet emerged at the time of inoculation. Infection threads appeared to originate in pockets formed by contact of the cell wall of the curled root hair with itself. Infection threads in the hairs were multiple and (or) branched. By 48 h, the infection thread(s) had progressed to the base of the root hair but had not yet penetrated into the cortex. Increases in cortical cell cytoplasm and in mitotic division occurred in advance of the penetrating infection thread. A nodule meristem developed in the outer cortex next to the infected root hair by 4 days and was accompanied by cell division across the cortex.

Cytoplasmic bridge formation in the nodule apex of actinorhizal root nodules

1999 ◽  
Vol 77 (9) ◽  
pp. 1351-1357 ◽  
Author(s):  
R Howard Berg
Keyword(s):  
Structural Changes ◽  
Root Nodules ◽  
Parenchyma Cell ◽  
Infection Process ◽  
Infection Thread ◽  
Cytoplasmic Bridge ◽  
Cortical Cells ◽  
Infection Threads ◽  
Related Plant ◽  
Cytoplasmic Bridges

High-pressure frozen - freeze-substituted actinorhizal root nodules of several distantly related plant genera were used to document the sequence of structural changes in cortical cells of the nodule apex that happened prior to their infection. The sequence of mobilization of the plant cell cytoplasm requisite to infection by Frankia was (i) penetration of the parenchyma cell vacuole by cytoplasmic strands, which contained microtubules; (ii) movement of the nucleus and other organelles (Golgi stacks, endoplasmic reticulum, mitochondria), involved later in growth of the infection thread, to the cell center on these strands; (iii) thickening of some of these strands generally located at midpoints of the wall, forming cytoplasmic bridges (preinfection threads); and (iv) infection of the cell by initiation of infection threads (containing Frankia) within the cytoplasmic bridges. The infection thread was caged in microtubules that were oriented along its axis, suggesting the cytoskeleton had a major role in the infection process, perhaps guiding the growth of the infection thread across the cell. The coalignment of cytoplasmic bridges, along several cells, towards the advancing microsymbiont suggested Frankia secretes a diffusible signal eliciting this host response.Key words: actinorhiza, cryofixation, development, infection, microtubules, symbiosis.

scholarly journals Isolation and characterization of the membrane envelope enclosing the bacteroids in soybean root nodules.

1978 ◽  
Vol 78 (3) ◽  
pp. 919-936 ◽  
Author(s):  
D P Verma ◽  
V Kazazian ◽  
V Zogbi ◽  
A K Bal
Keyword(s):  
Cell Wall ◽  
Plasma Membrane ◽  
Host Cell ◽  
Root Nodules ◽  
Infection Thread ◽  
Hypodermic Needle ◽  
Soybean Root ◽  
Isolation And Characterization ◽  
Infection Threads ◽  
Membrane Envelope

The membrane envelope enclosing the bacteroids in soybean root nodules is shown by ultrastructural and biochemical studies to be derived from, and to retain the characteristics of, the host cell plasma membrane. During the early stages of the infection process, which occurs through an invagination, Rhizobium becomes surrounded by the host cell wall and plasma membrane, forming the infection thread. The cell wall of the infection thread is degraded by cellulolytic enzyme(s), leaving behind the enclosed plasma membrane, the membrane envelope. Cellulase activity in young nodules increases two- to threefold as compared to uninfected roots, and this activity is localized in the cell wall matrix of the infection threads. Membrane envelopes were isolated by first preparing bacteroids enclosed in the envelopes on a discontinuous sucrose gradient followed by passage through a hypodermic needle, which released the bacteroids from the membranes. This membrane then sedimented at the interface of 34--45% sucrose (mean density of 1.14 g/cm3). Membranes were characterized by phosphotungstic acid (PTA)-chromic acid staining. ATPase activity, and localization, sensitivity to nonionic detergent Nonidet P-40 (NP-40) and sodium dodecyl sulfate (SDS) gel electrophoresis. These analyses revealed a close similarity between plasma membrane and the membrane envelope. Incorporation of radioactive amino acids into the membrane envelope proteins was sensitive to cycloheximide, suggesting that the biosynthesis of these proteins is primarily under host-cell control. No immunoreactive material to leghemoglobin antibodies was found inside or associated with the isolated bacteroids enclosed in the membrane envelope, and its location is confined to the host cell cytoplasmic matrix.

The occurrence of infected cells, with persistent infection threads, in legume root nodules

1987 ◽  
Vol 65 (3) ◽  
pp. 553-558 ◽  
Author(s):  
Sergio M. de Faria ◽  
Shona G. McInroy ◽  
Janet I. Sprent
Keyword(s):  
Root Nodules ◽  
Root Hairs ◽  
Infection Process ◽  
Wall Material ◽  
Cell Wall Material ◽  
Legume Trees ◽  
Infection Threads ◽  
Host Cell Wall ◽  
Infected Cells ◽  
Intercellular Spread

A survey of the structure of nodules from primitive legume trees was conducted. All genera examined in the subfamily Caesalpinioideae, some from the Papilionoideae, but none from the Mimosoideae had cells in the central, nitrogen-fixing region in which bacteria were confined by host cell wall material in structures resembling infection threads. However, infection of these cells occurred by intercellular spread of rhizobia rather than by infection threads. It is suggested that infection threads may have evolved in infected cells and later extended to early stages of the infection process including entry into root hairs.

Microscopic investigations of novel intracellular bodies of a Nocardia SP

1994 ◽  
Vol 52 ◽  
pp. 356-357
Author(s):  
J. L. Stites
Keyword(s):  
Uranyl Acetate ◽  
Electron Microscopic ◽  
En Bloc ◽  
Ribonucleic Acids ◽  
Transmission Electron ◽  
Nocardia Sp ◽  
Internal Constitution ◽  
Membrane Bound ◽  
Molecular Components ◽  
Protein Rich

A Nocardia sp.was found during an initial transmission electron microscopic (TEM) examination to have unusual intracellular bodies (ICB's) which do not appear to have been described previously in the literature. Most intracellular structures within bacteria have been classified as storage granules, a product of membrane invagination (i.e. mesosomes), or vacuoles. In bacteria there are no known intracellular membrane-bound organelles, and all internal membranes are invaginations of the unit membrane. Several microscopic-level examinations of the Nocardia sp. ICB's were initiated in order to determine their overall structure, classification, and internal constitution.Different TEM staining procedures were performed to determine possible molecular components of the ICB. In all of the staining protocols the ICB's showed a lack of electron density similar to the cell wall. Because the ICB's showed no affinity to any stain, it appeared they do not have strong positive charge (phosphotungstic acid), are not protein rich (en bloc uranyl acetate), lack glycogen and are not phosphate or sulphur rich (lead citrate), nor do they contain lipids or ribonucleic acids (osmium tetroxide).

Haustorial development of Peronospora tabacina infecting Nicotiana tabacum

1983 ◽  
Vol 61 (12) ◽  
pp. 3444-3453 ◽  
Author(s):  
R. N. Trigiano ◽  
C. G. Van Dyke ◽  
H. W. Spurr Jr.
Keyword(s):  
Cell Wall ◽  
Toluidine Blue ◽  
Mesophyll Cell ◽  
Wall Layer ◽  
Peronospora Tabacina ◽  
Host Cytoplasm ◽  
Transmission Electron ◽  
Mold Fungus ◽  
Host Cell Wall ◽  
Hyphal Wall

The development of haustoria in tobacco by the blue-mold fungus Peronospora tabacina was examined using light, scanning, and transmission electron microscopy. Electron-lucent, callose-like appositions were observed between the host plasmalemma and the host mesophyll cell wall prior to haustorial penetration. An electron-opaque penetration matrix was present between the apposition and the host cell wall. The intercellular hyphal wall consisted of two layers which differed in staining quality. The haustorial wall was also two layered, but was primarily composed of and continuous with the inner wall layer of the intercellular hypha. Haustoria were either finger-like or branched and were encased with callose-like material. Most encasements were thickened at the proximal regions of haustoria but were thinner along the distal portions. Vesicles were present in host cytoplasm and were occasionally attached to the invaginated host plasmalemma. These vesicles might contribute to the deposition of the encasement material. The encasement stained positively for callose using aniline blue; calcofluor and toluidine blue O tests for cellulose were inconclusive, and lignin was not detected using toluidine blue O or phloroglucinol–HCl.

The growth of the pollen tube wall in Oenothera organensis

1975 ◽  
Vol 18 (3) ◽  
pp. 519-532
Author(s):  
H.G. Dickinson ◽  
J. Lawson
Keyword(s):  
Pollen Tube ◽  
Tube Wall ◽  
Wall Material ◽  
Fibrillar Material ◽  
Stage Of Development ◽  
Large Numbers ◽  
Membrane Bound ◽  
Pollen Tube Wall ◽  
Different Sources ◽  
Dictyosome Vesicles

The growth of the pollen tube wall of Oenothera is effected by the expulsion of fibrillar material from the cytoplasm into the developing wall. This material may also be seen in the cytoplasm, contained in membrane-bound vesicles. It is not clear how the content of the vesicles is discharged, but it appears not to involve the participation of microtubules. The source of the cytoplasmic fibrillar bodies depends upon the stage of development of the pollen tube. The earilest growth is derived from the inclusion into the wall of vesicles containing pre-formed materials present in the grain on pollination. During the next stage of growth the wall is derived from the content of double-membraned inclusions also present in the pollen. The content of the former vesicles is not so similar to the wall as the latter, but intermediates between the 2 types of vesicle may be seen in the cytoplasm, indicating that the former are formed from the latter. Most of the tube wall is derived from the products of dictyosomes in the pollen grain or tube. These dicytosomes are few in number and they must be exceedingly active. This, and the observation that dictyosome vesicles are frequently associated with banked complexes of mitochondria, indicates that some steps in the metabolism of the vesicular content, perhaps phosphorylation, take place distant from the dicytosomes. These different sources of fibrillar material presumably permit the rapid starting of tube growth, without any attendant metabolism. However, it would be impossible to include enough pre-formed wall material in the grain to enable the full growth of the tube, so once started, it seems that the tube then relies on the elaboration of simple reserves for the contruction of its wall. These reserves are likely to be held in the pollen, and may be the large numbers of starch grains characteristic of the pollen cytoplasm.

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