Structural aspects of Ascochyta blight of lentil

1995 ◽  
Vol 73 (3) ◽  
pp. 485-497 ◽  
Author(s):  
S. J. Roundhill ◽  
B. A. Fineran ◽  
A. L. J. Cole ◽  
M. Ingerfeld

Ascochyta fabae Speg. f.sp. lentis (Gossen et al. 1986) causes lesions on the leaf, stem, and pod of lentil (Lens culinaris Medik.), thereby reducing seed quality and yield. Lesion formation was studied in two cultivars, Laird and Invincible, using light and electron microscopy of intact and excised leaves and stems inoculated with spore suspension. Spores germinated usually within 6 h of inoculation and germ tubes grew for varying distances along the leaf surface before forming an appressorium, sometimes within less than 10 h. A penetration peg then either directly entered the underlying epidermal cell, or grew as a subcuticular hypha for a short distance before entering the cell. The first response of epidermal cells to presence of the fungus was an aggregation of cytoplasm abutting the site of infection. This was followed closely by deposition of a papilla. Some relatively thick papillae were seen at 29 h postinoculation. The fungus then grew into the papilla and formed an infection vesicle. In susceptible host cells, the protoplasm became necrotic before hyphae grew into the lumen of the cell from the infection vesicle. In more resistant cells, the infection vesicle often became surrounded by electron-dense wall material developed by the host. The fungus remained in susceptible epidermal cells for up to 4 days, amongst remnants of the protoplast, before spreading to the adjacent mesophyll. Hyphae grew into intercellular spaces of the mesophyll and remained there for 2 – 3 days before penetrating the cells. The mesophyll reacted in a similar way to infection as did the epidermis, with only host cells close to the fungus becoming affected. Cultivar Laird was found to be less susceptible to infection than cv. Invincible. At the structural level, the infection process was found to be similar except that in cv. Laird the infection vesicle more frequently became surrounded by electron-dense wall material formed by the host. In stem tissue of cv. Laird the middle lamella was also occasionally thickened with electron-dense material deposited on either side of it. After the degeneration of host tissue, pycnidia-bearing spores were formed 10 – 14 days after inoculation of the leaf. Key words: Ascochyta, lentil, ultrastructure, infection process.

1992 ◽  
Vol 70 (1) ◽  
pp. 58-72 ◽  
Author(s):  
Jeffrey G. Duckett ◽  
Roberto Ligrone

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.


Botany ◽  
2010 ◽  
Vol 88 (10) ◽  
pp. 875-885 ◽  
Author(s):  
Angela Maria Schäfer ◽  
Martin Kemler ◽  
Robert Bauer ◽  
Dominik Begerow

The plant-parasitic genus Microbotryum (Pucciniomycotina) has been used as a model for various biological studies, but fundamental aspects of its life history have not been documented in detail. The smut fungus is characterized by a dimorphic life cycle with a haploid saprophytic yeast-like stage and a dikaryotic plant-parasitic stage, which bears the teliospores as dispersal agents. In this study, seedlings and flowers of Silene latifolia Poir. (Caryophyllaceae) were inoculated with teliospores or sporidial cells of Microbotryum lychnidis-dioicae (DC. ex Liro) G. Deml & Oberw. and the germination of teliospores, the infection process, and the proliferation in the host tissue were documented in vivo using light and electron microscopy. Although germination of the teliospore is crucial for the establishment of Microbotryum, basidium development is variable under natural conditions. In flowers, where the amount of nutrients is thought to be high, the fungus propagates as sporidia, and mating of compatible cells takes place only when flowers are withering and nutrients are decreasing. On cotyledons (i.e., nutrient-depleted conditions), conjugation occurs shortly after teliospore germination, often via intrapromycelial mating. After formation of an infectious hypha with an appressorium, the invasion of the host occurs by direct penetration of the epidermis. While the growth in the plant is typically intercellular, long distance proliferation seems mediated through xylem tracheary elements. At the beginning of the vegetation period, fungal cells were found between meristematic shoot host cells, indicating a dormant phase inside the plant. By using different microscopy techniques, many life stages of Microbotryum are illustrated for the first time, thereby allowing new interpretations of laboratory data.


2020 ◽  
Vol 110 (10) ◽  
pp. 1704-1712 ◽  
Author(s):  
Lijiang Liu ◽  
Li Qin ◽  
Zhuqing Zhou ◽  
Wilhelmina G. H. M. Hendriks ◽  
Shengyi Liu ◽  
...  

As a soilborne protist pathogen, Plasmodiophora brassicae causes the devastating clubroot disease on Brassicaceae crops worldwide. Due to its intracellular obligate biotrophic nature, the life cycle of P. brassicae is still not fully understood. Here, we used fluorescent probe-based confocal microscopy and transmission electron microscopy (TEM) to investigate the infection process of P. brassicae on the susceptible host Arabidopsis under controlled conditions. We found that P. brassicae can initiate the primary infection in both root hairs and epidermal cells, producing the uninucleate primary plasmodium at 1 day postinoculation (dpi). After that, the developed multinucleate primary plasmodium underwent condensing and cytoplasm cleavage into uninucleate zoosporangia from 1 to 4 dpi. This was subsequently followed by the formation of multinucleate zoosporangia and the production of secondary zoospores within zoosporangium. Importantly, the secondary zoospores performed a conjugation in the root epidermal cells after their release. TEM revealed extensive uninucleate secondary plasmodium in cortical cells at 8 dpi, indicating the establishment of the secondary infection. The P. brassicae subsequently developed into binucleate, quadrinucleate, and multinucleate secondary plasmodia from 10 to 15 dpi, during which the clubroot symptoms appeared. The uninucleate resting spores were first observed in the cortical cells at 24 dpi, marking the completion of a life cycle. We also provided evidence that the secondary infection of P. brassicae may represent the diploid sexual life stage. From these findings, we propose a refined life cycle of P. brassicae which will contribute to understanding of the complicated infection biology of P. brassicae.


1987 ◽  
Vol 65 (8) ◽  
pp. 1586-1598 ◽  
Author(s):  
L. Shain ◽  
U. Järlfors

The infection process in four clones of eastern cottonwood susceptible or resistant to leaf rust caused by Melampsora medusae was studied by light and electron microscopy. Infection was initiated by stomatal rather than direct entry. Typical dikaryotic haustoria were observed in all clones within 1 day of inoculation. Some healthy-appearing haustoria were observed in susceptible clones throughout the duration of the study, which was terminated during the initiation of uredial production. Incompatibility was expressed differently in the two resistant clones. In clone St 75, most haustoria and invaded host cells that were observed appeared necrotic within 2 days of inoculation. Cell wall appositions appeared during this time in cells adjoining necrotic host cells. Some infected cells disintegrated within 4 days of inoculation. Affected host cells of clone St 92, on the other hand, plasmolyzed during the first 2 to 3 days after inoculation. Necrotic host cells were not observed in this clone until the 4th day after inoculation. Hyphal ramification and host plasmolysis were extensive at 6 days after inoculation.


Botany ◽  
2008 ◽  
Vol 86 (5) ◽  
pp. 533-538 ◽  
Author(s):  
Charles W. Mims ◽  
Elizabeth A. Richardson

Hyphae of Gymnoconia peckiana (Howe in Peck) Trotter spread from infected Rubus argutus Link. stems into leaf primordia where they proliferated in an intercellular fashion as leaves differentiated. Hyphae were septate, and each compartment appeared to contain a single nucleus. Hyphae gave rise to numerous haustoria that resembled the monokaryotic haustoria of other rust fungi. Hyphae located immediately adjacent to the upper and lower leaf epidermis gave rise to spermogonial initials. Each initial consisted of a small group of tightly packed hyphae that developed in an intercellular space adjacent to the epidermis. As an initial enlarged, the proliferating hyphae pushed their way between, as well as into, epidermal cells. Invaded epidermal cells soon died. A layer of spermatiophores then developed within each young spermogonium and appeared to push the epidermal cell wall material and leaf cuticle covering the spermogonium out from the leaf surface. Once mature, spermatiophores gave rise to a succession of uninucleate spermatia that emerged from the tip of each spermatiophore. Spermatia initially accumulated beneath the layer of epidermal cell wall material and cuticle that covered the developing spermogonium and appeared to push this layer further out from the leaf surface until it ruptured. A few receptive hyphae were observed in mature spermogonia.


1984 ◽  
Vol 62 (6) ◽  
pp. 1232-1244 ◽  
Author(s):  
Pierre Mathieu Charest ◽  
G. B. Ouellette ◽  
F. J. Pauzé

A survey of the initial infection phases of Fusarium oxysporum Schlecht. f. sp. radicis-lycopersici Jarvis and Shoemaker in tomato roots demonstrated that the epidermis was colonized from 12 to 24 h after inoculation. Until 96 h the pathogen was usually limited to the outer cortical area, where fungal cells were found to be either intercellular or intracellular. Host cell wall thickenings and papilla formation were noticeable in the cortical cells but totally absent in the endodermis and the vascular stele. In the cortical area, cytoplasm and walls of affected host cells were mostly disintegrated when the whole root tissues were colonized about 144 h after inoculation. Between 96 and 120 h, hyphae were visible in the endodermis, and 24 h later the vascular stele was colonized. In the latter area, parenchyma cells generally reacted as in the inner cortical area and invasion of vessels proceeded directly through middle lamella and pit membranes. When colonized, vessels contained fibrillogranular material interspersed with bubbles and an osmiophilic coating material. This coating material lined the vessel secondary wall and pit cavities and appeared thicker in the more occluded vessels. The possible implications of these observations on symptoms expression in this and similar plant diseases are discussed.


1985 ◽  
Vol 63 (1) ◽  
pp. 36-42 ◽  
Author(s):  
D. M. Hinton ◽  
C. W. Bacon

The symptomless relationship of tall fescue grass (Festuca arundinacea Schreber) and its endophytic parasitic fungus, Sphacelia typhina (Fr.) Tul. (= Acremonium coenophialum fide Morgan-Jones et Gams), was studied with light and electron microscopy. The fungus was intercellular and was distributed throughout the plant with the exception of roots and leaf blades. The highest concentration of the fungus occurred in the base of the leaf sheath. In the seed the fungus was sequestered between the epithelial cells of the scutellum and the starchy endosperm. The seed embryo was not infected; infection was also not evident during the early stages of seed germination. At the ultrastructural level, the parasitic relationship was established in tissues where there were intercellular spaces. Micrographs indicated that the fungus did not separate the middle lamella of host cells, nor did it produce any obvious alteration in host cell morphology. Ultrastructural characteristics of the fungus depended on the age and type of fescue tissue. Hyphal cells possessed numerous vesicular and multivesicular structures, and in some sections, the hyphal walls were inordinately thickened and with or without an extracellular matrix. The fungus appeared to colonize the innermost areas of tissues in all infected organs and no signs of egress were ever detected.


1981 ◽  
Vol 59 (2) ◽  
pp. 251-263 ◽  
Author(s):  
X. Mourichon ◽  
G. Sallé

An electron microscopic study was performed on haustoria of Phytophthora cactorum (L. et C.) Schroeter developed in tissues of two cultivars of apple fruits: a susceptible variety ('Golden delicious') and a resistant one ('Belle de Boskoop'). Ultrastructure of intercellular hyphae and some aspects of their penetration between contiguous host cells were described. A light dissolution of the host cell walls was observed. Ontogenic investigations indicated that in the susceptible host, the wall of the fungal haustoria was covered with a dense-stained extrahaustorial matrix. Its origin and its polysaccharide nature were demonstrated. On the other hand, the resistant host developed, immediately after the inoculation, a papilla which gave rise, later on, to a sheath enclosing adult haustoria. The role of these callosic structures in the phenomenon of resistance was discussed.


2007 ◽  
Vol 88 (11) ◽  
pp. 2977-2984 ◽  
Author(s):  
Don Stoltz ◽  
Renée Lapointe ◽  
Andrea Makkay ◽  
Michel Cusson

Unlike most viruses, the mature ichnovirus particle possesses two unit membrane envelopes. Following loss of the outer membrane in vivo, nucleocapsids are believed to gain entry into the cytosol via a membrane fusion event involving the inner membrane and the plasma membrane of susceptible host cells; accordingly, experimentally induced damage to the outer membrane might be expected to increase infectivity. Here, in an attempt to develop an in vitro model system for studying ichnovirus infection, we show that digitonin-induced disruption of the virion outer membrane not only increases infectivity, but also uncovers an activity not previously associated with any polydnavirus: fusion from without.


2020 ◽  
Author(s):  
Lee Dolat ◽  
Raphael H. Valdivia

ABSTRACTOur understanding of how the obligate intracellular bacterium Chlamydia trachomatis reprograms the cell biology of host cells in the upper genital tract is largely based on observations made in cell culture with transformed epithelial cell lines. Here we describe a primary spherical organoid system derived from endometrial tissue to recapitulate epithelial cell diversity, polarity, and ensuing responses to Chlamydia infection. Using high-resolution and time-lapse microscopy, we catalogue the infection process in organoids from invasion to egress, including the reorganization of the cytoskeleton and positioning of intracellular organelles. We show this model is amenable to screening C. trachomatis mutants for defects in the fusion of pathogenic vacuoles, the recruitment of intracellular organelles, and inhibition of cell death. Moreover, we reconstructed a primary immune cell response by co-culturing infected organoids with neutrophils, and determined that the effector TepP limits the recruitment of neutrophils to infected organoids. Collectively, our model details a system to study the cell biology of Chlamydia infections in three dimensional structures that better reflect the diversity of cell types and polarity encountered by Chlamydia upon infection of their animal hosts.Summary statement3D endometrial organoids to model Chlamydia infection and the role of secreted virulence factors in reprogramming host epithelial cells and immune cell recruitment


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