A comparison of the death induced by fungal invasion or toxic chemicals in cowpea epidermal cells. II. Responses induced by Erysiphe cichoracearum

1988 ◽  
Vol 66 (4) ◽  
pp. 624-634 ◽  
Author(s):  
Susan L. F. Meyer ◽  
Michèle C. Heath

Cowpea leaves were inoculated with the plantain powdery mildew fungus, Erysiphe cichoracearum, and fresh epidermal cells overlying veins were examined by light microscopy before being cleared or prepared for electron microscopy. Fungal appressoria usually formed a haustorium in the underlying nonhost cell, but only after what appeared to be an unsuccessful penetration attempt that induced a transient cytoplasmic aggregate, a ring of autofluorescence in the plant wall (best seen in cleared tissue), and in two examples observed ultrastructurally, a small penetration peg embedded in a callose-like papilla. The haustorium developed from a different penetration peg and elicited the death of the invaded cell. As reported for the death of cowpea epidermal cells elicited by CuCl2, cytoplasmic changes that occurred rapidly in fresh tissue after cytoplasmic streaming had stopped correlated closely with changes in ultrastructure. Compared with the CuCl2 study, microtubules and Golgi bodies disappeared faster and membranes appeared more disorganized. These data suggest that in cowpea epidermal cells, ultrastructural changes accurately predict the onset of cell death and may also reflect differences in its modes of induction.

1988 ◽  
Vol 66 (4) ◽  
pp. 613-623 ◽  
Author(s):  
Susan L. F. Meyer ◽  
Michèle C. Heath

Death was induced in cowpea leaf epidermal cells by the application of the metal salts CuCl2, CuSO4, and HgCl2. When unfixed, salt-treated cells were observed by light microscopy, their degeneration and death were seen to follow a series of stages: (1) cessation of cytoplasmic streaming, (2) morphological changes in the cytoplasm such as the formation of large vesicles and the appearance of particles in the vacuole, and (3) protoplast collapse. The relative timing of these stages after salt application was affected by the nature and concentration of the chemical used. The application of fluorescein diacetate and plasmolysing sucrose solutions before or after the addition of the copper salts suggested that the semipermeability of the tonoplast was lost during stage 2 and that of the protoplast boundary at stage 3. Following the cessation of cytoplasmic streaming, the changes observed in unfixed cells occurred rapidly and correlated well with the ultrastructural changes observed in the same cells after fixation. First, microtubules appeared to decrease in abundance. Then coiled polyribosomes, endoplasmic reticulum, and Golgi bodies rapidly became undetectable, mitochondrial cristae became dilated, and the cytoplasm consisted of aggregates of ribosome-containing material interspersed with vesicles. However, the study also showed that the first observable sign of degeneration seen in unfixed cells (i.e., the cessation of cytoplasmic streaming) appears to have no clearly diagnostic feature detectable by electron microscopy.


1989 ◽  
Vol 67 (1) ◽  
pp. 201-210 ◽  
Author(s):  
H. B. Massicotte ◽  
C. A. Ackerley ◽  
R. L. Peterson

Ultrastructural features of the two symbionts in ectomycorrhizae formed between Alnus rubra and Alpova diplophloeus change with developmental stage. In the root cap – meristem zone, hyphae penetrate between vacuolated root cap cells and become appressed to epidermal cells containing small vacuoles, plastids with starch, numerous Golgi bodies, mitochondria, and endoplasmic reticulum cisternae. In the young Hartig net zone, hyphae with few vacuoles penetrate between vacuolated epidermal cells that still contain numerous Golgi bodies but now have plastids with small starch grains. Hartig net hyphae begin to branch and eventually form a complex branching system in the mature Hartig net zone. Hartig net hyphae in the basal portion of the ectomycorrhizae synthesize lipid and finally become vacuolated.


1991 ◽  
Vol 69 (6) ◽  
pp. 1199-1206 ◽  
Author(s):  
C. Y. Chen ◽  
Michele C. Heath

The cytological responses to the monokaryotic primary hyphae of the cowpea rust fungus (Uromyces vignae Barcl.) were observed in vein epidermal cells of a resistant and a susceptible cowpea cultivar. Unlike the previously examined response to haustoria of a nonpathogenic powdery mildew fungus, plant cell walls did not become autofluorescent in response to fungal penetration, and the primary hypha only rarely became encased. Following fungal penetration, the response of invaded cells of the resistant, intact plant could be divided into the following stages: (I) cytoplasmic streaming normal; (II) cytoplasmic streaming slow or stopped, Brownian motion of particles visible in the vacuole, granulated cytoplasm aggregated along the cell walls, some host nuclei disappeared; and (III) protoplast collapsed. Epidermal tissue of the resistant cultivar did not exhibit stages II–III when detached and mounted in water 12 h after inoculation and examined 9 h later. The frequency of stage III increased when the tissue was mounted in CaCl2, Ca(NO3)2, and KNO3, but only in a kinetin solution did it approximate that in attached tissue. Although kinetin inhibited fungal growth in both the resistant and the susceptible cultivar, the hypersensitive response occurred only in the former, suggesting that kinetin affects the hypersensitive response directly rather than through its inhibitory effect on the fungus. Key words: cowpea, Vigna unguiculata, cowpea rust fungus, Uromyces vignae (Barcl.), hypersensitivity.


1981 ◽  
Vol 59 (3) ◽  
pp. 292-300 ◽  
Author(s):  
L. Van Caeseele ◽  
J. T. Mills ◽  
M. Sumner ◽  
R. Gillespie

The development of mucilage in the epidermal cells of canola seeds (Brassica campestris L. cv. Candle) was studied with light and electron microscopy from 5 days after pollination to maturity. During the first 17 days starch was deposited in amyloplasts. At or near the 17th day mucilage appeared between the plasmalemma and the outer tangential wall of the epidermal cells. As the volume of mucilage increased, starch grains disappeared and were totally absent by 25 days. Membrane-bound structures and Golgi bodies were visible within the cytoplasm adjacent to the site of mucilage deposition. At maturity the seed epidermal cells were totally devoid of cytoplasm and engorged with mucilage.


1976 ◽  
Vol 54 (14) ◽  
pp. 1647-1655 ◽  
Author(s):  
W. R. Bushnell ◽  
R. J. Zeyen

Cytoplasmic aggregates that formed in susceptible barley epidermal cells 11-12 h after inoculation with Erysiphe graminis were examined by light microscopy in living specimens and by electron microscopy in fixed specimens. Rapid development of the aggregate (5–10 min) suggested that cytoplasm migrated to the site of each aggregation. The aggregate contained features generally associated with areas of high metabolic and synthetic activity: abundant mitochondria, rough endoplasmic reticulum (associated with smooth cisternae), Golgi bodies, and polyribosomes. Leucoplasts and nuclei were sometimes near aggregates but not consistently. Microbodies and osmiophilic spherosomes were not present.


Author(s):  
M. O. Magnusson ◽  
D. G. Osborne ◽  
T. Shimoji ◽  
W. S. Kiser ◽  
W. A. Hawk

Short term experimental and clinical preservation of kidneys is presently best accomplished by hypothermic continuous pulsatile perfusion with cryoprecipitated and millipore filtered plasma. This study was undertaken to observe ultrastructural changes occurring during 24-hour preservation using the above mentioned method.A kidney was removed through a midline incision from healthy mongrel dogs under pentobarbital anesthesia. The kidneys were flushed immediately after removal with chilled electrolyte solution and placed on a LI-400 preservation system and perfused at 8-10°C. Serial kidney biopsies were obtained at 0-½-1-2-4-8-16 and 24 hours of preservation. All biopsies were prepared for electron microscopy. At the end of the preservation period the kidneys were autografted.


Author(s):  
A. P. Lupulescu ◽  
H. Pinkus ◽  
D. J. Birmingham

Our laboratory is engaged in the study of the effect of different chemical agents on human skin, using electron microscopy. Previous investigations revealed that topical use of a strong alkali (NaOH 1N) or acid (HCl 1N), induces ultrastructural changes in the upper layers of human epidermis. In the current experiments, acetone and kerosene, which are primarily lipid solvents, were topically used on the volar surface of the forearm of Caucasian and Negro volunteers. Skin specimens were bioptically removed after 90 min. exposure and 72. hours later, fixed in 3% buffered glutaraldehyde, postfixed in 1% phosphate osmium tetroxide, then flat embedded in Epon.


2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Mutsumi Yokota ◽  
Soichiro Kakuta ◽  
Takahiro Shiga ◽  
Kei-ichi Ishikawa ◽  
Hideyuki Okano ◽  
...  

AbstractMitochondrial structural changes are associated with the regulation of mitochondrial function, apoptosis, and neurodegenerative diseases. PRKN is known to be involved with various mechanisms of mitochondrial quality control including mitochondrial structural changes. Parkinson’s disease (PD) with PRKN mutations is characterized by the preferential degeneration of dopaminergic neurons in the substantia nigra pars compacta, which has been suggested to result from the accumulation of damaged mitochondria. However, ultrastructural changes of mitochondria specifically in dopaminergic neurons derived from iPSC have rarely been analyzed. The main reason for this would be that the dopaminergic neurons cannot be distinguished directly among a mixture of iPSC-derived differentiated cells under electron microscopy. To selectively label dopaminergic neurons and analyze mitochondrial morphology at the ultrastructural level, we generated control and PRKN-mutated patient tyrosine hydroxylase reporter (TH-GFP) induced pluripotent stem cell (iPSC) lines. Correlative light-electron microscopy analysis and live cell imaging of GFP-expressing dopaminergic neurons indicated that iPSC-derived dopaminergic neurons had smaller and less functional mitochondria than those in non-dopaminergic neurons. Furthermore, the formation of spheroid-shaped mitochondria, which was induced in control dopaminergic neurons by a mitochondrial uncoupler, was inhibited in the PRKN-mutated dopaminergic neurons. These results indicate that our established TH-GFP iPSC lines are useful for characterizing mitochondrial morphology, such as spheroid-shaped mitochondria, in dopaminergic neurons among a mixture of various cell types. Our in vitro model would provide insights into the vulnerability of dopaminergic neurons and the processes leading to the preferential loss of dopaminergic neurons in patients with PRKN mutations.


Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 554
Author(s):  
Natália Salomão ◽  
Michelle Brendolin ◽  
Kíssila Rabelo ◽  
Mayumi Wakimoto ◽  
Ana Maria de Filippis ◽  
...  

Intrauterine transmission of the Chikungunya virus (CHIKV) during early pregnancy has rarely been reported, although vertical transmission has been observed in newborns. Here, we report four cases of spontaneous abortion in women who became infected with CHIKV between the 11th and 17th weeks of pregnancy. Laboratorial confirmation of the infection was conducted by RT-PCR on a urine sample for one case, and the other three were by detection of IgM anti-CHIKV antibodies. Hematoxylin and eosin (H&E) staining and an electron microscopy assay allowed us to find histopathological, such as inflammatory infiltrate in the decidua and chorionic villi, as well as areas of calcification, edema and the deposition of fibrinoid material, and ultrastructural changes, such as mitochondria with fewer cristae and ruptured membranes, endoplasmic reticulum with dilated cisterns, dispersed chromatin in the nuclei and the presence of an apoptotic body in case 1. In addition, by immunohistochemistry (IHC), we found a positivity for the anti-CHIKV antibody in cells of the endometrial glands, decidual cells, syncytiotrophoblasts, cytotrophoblasts, Hofbauer cells and decidual macrophages. Electron microscopy also helped in identifying virus-like particles in the aborted material with a diameter of 40–50 nm, which was consistent with the size of CHIKV particles in the literature. Our findings in this study suggest early maternal fetal transmission, adding more evidence on the role of CHIKV in fetal death.


1995 ◽  
Vol 23 (3) ◽  
pp. 200-206 ◽  
Author(s):  
P Carbognani ◽  
L Spaggiari ◽  
M Rusca ◽  
L Cattelani ◽  
P Solli ◽  
...  

During lung preservation, the vascular endothelium is probably the first site of damage and these lesions are considered the main limiting factor in solid-organ preservation. In the present study, the ultrastructural changes in the endothelial cells of human pulmonary artery hypothermically stored (at 4 °C) for 6 and 12 h in Euro-Collins, University of Wisconsin and Ringer-lactate solutions were compared. The arteries obtained from three patients who underwent pneumonectomy were divided into 20 segments and preserved in the three solutions mentioned. The specimens, which were fixed in osmic acid, were examined using transmission electron microscopy. Transmission electron microscopy indicated that the cells stored in the University of Wisconsin solution either for 6 or 12 h were the best preserved, while the most severely damaged cells were those stored in Euro-Collins solution, even after just 6 h. The cells stored in Ringer-lactate showed an intermediate level of damage. The data from an ultrastructural grading scale, which quantified the damage to the cytoplasm, mitochondria and nucleus, were in broad agreement with the general transmission electron microscopy observations. Analysis of variance of the grading scale data showed that there were statistically significant differences between the groups after both 6 and 12 h storage ( P < 0.05).


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