Microscopy studies of Tilletia controversa teliospore development in wheat ovaries and in culture

Author(s):  
W. M. Hess ◽  
E. J. Trione

Teliospore formation begins in wheat ovaries when they are about 0.5 mm diam. Tilletia controversa hyphal cells initially invade host cells intercellularly. During development a narrow band of sporogenous hyphal cells develop into reticulated teliospores without hyphal attachments. The host tissue is utilized in the formation of mature teliospores. Eventually a thin layer of host tissue surrounds the mature teliospores. In contrast, when the teliospores form in medium hyphal tips appear to enlarge into developing teliospores. Light microscopy, SEM, and thin sections with TEM were used to determine whether teliospores formed from enlarged hyphal tips or from sporogenous cells which fragment from hyphae.Teliospores were obtained from USDA ARS experimental plots at Logan, UT. Wheat plants were inoculated and grown in a greenhouse. Tissues were fixed for 1 h or longer in 2% glutaraldehyde, 3% acrolein, 0.1 M Na-cacodylate buffer, pH 7.3; rinsed several times in the same buffer; postfixed in 1% OsO4 in 0.1 M Na-cacodylate buffer, pH, 7.3 for 2 h in an ice bath; and rinsed in 0.1 M Na-cacodylate buffer prior to dehydration in a graded ethanol series.

Author(s):  
Patricia L. Jansma

The presence of the membrane bound vesicles or blebs on the intestinal epithelial cells has been demonstrated in a variety of vertebrates such as chicks, piglets, hamsters, and humans. The only invertebrates shown to have these microvillar blebs are two species of f1ies. While investigating the digestive processes of the freshwater microcrustacean, Daphnia magna, the presence of these microvillar blebs was noticed.Daphnia magna fed in a suspension of axenically grown green alga, Chlamydomonas reinhardii for one hour were narcotized with CO2 saturated water. The intestinal tracts were excised in 2% glutaraldehyde in 0.2 M cacodyl ate buffer and then placed in fresh 2% glutaraldehyde for one hour. After rinsing in 0.1 M cacodylate buffer, the sample was postfixed in 2% OsO4, dehydrated with a graded ethanol series, infiltrated and embedded with Epon-Araldite. Thin sections were stained with uranyl acetate and Reynolds lead citrate before viewing with the Philips EM 200.


Author(s):  
A. Singh ◽  
A. Dykeman ◽  
J. Jarrelf ◽  
D. C. Villeneuve

Hexachlorobenzene (HCB), a persistent and mobile organochlorine pesticide, occurs in environment. HCB has been shown to be present in human follicular fluid. An objective of the present report, which is part of a comprehensive study on reproductive toxicity of HCB, was to determine the cytologic effects of the compound on ovarian follicles in a primate model.Materials and Methods. Eight Cynomolgus monkeys were housed under controlled conditions at Animal facility of Health and Welfare, Ottawa. Animals were orally administered gelatin capsules containing HCB mixed with glucose in daily dosages of 0.0 or 10 mg/kg b.w. for 90 days; the former was the control group. On the menstrual period following completion of dosing, the monkeys underwent an induction cycle of superovulation. At necropsy, one-half of an ovary from each animal was diced into ca. 2- to 3-mm cubed specimens that were fixed by immersion in 2.5% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.3). Subsequent procedures followed to obtain thin sections that were examined in a Hitachi H-7000 electron microscope have been described earlier.


Author(s):  
M. Kessel ◽  
R. MacColl

The major protein of the blue-green algae is the biliprotein, C-phycocyanin (Amax = 620 nm), which is presumed to exist in the cell in the form of distinct aggregates called phycobilisomes. The self-assembly of C-phycocyanin from monomer to hexamer has been extensively studied, but the proposed next step in the assembly of a phycobilisome, the formation of 19s subunits, is completely unknown. We have used electron microscopy and analytical ultracentrifugation in combination with a method for rapid and gentle extraction of phycocyanin to study its subunit structure and assembly.To establish the existence of phycobilisomes, cells of P. boryanum in the log phase of growth, growing at a light intensity of 200 foot candles, were fixed in 2% glutaraldehyde in 0.1M cacodylate buffer, pH 7.0, for 3 hours at 4°C. The cells were post-fixed in 1% OsO4 in the same buffer overnight. Material was stained for 1 hour in uranyl acetate (1%), dehydrated and embedded in araldite and examined in thin sections.


Author(s):  
M. Ashraf ◽  
F. Thompson ◽  
S. Miki ◽  
P. Srivastava

Iron is believed to play an important role in the pathogenesis of ischemic injury. However, the sources of intracellular iron in myocytes are not yet defined. In this study we have attempted to localize iron at various cellular sites of the cardiac tissue with the ferrocyanide technique.Rat hearts were excised under ether anesthesia. They were fixed with coronary perfusion with 3% buffered glutaraldehyde made in 0.1 M cacodylate buffer pH 7.3. Sections, 60 μm in thickness, were cut on a vibratome and were incubated in the medium containing 500 mg of potassium ferrocyanide in 49.5 ml H2O and 0.5 ml concentrated HC1 for 30 minutes at room temperature. Following rinses in the buffer, tissues were dehydrated in ethanol and embedded in Spurr medium.The examination of thin sections revealed intense staining or reaction product in peroxisomes (Fig. 1).


Author(s):  
Y. R. Chen ◽  
Y. F. Huang ◽  
W. S. Chen

Acid phosphatases are widely distributed in different tisssues of various plants. Studies on subcellular localization of acid phosphatases show they might be present in cell wall, plasma lemma, mitochondria, plastid, vacuole and nucleus. However, their localization in rice cell varies with developmental stages of cells and plant tissues. In present study, acid phosphatases occurring in root cap are examined.Sliced root tips of ten-day-old rice(Oryza sativa) seedlings were fixed in 0.1M cacodylate buffer containing 2.5% glutaraldehyde for 2h, washed overnight in same buffer solution, incubated in Gomori's solution at 37° C for 90min, post-fixed in OsO4, dehydrated in ethanol series and finally embeded in Spurr's resin. Sections were doubly stained with uranyl acetate and lead citrate, and observed under Hitachi H-600 at 75 KV.


Author(s):  
Hsin-Kan Wu ◽  
Mei-Chu Chung

In one of our earlier papers (Wu et al. 1978), we suggested that glutelin is the major composition of the round storage protein bodies although they also contain relatively more prolamine than the angular one does. Immunochemical studies of Krishnan et al. (1986) later showed the presence of glutelin in the irregularly-shaped (angular) protein bodies while the prolamines were found in the round ones. Our recent experiment using protein A-gold technique found that prolamine is mainly deposited into the angular protein bodies.Small blocks (1 mm3) of 7 DAF (days after flowering) caryopsis of Orvza perennis were fixed with 3% paraformaldehyde and 3% glutaraldehyde in 0.1M sodium phosphate, pH7.4, dehydrated in a graded ethanol series and infiltrated with Spurr’s resin. Thin sections, after gold labeling, were stained with uranyl acetate and lead citrate. Rabbit antibodies were raised against purified prolamine. Protein A-gold sol complex was prepared based on the technique of Horisberger et al. (1977).


HortScience ◽  
1991 ◽  
Vol 26 (6) ◽  
pp. 719A-719
Author(s):  
Richard L. Harkess ◽  
Robert E. Lyons

A study was undertaken to determine the rate of floral initiation in Rudbeckia hirta. R. hirta plants were grown to maturity, 14-16 leaves, under short days (SD). Paired controls were established by placing half of the plants under long days (LD) with the remainder left under SD. Beginning at the start of LD (day 0), five plants were harvested daily from each photoperiod group for twenty days. Harvested meristems were fixed in 2% paraformaldehyde - 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer (pH 7.0) for 24 hrs, dehydrated in an ethanol series, embedded in paraffin and sectioned at 8 μm. Serial sections were stained with Methyl-green Pyronin, with adjacent sections treated with RNase for nucleic acid comparison. All events of floral initiation were identified, The results of limited inductive photoperiod indicate that 16-18 LD were required for flowering.


2006 ◽  
Vol 80 (5) ◽  
pp. 2437-2444 ◽  
Author(s):  
Florian Frohns ◽  
Anja Käsmann ◽  
Detlef Kramer ◽  
Britta Schäfer ◽  
Mario Mehmel ◽  
...  

ABSTRACT Previous studies have established that chlorella viruses encode K+ channels with different structural and functional properties. In the current study, we exploit the different sensitivities of these channels to Cs+ to determine if the membrane depolarization observed during virus infection is caused by the activities of these channels. Infection of Chlorella NC64A with four viruses caused rapid membrane depolarization of similar amplitudes, but with different kinetics. Depolarization was fastest after infection with virus SC-1A (half time [t 1/2], about 9 min) and slowest with virus NY-2A (t 1/2, about 12 min). Cs+ inhibited membrane depolarization only in viruses that encode a Cs+-sensitive K+ channel. Collectively, the results indicate that membrane depolarization is an early event in chlorella virus-host interactions and that it is correlated with viral-channel activity. This suggestion was supported by investigations of thin sections of Chlorella cells, which show that channel blockers inhibit virus DNA release into the host cell. Together, the data indicate that the channel is probably packaged in the virion, presumably in its internal membrane. We hypothesize that fusion of the virus internal membrane with the host plasma membrane results in an increase in K+ conductance and membrane depolarization; this depolarization lowers the energy barrier for DNA release into the host.


Author(s):  
J.F. David-Ferreira ◽  
K.L. David-Ferreira ◽  
M.H. Miranda ◽  
J.C. Lemos

The testis interstitial cells of the rat are closely associated forming small aggregates in the vicinity of the vessels. The finality of the present study is to analise the relationship between the interstitial cells and to describe some particularities of their junctions.The observations were made in thin sections from testis of 2 to 3 months old rats fixed by immersion or perfusion with 2 to 3% glutaral- dehyde in s-collidine or cacodylate buffer followed by 2% osmium tetro-xyde in the same buffers. Some specimens have been prepared following the technique of Shea. The thin sections obtained after embedding in Epon were double stained in uranyl acetate and lead citrate. For the cryofracture study the tissues were fixed in 3% glutaraldehyde in cacodylate buffer, impregnated in 28% glycerol, frozen in Freon 22 and stored in liquid nitrogen. Freeze fracture and platinum-carbon shadowing were done in a Balzers 300.


Author(s):  
E. C. V. Ooi ◽  
C. W. Chan

The histology of the reptilian alimentary tract has been described in numerous light microscopic studies. However, no information on surface morphology is available for any species of reptiles. In the present communication, the alimentary canal of the grass lizard, Takydromus sexlineatus, is investigated by means of light and scanning electron microscopy.Various portions of the alimentary canal were dissected and removed rapidly. While immersed in cacodylate buffer, the inner lining of each portion was exposed and rinsed. The tissues were then fixed in 2% glutaraldehyde in cacodylate buffer, dehydrated in graded ethanol series, and critical point dried with carbon dioxide. After coating with gold, the specimens were examined on a JOEL JSM-35 operated at 35 KV. Tissues for light microscopy were processed routinely.


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