Localization of acid phosphatases in root cap of rice plant

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):  
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):  
Kit W. Lee

The structure of the polycomplexes has been extensively studied in recent years (1,2,3). Theres structures are usually described as stacks or aggregates of the tripartite synaptonemal complexes. Although the presence of the synaptonemal complexes is a consistent feature in the late zygotene or pachytene stages of chlasmate meiosis and Its role in the processes of chromosome synapsis and crossing over has been suggested (4), the function of the polycomplexes remains obscure. Most of our understandings of the polycomplexes are obtained from the observations during the gametogenesis of the insects, and only a few examples of this structure in fungi and higher plants have been reported. The present study examines the occurrence of polycomplexes during the sporogenesis in the primitive vascular plant. Sporangia at different developmental stages were fixed with 3% glutaraldehyde in 0.1 M phosphate buffer, and postflxed in 2% osmium tetroxide. Dehydration was carried out with the ethanol series followed by embedding in Epon 812. Ultrathln sections were stained with uranyl acetate and lead citrate.


Author(s):  
R. J. Williams ◽  
N. R. Dollahon ◽  
E. Larsen ◽  
S. O’Neill ◽  
R. Chapman

In studies of several families of aquatic heteropterans we have found exoskeletal pits not described in the literature. These structures are associated with the lateral margins of the pronotum and/or the dorsal surface, posterior to the scutellum in notonectids, nepids, and corixids. The function of these pits is unknown, but we presumed that they might be either sensory or secretory in nature. We undertook this study of the microscopy of pits in the notonectid, Buenoa margaritacea to learn if either of these functions is consistent with the fine structure.For TEM, adult insects were submerged in 1.0% paraformaldeyde and 2.0% glutaraldehyde in 0.02M sodium cacodylate buffer with 0.01% calcium chloride at a pH of 7.2, then dissected with a razor blade cleaned with acetone. Tissues were fixed overnight at 4°C in paraformaldehyde and glutaraldehyde, then fixed in 1% osmium tetroxide in cacodylate buffer, dehydrated in a graded series of acetone, embedded in Spurr resin and polymerized at 70°C for 21 hours. Blocks were thin sectioned with a diamond knife, and sections were stained with uranyl acetate and lead citrate. Whole specimens for SEM were similarly fixed, dehydrated in an ethanol series and critical point dried. SEM micrographs of internal pit anatomy were produced by adhering 500nm sections to a glass coverslip, then removing the embedding resin by incubation for 5 minutes in a saturated solution of potassium hydroxide in absolute ethanol, followed by three 20 minute rinses in absolute ethanol and air drying. Cover slips were attached to an aluminum specimen stub and sputter coated for 60 seconds with gold/palladium as were whole insects .


1986 ◽  
Vol 34 (5) ◽  
pp. 561-567 ◽  
Author(s):  
C T Lin ◽  
D Sun ◽  
G X Song ◽  
J Y Wu

Calmodulin was purified from bovine brain by preparative SDS-polyacrylamide gel electrophoresis. The denatured, purified calmodulin was used to immunize rabbits to produce antiserum. This antiserum was used to study the distribution of calmodulin in plant tissues by indirect immunohistochemistry. The root tips from corn seeds, oat seeds, peanuts, spaghetti squash seeds, and the terminal buds of spinach were investigated. A method for plant tissue sectioning and inhibition of endogenous peroxide activity was developed. In the corn root section, reaction product from anti-calmodulin was found mainly in the root cap cells. Lesser but significant amounts of calmodulin were localized in metaxylem elements, in some stele cells surrounding metaxylem elements, in apical initials, and in the cortical cells. Similar findings were also observed in other root tips from oat seeds, peanuts, and spaghetti squash seeds. In the terminal buds of the spinach, calmodulin-stained cells were highly concentrated in the apical meristem and leaf primordium. These findings suggest that the high concentration of calmodulin in the root cap may be important in relation to gravitropism and growth development.


1997 ◽  
Vol 45 (1) ◽  
pp. 89-95 ◽  
Author(s):  
Stanislav Vitha ◽  
František Baluška ◽  
Miriam Mews ◽  
Dieter Volkmann

We developed a simple and reliable technique for immunofluorescence detection of F-actin on microtome sections of plant tissues. For the first time, large numbers of plant cells from various tissues that pass through their developmental stages could be consistently visualized on one section from plant organs. n-Maleimidobenzoic acid N-hydroxy-succinimide ester-pretreated and formalin-fixed segments of plant roots and shoots were embedded in low melting point ester wax at 37C and sectioned on a microtome. After dewaxing and rehydration, microfilaments were visualized by indirect immunofluorescence technique with a monoclonal anti-actin antibody. The technique has been successfully used for visualization of tissue- and development-specific F-actin arrays in cells of Zea mays and Lepidium sativum root tips and of maize stem nodes.


Author(s):  
Amankwah K.S. ◽  
A.D. Weberg ◽  
R.C. Kaufmann

Previous research has revealed that passive (involuntary inhalation) tobacco smoking during gestation can have adverse effects upon the developing fetus. These prior investigations did not concentrate on changes in fetal morphology. This study was undertaken to delineate fetal neural abnormalities at the ultrastructural level in mice pups exposed in utero to passive maternal smoking.Pregnant study animals, housed in a special chamber, were subjected to cigarette smoke daily from conception until delivery. Blood tests for determination of carbon monoxide levels were run at 15-18 days gestation. Sciatic nerve tissue from experimental and control animals were obtained following spontaneous delivery and fixed in 2.5% gluteraldehyde in 0.1M cacodylate buffer pH 7.3. The samples were post-fixed in osmium ferrocyanide (1:1 mixture of 1.5% aqueous OSO4 and 2.5% K4 Fe(CN)6). Following dehydration, the tissues were infiltrated with and embedded in Spurr. Sections were stained with uranyl acetate and lead citrate.


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):  
José A. Serrano ◽  
Hannah L. Wasserkrug ◽  
Anna A. Serrano ◽  
Arnold M. Seligman

As previously reported (1, 2) phosphorylcholine (PC) is a specific substrate for prostatatic acid phosphatase (PAP) as opposed to other acid phosphatases, e.g., lysosomal acid phosphatase. The specificity of PC for PAP is due to the pentavalent nitrogen in PC, a feature that renders PC resistant to hydrolysis by all other acid phosphatases. Detailed comparative cytochemical results in rat tissues are in press. This report deals with ultracytochemical results applying the method to normal and pathological human prostate gland.Fresh human prostate was obtained from 7 patients having transurethral resections or radical prostatectomies. The tissue was fixed in 3% glutaraldehyde- 0.1 M cacodylate buffer (pH 7.4) for 15 min, sectioned at 50 μm on a Sorvall TC-2 tissue sectioner, refixed for a total of 2 hr, and rinsed overnight in 0.1 M cacodylate buffer (pH 7.4)-7.5% sucrose.


Author(s):  
Li-Chu Tung ◽  
Yung-Reui Chen ◽  
Shiu-Nan Chen ◽  
Guang-Hsiung Kuo

In the present study, the ultrastructural changes of BPK cells, a fibroblast-like cell line, derived from the kidney of juvenile black porgy Acanthopagrus schlegeli, under heat shock treatment are described.The BPK cells were maintained in L-15 medium supplemented with 10% fetal calf serum and 0.15 M NaCl at 28|C2. The heating was carried out in precalibrated water baths. Monolayers of cells, grown on coverslips in parafilm-sealed petri dishes were submerged under water for 30 min at 40|C treatments. Cells were fixed in 2.5% glutaraldehyde in 0.1 M cacodylate buffer supplemented with 6.6% sucrose, postfixed in 1% OsO4 and flat embedded in Spurr’s resin. Silver section were cut parallel to the substratum, stained with uranyl acetate and Reynold’s lead citrate, and examined in a Hitachi H-600 electron microscope 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).


Sign in / Sign up

Export Citation Format

Share Document