The influence of experimentally induced changes in the (Mg + Ca):K balance on protein bodies formed in developing Cucurbita seeds

1994 ◽  
Vol 72 (3) ◽  
pp. 364-369 ◽  
Author(s):  
John N. A. Lott ◽  
Irene Ockenden ◽  
Patrice Kerr ◽  
Marcia West ◽  
Thelma Leech ◽  
...  

Phytate, the main mineral nutrient storage compound in seeds, is stored inside protein bodies. Phytate is usually concentrated in dense bodies called globoid crystals. In 1985, Lott and co-workers proposed that the balance of (Mg + Ca):K may be important in controlling globoid crystal formation and provided some experimental evidence to support this proposal. To test this hypothesis further, developing Cucurbita fruits were injected with sterile K salt solutions. Squash cotyledons generally have large globoid crystals and a relatively high (Mg + Ca):K ratio. We hypothesized that experimental reduction of the ratio by the addition of K would result in the alteration of the size and number of globoid crystals. Developing seeds were remarkably resistant to attempts to alter the elements taken up for storage. The elemental content of embryo tissues was measured quantitatively with neutron activation analysis. Controls plus those few samples showing a distinct shift in the ratio owing to elevated K content were analyzed further. Energy dispersive X-ray analysis of cryogenically prepared samples was used to study the elemental content of globoid crystals and transmission electron microscopy was used to study the ultrastructure of the protein bodies. The results provide additional support for the hypothesis being tested. Key words: phytate, globoid crystals, Cucurbita, seeds, mineral nutrients, protein bodies.

1997 ◽  
Vol 75 (7) ◽  
pp. 1137-1147 ◽  
Author(s):  
Tomikichi Wada ◽  
John N. A. Lott

To understand the differences in mineral nutrient storage within tissues and organs of rice (Oryza sativa L.) grains, the distribution of globoids in rice embryo and endosperm tissues was examined using light and transmission electron microscopy and energy dispersive X-ray microanalysis was used to study globoid composition. Globoids were found in most embryo tissues, including provascular cells, and their location and size in sections of protein bodies is described. While P, Mg, and K were commonly detected in all globoids, other elements such as Ca, Mn, Fe, and Zn were sometimes detected in globoids of specific tissues and (or) regions. High peak-to-background ratios for P were obtained in globoids of scutellar and aleurone cells, and moderately high values were detected in ground meristem regions of the mesocotyl and coleoptile. Relatively high K levels were found in globoids in parenchyma cells of the scutellum and coleorhiza; in provascular cells of the radicle; and in ground meristem cells from the mesocotyl, coleoptile, and plumule. Calcium was mainly detected in globoids of the aleurone layer. Iron was mostly found in radicle tissue globoids. Zinc was commonly found in globoids of the scutellar epithelium and in provascular tissues of the mesocotyl, coleoptile, and radicle. Manganese was distributed throughout most of the tissues examined, but the highest levels of Mn were detected in globoids from the coleoptile tip regions and the plumule. A novel finding was that, in the provascular tissues of the coleoptile tip, distinctive differences were found in Mn, Fe, and Zn storage between globoids in the future xylem and the future phloem. Key words: EDX analysis, embryos, globoids, mineral storage, phytate, Oryza sativa, rice.


Author(s):  
B. Craig ◽  
L. Hawkey ◽  
A. LeFurgey

Ultra-rapid freezing followed by cryoultramicrotomy is essential for the preservation of diffusible elements in situ within cells prior to scanning transmission electron microscopy and quantitative energy dispersive x-ray microanalysis. For cells or tissue fragments in suspension and for monolayer cell cultures, propane jet freezing provides cooling rates greater than 30,000°C/sec with regions up to 40μm in thickness free of significant ice crystal formation. While this method of freezing has frequently been applied prior to freeze fracture or freeze substitution, it has not been widely utilized prior to cryoultramicrotomy and subsequent x-ray microanalytical studies. This report describes methods devised in our laboratory for cryosectioning of propane jet frozen kidney proximal tubule suspensions and cultured embryonic chick heart cells, in particular a new technique for mounting frozen suspension specimens for sectioning. The techniques utilize the same specimen supports and sample holders as those used for freeze fracture and freeze substitution and should be generally applicable to any cell suspension or culture preparation.


1980 ◽  
Vol 58 (6) ◽  
pp. 699-711 ◽  
Author(s):  
Ernest Spitzer ◽  
John N. A. Lott

Protein bodies of dry seeds of tomato (Lycopersicon esculentum) from radicle, hypocotyl, cotyledon, and endosperm tissue were extensively studied using thin-sectioning, freeze-fracturing and energy dispersive x-ray (EDX) analysis. Protein bodies varied in size, were oval to circular in section, and generally consisted of a proteinaceous matrix, globoid crystal, and protein crystalloid components. Size, shape, and arrangements of globoid crystals and protein crystalloids varied even within the same cell. Globoid crystals were generally oval to circular in section. They were always surrounded by a proteinaceous matrix. In a given protein body the number present ranged from a few to numerous. A protein body generally contained only one protein crystalloid. In section, protein crystalloids were irregular or angular in shape. They were composed of substructural particles which formed lattice planes. EDX analysis of tomato seed globoid crystals revealed the presence of P, K, and Mg in all cases, a fact that is consistent with globoid crystals being phytin-rich. Rarely, small amounts of calcium were found along with P, K, and Mg in globoid crystals of each of the tissue regions considered. The distribution pattern of cells with Ca containing globoid crystals was random. Small amounts of Fe and Mn were also found in the globoid crystals of protein bodies from certain cell types. These two elements, unlike calcium, were specific in terms of their distribution. Globoid crystals from the protodermal cells often contained Mn and Fe. The globoid crystals from provascular tissue of radicle, hypocotyl, and cotyledon regions often contained Fe while globoid crystals in the first layer of large cells surrounding these provascular areas always contained Fe. Results from EDX analysis of the proteinaceous material from the protein bodies are presented and discussed as are variations in elemental content due to different fixations.


1995 ◽  
Vol 73 (6) ◽  
pp. 954-957 ◽  
Author(s):  
M. Marcia West ◽  
Derrick T. Flannigan ◽  
John N.A. Lott

The mature seeds of some angiosperms contain perisperm tissue derived from remains of the nucellus tissue of the ovule. In our transmission electron microscopy studies, the perisperm tissues of the seeds of Yucca brevifolia, Coffea arabica, Beta vulgaris, Piper nigrum, and Zostera capricorni contained naturally electron-dense globoids. Energy dispersive X-ray analysis of globoids from the perisperm tissues of these seeds revealed varying levels of phosphorus, potassium, magnesium, and calcium, results that are consistent with the presence of the mineral nutrient store called phytate. Phosphorus, potassium, magnesium, calcium, and other mineral nutrients, likely stored as phytate, have routinely been located in globoids of endosperm, female gametophyte, and embryo tissues of seeds. Key words: perisperm, globoids, mineral nutrient reserves, phosphorus, seeds, energy dispersive X-ray analysis.


1991 ◽  
Vol 69 (11) ◽  
pp. 2545-2554 ◽  
Author(s):  
Sara Maldonado ◽  
John N. A. Lott

The structure of protein bodies in the endosperm and embryo of Datura stramonium was studied with a variety of light-and electron-microscopic techniques. Protein bodies had one to several globoid crystals and one or two protein crystalloids in the proteinaceous matrix. Although the embryo protein bodies rarely had more than two globoid crystals, the endosperm protein bodies had varying sizes and numbers of globoid crystals, even within the same cell. Energy-dispersive X-ray analysis of globoid crystals revealed the presence of P, K, and Mg in all cases. Traces of Fe, Mn, and Zn were also found in globoid crystals of protein bodies from certain cell types. The distribution patterns of these three elements were quite specific; for example, Mn traces were found only in the globoid crystals of the protoderm. Neutron-activation analysis of endosperm and embryo tissues was used to quantitatively measure the concentration of Ca, Cl, Cu, I, K, Mg, Mn, Na, and S. The results from structural studies and the element analysis studies are discussed in the context of solanaceous seeds in particular but also with relation to seeds in general. Key words: protein bodies, Datura stramonium, seed, globoid crystals, energy-dispersive X-ray analysis, Solanaceae.


1995 ◽  
Vol 25 (8) ◽  
pp. 1313-1325 ◽  
Author(s):  
Minna Turunen ◽  
Satu Huttunen ◽  
Jaana Back ◽  
Jukka Lamppu

Seedlings of Scots pine (Pinussylvestris L.) and Norway spruce (Piceaabies (L.) Karst.) were subjected to acid rain irrigation at pH 7, pH 4, and pH 3 three times a week during the growing seasons of 1986–1989 in a field experiment. Scanning and transmission electron microscopy, energy dispersive spectrometry, contact angle measurements, and chloroform extraction of waxes were used to detect physicochemical changes in the needle cuticles. The first detectable symptoms of acid rain were observed after 5 weeks of acid rain treatment at pH 3 and pH 4, which resulted in few CaSO4 crystallites on visibly undamaged pine and spruce needle surfaces. After 7 weeks of acid rain treatment there were CaSO4 crystallites scattered over the whole needle surface area and erosion of the epicuticular waxes could be observed occasionally. CaSO4 crystal formation later decreased, especially on the needles of seedlings treated at pH 3. Ca concentrations in the needles and roots of the seedlings and in the soil in the boxes were higher in the pH 3 treatments than elsewhere. The more abundant deposition of Ca oxalate crystallites on the inner walls of the epidermal and hypodermal cells of the spruce needles than on their outer walls was probably also connected with Ca leaching, caused by acid rain. Acid rain also delayed wax synthesis, as 2-month-old pine needles exposed to pH 3 and pH 4 had about 50% less wax than the water controls in early August. The needle surfaces of the southern provenances of spruce and pine seedlings were slightly less wettable after pH 4 treatment than after the control water treatment, because they probably benefited from N and S compounds in the irrigation water. The needle surfaces were more wettable in the pH 3 and water control seedlings than in the other treatments.


1984 ◽  
Vol 11 (6) ◽  
pp. 459 ◽  
Author(s):  
JNA Lott ◽  
DJ Goodchild ◽  
S Craig

Most of the phytin in pea (Pisum sativum) cotyledons is water soluble. In order to determine where K and P are located it was necessary to use anhydrous or low water content tissue preparation procedures to obtain samples suitable for energy dispersive X-ray analysis studies using a transmission electron microscope. While some protein bodies do contain electron-dense globoid crystals, most do not. Globoid crystals are more prevalent in the abaxial part of the cotyledon where the provascular network is located. When present, globoid crystals contain considerable Mg, and/or Ca along with P and K. Protein bodies that lack globoid crystals still contain considerable P and K with lesser amounts of elements such as S, Cl and Mg. This is consistent with these protein bodies containing K-phytate in the proteinaceous matrix. While there is a lot of K inside the protein bodies, K is widespread in pea cotyledon tissue and could be detected in starch grains, cell walls and the cytoplasmic matrix.


1993 ◽  
Vol 309 ◽  
Author(s):  
Thomas J. Licata ◽  
Timothy D. Sullivan ◽  
Roy S. Bass ◽  
James G. Ryan ◽  
David B. Knorr

AbstractIncreasing circuit densities produce higher metal wiring aspect ratios, and more difficult feature fill for damascene processing. One method of extending the use of sputter deposition to challenging aspect ratios is to collimate the sputtered flux using a collimator plate, and to avoid randomizing the collimated flux by using low process pressures corresponding to long sputtered atom mean free paths. In this paper, we discuss our fabrication of damascene AI-0.5Cu-2Si and AI-2Mg wiring using both collimated and uncollimated sputtering, and our observations of collimation-induced changes in Al alloy electromigration and microstructure. Our experiments show that collimation has only a small effect on AlCuSi, but a large effect on AIMg. Specifically, the median time to electromigration failure for collimated AIMg was ∼10X the value for uncollimated AlMg and ∼6X the values for collimated and uncollimated AlCuSi. Transmission electron microscope and x-ray diffraction analyses of these films show that the collimation-induced improvement in AIMg t50 is associated with the formation of smaller, lower strain grains which are clustered in very well-oriented (111) domains. We propose that the advantageous AlMg microstructure results from enhanced texture produced by aspects of the collimated deposition active in the absence of incoherent precipitates.


Author(s):  
F. P. Ottensmeyer

Microanalysis by electron energy loss spectroscopy has gained momentum in the last few years with the utilization of more and more magnetic electron spectrometers coupled to dedicated or hybrid scanning transmission electron microscopes. Two approaches to analysis are the spectral evaluation of a single spot and the mapping of a single element over the entire image. In the first method, as in x-ray microanalysis, a finely focused electron beam is placed on a small area of the specimen. The impinging electrons traversing this spot subsequently provide a spectrum characteristic of the elemental content of the area.


2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Tong Xu ◽  
Hong Xu ◽  
Yi Zhong ◽  
Linping Zhang ◽  
Di Qian ◽  
...  

Abstract A kind of organic/inorganic composite material composed of waterborne polyurethane and sepiolite was prepared in this work. Sepiolite was organically modified by three kinds of silane coupling agents, and then compounded with waterborne polyurethane through layer-by-layer method in order to prepare composite materials. X-ray diffraction (XRD) and Fourier transform infrared (FTIR) show the crystal and chemistry structure of sepiolite samples, and confirmed the preparation of organic sepiolite. Scanning electron microscope (SEM) and energy dispersive X-ray spectroscopy (EDS) showed the surface microstructure and elemental content of sepiolite and organic sepiolite, and was consistent with the XRD results. Transmission electron microscope (TEM) examination of waterborne polyurethane composites surfaces showed that sepiolite particles were regularly dispersed in the waterborne polyurethane matrix. Thermal resistance of waterborne polyurethane composites was determined by thermogravimetry analyzer (TG) and derivative thermogravimetry analyzer (DTG), differential scanning calorimetry (DSC), gas chromatography (GC), and mass chromatography (MS). Mechanical behavior was examined by tensile strength tester, showed higher break strength than that of the control waterborne polyurethane. Therefore, organically modified sepiolite was considered to be a kind of wonderful inorganic material that could be used to improve the thermal stability and mechanical property of polymer.


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