Thin sectioning, freeze fracturing, energy dispersive x-ray analysis, and chemical analysis in the study of inclusions in seed protein bodies: almond, Brazil nut, and quandong

1978 ◽  
Vol 56 (17) ◽  
pp. 2050-2061 ◽  
Author(s):  
John N. A. Lott ◽  
Mark S. Buttrose
Keyword(s):  
Chemical Analysis ◽  
Freeze Fracture ◽  
Prunus Dulcis ◽  
Energy Dispersive ◽  
Protein Bodies ◽  
Brazil Nut ◽  
Fixed Tissue ◽  
Thin Sections ◽  
X Ray ◽  
Freeze Dried

Protein bodies from almond (Prunus dulcis), Brazil nut (Bertholletia excelsa), and quandong (Santalum acuminatum) have been studied in thin sections of fixed and embedded tissue, in freeze-fracture replicas of unfixed tissue, by chemical analysis of tissue for P, K, Mg, and Ca, and by energy dispersive x-ray (EDX) analysis of both sections of glutaraldehyde-fixed tissue and freeze-dried tissue powders. The protein bodies in all three species contained globoid crystals, protein crystalloids, and proteinaceous matrix regions. Results of EDX analyses were consistent with globoid crystals being rich in phytin. Variation in both the structure and the elemental composition of globoids was common. In almond some globoids were lobed rather than spherical, and large globoid crystals often contained considerable calcium whereas small globoid crystals contained little if any calcium. The globoid crystals of Brazil nut often contained barium in addition to P, K, Ca, and Mg. Protein crystalloids of Brazil nut were compound crystals. Protein bodies of quandong seed, which is largely endosperm rather than embryo, were unexceptional.

Thin-section, freeze-fracture, and energy dispersive x-ray analysis studies of the protein bodies of tomato seeds

1980 ◽  
Vol 58 (6) ◽  
pp. 699-711 ◽  
Author(s):  
Ernest Spitzer ◽  
John N. A. Lott
Keyword(s):  
Freeze Fracture ◽  
Protein Body ◽  
Cell Types ◽  
Energy Dispersive ◽  
Protein Bodies ◽  
Elemental Content ◽  
Tomato Seed ◽  
Endosperm Tissue ◽  
X Ray ◽  
Edx Analysis

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.

Manganese and Iron in Globoid Crystals of Protein Bodies From Avena and Casuarina

1978 ◽  
Vol 5 (5) ◽  
pp. 631 ◽  
Author(s):  
MS Buttrose
Keyword(s):  
Chemical Analysis ◽  
Avena Sativa ◽  
Storage Protein ◽  
Major Elements ◽  
Energy Dispersive ◽  
Protein Bodies ◽  
X Ray ◽  
Plant Seeds

The storage protein bodies of plant seeds usually contain globoid crystals with a high content of phytin, a rich store of Mg, P, K and Ca. By energy dispersive X-ray analysis, Mn and Fe have now been located in the globoid crystals of protein bodies in the seed embryos of Avena sativa and Casuarina species. Their levels in sections of globoids, relative to the levels of the major elements present, is consistent with their relative levels stored in whole seeds of various species as determined by chemical analysis.

ENERGY DISPERSIVE X-RAY ANALYSIS OF FIELD PEAS WITH DIFFERENT COOKING QUALITY

1983 ◽  
Vol 63 (4) ◽  
pp. 1071-1074 ◽  
Author(s):  
J. CHONG ◽  
S. T. ALI-KHAN ◽  
B. B. CHUBEY ◽  
G. H. GUBBELS
Keyword(s):  
Statistical Analysis ◽  
Pisum Sativum ◽  
Cooking Quality ◽  
Energy Dispersive ◽  
Protein Bodies ◽  
X Ray ◽  
Pisum Sativum L ◽  
Field Peas ◽  
Freeze Dried ◽  
High Concentrations

An energy dispersive X-ray (EDX) analytical method was used to study the freeze-dried powder of seeds of field peas (Pisum sativum L.) with good and poor cooking quality. EDX analysis of the electron-dense particles in the freeze-dried powder revealed the presence of high concentrations of Mg, P, and K, suggesting that the particles were protein bodies. Seeds with different cooking quality were compared with respect to the ratios of these elements in the dense particles. Statistical analysis indicated a significant correlation between these ratios and cooking quality.Key words: Pisum sativum, protein bodies, elemental analysis

An energy dispersive x-ray analysis study of elemental loss from globoid crystals in protein bodies as a result of osmium tetroxide fixation

1978 ◽  
Vol 56 (19) ◽  
pp. 2408-2414 ◽  
Author(s):  
J. N. A. Lott ◽  
J. S. Greenwood ◽  
C. M. Vollmer
Keyword(s):  
Osmium Tetroxide ◽  
Juglans Regia ◽  
Energy Dispersive ◽  
Protein Bodies ◽  
X Ray ◽  
Edx Analysis ◽  
Freeze Dried ◽  
Major Loss ◽  
Degree Of Extraction ◽  
Helichrysum Bracteatum

This study was undertaken to discover what elemental losses, if any, were occurring from globoid crystals in seed protein bodies during glutaraldehyde – osmium tetroxide fixation. Unfixed cotyledon and radicle tissue of Cucurbita maxima seed, or tissue after glutaral–dehyde–OsO4 treatment, was quick frozen in liquid N2, ground with a cold mortar and pestle, and low-temperature freeze-dried. Globoid crystals in the freeze-dried powder were subjected to energy dispersive x-ray (EDX) analysis. OsO4 fixation resulted in a major loss of P, Mg, and K from cotyledon globoid crystals and a major loss of P, Mg, K, and Ca from radicle globoid crystals. Despite the loss of elements, the OsO4-fixed globoid crystals were still electron dense. When globoid crystals from glutaraldehyde-fixed, dehydrated, and embedded cotyledon tissue were compared with globoid crystals from glutaraldehyde–OsO4-fixed, dehydrated, and embedded tissue, some extraction was found. The degree of extraction varied from complete loss of P, K, and Mg to loss of K only.Effects of glutaraldehyde–OsO4 fixation upon elemental composition of globoid crystals in several other species was also determined. Brazil nut (Bertholletia excelsa) radicle tissue or cotyledon tissue from walnut (Juglans regia), hazelnut (Corylus avellana), sunflower (Helianthus annuus), golden everlasting daisy (Helichrysum bracteatum), cashew (Anacardium occidentale), pistachio (Pistacia vera), and the Western Australian red-capped gum (Eucalyptus erythrocorys) were fixed either in glutaraldehyde or in glutaraldehyde–OsO4. In these species, EDX analysis of globoid crystal sections showed that OsO4 fixation results in major loss of Mg, K, and Ca. Generally, phosphorus levels were reduced from control values as well. When carrying out EDX analysis studies of globoid crystals, we recommend (1) avoiding any use of OsO4, (2) keeping all fixation, washing, and dehydration times as short as possible, and (3) checking all observations with freeze-dried powders.

scholarly journals Ultrastructure and energy-dispersive x-ray microanalysis of cartilage after rapid freezing, low temperature freeze drying, and embedding in Spurr's resin.

1985 ◽  
Vol 33 (10) ◽  
pp. 1073-1079 ◽  
Author(s):  
J Appleton ◽  
R Lyon ◽  
K J Swindin ◽  
J Chesters
Keyword(s):  
Low Temperature ◽  
Freeze Drying ◽  
Energy Dispersive ◽  
Freezing Process ◽  
Rapid Freezing ◽  
Thin Sections ◽  
X Ray ◽  
Freeze Dried ◽  
Rapid Production ◽  
Ideal Method

In order to undertake meaningful high-resolution x-ray microanalysis of tissues, methods should be used that minimize the introduction of artefacts produced by loss or translocation of ions. The most ideal method is rapid freezing but the subsequent sectioning of frozen tissues is technically difficult. An alternative method is to freeze dry the tissues at a low temperature, and then embed them in resin. This facilitates the rapid production of reproducible thin sections. With freeze-dried, embedded hypertrophic cartilage, the morphology was similar to that seen using aqueous fixatives even when no additional electron density is introduced by the use of osmium vapor. Energy-dispersive analysis of specific areas show that little or no loss or migration of ions occurs from structures such as mitochondria. Mitochondrial granules consisting of calcium and phosphorus precipitates were not observed except where the cells were damaged as a result of the freezing process. This may suggest that these granules only appear when tissue is damaged because of inadequate preservation.

Calcium oxalate druse crystals and other inclusions in seed protein bodies: Eucalyptus and jojoba

1978 ◽  
Vol 56 (17) ◽  
pp. 2083-2091 ◽  
Author(s):  
Mark S. Buttrose ◽  
John N. A. Lott
Keyword(s):  
Calcium Oxalate ◽  
Structural Variation ◽  
Seed Protein ◽  
Freeze Fracture ◽  
Protein Body ◽  
Protein Bodies ◽  
X Ray ◽  
Edx Analysis ◽  
Plant Seeds ◽  
Freeze Dried

Druse crystals of calcium oxalate are known to occur in some protein bodies of some plant seeds. This paper reports observations on crystals, and other contents, of protein bodies of Eucalyptus erythrocorys and Simmondsia chinensis (jojoba). Results are presented from thin section studies of glutaraldehyde–OsO4 fixed, dehydrated, and embedded tissue; freeze-fracture studies; energy dispersive x-ray (EDX) analysis studies of freeze-dried tissue powders (Eucalyptus only); and chemical analysis studies of P, Mg, K, and Ca content (Eucalyptus only). Many Eucalyptus protein bodies contained large druse crystals rich in Ca but devoid of P, K, and Mg, and an occasional protein body from jojoba contained some dispersed crystals. Eucalyptus seeds were exceptionally high in Ca content. EDX analysis results provide evidence favouring the phytin-rich nature of globoid crystals in the two species. Structural variation in the globoid crystals was great, especially in jojoba. Eucalyptus, whose protein bodies contained very large globoid crystals rich in P, Mg, and K, was found to have higher levels of these elements than most species investigated to date.

Protein bodies in Datura stramonium seeds: structure and mineral nutrient composition

1991 ◽  
Vol 69 (11) ◽  
pp. 2545-2554 ◽  
Author(s):  
Sara Maldonado ◽  
John N. A. Lott
Keyword(s):  
Distribution Patterns ◽  
Cell Types ◽  
Datura Stramonium ◽  
Energy Dispersive ◽  
Protein Bodies ◽  
Mineral Nutrient ◽  
Electron Microscopic ◽  
Element Analysis ◽  
X Ray ◽  
Endosperm Protein

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.

Globoids in Protein Bodies of Legume Seed Cotyledons

1978 ◽  
Vol 5 (1) ◽  
pp. 89 ◽  
Author(s):  
JNA Lott ◽  
MS Buttrose
Keyword(s):  
Critical Evaluation ◽  
Protein Body ◽  
Protein Bodies ◽  
Fixed Tissue ◽  
Edx Analysis ◽  
Freeze Dried ◽  
Thin Sectioning ◽  
Legume Protein ◽  
Structure Lead ◽  
Published Research

Inclusions were found in protein bodies in cotyledons of seeds of each of five legume species (Acacia conferta, Cassia artemisioides, Clianthus formosus, Glycine max, Vicia faba). They were studied by a number of techniques: thin sectioning of fixed, embedded tissue; freeze-fracturing of unfixed tissue; chemical analysis of P, K, Mg and Ca content; and energy dispersive X-ray (EDX) analysis of both sections of glutaraldehyde-fixed tissue and freeze-dried tissue powders. The results of the studies presented in this paper, combined with a critical evaluation of the published research on legume protein body structure, lead us to believe that globoid crystals are a frequent occurrence in legume protein bodies. EDX analysis results indicate that electron-dense globoid crystals are rich in phytin and that phytin may also be present throughout the proteinaceous matrix portions of some legume protein bodies.

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