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.

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.

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.

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.

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.

Cruciferous Oilseed Proteins: the Protein Bodies of Sinapis alba Seed

1976 ◽  
Vol 3 (6) ◽  
pp. 731 ◽  
Author(s):  
JTO Kirk ◽  
NA Pyliotis
Keyword(s):  
Proteolytic Activity ◽  
Storage Proteins ◽  
Protein Body ◽  
Protein Bodies ◽  
Seed Meal ◽  
Mustard Seed ◽  
Major Protein ◽  
Intense Band ◽  
Ungerminated Seed ◽  
Protein Classes

The solubility properties of the proteins of oil-free meal of white mustard seed (S. alba) in various aqueous extraction media are described. Electrophoresis on cellulose acetate of a salt extract of the seed meal at pH 7.0 shows the presence of two positively charged protein bands: a slow moving intense band (I) and a less intense band with higher mobility (II). On the basis of Sephadex G100 chromatography and sedimentation behaviour, these bands are deemed to be identical with the two major protein classes (12 S and 1.7 S, respectively) present in this and other Brassica-related species, as described by other workers. Centrifugation after filtration of a seed meal homogenate yields a preparation that is completely soluble in salt solution, and can be shown by electron microscopy to consist entirely of protein body fragments. Only the 12 S protein can be detected in significant quantity in this preparation: this protein at least we may assume to be present in the aleurone (protein) grains observed in micrographs of the cotyledon cells. In germinating seeds, disappearance of protein bodies is accompanied by a diminution in total salt-soluble protein and in the amounts of the 12 S and 1.7 S proteins, supporting their identification as storage proteins. The rate of utilization is the same in the light and in the dark. Proteolytic activity was detected in the ungerminated seed. The level of activity was more than sufficient to account for the subsequent observed rate of protein utilization. Proteolytic activity per seed increased by only 40-70% during 4 days germination.

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

Structural changes in whole milk during the production of sterile concentrates: an electron microscope study

1970 ◽  
Vol 37 (3) ◽  
pp. 513-522 ◽  
Author(s):  
P. W. Board ◽  
Joan M. Bain ◽  
D. W. Gove ◽  
J. T. Mullett
Keyword(s):  
Electron Microscopy ◽  
Structural Changes ◽  
Physical Stability ◽  
Protein Bodies ◽  
Large Protein ◽  
Casein Micelles ◽  
Fat Globules ◽  
Whole Milk ◽  
Freeze Dried ◽  
Average Size

SummaryElectron microscopy was used to study the effect of addition of NaOH, forewarming, homogenization, concentration, and heat sterilization on the structure of whole milk during the production of 4 heat-sterilized concentrates that showed different degrees of physical stability. The samples were prepared for electron microscopy by adding fixative either to liquid or freeze-dried material, and then embedding for sectioning.Noticeable structural changes occurred during forewarming of samples containing added NaOH, during homogenization, and during heat sterilization. Addition of NaOH and forewarming decreased the closeness of packing of the subunits in the casein micelles. Homogenization reduced the average size of the fat globules, and protein became attached to their surfaces. Heat sterilization caused coalescence of protein. In non-homogenized sterilized concentrate with added NaOH, protein bodies of about 100 times the volume of the original casein micelles were formed; these were free-floating and the concentrate was stable. In non-homogenized sterilized concentrate with no added NaOH the protein bodies were about 15 times the size of the casein micelles and bridged to each other, thereby forming a sediment consisting of large irregular particles. Very large protein bodies containing fat globules formed during heat sterilization of the homogenized samples, both in the presence and in the absence of NaOH, and were responsible for the formation of sediment in these 2 products.Needle crystals observed in most samples were identifed as CaCO3. H2O by selected area diffraction; KC1 crystals were also detected.

scholarly journals Subunit structure of junctional feet in triads of skeletal muscle: a freeze-drying, rotary-shadowing study.

1984 ◽  
Vol 99 (5) ◽  
pp. 1735-1742 ◽  
Author(s):  
D G Ferguson ◽  
H W Schwartz ◽  
C Franzini-Armstrong
Keyword(s):  
Three Dimensional ◽  
Freeze Fracture ◽  
Subunit Structure ◽  
Thin Sections ◽  
Large Structures ◽  
Foot Structure ◽  
Freeze Dried ◽  
Thin Sectioning ◽  
En Face ◽  
A Subunit

Isolated heavy sarcoplasmic reticulum vesicles retain junctional specializations (feet) on their outer surface. We have obtained en face three-dimensional views of the feet by shadowing and replicating the surfaces of freeze-dried isolated vesicles. Feet are clearly visible as large structures located on raised platforms. New details of foot structure include a four subunit structure and the fact that adjacent feet do not abut directly corner to corner but are offset by half a subunit. Feet aligned within rows were observed to be rotated at a slight angle off the long axis of the row creating a center-to-center spacing (32.5 nm) slightly less than the average diagonal of the feet (35.3 nm). Comparison with previous information from thin sections and freeze-fracture showed that this approach to the study of membranes faithfully preserves structure and allows better visualization of surface details than either thin-sectioning or negative-staining.

The aspartic proteinase is expressed in Arabidopsis thaliana seeds and localized in the protein bodies

1999 ◽  
Vol 9 (1) ◽  
pp. 75-84 ◽  
Author(s):  
Asuman Mutlu ◽  
Xia Chen ◽  
Sridhar M. Reddy ◽  
Susannah Gal
Keyword(s):  
Arabidopsis Thaliana ◽  
Seed Storage Protein ◽  
Protein Body ◽  
Seed Storage ◽  
Cell Types ◽  
Aspartic Proteinase ◽  
Protein Bodies ◽  
Marker Enzyme ◽  
Antigenic Peptides ◽  
2S Albumin

AbstractWe have been studying a seed aspartic proteinase, termed AtAP, from Arabidopsis thaliana. In previous work, we purified the proteinase, analysed its activity and isolated the cDNA sequence. In this paper, the expression of the mRNA for the aspartic proteinase was analysed in seed tissues both by Northern blots for overall regulation and by in situ hybridization to follow cell-specific localization of message. We found a 1.9 kb aspartic proteinase message in dry seeds and seed pods. This message was expressed in many different cell types of the mature dry seed. The localization of the protein within these cells was also determined. Antibodies were raised against the AtAP and purified using affinity chromatography on an AtAP–immobilized-pepstatin A–agarose column. This purified antibody recognized several AtAP peptides in seeds. To localize the enzyme in cells, we isolated protein bodies from the dry seeds of Arabidopsis using a non-aqueous isolation method. The AtAP activity and antigenic peptides were found to be highest in the protein body fraction and co-localized with the seed storage protein 2S albumin and the vacuolar marker enzyme α-mannosidase. This protein body localization of the AtAP was confirmed with immunocytochemical localization by electron microscopy and shows that the protein is not secreted by these cells.

Location of reserves of mineral elements in seed protein bodies: macadamia nut, walnut, and hazel nut

1978 ◽  
Vol 56 (17) ◽  
pp. 2072-2082 ◽  
Author(s):  
John N. A. Lott ◽  
Mark S. Buttrose
Keyword(s):  
Juglans Regia ◽  
Matrix Material ◽  
Freeze Fracture ◽  
Mineral Elements ◽  
Protein Bodies ◽  
Large Protein ◽  
Edx Analysis ◽  
Macadamia Integrifolia ◽  
Macadamia Nut ◽  
Very High

This paper presents the results of thin-section, freeze-fracture, energy dispersive x-ray (EDX) analysis, and chemical analysis studies of cotyledon tissue from dry seeds of macadamia nut (Macadamia integrifolia), walnut (Juglans regia), and hazel nut (Corylus avellana). Studies concentrated on mineral elements in the storage protein bodies. Protein bodies of macadamia nut contained no protein crystalloids but did contain some globoid crystals. Aqueous fixation of macadamia tissue appeared to result in a rapid solubilization of proteinaceous matrix material. EDX analysis of macadamia nut globoid crystals revealed the presence of P, K, and Mg in most cases but the occasional crystal was very high in calcium. Walnut protein bodies often contained many large protein crystalloids. Globoid crystals were present between the protein crystalloids and in the proteinaceous matrix. EDX analysis indicated the presence of P, K, and Mg as the main elements in walnut globoid crystals. Hazel nut protein bodies contained one or more small protein crystalloids, globoid crystals of various sizes, and proteinaceous matrix. Some protein bodies contained druse crystals. EDX analysis showed that the druse crystals were rich in calcium and thus are presumably calcium oxalate. EDX analysis of globoid crystals revealed the presence of elements consistent with the phytin-rich nature of globoid crystals. Variations in globoid crystal composition are discussed.

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