In 1980s, the fragmentation or subdivision of protein deposits at the periphery of protein storage vacuole was suggested as the only route of PB development in pea cotyledon cells. Since then, other independant processes such as terminal dilation , transformation and de novo development have been discussed as alternative routes for PB development, and today, these multiple mechanisms of PB development are accepted as a result of active investigations. For analysis of the protein accumulations in the ER cisternae during seed development, immunocytochemical gold labellings were applyed on the single cells separated by enzymatic digestion from cotyledon tissue. Anti-legumin labellings at the early stage, and anti-vicilin labellings at the intermediate stage were observed on the protein-filled ER. The $\alpha-Tip$, which is the ER retention protein, was labelled somewhat at late stage, and PPase, a sort of tonoplast membrane protein, was labelled at early stage.
This study was carried out to investigate the effects of grape seed oil on quality characteristics of pressed ham. Five different treatments were tested based on differences in the amount of grape seed oil added to the pressed ham. As a control, 10% back fat was added without any grape seed oil. For the first treatment, 10% grape seed oil replaced a portion of the lard component added to the pressed ham. For the 2nd, 3rd and 4th treatments, 20%, 30% and 40% of grape seed oil were substituted for lard, respectively. Pressed ham manufactured with grape seed oil was vacuum packaged and stored for 1, 7, 14,21 and 28 d at $4^{\circ}C$. Samples were analyzed for chemical composition, physico-chemical properties (pH, color) and texture characteristics. Typical chemical composition characteristics (crude protein, crude fat, crude ash) were not significantly different between control and grape seed oil treatment groups. Moisture content of grape seed oil treatment groups (GSO 30% and 40%) was significantly lower than that of controls (p<0.05). There was a not clear difference in pH between control and grape seed oil treatment groups. In the 21 d of storage, pH values of all treatments were significantly higher than those of other storage days. Meat color $(CIEL^*\;and\;b^*)$ of grape seed oil treatment group (GSO 40%) was significantly higher than that of control. Whereas meat color a value of GSO 40% treatment was significantly lower than that of control. It was not clearly changed as storage time increased. There was no significant difference in texture between control and grape seed oil treatment groups, and appeared to be unaffected by storage length. Based on these findings, we conclude that the chemical composition and texture characteristics of manufactured pressed ham were not affected by grape seed oil addition. These results also indicate that high-quality pressed ham can be manufactured with increased polyunsaturated fatty acid content.
Abundant proteins often cause problems in proteome study. Glutelin family proteins (hereafter referred to glutelin) are present in rice proteome sample as over-whelming constituents with very high abundance. In order to increase the number of identified proteins in rice proteome study, we developed a newly improved method for sample preparation through the removal of glutelin. When the protein samples from rice seed were extracted by the conventional trichloroacetic acid (TCA) extraction method, glutelin accounts for about 60% of total rice seed proteins in SDS gels. Using our new water extraction method, glutelin consists of only about 10% of total proteins. After analyzing on a two-dimensional gel electrophoresis (2-DE), 937 protein spots were detected using the conventional TCA extraction method. On the other hand, 1240 proteins could be seen using the new water extraction method. The selectivity for non-glutelin and less abundant protein by the water extraction method was also confirmed by ESI-Q/TOF mass spectrometry analysis. Thus, the new water extraction method developed here can be efficiently used to study the proteome analysis of rice storage seed.
Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
/
2001.06a
/
pp.1158-1158
/
2001
Fast and dynamic biochemical, enzymatic and morphological changes occur during the so-called generative development and during the vegetative processes in seeds. The most characteristic biochemical and compositional changes of this period are the formation and decline of storage components or their precursors, the change of their degree in polymerization and an extensive change in water content. The aim of the present study was to detect the maturation processes in seed nondestructively and to verify the applicability of near infrared spectroscopic methods in the measurement of physiological, chemical and biochemical changes in wheat seed. The amount and variation of different water “species” has been changed intensively during maturation. Characteristic changes of three water absorption bands (1920, 1420 and 1150 nm) during maturation were analysed. It was concluded that the free/bound transition of water molecules could be followed sensitively in different region of NIR spectra. Kinetic changes of carbohydrate reserves were characteristic during maturation. An intensive formation and decline of carbohydrate reserves were observed during early stage of maturation (0 -13 days, high energy demand). An accelerated formation of storage carbohydrates (starch) was detected in the second phase of maturation. Five characteristic absorption bands were analysed which were sensitive indicators the changes of carbohydrates occurred during maturation. Precursors of protein synthesis and the synthesis of reserve proteins and their kinetic changes during maturation were followed from NIR spectra qualitative and qualitatively. Dynamic formation of amino acids and the changes of N forms were detected by spectroscopic, chromatographic and by capillary electrophoresis methods. Calibration equations were developed and validated in order to measure the optimal maturation time protein and moisture content of developing wheat seeds. The spectroscopic methods are offering chance and measurement potential in order to detect fine details of physiological processes. The spectra have many hidden details, which can help to understand the biochemical background of processes.
The purpose of this study is to investigate the hemp (Cannabis sativa cv. Chungsam) seed structure and ultrastructure of food reserves by scanning and transmission electron microscopy. We examined the seed coat and embryo consisting of a hypocotyl-radicle axis and two cotyledons. The seed coat consisted of exotesta and endotesta. The exotesta was a mechanical layer with lignified and elongated cells, while endotesta of the underlying layers of the exotesta was consisted of two separated cell layers. The collapsed outer layer of endotesta showed the unique reticulate structures. In cotyledon cells, protein and lipid bodies occupied most of cytoplasm. Protein bodies varied in diameter from 1.8 to $5.0{\mu}m$ and possessed a protein matrix containing electron-dense globoid crystals. Numerous lipid bodies ranged from 0.8 to $3.0{\mu}m$ in diameter were distributed around the protein bodies. During the early stages of breakdown, protein bodies rapidly changed their shape into the granular feature, however, lipid bodies were gradually degradated and fused each other. The degeneration process of protein bodies and lipid bodies of cotyledon cells might be correlated with the reports which hemp seeds rapidly lose their ability to germinate.
Objective: The objective of this study was to investigate the effect of dietary pumpkin (Cucurbita moschata) seed meal (PSM) on laying hens' performance, quality, fatty acids, cholesterol, antioxidant compounds and shelf life of eggs. Methods: Eighty Tetra SL laying hens, 50-week-old, were randomly divided into two equal groups, having 10 replicates with 4 birds in each. The control (CON) treatment was fed with basal diet, while experimental treatment was fed a diet with 9% PSM, for a 6 week period. Results: Dietary PSM significantly decreased average daily feed intake (p<0.05), with no significant effect on other performance parameters. The PSM, enriched the eggs with polyunsaturated fatty acids, especially α linolenic acid (0.33 vs 0.21 g/100 g) and linoleic acid (20.65 vs 18.37 g/100 g), whereas it reduced the amount of arachidonic acid with 3.91% and n-6/n-3 ratio in PSM eggs compared with CON. The inclusion of 9% PSM significantly (p<0.05) diminished the cholesterol concentration in yolk with 11.31% and in egg with 10.38%, in respect to the CON samples. The significantly (p<0.05) higher concentration of polyphenols and antioxidant compounds, determined in PSM eggs, proved to be effective on shelf life of eggs preserved at refrigerator (5℃) and room temperature (21℃) for 28 days, by delaying the lipid oxidation and protein denaturation. This effect was reflected in significantly (p<0.05) higher Haugh unit in eggs stored 28 days at 21℃ and lower albumen pH values for the overall storage time, both at 5℃ and 21℃, proving the antioxidant effect of pumpkin. Conclusion: Dietary PSM supplementation was significantly effective on average daily feed intake and egg quality by increasing some fatty acids while lowering the cholesterol concentration. Also, PSM proved to be effective improving shelf life of eggs for 28 days storage time.
Experiments were conducted to obtain information on separation of nonviable seeds from seed lots of rape, Chinese cabbage and radish, by nondestrutive ways. Seeds were artificially aged at 90% relative humidity(RH) and 45$^{\circ}C$ to get different seed qualities. Large amounts of amino acids, proteins and sinapine were leaked from the dead seeds into water in the course of 4~8 hours soaking, while high quality seeds did not leak in all crops. Percentage of normal seedlings from nonfluorescent cellulose coated seeds of rape, Chinese cabbage and radish under ultraviolet light was 96, 96 and 74%, while that of fluorescent seeds was 8, 9 and 1%, respectively. Cellulose coating and storage of Chinese cabbage and rape seeds at 60 and 75% RH for 3 months and 90% RH for 2 months did not deteriorate seed quality. But percentage of normal seedlings from nonfluorescent radish seeds stored at 60, 75 and 90% RHs for 3 months was 63, 64 and 2%, respectively.
The expressed sequence tags(ESTs) from immature seed of rice, Oryza sativa cv Milyang 23, were partially sequenced and analyzed by homology. As of 1998, the partial sequences of about 6,600 cDNA clones were analyzed from normal and normalized immature seed cDNA libraries. About 2,200 ESTs were putatively identified by BLASTX deduced amino acid sequence homology analysis. About 20% of them were putatively identified as storage proteins. Also the clones were highly homologous to genes involved particularly in starch biosynthesis, glycolysis, signal transduction and defenses. Compared to 35% of redundancy in the ESTs of normal cDNA library, that from the substracted library was 15%. The Korea Rice Genome Network is maintained to provide the updated information of sequences, their homologies and sequence alignments of ESTs. For the stable expression of transgene in rice, diverse vectors were developed for overexpression, targeting and gene dosage effect with transit peptides (Tp) and matrix attachment region (MAR) sequence from chicken lysozyme locus. The rice calli were transformed via Agrobacterium tumefaciens LBA4404(pSB1) with the triparental mating technique and selected by herbicide resistance. The green fluorescent protein(GFP) gene in expression vector under the control of rbcS promoter-Tp was overexpressed upto 10 % of the total soluble protein. In addition, the Tp-sGFP fusion protein was properly processed during translocation into chloroplast. The expression of sGFP in the presence of MAR sequences was analyzed with Northern and immunoblot analysis. All the lines in which sGFP transgene with MAR sequence, showed position independent and copy number-dependent expression, while the lines without MAR showed the varied level of expression with the integration site. Thus the MAR sequence significantly reduced the variation in transgene expression between independent transformants.
This study has been carried out to investigate the ultrastructural changes in the associated with the disintegration of the storage materials in endosperm cell of ginseng (Panax ginseng C.A. Meyer) seed during after-ripening with light and electron microscope. The protein body of endosperm cells near the umbiliform layer showed various degenerative patterns, and so electron density of proteinaceous matrix was gradualJy decreased during afterripening. These results indicate that the decomposition of endosperm was already initiated during after-ripening. As the degeneration of endosperm was more progressed after the dehiscence of seed, non-decomposed part of protein body appeared amorphously with high electron density. Decomposed protein bodies were vacuolized with the loss of their matrix and gradually expanded by fusion. Also, spherosomes were gradually dissolved with the lowered electron density during the degeneration of endosperm. The vesicles of dictyosomes near the cell wall are observed in endosperm contacting with umbiliform layer and are fused with plasma membrane. Umbiliform layer which was the complex of the decomposed remnants of lysis and materials has strong stainability for toluidine blue and basic fuchsin.
Proceedings of the Botanical Society of Korea Conference
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1987.07a
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pp.261-282
/
1987
Plants store a significant amount of their nitrogen, sulfur and carbon reserves as storage proteins in seed tissues. The major proteins present in rice seeds are the glutelins. Glutelins are initially synthesized at 4-6 days postanthesis and deposited into protein bodies via Golgi apparatus. Based on nucleic acid sequences and Southern blot analysis, the three isolated glutelin genomic clones were representative members of three gene subfamilies each containing 5 to 8 copies. A comparison of DNA sequences displayed by relevant regions of these genomic clones showed that two subfamilies, represented by clones, Gt1 and Gt2, were closely, related and probably evolved by more recent gene duplication events. The 5' flanking and coding sequences of Gt1 and Gt2 displayed at least 87% homolgy. In contrast, Gt3 showed little or no homolgy in the 5' flanking sequences upstream of the putative CAAT boxes and exhibited significant divergence in all other portions of the gene. Conserved sequences in the 5' flanking regions of these genes were identified and discussed in light of their potential regulatory role. The derived primary sequences of all three glutelin genomic clones showed significant homology to the legume 11S storage proteins indicating a common gene origin. A comparison of the derived glutelin primary sequences showed that mutations were clustered in three peptide regions. One peptide region corresponded to the highly rautable hypervariable region of legume peptide region of legume 11S storage proteins, a potential target area for protein modification. Expression studies indicated that glutelin mRNA transcripts are differentially accumulated during endosperm development. Promoterss of Gt2 and Gt3 were functional as they direct transient expression of chloramphenicol acetyltransferase in cultured plant cell.
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