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A vacuolar processing enzyme, deltaVPE, is involved in seed coat formation at the early stage of seed development.

Vacuolar processing enzyme (VPE) is a Cys proteinase responsible for the maturation of vacuolar proteins. Arabidopsis thaliana deltaVPE, which was recently found in the database, was specifically and transiently expressed in two cell layers of the seed coat (ii2 and ii3) at an early stage of seed development. At this stage, cell death accompanying cell shrinkage occurs in the ii2 layer followed by cell death in the ii3 layer. In a deltaVPE-deficient mutant, cell death of the two layers of the seed coat was delayed. Immunocytochemical analysis localized deltaVPE to electron-dense structures inside and outside the walls of seed coat cells that undergo cell death. Interestingly, deltaVPE in the precipitate fraction from young siliques exhibits caspase-1-like activity, which has been detected in various types of plant cell death. Our results suggest that, at the early stage of seed development, deltaVPE is involved in cell death of limited cell layers, the purpose of which is to form a seed coat.

Arabidopsis↗

Proteomic and transcriptomic analysis of Arabidopsis seeds: molecular evidence for successive processing of seed proteins and its implication in the stress response to sulfur nutrition.

Seed storage proteins are synthesized as sources of carbon, nitrogen and sulfur for the next generation of plants. Their composition changes according to nutritional conditions. Here, we report the precise molecular identification of seed proteins by proteomic analysis of wild-type Arabidopsis thaliana and methionine-over-accumulating mutant mto1-1 plants. The identities of 50 protein spots were determined in the protein extract of mature Arabidopsis seeds by two-dimensional (2D) gel electrophoresis and subsequent mass spectrometric analysis. Of these protein spots, 42 were identified as derived from 12S globulins or 2S albumins. These results indicate that approximately 84% of protein species in Arabidopsis seeds are derived from a few genes coding for 12S globulins and 2S albumins. Extensive mass spectrometric analysis of the 42 spots revealed that successive C-terminal degradation occurred on the 12S globulins. The feasibility of this C-terminal processing was rationalized by molecular modeling of the three-dimensional structure of 12S globulins. The C-terminal degradation at glutamic acid residues of the 12S globulin subunits was repressed under sulfur-deficient conditions. Transcriptome analysis was combined with proteomic analysis to elucidate the mechanism of changes in seed protein composition in response to sulfur deficiency. The results suggest that seed storage proteins in Arabidopsis undergo multi-layer regulation, with emphasis on post-translational modifications that enable the plant to respond to sulfur deficiency.

Arabidopsis↗

Cross-sensitization between poppy seed and buckwheat in a food-allergic patient with poppy seed anaphylaxis.

The opium poppy, Papaver somniferum L., is the source of both poppy seeds and opium. The commercially available seeds are widely used as ingredients for various kinds of food. IgE-mediated sensitization to poppy seeds is rare, but, if present, clinical symptoms are usually severe. Cross-sensitizations between poppy seeds and other food allergens have been described with sesame, hazelnut, rye grain and kiwi fruit. We report the case of a 17-year-old female with an apparently food-allergic reaction after ingestion of a poppy seed cake. Allergological workup revealed a poppy seed anaphylaxis and led to the identification of a novel cross-sensitization with buckwheat.

Adolescent↗

Gibberellin induces alpha-amylase gene in seed coat of Ipomoea nil immature seeds.

Two full-length cDNAs encoding gibberellin 3-oxidases, InGA3ox1 and InGA3ox2, were cloned from developing seeds of morning glory (Ipomoea nil (Pharbitis nil) Choisy cv. Violet) with degenerate-PCR and RACEs. The RNA-blot analysis for these clones revealed that the InGA3ox2 gene was organ-specifically expressed in the developing seeds at 6-18 days after anthesis. In situ hybridization showed the signals of InGA3ox2 mRNA in the seed coat, suggesting that active gibberellins (GAs) were synthesized in the tissue, although no active GA was detected there by immunohistochemistry. In situ hybridization analysis for InAmy1 (former PnAmy1) mRNA showed that InAmy1 was also synthesized in the seed coat. Both InGA3ox2 and InAmy1 genes were expressed spatially overlapped without a clear time lag, suggesting that both active GAs and InAmy1 were synthesized almost simultaneously in seed coat and secreted to the integument. These observations support the idea that GAs play an important role in seed development by inducing alpha-amylase.

Amino Acid Sequence↗

[Novel application of FTIR to the analysis of sweet pepper seeds and tomato seeds with space mutation].

Comparative study of the sweet pepper and tomato seeds lines with space mutation and corresponding conventional seeds was carried out with FTIR, in an attempt to search for the influence of space environment upon mutation. The similarities among space seeds lines are: the absorbance at near 1,160-1,061 cm(-1) increased. This result shows that the stretching vibration of C-O on the structure of carbohydrates was enhanced by space mutation. The special mutations are: (1) The absorbance at 2,854 cm(-1) of space sweet pepper was enhanced, showing that the stretching vibration of CH2 was stronger; (2) The absorbance at 1,160 cm(-1) of cherry tomato seeds lines was increased notably, indicating that the stretching vibration of C-O on the structure of carbohydrates was stronger. The results show that the space environment could notably increase the carbohydrates in the tomato seeds. The space sweet pepper and tomato seeds lines, which produced similar mutation, could be influenced by microgravity in space. The special mutations are the results of the influence of universe radiation in space.

Capsicum↗

In vitro characterization of mesenchymal stem cell-seeded collagen scaffolds for tendon repair: effects of initial seeding density on contraction kinetics.

Mesenchymal stem cells (MSCs) were isolated from bone marrow, culture-expanded, and then seeded at 1, 4, and 8 million cells/mL onto collagen gel constructs designed to augment tendon repair in vivo. To investigate the effects of seeding density on the contraction kinetics and cellular morphology, the contraction of the cell/collagen constructs was monitored over time up to 72 h in culture conditions. Constructs seeded at 4 and 8 million cells/mL showed no significant differences in their gross appearance and dimensions throughout the contraction process. By contrast, constructs seeded at 1 million cells/mL initially contracted more slowly and their diameters at 72 h were 62 to 73% larger than those seeded at higher densities. During contraction, MSCs reoriented and elongated significantly with time. Implants prepared at higher seeding densities showed more well aligned and elongated cell nuclei after 72 h of contraction. Changes in nuclear morphology of the MSCs in response to physical constraints provided by the contracted collagen fibrils may trigger differentiation pathways toward the fibroblastic lineage and influence the cell synthetic activity. Controlling the contraction and organization of the cells and matrix will be critical for successfully creating tissue engineered grafts.

Animals↗

Selective identification of different brachytherapy sources, ferromagnetic seeds, and fiducials in the prostate using an automated seed sorting algorithm.

PURPOSE: Routine permanent prostate brachytherapy (PPB) includes CT-based postimplant dosimetry (PID). A method of identifying different source types from CT data in the same implant volume is described. METHODS AND MATERIALS: A previously described automatic method for seed localization using CT data is used in this study. Two cases were analyzed: a PPB case with (103)Pd followed by salvage (125)I implantation, both performed at another institution, and a cadaver case where 4 different seed types, including ferromagnetic seeds, and fiducials were implanted. RESULTS: Automatic segregation of different seed types with minimal manual correction is demonstrated using the described localization algorithm. The process is confirmed accurate by comparison of plain film radiographs to CT data and digitally reconstructed radiographs. CONCLUSION: Unique identification of different source types, including PPB seeds, fiducial markers, and ferromagnetic seeds in permanent implants is possible and permits dosimetric analyses that are spatially coincident.

Aged↗

Flax seed oil and flax seed meal reduce the formation of aberrant crypt foci (ACF) in azoxymethane-induced colon cancer in Fisher 344 male rats.

Flax seed oil and flax seed meal are good sources of omega-3 fatty acids. The objective of this study was to explicate the effects of feeding flax seed oil and flax seed meal on AOM-induced aberrant crypt foci (ACF) in Fisher 344 male rats. Following an acclimatization period, rats were divided into six groups and fed AIN 93G diet Control (C), C+7 and 14% soybean oil (SBO), C+7 and 14% flax seed oil (FSO) and C+10 and 20% flax seed meal (FSM). All rats received 16 mg/kg body weight of AOM at 7 and 8 weeks of age. The rats were euthanized with CO2 at 17 weeks of age. FSM and FSO reduced the incidence of ACF which are putative precursor lesions in the development of colon cancer in the distal colon by 88% and 77%, in the proximal colon by 86% and 87% with a total reduction of 87.5% and 84%, respectively. Glutathione-S-transferase (GST) activities were significantly (P<0.05) higher in rats fed C+7 and 14% FSO and C+10 and 20% FSM, as compared to rats fed C+SBO diets. Results of this study showed that FSO and FSM reduced the incidence of AOM-induced ACF formation and may therefore be effective chemopreventive agents.

Animals↗

Endothelial cell seeding of (heparinized) collagen matrices: effects of bFGF pre-loading on proliferation (after low density seeding) and pro-coagulant factors.

Endothelial cell seeding to improve the performance of small-diameter vascular grafts requires a suitable substrate, such as crosslinked collagen. In addition to providing a suitable substrate for adhesion and growth of endothelial cells, proliferation of seeded endothelial cells can be enhanced by local, sustained release of basic fibroblast growth factor (bFGF, a heparin-binding growth factor for endothelial cells). We have previously shown that collagen crosslinked using N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS) supports adhesion and proliferation of human umbilical vein endothelial cells (HUVECs). In the present study, HUVECs were seeded on (heparinized) EDC/NHS-crosslinked collagen, pre-loaded with bFGF. Proliferation of HUVECs on (heparinized) crosslinked collagen increased with increasing amounts of pre-loaded bFGF. The minimal cell-seeding density required for proliferation proved to be very low after pre-loading the substrates with bFGF, and was 4-fold lower for heparinized crosslinked collagen compared to crosslinked collagen (250 versus 1000 cells/cm(2)). Pro-coagulant properties (von Willebrand factor secretion and tissue factor expression) of HUVECs seeded on (heparinized) crosslinked collagen, with or without pre-loading of bFGF, were comparable to those of HUVECs on TCPS. It is concluded that heparinized, EDC/NHS-crosslinked collagen pre-loaded with bFGF is a candidate matrix for in vivo endothelial cell seeding of synthetic vascular graft materials.

Cell Division↗

Automated seed detection and three-dimensional reconstruction. I. Seed localization from fluoroscopic images or radiographs.

An automated procedure for the detection of the position and the orientation of radioactive seeds on fluoroscopic images or scanned radiographs is presented. The extracted positions of seed centers and the orientations are used for three-dimensional reconstruction of permanent prostate implants. The extraction procedure requires several steps: correction of image intensifier distortions, normalization, background removal, automatic threshold selection, thresholding, and finally, moment analysis and classification of the connected components. The algorithm was tested on 75 fluoroscopic images. The results show that, on average, 92% of the seeds are detected automatically. The orientation is found with an error smaller than 50 for 75% of the seeds. The orientation of overlapping seeds (10%) should be considered as an estimate at best. The image processing procedure can also be used for seed or catheter detection in CT images, with minor modifications.

Algorithms↗

[Seed loss through the urinary tract and retrieval after prostate seed implant].

This study describes our experience with seed loss and retrieval through the urinary tract following seed implants for prostate cancer, and offers Japanese guidelines for safety and management. Two hundred consecutive patients were analyzed. All patients were preplanned with a modified peripheral loading technique and implanted with a Mick applicator under ultrasound guidance. All patients were instructed to return excreted seeds, if any, to our center. Seed loss occurred in 6% of patients and 0.13% of seeds. Seed loss tended to occur in the early period through either urine or ejaculation.

Brachytherapy↗

Silencing of chaperonin 21, that was differentially expressed in inflorescence of seedless and seeded grapes, promoted seed abortion in tobacco and tomato fruits.

Vitis vinifera L. cv. 'Thompson Seedless' presents a type of stenospermocarpy in grape where fertilization occurs but seeds abort and fail to develop. To unravel the molecular basis for stenospermocarpy in grapes, subtractive hybridization was carried out in order to isolate differentially regulated genes that participate in the seedlessness machinery. Two 'Thompson' lines, a seeded and a seedless, were screened during different flower developmental stages. One of the genes, that was differentially expressed between the seeded and seedless lines, was the chloroplast chaperonin 21 (ch-Cpn21). ch-Cpn21 is a 21-kDa co-chaperonin polypeptide formed by two GroES-like domains fused together in tandem. Silencing of ch-Cpn21 in Nicotiana benthamiana plants resulted in leaf stunting, chlorosis, as well as ovary necrogenesis leading to seed abortion. Moreover, organ-specific silencing of ch-Cpn21 only in Lycopersicum esculentum fruits resulted in the development of seedless tomatoes. These results suggest that ch-Cpn21 may play a role in seed abortion in stenospermocarpic grapes.

Chaperonins↗

Warm temperatures or drought during seed maturation increase free alpha-tocopherol in seeds of soybean (Glycine max [L.] Merr.).

Soybean seeds are an important source of dietary tocopherols, but like seeds of other dicotyledonous plants, they contain relatively little alpha-tocopherol, the form with the greatest vitamin E activity. To evaluate potential effects of environmental stress during seed maturation on tocopherols, soybeans were raised in greenhouses at nominal average temperatures of 23 degrees C or 28 degrees C during seed fill, with or without simultaneous drought (soil moisture at 10-25% of capacity), during normal growing seasons in 1999 (cvs. Essex and Forrest) and 2000 (cvs. Essex, Forrest, and Williams). Total free (nonesterified) tocopherols increased slightly in response to drought in Essex and Forrest. All three lines responded to elevated temperature and, to a lesser extent, drought with large (2-3-fold) increases in alpha-tocopherol and corresponding decreases in delta-tocopherol and gamma-tocopherol. The results suggest that weather or climate can significantly affect seed tocopherols. It may be possible to breed for elevated alpha-tocopherols by selecting for altered plant response to temperature.

Environment↗

Effect of seed development stage on sphingolipid and phospholipid contents in soybean seeds.

Glucosylceramide (GlcCer) and ceramide (Cer) are the predominant sphingolipids (SL) in soybeans. They have been recognized as functional components in plants and may have health benefits for humans. The objective of this study was to evaluate the changes in SL and phospholipid (PL) contents that occurred during seed development. Soybean seeds of three cultivars (IA1008, IA1010, and IA1014) were harvested at 5-day intervals from 28 days after flowering (DAF) to 68 DAF (mature seed). SL and PL contents of seeds were quantified using high-performance liquid chromatography (HPLC) with an evaporative light scattering detector (ELSD). SL and PL contents decreased significantly during seed development. Averaged across cultivars, Cer content on a dry weight basis decreased from 51.4 nmol/g at 28 DAF to 22.2 nmol/g at 68 DAF, whereas GlcCer content decreased from 522.8 nmol/g at 28 DAF to 135.8 nmol/g at 68 DAF. PL percentage of the total lipid decreased from 9.1% at 28 DAF to 3.5% at 68 DAF.

Ceramides↗

Incidence of seed-borne fungi and aflatoxins in Sudanese lentil seeds.

Thirteen seed samples of lentil (Lens esculenta) were incubated on agar plate and moist filter papers (Moist Chambers) at 28 +/- 2 degrees C for determination of the incidence of seed-borne fungi. Aflatoxins content of the seeds was measured using the bright greenish- yellow fluorescence test (BGYF) and thin-layer chromatography (TLC). Sixty-nine species and seven varieties, which belong to 24 genera of fungi, were isolated from this crop. Of these fungi, 51 species and two varieties are considered new for this crop, whereas seven genera and 13 species are new to the mycoflora of the Sudan. The genus Aspergillus (13 species and 6 varieties) which comprising 44% of the total colony count was the most prevalent genus followed by Rhizopus (2 species, 19%), Penicillium (6 species) and Fusarium (8 species) (12%), Chaetomium (3 species) and Cladosporium (5 species) (6%), where the 18 genera (1-4 species) showed very low level of incidence (19%). Of the possible pathogens of lentil plants, F. oxysporum the main cause of vascular wilt was recovered from seeds of this crop. Thin layer chromatographic analysis of chloroform extracts of 13 seed samples showed that only one samples was naturally contaminated with aflatoxins B1, B2, G1 and G2 (14.3 micrograms/kg).

Aflatoxins↗

Control of seed mass and seed yield by the floral homeotic gene APETALA2.

APETALA2 (AP2) is best known for its role in the regulation of flower meristem and flower organ identity and development in Arabidopsis. We show here that AP2 also plays an important role in determining seed size, seed weight, and the accumulation of seed oil and protein. We demonstrate genetically that AP2 acts through the maternal sporophyte and endosperm genomes to control seed weight and seed yield. Thus, AP2 functions outside the boundaries of flower meristem and flower organ development to affect agronomically relevant traits in Arabidopsis.

Arabidopsis↗

Electrodiffusional uptake of organic cations by pea seed coats. Further evidence for poorly selective pores in the plasma membrane of seed coat parenchyma cells.

In developing seeds, the permeability of the plasma membrane of seed coat parenchyma cells is crucial for the supply of nutrients to the embryo. Here, we report characteristics of the transport of the organic cation choline and the basic amino acid L- histidine (His; cation at pH 5, electroneutral at pH 7) into isolated seed coats of pea (Pisum sativum). Supplied at sub-micromolar concentrations, choline(+) accumulated in the seed coat tissue 5.1 +/- 0.8-fold, His(+) 2.4 +/- 0.3-fold, and His(0) 1.3 +/- 0.2-fold. Taking into consideration that at pH 5 His influxes as a cation but effluxes as a neutral molecule, these accumulations are in reasonable agreement with (electro) diffusional uptake at the prevailing membrane potential of -55 +/- 3 mV. At a concentration of 100 mM, choline(+) and His(+), but not His(0), depolarized the membrane of the parenchyma cells and neither of the substrates was accumulated. At this concentration, the relative influx (the ratio of influx and external concentration, a measure for membrane permeability) of choline and His was approximately 10 micromol g(-1) fresh weight min(-1) M(-1), similar to that found for neutral amino acids, sucrose, glucose, and mannitol. At lower concentrations, the relative influx of choline(+) and His(+) increased because of increasingly more negative membrane potentials, giving rise to apparent saturation kinetics. It is suggested that transport of organic cations can proceed by a general, poorly selective pore in the plasma membrane of seed coat parenchyma cells. This pore is thought to be responsible for the unloading of a range of solutes that serve as nutrients for the embryo.

Algorithms↗

The role of research institutions in seed-related disaster relief: Seeds of Hope experiences in Rwanda.

The article describes the efforts of a coalition of agricultural research centres, Seeds of Hope (SOH) in the rebuilding, of Rwanda, after the genocide and war of 1994. Research involvement in emergency relief and rehabilitation was unusual at the time and SOH had to forge its unique complementary role. Focusing on crop and variety development and conservation it: provided technical advice to relief agencies on seed procurement; used its baseline ken to assess the effects of war on seed diversity and seed security; made preparations to restore specific germplasm (which, fortunately, proved unnecessary) and spent substantial effort on rebuilding human resource capacity in research as well as basic scientific facilities. The involvement of SOH highlighted the critical, yet very different, roles for research during emergency versus rehabilitation periods and demonstrated the cost effectiveness of building in a diagnostic component--before massive seed or germplasm distributions are programmed.

Academies and Institutes↗