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Water Relations of Seed Development and Germination in Muskmelon (Cucumis melo L.) : I. Water Relations of Seed and Fruit Development.

Total water potential (psi), solute potential, and turgor potential of field-grown muskmelon (Cucumis melo L.) fruit tissue (pericarp) and seeds were determined by thermocouple psychrometry at 5-day intervals from 10 to 65 days after anthesis (DAA). Fruit maturity occurred between 44 and 49 DAA, and seed germination ability developed between 35 and 45 DAA. Pericarp psi was essentially constant at approximately -0.75 megapascal (MPa) from 10 to 25 DAA, then decreased to a minimum value of -1.89 MPa at 50 DAA before increasing to -1.58 MPa at 65 DAA. Seed psi remained relatively constant at approximately -0.5 MPa from 10 to 30 DAA then decreased to -2.26 MPa at 50 to 60 DAA before increasing to -2.01 MPa at 65 DAA. After a rapid increase to 20 DAA, seed fresh weight declined until 30 DAA due to net water loss, despite continuing dry weight gain. As fruit and seed growth rates decreased, turgor potential initially increased, then declined to small values when growth ceased. A disequilibrium in psi was measured between seeds and pericarp both early and late in development. From 20 to 40 DAA, the psi gradient was from the seed to the tissue, coinciding with water loss from the seeds. From 50 to 65 DAA, seed psi decreased, causing a reversal of the psi gradient and a slight increase in seed water content. The partitioning of solutes between symplast and apoplast may create and maintain psi gradients between the pericarp and seed. The low solute potential within the pericarp due to solute accumulation and loss of cellular compartmentation during ripening and sensecence may be involved in prevention of precocious germination of mature seeds.

Journal Article↗

Dosimetry of a thyroid uptake detected in seed migration survey following a patient's iodine-125 prostate implant and in vitro measurements of intentional seed leakages.

As a quality control procedure, a post-implant seed migration survey has been accomplished on 340 prostate cancer patients since November 2001. Pulmonary seed embolization and intracardiac seed embolization have been detected. A case of thyroid uptake due to leaking iodine-125 (I-125) sources was also seized. In order to determine the dose to the thyroid, a dosimetry method was developed to link in vivo measurements and the cumulated dose to the thyroid. The calculated source leakage half-life in the case was approximately 15 days based on the measurements and the estimated cumulated dose to thyroid was 204 cGy. It is concluded that one seed was leaking. In order to verify the in vivo measurements, intentional in vitro seed leakage tests were performed. A seed was cut open and placed in a sealed glass container filled with a given volume of saline. The I-125 concentration in the saline was subsequently measured over a period of six months. Consistent in vivo and in vitro results were obtained. Recent incidents of seed leaks reported from other centers have drawn practitioners' attention to this problem. In order to make the measurements more useful, the seed leakage tests were expanded to include I-125 seeds from six other vendors. The results show that the leakage half-lives of those seeds varied from nine days to a half-year. Two seed models demonstrated least leakage. Since the measurements lasted for six months, the escape of iodine resulted from oxidation of iodide in the saline was a concern for the measurement accuracy. As a reference, another set of leakage tests were performed by adding sodium thiosulfate salt (Na2S2O3 x 5 H2O) to the saline. Sodium thiosulfate is a reducing agent that prevents the conversion of iodide to iodate so as to minimize I-125 evaporation. As a result, significantly shortened leakage half-lives were observed in this group. Seed agitation was also performed and no significant deviations of the leakage rates were observed. Considering the body fluid is more complex than saline, the in vivo leakage half-life, in case a source leak is encountered, may vary significantly from what is presented in this paper due to chemical reactions. In vivo measurements thus may produce a more accurate estimation of leakage half-life and thyroid uptake dose.

Brachytherapy↗

Chronic herbivory negatively impacts cone and seed production, seed quality and seedling growth of susceptible pinyon pines.

Although herbivory often reduces the reproduction of attacked trees, few studies have examined how naturally occurring insect-resistant and susceptible trees differ in their reproduction, nor have these effects been experimentally examined through long-term herbivore removals. In addition, few studies have examined the effects of herbivory on the quality of seeds produced and the implications of reduced seed quality on seedling establishment. We evaluated the impact of chronic herbivory by the stem-boring moth, Dioryctria albovittella, on cone and seed production of the pinyon pine (Pinus edulis) during two mast years. Three patterns emerged. First, moth herbivory was associated with reductions in cone production, viable seed production and seed mass. Specifically, pinyons susceptible to moth attack had 93-95% lower cone production, and surviving cones produced 31-37% fewer viable seeds, resulting in a 96-97% reduction in whole tree viable seed production. In addition, surviving seeds from susceptible trees had 18% lower mass than resistant trees. Second, long-term experimental removal of the herbivore resulted in increased rates of cone and seed production and quality, indicating that moth herbivory was the driver of these reductions. Third, seed size was positively associated with seed germination and seedling biomass and height, suggesting that trees suffering chronic herbivory produce poorer quality offspring. Thus, the resistance traits of pinyons can affect the quality of offspring, which in turn may affect subsequent seedling establishment and population dynamics.

Analysis of Variance↗

Developmental patterns of free and protein-bound biotin during maturation and germination of seeds of Pisum sativum: characterization of a novel seed-specific biotinylated protein.

Mature dry pea seeds contain three major biotinylated proteins. Two of these of subunit molecular mass about 75 kDa and 200 kDa are associated with 3-methylcrotonyl-CoA carboxylase (EC 6.4.1.4) and acetyl-CoA carboxylase activities (EC 6.4.1.2) respectively. The third does not exhibit any of the biotin-dependent carboxylase activities found in higher organisms and represents the major part of the total protein-bound biotin in the seeds. This novel protein has been purified from a whole pea seed extract. Because in SDS/polyacrylamide gels the protein migrates with an apparent molecular mass of about 65 kDa, it is referred to as SBP65, for 65 kDa seed biotinylated protein. The molecular mass of native SBP65 is greater than 400 kDa, suggesting that the native protein assumes a polymeric structure, resulting from the association of six to eight identical subunits. The results of CNBr cleavage experiments suggest that biotin is covalently bound to the protein. The stoichiometry is 1 mol of biotin per 1 mol of 65 kDa polypeptide. The temporal and spatial pattern of expression of SBP65 is described. SBP65 is specifically expressed in the seeds, being absent from leaf, root, stem, pod and flower tissues of pea plants. The level of SBP65 increases dramatically during seed development. The protein is not detectable in very young seeds. Its accumulation pattern parallels that for storage proteins, being maximally expressed in the mature dry seeds. SBP65 disappears at a very high rate during seed germination. The level of free biotin has also been evaluated for various organs of pea plants. In all proliferating tissues examined (young developing seeds, leaf, root, stem, pod and flower tissues), free biotin is in excess of protein-bound biotin. Only in the mature dry seeds is protein-bound biotin (i.e. that bound to SBP65) in excess of free biotin. These temporal expression patterns, and the strict organ specificity for expression of SBP65, are discussed with regard to the possibility that in plants, as in mammals, biotin plays a specialized role in cell growth and differentiation.

Binding Sites↗

Early embryo invasion as a determinant in pea of the seed transmission of pea seed-borne mosaic virus.

Seed transmission of an isolate of pea seed-borne mosaic virus (PSbMV) in several pea genotypes has been studied. Cross-pollination experiments showed that pollen transmission of PSbMV did not occur and accordingly, virus was not detected in pollen grains by ELISA or electron microscopy. Comparative studies between two pea cultivars, one with a high incidence of seed transmission and one with none, showed that PSbMV infected the floral tissues (sepals, petals, anther and carpel) of both cultivars, but was not detected in ovules prior to fertilization. Virus was detected equally well in seed coats of the progeny in both cultivars. Analysis of virus incidence and concentration in pea seeds of different developmental stages demonstrated that in the cultivar with a high incidence of seed transmission, PSbMV directly invaded immature embryos, multiplied in the embryonic tissues and persisted during seed maturation. In contrast, the cultivar without seed transmission did not show invasion of immature embryos by the virus; there was no evidence for virus multiplication or persistence during embryo development and seed maturation. Hence seed transmission of PSbMV resulted from direct invasion of immature pea embryos by the virus and the block to seed transmission in the non-permissive cultivar probably occurred at this step.

Blotting, Northern↗

[The influence of seed maturity and seed coat on the germination of Bupleurum chinense and B. falcatum].

OBJECTIVE: To investigate the factors influencing the seed germination of Bupleurum chinense and B. falcatum. METHOD: The seeds of different maturity, and the seeds whose coat were removed or nicked before dipping in water, were germinated in a light growth chamber. RESULT: The different inflorescence level represents different development period. Seeds harvested from cultivation have different maturity. The germination rate of the seeds in period I to III was nearly zero, and it was significant different from period IV to VII. The seed coat is not an obstacle for water absorption or gas exchange, but its removal led to significant increase of seed germination rate. However, to dip the seeds without coat into water for 48 h had no more effect on germination. CONCLUSION: The seed maturity and germination inhibitors in seed coat are two important factor influencing seed germination in B. chinense and B. falcatum.

Bupleurum↗

Treatment planning dosimetric parameters for 192Ir seed at short distances: effects of air channels and neighboring seeds based on Monte Carlo study.

The dose distributions around two different arrangements of a single radioactive 192Ir seed in water, (1) with air channels at the ends, and (2) surrounded by two nonactive ("dummy") seeds on both longitudinal ends, were calculated using MCNP4C Monte Carlo simulations at distances up to 1 cm. The contributions from beta particles and electrons emitted by 192Ir were included in the calculations. The effects of (a) the air channels at the seed ends and (b) the interference effect of the dummy seeds on the dose distribution were quantified and compared. It was found that the dummy seeds do not cause significant dose reduction for radial distances beyond 0.05 cm from the seed center. It is decided to report the dose rate values and the dosimetric parameters in TG43 format for a single seed with air channels for use in treatment planning computer systems. The dose rate constant (at 1 cm) of 192Ir seed, lambda, is 1.108 cGyU(-1) h(-1). The values of radial dose function, g(r), are within 1% from the TG43 recommended polynomial fit, except for distances within 0.08 cm. The anisotropy function, F(r, theta), attains large values near the seed ends and shallow angles (up to 8), as well as many values greater than 2 at the 20 degrees polar angle. Treatment planning systems involving intravascular brachytherapy do not compromise the accuracy for dosimetry of multiple seed trains by summing single seed values in water.

Air↗

The effect of seed source, light during germination, and cold-moist stratification on seed germination in three species of Echinacea for organic production.

Organic production of one of the most popular botanical supplements, Echinacea, continues to expand in the U.S. Echinacea seeds typically show a high degree of dormancy that can be broken by ethephon or gibberelic acid (GA), but these methods are currently disallowed in organic production. In order to determine the efficacy of non-chemical seed treatments, we evaluated the effect of varying seed source and supplying light, with and without cold-moist stratification, on seed germination of the three most important medicinal species of Echinacea, E. angustifolia DC, E. purpurea (L) Moench, and E. pallida (Nutt.) Nutt. Treatments included cold-moist stratification under 24 h light, 24 h dark, and 16/8 h light/dark to break seed dormancy. We found that germination was greater in the E. purpurea and E. pallida seeds from a commercial organic seed source compared to a public germplasm source. When seeds were not cold-moist stratified, 16-24 h light increased germination in E. angustifolia only. Echinacea angustifolia, E. purpurea, and E. pallida seeds that were cold-moist stratified under 16-24 h of light for 4 wk had a significantly greater percentage and rate of germination compared to seeds germinated in the dark. Therefore, cold-moist stratification under light conditions is recommended as a method to break seed dormancy and increase germination rates in organic production of Echinacea.

Journal Article↗

Accumulation of a Brazil nut albumin in seeds of transgenic canola results in enhanced levels of seed protein methionine.

We have increased the methionine content of the seed proteins of a commercial winter variety of canola by expressing a chimeric gene encoding a methionine-rich seed protein from Brazil nut in the seeds of transgenic plants. Transgenic canola seeds accumulate the heterologous methionine-rich protein at levels which range from 1.7% to 4.0% of the total seed protein and contain up to 33% more methionine. The precursor of the methionine-rich protein is processed correctly in the seeds, resulting in the appearance of the mature protein in the 2S protein fraction. The 2S methionine-rich protein accumulates in the transgenic seeds at the same time in development as the canola 11S seed proteins and disappears rapidly upon germination of the seed. The increase in methionine in the canola seed proteins should increase the value of canola meal which is used in animal feed formulations.

Gene Expression↗

Determining 'threshold' levels for seed-borne virus infection in seed stocks.

There have been many advances in testing procedures to detect seed-borne virus infection in seed samples. However, scant attention has been given to the implications of improved test results in terms of the economic losses resulting from sowing seed stocks with different amounts of infection. For agricultural and horticultural industries to use the results of tests on representative samples, defined 'threshold' values for percentage seed infection are required that identify acceptable levels of risk of economic losses resulting from sowing the virus-infected seed stocks. Such information is provided by field experiments in which infected seed is sown and the consequences are followed in terms of virus spread, yield losses and infection of newly produced seed. These field experiments need to continue over several years at diverse sites so that they represent a wide range of infection scenarios. Extensive surveys to determine seed-borne virus occurrence in different regions are also required to define areas of greater or lesser risk of economic losses. In this paper, an example is described of how field experiments and surveys were used to define 'threshold' values of seed-borne Cucumber mosaic virus infection in an annual crop (lupin: Lupinus angustifolius) and two such examples are given for pasture species: Cucumber mosaic virus in subterranean clover (Trifolium subterraneum), and Alfalfa mosaic virus in annual burr medic (Medicago polymorpha). The aim of this paper is to encourage others to address the urgent need for similar 'threshold' information with other economically important combinations of seed-borne viruses and host plant.

Australia↗

Pollination, seed set and seed predation on a landscape scale.

We analysed the combined effects of pollination and seed predation on seed set of Centaurea jacea in 15 landscapes differing in structural complexity. In the centre of each landscape, a patch of Centaurea plants was established for standardized measurements of flower visitation, seed predation and seed set. Both the number of flower-visiting bees and the proportion of flower heads damaged by seed predators increased with landscape complexity, which was measured as the proportion of semi-natural habitats. The mean number of seeds per flower head did not increase with the proportion of semi-natural habitats, presumably because of the counterbalancing effects of pollination and seed predation. For a subset of undamaged flower heads, the number of seeds per flower head was positively correlated with the number of flower visits. Further reasons for the unexpected failure to detect a correlation between landscape complexity and seed set appeared to be changes in flower-visitor behaviour and the contrasting responses of honeybees and wild bees to habitat context. Landscape analyses at eight spatial scales (radius of landscape sectors, 250-3000 m) showed that different groups perceived the landscape at different spatial scales. Changes in pollinator numbers could be explained only at small scales (up to 1000 m), while the seed predators also responded to large scales (up to 2500 m).

Animals↗

Endothelial cell seeding of de-endothelialised human arteries: improvement by adhesion molecule induction and flow-seeding technology.

OBJECTIVES: To assess re-endothelialisation of denuded human arteries by two different seeding techniques using adhesion molecule induction and a dynamic flow-seeding. DESIGN: Prospective, open study. SETTING: University Department of Cardiovascular Surgery. MATERIALS AND METHODS: In the first group (I) segments of human common carotid arteries (n = 4) were balloon-denuded, short-time seeded with cultured adult human venous endothelial cells (EC) and exposed to a mock circulation. In the second group (II) (n = 4), EC were incubated with a synthetic RGD peptide (arginine-glycine-aspartate) prior to seeding with the aim of upregulating the cellular adhesion molecules and increasing EC attachment. In the third group (III) (n = 4), EC were seeded not using the common technique of instillating cells and sequentially rotating the graft but by a dynamic flow application. The percentage of EC-covered luminal surface was assessed by image analysis of scanning electron micrographs. RESULTS: EC attachment was significantly increased in groups II (73%) and III (94%) compared with group I (34%). In group III, a preconfluent monolayer could be established immediately after seeding. One hour of artificial perfusion resulted in no significant EC loss in any of the study groups. CONCLUSIONS: RGD-peptide preincubation improves EC seeding of biological surfaces. Because of accelerated seeding times it may have good potential for clinical applications. The flow-seeding technology may be indispensable if EC seeding of the vascular surface of complete organ systems is required.

Adult↗

Physiology of Oil Seeds: IV. Role of Endogenous Ethylene and Inhibitory Regulators during Natural and Induced Afterripening of Dormant Virginia-type Peanut Seeds.

To further elucidate the regulation of dormancy release, we followed the natural afterripening of Virginia-type peanut (Arachis hypogaea L.) seeds from about the 5th to 40th week after harvest. Seeds were kept at low temperature (3 +/- 2 C) until just prior to testing for germination, ethylene production, and internal ethylene concentration. Germination tended to fluctuate but did not increase significantly during the first 30 weeks; internal ethylene concentrations and ethylene production remained comparatively low during this time. When the seeds were placed at room temperature during the 30th to 40th weeks after harvest, there was a large increase in germination, 49% and 47% for apical and basal seeds, respectively. The data confirm our previous suggestion that production rates of 2.0 to 3.0 nanoliters per gram fresh weight per hour are necessary to provide internal ethylene concentrations at activation levels which cause a substantial increase of germination. Activation levels internally must be more than 0.4 microliter per liter and 0.9 microliter per liter for some apical and basal seeds, respectively, since dormant-imbibed seeds containing these concentrations did not germinate. Abscisic acid inhibited germination and ethylene production of afterripened seeds. Kinetin reversed the effects of ABA and this was correlated with its ability to stimulate ethylene production by the seeds. Ethylene also reversed the effects of abscisic acid. Carbon dioxide did not compete with ethylene action in this system. The data indicate that ethylene and an inhibitor, possibly abscisic acid, interact to control dormant peanut seed germination. The inability of CO(2) to inhibit competitively the action of ethylene on dormancy release, as it does other ethylene effects, suggests that the primary site of action of ethylene in peanut seeds is different from the site for other plant responses to ethylene.

Journal Article↗

Water Relations of Seed Development and Germination in Muskmelon (Cucumis melo L.) : IV. Characteristics of the Perisperm during Seed Development.

We previously reported that an apparent water potential disequilibrium is maintained late in muskmelon (Cucumis melo L.) seed development between the embryo and the surrounding fruit tissue (mesocarp). To further investigate the basis of this phenomenon, the permeability characteristics of the tissues surrounding muskmelon embryos (the mucilaginous endocarp, the testa, a 2- to 4-cell-layered perisperm and a single cell layer of endosperm) were examined from 20 to 65 days after anthesis (DAA). Water passes readily through the perisperm envelope (endosperm + perisperm), testa, and endocarp at all stages of development. Electrolyte leakage (conductivity of imbibition solutions) of individual intact seeds, decoated seeds (testa removed), and embryos (testa and perisperm envelope removed) was measured during imbibition of freshly harvested seeds. The testa accounted for up to 80% of the total electrolyte leakage. Leakage from decoated seeds fell by 8- to 10-fold between 25 and 45 DAA. Presence of the perisperm envelope prior to 40 DAA had little effect on leakage, while in more mature seeds, it reduced leakage by 2- to 3-fold. In mature seeds, freezing, soaking in methanol, autoclaving, accelerated aging, and other treatments which killed the embryos had little effect on leakage of intact or decoated seeds, but caused osmotic swelling of the perisperm envelope due to the leakage of solutes from the embryo into the space between the embryo and perisperm. The semipermeability of the perisperm envelope of mature seeds did not depend upon cellular viability or lipid membrane integrity. After maximum seed dry weight is attained (35-40 DAA), the perisperm envelope prevents the diffusion of solutes, but not of water, between the embryo and the surrounding testa, endocarp, and mesocarp tissue.

Journal Article↗

Synthetic vascular grafts seeded with genetically modified endothelium in the dog: evaluation of the effect of seeding technique and retroviral vector on cell persistence in vivo.

Unique characteristics of endothelium make it an attractive target cell for gene transfer. Genetically modified endothelial cells (ECs) seeded on synthetic vascular grafts offer the potential to control neointimal hyperplasia, decrease graft thrombogenicity and improve small diameter graft patency. This study addresses the issue of synthetic vascular graft colonization with endothelial cells transduced with noninducible retroviral marker genes in the dog. Autologous endothelial cells were enzymatically harvested and transduced with either the bacterial NeoR gene or human growth hormone gene using retroviral vectors. All transduced cells were positive by polymerase chain reaction (PCR) amplification for the transduced gene sequence prior to graft seeding. Transduced ECs were seeded on Dacron grafts (n = 3) preclotted with autologous blood. These grafts exhibited complete endothelialization at times from 250 to 360 days. Recovered DNA, however, was negative for the transduced gene sequence when analyzed by PCR and Southern blotting. Expanded polytetrafluoroethylene (ePTFE) was evaluated (n = 8) using several different cell seeding protocols. Grafts were seeded at 3 densities (ranging from 6 x 10(3) to 1.5 x 10(5) cells/cm2) and 2 different adherence times. Seeding substrate was also evaluated. Grafts were either preclotted with whole blood or incubated with 20 or 120 micrograms/ml fibronectin for 60 min. Graft biopsies were evaluated from 2 to 52 wk. Limited endothelialization was present in 4 dogs as early as 2 wk, but never progressed to full luminal coverage. The remaining dogs failed to ever exhibit any luminal EC adherence. Two dogs with limited EC coverage had positive DNA by PCR for the NeoR gene sequence at 2 and 3 wk. In contrast to transduced EC's, nontransduced EC colonization of ePTFE was complete at 2 wk when seeded under conditions that transduced cells had failed to persist. Neither seeding density, adherence time, seeding substrate or retroviral vector used influenced the uniformly poor graft coverage seen with transduced cells. Results of this study indicate that despite successful gene transfer using 4 different retroviral vectors, transduced endothelial cells seeded under varying conditions appear altered in their ability to stably adhere and colonize synthetic vascular grafts in vivo.

Animals↗

Polyembryony in Citrus. Accumulation of seed storage proteins in seeds and in embryos cultured in vitro.

Citrus exhibits polyembryonic seed development, an apomictic process in which many maternally derived embryos arise from the nucellus surrounding the developing zygotic embryo. Citrus seed storage proteins were used as markers to compare embryogenesis in developing seeds and somatic embryogenesis in vitro. The salt-soluble, globulin protein fraction (designated citrin) was purified from Citrus sinensis cv Valencia seeds. Citrins separated into two subunits averaging 22 and 33 kD under denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A cDNA clone was isolated representing a citrin gene expressed in seeds when the majority of embryos were at the early globular stage of embryo development. The predicted protein sequence was most related to the globulin seed storage proteins of pumpkin and cotton. Accumulation of 33-kD polypeptides was first detected in polyembryonic Valencia seeds when the majority of embryos were at the globular stage of development. Somatic Citrus embryos cultured in vivo were observed to initiate 33-kD polypeptide accumulation later in embryo development but accumulated these peptides at only 10 to 20% of the level observed in polyembryonic seeds. Therefore, factors within the seed environment must influence the higher quantitative levels of citrin accumulation in nucellar embryos developing in vivo, even though nucellar embryos, like somatic embryos, are not derived from fertilization events.

Amino Acid Sequence↗

Seed drops and caches by the harvester ant Messor barbarus: do they contribute to seed dispersal in Mediterranean grasslands?

To determine whether the harvester ant Messor barbarus acts as a seed disperser in Mediterranean grasslands, the accuracy level of seed processing was assessed in the field by quantifying seed drops by loaded foragers. In the vicinity of exploited seed patches 3 times as many diaspores were found as in controls due to seed losses by foragers. Over trails, up to 30% of harvested seeds were dropped, singly, by workers but all were recovered by nestmates within 24 h. Seeds were also dropped within temporary caches with very few viable diaspores being left per cache when ants no longer used the trail. Globally, ant-dispersed diaspores accounted for only 0.1% of seeds harvested by M. barbarus. We discuss the possible significance for grassland vegetation of harvester-ant-mediated seed dispersal.

Animals↗

Methods of estimating seed banks with reference to long-term seed burial.

We compared two standard seed-bank estimation techniques using buried seed populations that had been covered to depths of >1 m by volcanic deposits for 20 years. Some seeds were germinated in a greenhouse (germination method [GM]), and other seeds were extracted by flotation using 50% K(2)CO(3) solution (flotation method [FM]). In total, FM could detect more species and seeds in the soils than GM. However, many species that were extracted by FM did not germinate by GM and smaller seeds were extracted to a lesser extent by FM. FM and GM have distinct advantages and disadvantages. We concluded that the application of a single method should be avoided in estimating seed banks, in particular for long-lived seed banks, because the seeds cannot be readily germinated and are structurally weak.

Agriculture↗