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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 85 (1992), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Development of galactomannan hydrolyzing activity was followed in seeds of tomato [Lycopersicon esculentum (L.) Mill. cv. Toyonishiki] during priming and germination. The activity developed in seeds that were being primed in polyethylene glycol (-0.8 MPa). The activity was detected exclusively in the endosperm portion just adjacent to the radicle tip. Part of the activity remained active after desiccation of the primed seeds. After transfer to water, the activity in the primed seeds immediately began to increase, while in unprimed seeds the beginning of the increase in activity was delayed by about 1 day. In scanning electron microscopy, the inner surface of the cell walls of the micropylar endosperm portion appeared eroded in primed seeds that had been imbibed in water for 16 h (before germination), but not in unprimed seeds imbibed for the same period. These results support the hypothesis that galactomannan hydrolyzing enzyme, which is believed to be responsible for breakdown of tomato endosperm cell walls and hence for the weakening of mechanical restraint against radicle growth, may be involved in the improved germination of primed tomato seeds.
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  • 2
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The development of mitochondrial NAD+-malate dehydrogenase (EC 1.1.1.37) in mung bean and cucumber cotyledons was followed. using the antibody raised against it, during and following germination. The developmental patterns were quite different between the two. In cucumber, the content of mitochondrial malate dehydrogenase continued to increase through 3–4 days after the beginning of imbibition. This was, at least in part, due to active synthesis of the enzyme protein, and the synthesis seemed to be regulated by the availability of the translatable mRNA for the enzyme. In mung bean, on the other hand, the enzyme was present in dry cotyledons at a rather high concentration, and remained at a constant level between day 1 and day 3 after the reduction of the content to one-half its initial level during the first day. De novo synthesis of the enzyme could not be detected in mung bean cotyledons by pulse-labeling experiment.
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 89 (1993), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Mitochondria isolated from cotyledons of dark-grown cucumber (Cucumber sativus L., cv. Shimotsuki-Aonaga) seedlings after illumination with continuous far-red light showed an increased capacity for oxidation of malate or α-ketoglutarate, as compared with those from cotyledons of non-illuminated seedlings. This increase is supposed to be caused by phytochrome action (high irradiance response). Exogenous NAD+ had no effect on the rate of the oxidation of α-ketoglutarate or malate by mitochondria isolated from far-red light-treated cotyledons, but it enhanced the oxidation rate of mitochondria from control cotyledons to the level of mitochondria from light-treated ones. The NAD (NAD++ NADH) content was higher in mitochondria isolated from continuously far-red light-treated cotyledons than in mitochondria from controls. The NAD content was also increased by the treatment with a red light pulse and this response was reversed by a subsequent far-red light pulse. It is proposed that phytochrome controls respiratory activities of cucumber mitochondria by changing the size of the NAD pool in the mitochondria.
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  • 4
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Electron microscopic observations of the endosperm of tomato (Lycopersicon esculentum Mill.) seeds revealed that changes in the cell wall structures along with the vacuolation of protein bodies occurred in the micropylar portion of the endosperm prior to germination. No changes were detected at that time in the rest of the endosperm. Endo-β-mannanase activity was restricted to the micropylar region of the endosperm prior to germination. Cell wall digestion by this pregerminative mannanase seemed to be associated with the changes in cell wall structures occurring in the micropylar region prior to germination. The protein content in the micropylar part of the endosperm began to decrease shortly after imbibition and attained about 40% of the initial level by the time of radicle protrusion (38 h after imbibition). On the other hand, only slight changes in the content were detected in the lateral endosperm during the same time; the protein content in the lateral endosperm decreased only after germination started. In conformity with the results on protein contents, proteolytic activity began to develop first in the micropylar portion prior to germination, and then in the lateral portion after germination. Thus, the timing of the biochemical activation of the endosperm after imbibition differed between the micropylar and the lateral region. Some qualitative differences in patterns of polypeptides synthesized in vivo were detected, as analyzed by pulse-labeling and fluorography, between the micropylar and the lateral portions of the endosperm of seeds imbibed for 25 h. This suggests that processes of the biochemical activation of the endosperm may be qualitatively, as well as quantitatively, different depending on the regions of the endosperm.
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 94 (1995), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Three forms of galactomannan-hydrolyzing enzymes (dyed galactomannan as substrate) were partially purified from germinated tomato [Lycopersicon esculentum (L.) Mill.] seed. Two of the enzymes were of the same molecular mass, 38 kDa, as judged by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), but the points of elution from a CM-Toyopearl column by a pH-gradient were different between the two (pH 5.15 and 5.45. respectively). The molecular mass of the third form was slightly less (37.5 kDa) than that of the other two. These 3 enzymes showed no α-galactosidase (EC 3.2.1.22) or β-mannosidase (EC 3.2.1.25) activity. Thin-layer chromatography (TLC) revealed that the products of the reaction were oligosaccharides and that free galactose and mannose were not released. These results indicate that the 3 galactomannan-hydrolyzing enzymes are endo-β-mannanases (EC 3.2.1.78). Polyclonal antibodies raised against the 37.5-kDa polypeptide cross-reacted with the two 38-kDa polypeptides, indicating that the 3 endo-β-mannanases are immunologically homologous. Activity staining and immunoblotting of native PAGE of endosperm extracts revealed that only two (38-kDa. elution point pH 5.15 and 37.5-kDa proteins) of the 3 forms were major endo-β-mannanases present in the endosperm of germinated tomato seeds.
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  • 6
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Starch debranching enzyme was purified from mung bean (Vigna radiata) cotyledons to investigate its properties and developmental pattern during and following germination. A debranching enzyme was purified up to the step where only a doublet of polypeptides with molecular masses of 99 and 101 kDa, respectively, was detected by SDS-PAGE. The enzyme is thought to be a single chain monomer, as the molecular mass of the enzyme determined by gel filtration was 72 kDa. Monoclonal antibodies raised against the purified preparation recognized the doublet, indicating that the two polypeptides have immunological homology to each other. The enzyme preparation showed a high activity with pullulan as a substrate, low activity with soluble starch and amylopectin, and no activity with glycogen. These substrate specificities indicate that the debranching enzyme from mung bean cotyledons is of the pullulanase type. Immunoblotting profiles revealed that the enzyme is present in dry seeds and decreases gradually after imbibition, suggesting the possibility that the pullulanase plays a role in developing mung bean cotyledons.
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  • 7
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 83 (1991), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Treatment of cotyledons of 4-day-old cucumber (Cucumis sativus L. cv. Tokiwa-jibae) seedlings with cycloheximide (0.3 mM) inhibited protein synthesis in the cotyledons by 80%. In spite of this inhibition, the cycloheximide treatment induced a marked increase in the capacity of the alternative respiration in mitochondria, accompanied by an increase in the contribution of the pathway to the total respiration. In contrast, the activity of the cytochrome pathway was reduced by cycloheximide treatment. As a result, the total respiration was almost the same in mitochondria from cycloheximide-treated cotyledons and untreated cotyledons. Activities of some mitochondrial enzymes examined were also similar. Mitochondrial proteins synthesized during the treatment were separated by two-dimensional gel-electrophoresis and examined by fluorography. No new spots appeared and no spots disappeared on the fluorograms, but the labeling intensity of some polypeptides showed a relative increase or decrease as the result of cycloheximide treatment.
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  • 8
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 90 (1994), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: An aminopeptidase (EC 3.4.11.-) was purified to homogeneity, as judged by SDS-PAGE. from mung bean (Vigna radiata) cotyledons. The molecular mass of this peptidase was estimated as 75 kDa by gel filtration. When an oligopeptide consisting of 5 amino acid residues was used as substrate, amino acids were released in the order of the N-terminal sequence of the oligopeptide chain. This enzyme apparently requires free sulfhydryl for its activity, as judged by the effects of various proteinase inhibitors. Among aminoacyl-p-nitroanilides examined for the availability as substrates of the enzyme, p-nitroanilides with hydrophobic amino acids were preferred substrates. According to western immunoblot profiles, the enzyme level in cotyledons was high at the early stage of imbibition and declined rapidly after germination.
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  • 9
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 94 (1995), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Development of galactomannan-hydrolyzing activity, that is involved in the weakening of the mechanical restraint of the endosperm, was followed at pre-germinative stages in tomato (Lycopersicon esculentum) seed. Prior to germination the activity developed exclusively in the endosperm portion just adjacent to the radicle tip. In other parts of the endosperm, the activity developed only after germination occurred. Under the conditions where germination was suppressed (far-red light- or ABA-treatment). no activity was detected in the endosperm at the pre-germinative stages. Under the conditions where the inhibition of germination was alleviated (far-red + red or ABA + GA3), the activity developed prior to germination in the endosperm part in front of the radicle tip. Thus, a clear parallel relationship was observed between germinability of the seed and the pre-germinative development of activity in the part of the endosperm portion adjacent to the radicle tip.
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  • 10
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 81 (1991), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: When cotyledons of mung bean [Vigna radiata (L.) Wilczek] were treated with spermidine (3 mM) during the first 6 h of imbibition, the development of α-amylase activity in cotyledons during the following 3 days was severely inhibited (75%) This inhibition was due to a slower accumulation of α-amylase protein, which in turn resulted from an inhibition of α-amylase synthesis. The rise in the level of α-amylase mRNA in cotyledons was also inhibited by spermidine treatment. However, the degree of inhibition of mRNA accumulation (40%) was not so marked as that of the activity of α-amylase synthesis (80%). These results are discussed in relation to the mode of action of spermidine on α-amylase expression.
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