Reversal by pressure of seed germination promoted by anesthetics

Germination of Panicum capillare L. caryopses induced by solutions of ethanol and ethyl ether was prevented by application of pressure >1 MPa during the period of exposure to the anesthetic. This effect of pressure indicates that germination is correlated with expansion at a site of anesthetic action in a cell membrane. The effects of several other anesthetics were measured on germination of P. capillare seeds. Ethanol, chloroform, and ethyl ether had the highest activity. Methanol and isopropanol were inactive. The effective compounds are thought to distribute preferentially to lipid-solution interfaces in cell membranes of the seeds.     

Action spectra for light-induced germination in dormant lettuce seeds

Fluence response curves for red light-induced germination of thermodormant (TD) seeds of Lactuca sativa L. show two regions that differ in their light sensitivity. In the region of high sensitivity, the germination responses differ between seed batches and can be altered by dark storage or far red irradiation. Induction of germination in far red dormant (FRD) seeds requires far higher fluences. Action spectra for induction to 60% germination were determined for these various response types. Spectra for the regions of low sensitivity response are similar for TD and FRD seeds. In comparison, the action spectrum for the highly sensitive response in TD seeds is significantly shifted to longer wavelengths. Analogous differences exist in the action spectra for far red reversal of the red induced germination responses. Germination induction in the low sensitivity region shows repeated red-far red reversibility. Far red reversal of red induction in the high sensitivity region does not saturate even at the highest far red fluences available and requires increased red fluences for subsequent reinduction. A model quantitatively accounting for these observations is presented. It is pointed out that action spectra of processes involving photoreversible pigments with partly overlapping absorption spectra in general are not identical with the absorption spectra of the partners. They should depend upon the degree of phototransformation required to elicit a given physiological response. In the case of induction of lettuce seed germination the observed action spectra can be interpreted as reflecting different requirements for P _fr of the various response types. Our results do not necessitate the assumption of spectroscopically different forms of phytochrome in these seeds.Abbreviations TD thermodormant - FRD far red dormant - P phytochrome - P _r red absorbing form of P - P _fr far red absorbing form of P     

The dual role of temperature in the regulation of the seasonal changes in dormancy and germination of seeds of Polygonum persicaria L.

Summary The role of temperature in the regulation of seasonal changes in dormancy and germination was studied in seeds of Polygonum persicaria. Seeds were buried in the field and under controlled conditions. Portions of seeds were exhumed at regular intervals and germination was tested over a range of conditions. Seeds of P. persicaria exhibited a seasonal dormancy pattern that clearly showed the typical features of summer annuals, i.e. dormancy was relieved at low winter temperatures, the germination peak occurred in spring and dormancy was re-induced in summer. The expression of the dormancy pattern was influenced by the temperature at which germination was tested. At 30°C exhumed seeds germinated over a much longer period of the year than at 20° or 10°C. Nitrate added during the germination test occasionally stimulated germination. The seasonal changes in dormancy of buried seeds were regulated by the field temperature. Soil moisture and nitrate content did not influence the changes in dormancy. The fact that, on the one hand, field temperature determined the changes in dormancy and, on the other hand, germination itself was influenced by temperature, was used to describe the seasonal germination pattern of P. persicaria with a model. Germination of exhumed seeds in Petri dishes at field temperature was accurately described with this model. Germination in the field was restricted to the period where the range of temperatures over which germination could proceed (computed with the model) and field temperature overlapped.     

Lipases in the storage tissues of peanut and other oil seeds during germination

The castor-bean endosperm-the best-studied material of reserve lipid hydrolysis in seed germination-was previously shown to have an acid lipase and an alkaline lipase having reciprocal patterns of development during germination. We studied oil seeds from 7 species, namely castor bean (Ricinus communis L.), peanut (Arachis hypogaea L.), sunflower (Helianthus annus L.), cucumber (Cucumis sativus L.), cotton (Gossypisum hirsutum L.), corn (Zea mays. L.) and tomato (Lycopersicon esculentum Mill.). The storage tissues of all these oil seeds except castor bean contained only alkaline lipase activity which increased drastically during germination. The pattern of acid and alkaline lipases in castor bean does not seem to be common in other oil seeds. The alkaline lipase of peanut cotyledons was chosen for further study. On sucrose gradient centrifugation of cotyledon homogenate from 3-d-old seedlings, about 60% of the activity of the enzyme was found to be associated with the glyoxysomes, 15% with the mitochondria, and 25% with a membrane fraction at a density of 1.12 g cm^-3. The glyoxysomal lipase was associated with the organelle membrane, and hydrolyzed only monoglyceride whereas the mitochondrial and membrane-fraction enzymes degraded mono-, di- and triglycerides equally well. Thus, although the lipase in the glyoxysomes had the highest activity, it had to cooperate with lipases in other cellular compartments for the complete hydrolysis of reserve triglycerides.     

The levels of GA3 and GA20 may be associated with dormancy release in Onopordum nervosum seeds

Endogenous levels of two gibberellins, GA₃ andGA₂₀, were quantified in unimbibed Onopordumnervosum seeds collected from two different populations, whichshoweddifferences in their germination capacity. After purifying the seed extracts,gibberellin levels were evaluated by gas chromatography mass spectrometry byusing selected ion monitoring (GC-MS-SIM) adding deuterated gibberellins asinternal standards. The intraspecific differences in germination capacity wereassociated with differences in the endogenous levels of both gibberellins. Thecontents of GA₃ and GA₂₀ in seeds with high germinationrate were twice and five times higher, respectively, than those from seeds witha low germination rate, indicating a possible role of gibberellins in dormancyrelease in this plant species.     

The growth physics and water relations of red-light-induced germination in lettuce seeds

Using lettuce (Lactuca sativa L., cv. Grand Rapids) embryos in osmotica, we have demonstrated that when the growth rates of the embryonic axes of seeds treated with red (R) or far-red (FR) light are equalized, the axes of R-treated seeds develop a 3.4-bar decrease in water potential (paper No. III).As axial growth begins, reserve protein and phytin decrease rapidly, concomitant with increases in reducing sugars, -amino nitrogen, and inorganic and esterified soluble phosphates. However, no differences between the axes of R-and FR-treated seeds are found with respect to the changes in these compounds, indicating that these changes arise as a result of growth and are not under immediate phytochrome control. Little change in the total lipid content is found in either treatment. The axes of FR-treated seeds hydrolyze endogenous sucrose at a greater rate thant those of R-treated seeds. Axes of R-treated seeds accumulate K⁺ and Na⁺ to a greater extent than those of FR-treated seeds. When potassium salts are added to the incubation medium, R induces increased K⁺ uptake by the axis and greater medium acidification by the axis. Malate and other organic acids and acidic amino acids increase at equal rates in both treatments, indicating that inorganic anions may also be taken up to balance the ionic charges. The results are compatible with the assumption that changes in the osmotic and pressure potentials of the embryonic axes of R-treated seeds are the result of a phytochrome-stimulated proton pump which, in whole dormant seeds, would initiate water-potential changes allowing the embryos to overcome the mechanical restraint of the surrounding seed layers, resulting in germination.Abbreviations FR far-red light - PEG polyethylene glyeol 4000 - Pfr far-red-absorbing form of phytochrome - R red light III=Carpita et al. 1979     

The influence of plant growth regulators on the growth of the embryonic axes of red- and far-red-treated lettuce seeds

The influence of several plant growth regulators on the growth of the embryonic axes from red- and far-red-(R- and FR-)treated lettuce (Lactuca sativa L., cv. Grand Rapids) seeds was examined; as shown previously, the water potential of the axes from R-treated seeds has been lowered by 3.5–5.6 bars compared to that in axes from FR-treated ones. Kinetin and abscisic acid (ABA), when included in the incubation medium, reduced the elongation of the axes whereas fusicoccin stimulated it; however, these effects were the same in axes of both R- and FR-treated seeds. In contrast, elongation of axes from FR-treated seeds was stimulated by gibberellic acid (GA₃, but elongation of axes from R-treated ones was not affected by this hormone. This latter result indicates that gibberellins may be involved in the phytochrome-mediated growth responses in lettuce axes.When the root caps of the embryos were removed prior to light treatment, R was still able to induce a water-potential decrease in the embryonic axes, indicating that at least a portion of the active Pfr resides in the axis and not the root cap.Abbreviations ABA abscisic acid - FR far red light - GA₃ gibberellic acid - PEG polyethylene glycol - Pfr far-red-absorbing form of phytochrome - R red light     

Dormancy in somatic embryos and seeds ofVitis: changes in endogenous abscisic acid during embryogeny and germination

Abscisic acid (ABA) in extracts of somatic embryos and seeds of Gloryvine (Vitis vinifera L.xV. rupestris Scheele) was measured by gas chromatography-mass spectrometry-selected ion monitoring using deuterated ABA, (±)-[C-3Me-²H₃]ABA, ([²H₃]ABA) as internal standard. The ABA content increased rapidly during embryogeny (0.035 ng/embryo at the globular stage to 0.22 ng/embryo at the mature stage). The level of ABA in the tissues of somatic embryos, expressed in ng/mg dry weight, decreased from the globular stage (0.76 ng/mg) to the mature stage (0.25 ng/mg). Chilling (4° C) induced normal germination of seeds and mature somatic embryos and precocious germination of globular, heart-shaped and torpedoshaped somatic embryos. In all cases chilling led to a marked reduction in endogenous ABA. Exogenous (±)-ABA inhibited the germination of chilled somatic embryos.Abbreviations ABA abscisic acid - [²H₃]ABA (±)-[C-3Me-²H₃]-abscisic acid - BHT 2,6-di-t-butyl-4-methylphenol - GC-MS gas chromatography-mass spectrometry - Me-ABA and Me-[²H₃]ABA methyl esters of ABA and [²H₃]ABA, respectively - SIM selected ion monitoring     

Germination inhibitors in the pine seed coat

Inhibitors from (Pinus pinea L.) seed coats were separated using paper chromatography, thin layer chromatography, and a Sephadex G-10 column. The inhibitory activity was resolved into several fractions. One of these behaved similarly to abscisic acid. It has exhibed the same properties as ABA in thin-layer chromatography, paper chromatography, and Sephadex G-10 chromatography and in UV absorption and fluorescence spectra. These germination inhibitors, present in the seed coats, are involved in the regulation of P. pinea seed germination.Abbreviations ABA abscisic acid - TLC thin-layer chromatography     

Light effects upon dry lettuce seeds

Germination of certain dry seeds (achenes) of Lactuca sativa L. cv. Grand Rapids was increased to ca. 75% after irradiation with 665 nm red light (R; 1x10³ J m^-2); this response was eliminated by far-red light (FR) following the R. The response of dry seeds required an order of magnitude more light than that of wetted seeds, and was not maximal until 48 h after irradiation. Other seeds, which could not be stimulated by R in dry state, showed a partial response after 10 min hydration. Irradiation of dry seeds (or seeds wetted 1 h) with FR (1x10³ J m^-2) reduced dark germination from 26% to 2%. Seeds dehydrated in an oven (60°C, 90 min) showed a decrease in germination if irradiated with R (1x10⁵ J m^-2) before wetting. The results show that phytochrome is present in dry lettuce seeds (and functional in some seed lots) prior to wetting; and that in other seed lots the molecule becomes functional within minutes after wetting the seeds. Transformation of the FR absorbing from of phytochrome (P_FR) to the inactive from (P_R) occurs at lower seed moisture content than the reverse reaction. It appears that dormancy in seeds ripened in sunlight might be assured during seed drying and maturation by the more effective transformation of P_FR to P_R than vice versa as phytochrome is dehydrated.Abbreviations FR far-red - R red - CAL seeds from California - NC seeds from North Carolina (see text)     

The control of seed germination in Trollius ledebouri: The breaking of dormancy

Seeds of Trollius ledebouri exhibit low germination when maintained on moistened filter paper. Dormancy can be overcome by the application of gibberellins A₄+A₇ or by testa removal. Germination is characterised by a change in the anatomy of the seed and by specific alterations in the protein complement of the endosperm tissue. These anatomical and biochemical changes are also exhibited by isolated endosperm tissue maintained in the absence of the embryo. The observations described are discussed in relation to the interaction between the endosperm and the embryo in the control of seed germination in T. ledebouri.Abbreviation GA gibberellin     

The control of seed germination in Trollius ledebouri A model of seed dormancy

Treatment of Trollius ledebouri seeds with gibberellins A₄+A₇ promotes germination. The efficacy of the treatment is dependent upon the duration of imbibition in distilled water prior to GA₄₊₇ application. Presoaking increases both the final percentage germination attained and also its rate of achievement. No presoaking effect is exhibited by seeds induced to germinate by testa removal in the absence of GA₄₊₇. Active washing of Trollius seeds enhances the presoaking effect and the eluent from washed seeds is inhibitory to germination. The results support the hypothesis that the presoaking effect exhibited by Trollius is the result of the leaching of a germination inhibitor from the seeds which is antagonistic to GA₄₊₇. Additionally, treatment of Trollius seeds with the gibberellin-biosynthesis inhibitor (2-chloroethyl)-trimethylammonium chloride (CCC) prior to testa removal retards germination. The inhibitory effect of CCC on germination is overcome by GA₄₊₇. Although CCC inhibits embryo growth during the presoaking of intact seeds, it does not affect the increased sensitivity of presoaked seeds to GA₄₊₇. Therefore, although endogenous gibberellins may be involved in the germination process, they do not contribute to the presoaking phenomenon. The expansion of isolated endosperm tissue is not affected by CCC. However, the chemical markedly inhibits endosperm expansion in intact seeds and implicates the embryo as both the site of production of the germination inhibitor and of gibberellin. These results are discussed in relation to previous studies and a model is presented to account for the characteristics of germination in Trollius.Abbreviations GA gibberellin - CCC (2-chloroethyl)-trimethylammonium chloride     

Induction of secondary dormancy in Amaranthus caudatus seeds

Nondormant A. caudatus seeds germinated in the darkat temperatures between 20 and 35° but not at 45 °C.Incubation at this temperature for at least 10 h inhibited seedgermination over the temperature range 20 to 35 °C,temperatures previously suitable for germination. Thus incubation at 45°C induced secondary dormancy. Mechanical or chemicalscarification or exposure to pure oxygen caused complete or almost completegermination of dormant seeds although more slowly in comparison to nondormantseeds. Secondary dormant scarified seeds required a lower concentration of ABAthan nondormant seeds to inhibit germination. The high temperature, whichinduced dormancy, 45 °C, caused the seed coat to be partiallyresponsible for secondary dormancy. Involvement of ABA (synthesis orsensitivity) in the induction and/or maintenance of this dormancy should beconsidered.     

Abscisic acid relationships in the chill-related dormancy mechanism in apple seeds

Abscisic acid (ABA) levels in seeds from three cultivars of apple (Malus domestica Borkh.) which have substantially different chilling requirements were investigated by gas chromatography mass-spectrometry selected ion monitoring (GCMS-SIM) during stratification. The ABA content of dormant unchilled seeds was similar in the three cultivars, suggesting no relationship between the chilling requirement of those seeds and their ABA status. That chilling is not related to ABA changes during stratification was confirmed by warm (20°C) and cold (5°C) stratification experiments. ABA content dropped rapidly and nearly identically under both temperature regimes, but only cold stratification promoted germination. The decline in ABA during stratification was due in large part to leaching from the seed coat and nucellar membrane; the ABA content of the embryo remained nearly constant. The radicle in intact seeds stratified at 5°C began growing 20–30 days after the ABA in the seed coat and nucellar membrane had nearly disappeared. Radicle growth did not occur in unchilled seeds, even though ABA had leached from them as well. It is possible that the leaching of ABA from the seed allows certain promotive forces to develop, but if so, these can develop only at chilling temperatures. Studies were also conducted on 2-trans ABA relationships to apple seed dormancy, but no association was evident.Report No. 12, Department of Fruit and Vegetable Science, Cornell University.     

Rate of synthesis of spermine and spermidine in germinating seeds of Glycine,Helianthus and Triticum

The spermine, spermidine, and the total protein content of embryos or embryonic axes from Triticum durum, Helianthus annuus, and Glycine max seeds at different times of early germination was evaluated. Mitotic activity of root-tip meristems from germinating seeds was also determined. The hypothesis is suggested that differences in polyamine and protein pattern during early germination could be correlated with the onset of mitotic activity and with the different characteristics of the seeds assayed.Abbreviations SPM spermine - SPD spermidine     

The control of food mobilization in seeds of Cucumis sativus L.

An analysis of the in vitro activities of proteolytic enzymes from cotyledons of germinating cucumber seeds has been carried out and the effects of protein degradation products on such activities monitored. Aminopeptidase activity is substantially inhibited with either L-leucine or L-phenylalanine and trypsin activity with L-arginine. Aminopeptidase activity was also markedly reduced in the presence of individual di- and tripeptides. Of the peptides tested, however, only L-tryptophyl-L-phenylalanine inhibited the degradation of native cucumber seed protein by the endogenous cucumber seed protease(s) (autodigestive activity).Abbreviations TCA trichloroacetic acid - L-leuglygly L-leucylglycylglycine - L-pheglygly L-phenylalanylglycylglycine - L-phe-L-leu L-phenylalanyl-L-leucine - L-leu-L-phe L-leucyl-L-phenylalanine - L-tryp-L-phe L-tryptophyl-L-phenylalanine - LPA L-leucine-p nitroanilide - BAPNA -N-benzoyl-DL-arginine-p nitroanilide - ADA autodigestive activity     

Genes associated with the end of dormancy in grapes

A grape bud EST library was constructed and 4,270 ESTs sequenced. The library clones were arrayed for the purpose of investigating the level of gene expression over time, particularly leading up to the buds release from dormancy. The arrays were hybridized with P³³-labeled probes produced from samples of buds collected at weekly intervals. These probes covered the time from 9 weeks prior to bud burst until just after the emergence of the shoots. Expression patterns from these genes have been examined. It was found that 74% of the genes in the data set were homologous to known proteins. Genes were then assigned to functional categories according to their primary BLAST match. Of these 13% were involved with photosynthesis, 13% with disease resistance and defense, 5% energy, 12% metabolism, 20% protein production and processing, 25% cell structure and plant growth and the remaining 12% were unclassified The expression pattern of a selection of candidate genes retrieved from literature previously reporting an association with dormancy changes was assessed. On closer examination most of these genes relate to the oxidative processes and stress responses within the cell. The results of this study show that even in the dormant state, gene expression in the buds is high.     

An ATP-synthesizing system in seeds

Using onion seed powder, a semi-in vitro system for ATP synthesis in seeds has been developed. The system requires AMP, phosphoenolpyruvate (PEP) and orthophosphate with apparent Km values of 0.8, 1.5 and 3.0 mM, respectively. ATP synthesis is pH-dependent with a sharp optimum at pH 6.4, it exhibits linearity with time up to 40 min, and with a seed powder concentration between 25 and 150 mg ml^-1. The system is stimulated by low concentrations (<25 mM) of K⁺ and Mg²⁺ but is inhibited by higher concentrations of K⁺ and Mg²⁺ as well as by low concentrations of Li⁺, Na⁺ and especially Ca²⁺. The maximal rate is about 5 pmol min^-1 mg seed powder^-1 in dry onion seeds. During seed imbibition the rate of activity increases by about 120% after 3 h, reaching a plateau which is steady up to 18 h, when the radicle emerges. A comparison of the ATP content in seeds during the early period of imbibition with the capacity of ATP synthesis at this stage reveals that the described system could provide, during germination, 100 times more ATP than that found in imbibed seeds. The system is shown to be present in ten different types of seeds.     

Note on organic dormancy of estuarine Ruppia maritima L. seeds

Information on seed dormancy is one of the primary requirements for successful seedling propagation of submerged aquatics and seagrass. Studies on Ruppia maritima seed germination have been done, but the presence, requirements, and the types of dormancy have not been well understood. A laboratory study was conducted to understand presence and types of organic dormancy of the seeds of estuarine R. maritima collected from Lake Pontchartrain, Louisiana, USA. Our study results indicate that the brackish estuarine R. maritima population produces seeds that do not have any noticeable initial morphological, physical, and physiological dormancy. Although dry stratification reduced seed viability and final germination rates, drying seems to induce an earlier germination in R. maritima. Desiccation also appears to induce an environmental dormancy that can be disrupted by exposure to water. Further study on environmental dormancy is needed to provide information to develop methods for long-term seed storage that can be employed in greenhouse seedling propagation. Handling editor: P. Viaroli