Proteolytic Activities in x Haynaldoticum sardoum Seeds of Different Ages

Carboxypeptidase, aminopeptidase and proteinase activities weremeasured in endosperms from dry and germinating x Haynaldoticumsardoum naturally aged seeds. Carboxypeptidase activity, presentin dry seeds, decreased slightly during germination and remainednearly unchanged during the storage period. Aminopeptidase activityincreased during germination in younger seeds, but decreasedin non-viable seeds. Proteinase activity was absent in dry seeds,increased during germination in younger seeds and disappearedin the older ones. Proteinase activity was not recovered in old endosperms followingtransplantation of young embryos, and was recovered only toa very small extent in young endosperms following transplantationof old embryos. Young endosperms onto which young embryos hadbeen transplanted gave maximum recovery of enzyme activity,although this was lower than in young intact seeds. These results suggest that the reduced or delayed availabilityof nutrients to the embryo axis is not the only factor causingthe failure of seeds to germinate, the ageing process beinga progressive phenomenon affecting both embryo and endosperm. x Haynaldoticum sardoum, Denti de cani, seed ageing, proteolytic activities, embryo-transplantation     

Changes in RNA and Protein Metabolism Associated with Alterations in the Germination Efficiency of Sunflower Seeds

Precise knowledge of seed quality after harvest and during storageis of particular importance for seed producers. We analyseddifferent sunflower seed lots (Helianthus annuusL.) characterizedby extremes of germination ability. We used RNA analysis tostudy possible changes in gene expression in seeds unable togerminate. Total RNA content was very small in dry seeds showinga low germination ability. Capacity for total RNA synthesisat the onset of imbibition was also reduced in these seeds.In addition, correlations were found between these parametersand germination ability at 19 °C. We demonstrated a highcorrelation between the amount of total RNA in the dry seed,the capacity of RNA synthesis at the onset of imbibition andthe seed moisture content at the time of the harvest. The abilityof dry seed mRNAs to be translatedin vitrowas also reduced andseven polypeptides, from stored mRNAs, were characteristic ofthe cotyledons from high germinability seeds. Germination canthus be affected at several levels including membrane, enzymaticand nucleic acid deteriorations. Gene expression; germination ability; Helianthus annuusL.; marker; protein; RNA; seed; sunflower     

Recalcitrant Seeds: Short-term Storage Effects in Avicennia marina (Forsk.) Vierh. may be Germination-associated

When recalcitrant propagules (seeds) of Avicennia marina werestored in a dry air stream there was no significant change intheir moisture content for 10 days, after which it declinedrapidly. Seed viability remained high during this 10-d period,only then declining. Ultrastructural changes in embryonic rootprimordium cells during the 10-d storage period were similarto those characterizing germination. It is suggested that thecontinuation of germination processes after shedding may beresponsible for the deterioration in stored recalcitrant seeds. Avicennia marina, recalcitrance, seed storage, germination     

Environmental Factors Influencing the Expression of Dormancy Patterns in Weed Seeds

Buried seeds often show seasonal periodicity of dormancy. Dormancypatterns of Chenopodium album, Polygonum persicaria, Sisymbriumofficinale and Spergula arvensis were studied by burying seedsunder field conditions in sandy loam in December, 1986. Seedswere exhumed at regular intervals and germination was subsequentlytested in the laboratory. It was shown that the conditions ofthe germination test influenced the expression of the dormancypattern. Germination of C. album and 5. arvensis always dependedon the presence of light, whereas seeds of S. officinale completelylost their light dependency during the first winter. Applicationof nitrate during the germination test in light improved germinationof all species. Dark germination was not stimulated by nitratealone. Desiccation of the exhumed seeds at a r.h. of approx.15% enhanced germination under all conditions. A combinationof several stimulating factors revealed breaking of dormancymuch earlier in the season. During induction of secondary dormancythe effect of the test conditions was even more pronounced.Dormancy induction could be overlooked for several months whenseeds were desiccated and/or given nitrate during the germinationtest in light. It is hypothesized that in the field both desiccation- due to cultivation and dry spells - and nitrate enrichmentof the soil will influence the expression of the seasonal patternof dormancy and therefore enlarge the period of possible seedlingemergence Desiccation, dormancy pattern, germination, light, nitrate, seeds, weeds, Chenopodium album, Polygonum persicaria, Sisymbrium officinale, Spergula arvensis     

Germination Preventing Mechanisms in Iris Seeds

The different germination behaviour of the seeds of two irises,Iris lorteti and I. atropurpurea was found to be due to thedifferent mechanical resistance of the integument, at the micropylarend, to radicle protrusion. A pressure of 135 atm was necessaryin l. lorteti seeds for radicle protrusion, while in I. atropurpurea77 atm was sufficient. In contrast Pancratium maritimum requireda pressure of only 10 atm. The outer integument of seeds ofI. lorteti was found to contain a compound which was toxic tothe germinated embryo but did not act as a germination inhibitor.Extracts of the endosperm also had a slight germination inhibitingeffect. An interaction between this weak inhibitor and the effectof the testa could not be ruled out completely. A test assayfor germination using excised embryos was developed. A methodfor germination of Iris seeds, by cutting off the outer integumentat the micropylar end, was developed and is being exploitedcommercially. Iris lorteti, Iris atropurpurea, germination, germination inhibition, embryo culture, seed coat mechanical resistance     

Dormancy in Seeds of Anona crassiflora Mart

The seeds of Anona crassiflora Mart., an important tree of theBrazilian cerrado, have been found to take about 8 months forgermination to commence, and some 35-70 d more to achieve itfully; such seeds must remain in moist ground all this time.A poorly developed embryo, little more than a hyaline mass ofcells, was all that could account for the delayed germination.Inhibition by extracts of seed parts was negligible. In A. crassifloraafter-ripening is related more to morphological changes in theembryo than to chemical changes. As the seed cannot endure evena brief sojourn in dry soil, vegetative reproduction is themeans by which the tree spreads into new areas.     

Methanol Accumulation in Maturing Seeds

During in vitro growth and maturation of soybean seeds, cessationof embryo growth and dry weight accumulation occurred in thepresence of abundant C and N nutrients. Axis followed by cotyledontissues changed from green to yellow, and post-harvest germinationpotential declined if cultured after yellowing of axis tissues.A tissue specific accumulat;on of methanol occurred during thein vitro culture of immature seeds (i.e. initially 50 to 70mg fresh weight) to maturity in liquid medium. Methanol accumulatedto 3.0 g m^–3 or 50 µg seed^–1 in the medium,while methanol decreased from 37 to about 3.0 µg g^–1fresh weight in cotyledons. By contrast, axis tissues increased20-fold in methanol concentration to 90 µg g^–1 during20 d in culture. Ethanol was present only in trace amounts inaxis tissues and medium. Addition of exogenous methanol vapourto in situ grown seeds during precocious maturation decreasedsubsequent seedling vigour and germination with increasing levelsof exposure. Methanol accumulation in axis tissues during thegermination phase was not correlated with high temperature andtissue water content treatments which simulated pre-harvestdeterioration of seeds. However, the accumulation of methanolduring in vitro seed development and maturation in liquid culturemay contribute to reduced post-harvest germination performance. Key words: Soybean, Glycine max, seed maturation, in vitro, methanol     

Degradation of Protein Bodies in Germinating Seeds of Brassica campestris L. var. sarson Prain

Histochemical investigations were carried out on dry and germinatingseeds of Brassica campestris var. sarson, to study the degradationof protein bodies with globoidal inclusions. During germination,a wave of protein body degradation sets in from the radicularend of the embryo, passing through the hypocotyl and shoot apex,and ending in the cotyledons. The digestion of protein bodiesis of the internal type. The various isolated parts of the embryoshowed a similar pattern of protein body digestion to that ofthe whole embryo, except that in some cells the globoids persistedeven after complete digestion of protein bodies; the rate wasfaster in the comparatively more expanded part of the isolatedorgans. No specific factor controlling the initiation of thewave of protein body digestion could be ascertained. Brassica campestris var. sarson, yellow sarson, seed, germination, protein bodies, degradation     

Germination Inhibitors in some Umbellifer Seeds

Presence of inhibitors of germination in seeds of five umbellifercrops viz. fennel (Foeniculum vulgare Mill.), cumin (Cuminumcyminum L.), carum (Carum copticum Benth. and Hook.), carrot(Daucus carota L.), and coriander (Coriandrum sativum L.) wasdemonstrated. The effect of seed exudates on the pattern ofinhibition of germination of these seeds was also studied. Presence of exudates in general delayed germination althoughthe time of maximum inhibition was different with differentcrop seeds. The inhibitory effect was high at an early stagein the case of carum and carrot and persisted in the latterbut disappeared in the former. The inhibitory effect on cuminand fennel increased up to the seventh day and then began todecrease. Growth of root and hypocotyl of 16-day-old seedlingswas also inhibited by the exudates.     

Changes in Phenolic Inhibitors in Seeds of Acer saccharum during Stratification

Phenolics in sugar maple seeds were identified by colour reactions,u.v. spectroscopy and gas-liquid chromatography. Changes intheir inhibitory activity during stratification were also followedusing several bioassays. p-Coumaric, o-coumaric, and ferulicacids were found in all tissues. The highest concentrationswere noted in the embryonic axis and testa, but total amountsper fruit were highest in cotyledons and pericarp. p-Coumaricacid was the principal phenolic in dry seeds, and it declinedsubstantially in all tissues during the first half of stratificationwith little subsequent change thereafter. Decreases in o-coumaricand ferulic acids per seed were small by comparison, neverthelesslosses ranged between 36 and 68% of the original concentrations.Bioassays confirmed that the three endogenous phenolics possessedmarked inhibitory properties that diminished with the progressof stratilication, but only the compound identified as p-coumaricacid showed major inhibition in seed germination tests.     

Seasonal Periodicity in Germination of Seeds of Chenopodium album L.

Seeds of Chenopodium album L. were buried under field and controlledconditions. The germination capacity of these seeds was testedover a range of conditions at regular intervals. Seed buriedin the field only showed small seasonal changes in germinationcapacity when tested at constant temperatures in incubators.However, when germination was tested at field temperatures,seasonal changes in germination were more obvious. Nitrate andlight always promoted germination. There was a strong positiveinteraction between the effects of the two factors. When nitrateand light were combined, exhumed seeds germinated over a muchlonger period of the year than in water with or without light.Desiccation only stimulated under particular conditions, forexample, when germination was tested in nitrate in darkness.A regression model was developed with the data from the germinationtests in incubators. The model describes the changes in dormancyand germination and estimates germination at field temperaturesaccurately throughout the year. Despite the absence of clearseasonal changes in the temperatures suitable for germination(computed with the model), germination in the field showed seasonalperiodicity, because the field temperature and the germination-temperaturerange only overlapped from spring to late summer.Copyright 1993,1999 Academic Press Chenopodium album, lamb's quarters, dormancy pattern, germination, regression model, temperature, light, nitrate, desiccation     

Cytochemical Localization of Phenolic Compounds in Columella Cells of the Root Cap in Seeds of Brassica napus--Changes in the Localization of Phenolic Compounds during Germination

Changes in the localization of phenolic compounds were investigatedin columella cells of embryonic roots in 1-year-old Brassicanapus seeds during 48 h of imbibition and germination. In dry,dormant seeds, phenolic compounds were located in the apoplasticcompartment between the cell wall and plasmalemma in the outermostlayer of the columella. These apoplastic phenolic deposits disappearedduring the activation processes associated with imbibition andgermination, but new deposits appeared successively in the nucleus,ER cisternae, protein bodies and on the outer surface of theroot cap. A large number of phenolic deposits were observedin the outermost part of the columella, becoming less frequenttowards the initial centre. Their appearance coincided withrestoration of the ER and immediately preceded cytological activation.After the primary root had emerged from the seed coat, depositsof phenolic compounds disappeared from the cytoplasm, but simultaneouslyappeared in the vacuoles. Copyright 1999 Annals of Botany Company Brassica napus var. oleifera, root columella, germination, phenolic compounds localization.     

Changes in Seed Constituents During Germination and Seedling Growth of Precociously Matured Soybean Seeds(Glycine max)

During 7 d of precocious maturation of soybean seed (Glycinemax), the starch content declined and soluble sugar levels increasedin patterns similar to natural seed dehydration and maturation.Total seed protein content and total seed dry weight increasedwhereas oil content remained relatively unchanged. Overall,the proportions of the constituents in precociously maturedseeds were comparable to naturally mature seeds. Precociouslymatured soybean seeds showed much the same germination and seedlinggrowth frequency patterns as naturally matured seeds. Duringgermination and seedling growth of precociously matured seeds,starch, soluble sugar, protein and oil levels followed patternssimilar to naturally mature, germinating seeds and seedlings.Therefore, precocious maturation may be used as a model systemto investigate the control of the physiological and biochemicalevents occurring during seed maturation which lead to germinationand subsequently, seedling growth. Glycine max (L.) Merr., soybean, cotyledons, maturation, germination/seedling growth     

Ethylene Involvement in Dormancy Release of Ricinodendron rautanenii Seeds

Ricinodendron rautanenii (manketti) seeds respond to as littleas 10^–2 µl I^–1 ethylene. Both dry and imbibedseeds respond to ethylene treatments. The length of the imbibitionperiod prior to the ethylene treatment is not important as seedswhich were simultaneously gassed and imbibed germinated successfully.Seeds were sensitive to the gas at temperatures between 25 and35 °C. No temperature effect was observed in seeds whichwere transferred from temperatures below 20 °C, or above35 °C, to 30 °C. There is no period of carbon dioxidesensitivity associated with the ethylene treatment and the effectsof the ethylene treatment persisted in both dry and imbibedseeds when subsequently maintained (desiccated) at room temperaturefor 8 d Ricinodendron rautanenii, manketti nut, ethylene, germination     

Cytokinins in Germinating Seeds of Phaseolus vulgaris L. I. Changes in Endogenous Levels Within the Cotyledons

Ethanolic extracts from the cotyledons of mature dry Phaseolusvulgaris L. seed yielded cytokinin-like activity which co-chromatographedwith zeatin and ribosylzeatin. Under conditions which stimulatedgermination and cotyledon expansion, the level of these cytokininsdecreased rapidly in both intact embryos and excised cotyledons.In the excised cotyledons the decrease was continuous, resultingin very low levels of cytokinin being detected after 4 daysof incubation. With the embryonic axis present, however, theinitial decrease was arrested and reversed after 3 days. Thissuggests that the cotyledons do not synthesize cytokinins butthat these hormones are imported from the embryonic axis, particularlyonce radicle growth is well under way. Phaseolus vulgaris, bean, cotyledons, cytokinins, germination     

Flow Cytometric Determination of Nuclear Replication Stages in Tomato Seeds during Priming and Germination

Flow cytometric determination of DNA levels in embryos of fullymatured dry tomato (Lycopersicon esculenium) seeds revealedlarge amounts of 2C DNA signals, indicating that most cellshad arrested in the cell cycle at the presynthetic G₁ phaseof nuclear division. After imbibition in water, an augmentationof the 4C signal in the embryonic root tip region was found.This increase could be ascribed to cells entering the syntheticphase of nuclear division leading towards the doubling of chromosomalmaterial. In the root tip cells, 4C:2C ratios increased I dafter imbibition in water though radicle emergence started 2d later. Apparently, DNA synthesis preceded germination. Onlya small increase in the number of cells with 4C DNA levels wasfound in the rest of the embryonic tissues. In whole dry seeds,DNA histograms revealed both a 2C signal and a considerable6C peak, the latter originating from the endoreduplicated endosperm. A priming period of 14 d in PEG-6000 considerably enhanced therate and uniformity of germination. In the ungerminated seeds,the 4C DNA signal of root tip cells started to increase after3 d incubation in PEG. The ratio of 4C:2C steadily increasedduring the 14 d priming period, though did not reach the levelobtained after hydration in water. Upon priming, the 4C:2C ratiowas constant after redrying the seeds towards the original moisturecontent, indicating that the chromosomal material in the rootcells had stably ceased cell cycle activity at the G² phase.The present results indicate that the beneficial effects ofpriming on seedling performance are associated with the actionof replicative DNA synthetic processes prior to germination. Lycopersicon esculeniumMill, tomato, DNA content, flow cytometry, priming, seed, nuclear replication stage, C levels     

Oxidative Phosphorylation in Germinating Lettuce Seeds

The effect of thiourea and coumarin, in vivo and in vitro, onthe phosphorylating activity of lettuce mitochondria was investigated,as well as the effect of coumarin on the P/O ratio. It was shownthat both substances in vitro inhibit phosphorylation; whilein vivo coumarin inhibits but thiourea under certain circumstancesstimulates. Coumarin was also shown to decrease the P/O ratioand therefore may be considered as an uncoupler. The difficulties in considering these effects of the substancesas a primary mechanism controlling germination are pointed outand discussed.     

Studies on the Germination of the Seeds of Striga hermonthica: III. On the Nature of Pretreatment and after-ripening

An attempt has been made to elucidate some of the changes whichtake place in seeds of Striga hermonthica (immature, ripe andoverripe), (a) during moisture-treatment at three temperatures(15°C., 22°C., and 32°C.), and (b) during after-ripeningin dry storage. Observations on the drift of respiration of immature seed throughoutpretreatment at 15°C. show that an inverse relationshipexists between germination and respirational activity. Thissuggests that germination depends on the accumulation of somemetabolite during pretreatment. It would seem that this metabolitemay also be used as a substrate for respiration during pretreatment. After-ripening of the immature seeds has been shown to changethe nature of the seeds in some way so that their peak respirationon exposure to moisture is at first progressively reduced. Thisreduction has been ascribed to the presence of some barriercapable of restricting gaseous diffusion. The data presented, togetherwith those of the previous paperof this series, suggest that the specific metabolite requiredfor germination might be proteinaceous. Certain other data appearto rule out the suggestion that germination capacity is conditionedby the sugar content of the seeds. Reasons are given which supportthe view that one of the main effects of the stimulating solutionon the seed is to increase the permeability of the seeds togaseous diffusion.     

Storage Behaviour of Salix alba and Salix matsudana Seeds

Effects of dehydration, storage temperature and humidificationon germination of Salix alba andS. matsudana seeds were studied.Newly released seeds showed 100% germination before and afterdehydration to 11–12% moisture content. Germination ofthe high vigour lot (100% initial normal germination) was notaffected by dehydration to 6.7% moisture content but germinationdecreased with further dehydration to 4.3%. The lower vigourlot (75% initial normal germination) was more susceptible todehydration and germination decreased following dehydrationto 6.7% moisture content. Dry seeds of both species survivedimmersion in liquid nitrogen without loss of viability. Thegermination of seeds stored with 9% moisture content decreasedto 35–40% in 5 months at -20°C or in 2 months at 5°C.However, at 25°C seeds entirely lost viability within 2weeks. Seeds showed improved performance when stored at -70°C> - 20°C > 5°C > 25°C and tolerated dehydrationto a moisture content in equilibrium with 15% relative humidity.Results suggest that they are orthodox in storage behaviouralthough they are short-lived. Humidification treatment of lowvigour seed lots resulted in a remarkable increase in germinationpercentage. Copyright 2000 Annals of Botany Company Salix alba, Salix matsudana, willow, seed storage behaviour, dehydration, humidification, cryopreservation     

Effects on Phytochrome Controlled Germination Produced by Far-Red Irradiation of Seeds Before and During Rehydration

Seeds irradiated with red light and then re-dried will respondto this light treatment on subsequent rehydration in the dark.If such high-Pfr seeds are irradiated in the dry state withfar-red light immediately before rehydration the percentagegermination is significantly reduced in the case of Plantagomajor and Sinapis arvensis but increased in Bromus steriliswhere Pfr inhibits germination. This effect of far-red lightcan be reversed by red light despite the fact that red lightalone has no effect on dry seed. This is due to the interconversionof Pfr and the red light absorbing phytochrome intermediatecomplex meta-Fa. If there is a delay between far-red irradiationand rehydration of Sinapis seeds, the inhibitory effect of thefar-red irradiation becomes progressively less the longer thedelay. This reduction in effectiveness of far-red is interpretedin terms of a dark reversal of meta-Fa to Pfr with a half-lifeof about 4–6 h. The reappearance of Pfr is either veryslow or docs not occur in dehydrated Plantago seeds, as far-redtight given 96 h prior to hydration is just as inhibitory asfar-red light given immediately before hydration. Meta-Fa doesappear to revert to Pfr in darkness in Bromus seeds, but onlyvery slowly. The rapid increase in effectiveness of red irradiationduring rehydration of high-Pfr Plantago seeds suggests that,in this species, the pre-treatment used in preparation of high-Pfrseeds may increase the receptivity or amount of the Pfr reactionpartner. Key words: Phytochrome intermediates, Seeds, Germination