Seed Coat Dormancy in Two Species of Grevillea(Proteaceae)

The role played by the seed coat in seed dormancy of Grevillealinearifolia(Cav.) Druce and G. wilsonii(A. Cunn.) was testedby a series of manipulations in which the seed coat was dissectedand removed, dissected and returned to the decoated seed, ordissected, removed and given a heat shock, and returned to thedecoated seed. Germination of intact seeds of both species wasalso examined after exposure to heat shock, smoke, or heat shockand smoke combined. Water permeability of the seed coat wasinvestigated by examining imbibition. For intact seeds, virtuallyno germination occurred under any treatment (G. wilsonii), orgermination was increased by exposure to either heat or smoke(G. linearifolia). Removal of the seed coat led to germinationof all decoated seeds for G. linearifolia, or a proportion ofdecoated seeds for G. wilsonii. Inclusion of smoked water inthe incubation medium led to a higher proportion of decoatedseeds germinating for G. wilsonii. Returning the seed coat,either with or without heat shock to the seed coat, did notsignificantly affect germination in either species. Seed coatswere permeable to water in both species. For the two Grevilleaspecies, there were different dormancy mechanisms that werecontrolled by the seed coat (G. linearifolia) or by both theseed coat and embryo (G. wilsonii). Copyright 2000 Annals ofBotany Company Grevillea linearifolia, Grevillea wilsonii, dormancy, seed coat dormancy, seed coat permeability, smoke, heat shock, germination     

'Callose' in the Impermeable Seed Coat of Sesbania punicea

Dry legumes of Sesbania punicea (Cav.) Benth. contain impermeableseeds which are either beige-greenish or reddish. When keptin water for over one year they do not swell. Quick imbibitionwas induced by a deep cut, abrasion with coarse abrasive paper,immersion in concentrated sulphuric acid, in boiling water andin boiling KOH solutions. Neither lipid solvents (acetone, chloroform,cyclohexane, diethyl ether, ethanol, xylene), nor, at room temperature,KOH solutions up to 50% per cent, permeabilized the seeds. Astrong barrier to water entry appeared to be located in thepalisade layer which showed ‘light points’ in places,and which seemed to be transversal thickenings. In the testa,‘ callose’, which may play a role in the impermeabilityof the Sesbania punicea seed, was detected. Sesbania punicea, germination, seed dormancy, seed coat impermeability, ‘callose’     

Seed Water Relations and the Regulation of the Duration of Seed Growth in Soybean

Soybean [Glycine max (L.) Merrill] seeds and cotyledons weregrown in an in vitro culture system to investigate the relationshipsbetween cell expansion (net water uptake by the seed) and drymatter accumulation. Seeds or cotyledons grown in a completenutrient medium containing 200 mol m^–3 sucrose continueddry matter accumulation for up to 16 d after in planta seedsreached physiological maturity (maximum seed dry weight). Seedor cotyledon water content increased throughout the cultureperiod and the water concentration remained above 600 g kg^–1fresh weight. These data indicate that the cessation of seeddry matter accumulation is controlled by the physiological environmentof the seed and is not a pre-determined seed characteristic.Adding 600 mol m^–3 mannitol to the medium caused a decreasein seed water content and concentration. Seeds in this mediumstopped accumulating dry matter at a water concentration ofapproximately 550 g kg^–1. The data suggest that dry matteraccumulation by soybean seeds can continue only as long as thereis a net uptake of water to drive cell expansion. In the absenceof a net water uptake, continued dry matter accumulation causesdesiccation which triggers maturation. Key words: Glycine max (L.) Merrill, solution culture, duration of seed growth, water content, dry matter accumulation     

Germination Inhibitors in Fruit Extracts of Red Beet (Beta vulgaris cv. rubra

Germination inhibitors in methanol and water extracts of redbeet fruits (Beta vulgaris cv. rubra L.) have been studied usinglettuce and red beet seed germination as bioassays. The methanolextracts contained substances which inhibited lettuce seed germination,but had no effect on the germination of red beet seeds. Germinationof both lettuce seeds and of water-leached or sulphuric acid-treatedred beet seed balls were inhibited by the water extracts. Theconcentrations of ammonia, ferulic acid, and oxalate in thewater extracts were much lower than required for inhibitionof red beet seed germination. The water extracts contained,however, large amounts of inorganic ions, and the results clearlydemonstrated that the inhibitory effect of the water extractson red beet seed germination was mainly due to the content ofsuch inorganic ions.     

Winterfat (Eurotia lanata(Pursh) Moq.) Seedbed Ecology: Low Temperature Exotherms and Cold Hardiness in Hydrated Seeds as Influenced by Imbibition Temperature

Thermal analyses of freezing events in hydrated lettuce (LactucasativaL.) seeds show a correlation between low temperature exotherms(LTEs) (evidence of ice crystal formation) and seed death. Yet,weather patterns common to the Northern Great Plains of NorthAmerica regularly create conditions where non-dormant seedsof native plants hydrate with snow melt and are subsequentlyexposed to -30 °C or colder conditions. To determine ifsuch weather patterns decimate dispersed seeds, we measuredthe effects of freezing on fully hydrated winterfat (Eurotialanata(Pursh) Moq.) seeds harvested from the Northern Plainsat two USA and one Canadian location. Survival of hydrated seedsto -30 °C at a cooling rate of 2.5 °C h^-1was similarto that of seeds not subjected to cooling, even though botha high temperature exotherm (HTE) and an LTE were observed.Although the LTE was not related to winterfat seed survival,freeze-stressed seeds had reduced germination rates and reducedseedling vigour, particularly for the collection with the lightestseeds. The temperature of LTEs was similar among seed collectionswith a mean of -17.6 °C, but was warmer when the seeds wereimbibed at 0 °C compared to 5, 10 or 20 °C. We founda significant correlation between the HTE and LTE temperatures.The difference and the correlation may be due to the highermoisture content of seeds imbibed at 0 °C. After pericarpremoval, only one exotherm in the range of the LTE was observed.This was also true for the naked embryo. We conclude that anLTE indicates ice formation in the embryo, but that it doesnot signal the death of a winterfat seed.Copyright 1998 Annalsof Botany Company Eurotia lanata(Pursh) Moq.,Krascheninnikovia, Ceratoides,winterfat, exotherm, freezing tolerance, freezing avoidance, seedbed ecology, germination, D50, seedling vigour, seed collection     

The Role of the Lens in Seed Imbibition and Seedling Vigour of Sesbania punicea (Cav.) Benth. (Leguminosae: Papilionoideae)

Seeds of Sesbania punicea (Cav.) Benth. are largely impermeable,and the natural, low, percentage which germinate are permeableat the lens. Scarifying seed in acid stimulates germinationand causes various types of damage to the testa, including thelens. Placing seeds in boiling water stimulates germinationby rendering the seeds permeable at the lens. Seeds were mechanicallyscarified at precise sites on the testa to assess influenceof the site and degree of damage on germination. Results indicatethat both the site of initial water entry and the rate of waterentry are important in determining seed and seedling vigour.Damaging the lens is more effective in stimulating germinationand reducing seedling abnormalities than complete excision ofthe lens. This effect can be reversed if seeds from which thelens was excised are allowed to imbibe gradually in inert osmotica.The leguminous lens thus acts as the site of initial water entryand regulates the rate of water uptake, thereby increasing seedand seedling vigour. Sesbania punicea (Cav.) Benth., hardseededness, lens, seed germination, imbibition damage, seedling vigour     

Phylogeny of seed dormancy in Convolvulaceae, subfamily Convolvuloideae (Solanales)

Background and Aims: The water gap is an important morphoanatomical structure inseeds with physical dormancy (PY). It is an environmental signaldetector for dormancy break and the route of water into thenon-dormant seed. The Convolvulaceae, which consists of subfamiliesConvolvuloideae (11 tribes) and Humbertoideae (one tribe, monotypicHumberteae), is the only family in the asterid clade known toproduce seeds with PY. The primary aim of this study was tocompare the morphoanatomical characteristics of the water gapin seeds of species in the 11 tribes of the Convolvuloideaeand to use this information, and that on seed dormancy and storagebehaviour, to construct a phylogenetic tree of seed dormancyfor the subfamily. Methods: Scanning electron microscopy (SEM) was used to define morphologicalchanges in the hilum area during dormancy break; hand and vibratomesections were taken to describe the anatomy of the water gap,hilum and seed coat; and dye tracking was used to identify theinitial route of water entry into the non-dormant seed. Resultswere compared with a recent cladogram of the family. Key Results: Species in nine tribes have (a) layer(s) of palisade cells inthe seed coat, a water gap and orthodox storage behaviour. Erycibe(Erycibeae) and Maripa (Maripeae) do not have a palisade layerin the seed coat or a water gap, and are recalcitrant. The hilarfissure is the water gap in relatively basal Cuscuteae, andbulges adjacent to the micropyle serve as the water gap in theConvolvuloideae, Dicranostyloideae (except Maripeae) and theCardiochlamyeae clades. Seeds from the Convolvuloideae havemorphologically prominent bulges demarcated by cell shape inthe sclereid layer, whereas the Dicranostyloideae and Cardiochlamyeaehave non-prominent bulges demarcated by the number of sub-celllayers. The anatomy and morphology of the hilar pad follow thesame pattern. Conclusions: PY in the subfamily Convolvuloideae probably evolved in theaseasonal tropics from an ancestor with recalcitrant non-dormantseeds, and it may have arisen as Convolvulaceae radiated tooccupy the seasonal tropics. Combinational dormancy may havedeveloped in seeds of some Cuscuta spp. as this genus movedinto temperate habitats.     

Differentiation of a Functional Aleurone Layer Within the Seed Coat of Sinapis alba L.

The hitherto unresolved ontogenetic origin of the aleurone layerin mustard (Sinapis alba L.) seeds was investigated with lightand electron microscopy. Contrary to previous views, this layerof storage cells is neither derived from the endosperm nor fromthe nucellus, but from a particular cell layer within the innerintegument of the seed coat. These cells differentiate and becomefilled with storage protein and fat concurrently with the maturationof the embryo. They survive seed desiccation and become depletedof storage materials during seed germination. Temporally correlatedwith the germinating embryo, the aleurone cells produce microbodyenzymes, which are controlled by light in a similar fashionin both tissues. Sinapis alba L., mustard, aleurone layer, seed coat, seed formation, germination     

Seed Storage and Germination and Seedling Proliferation in Taro, Colocasia esculenta (L) Schott

Taro seeds maintained under c. 40 per cent r.h. and 22 ±2°C retained higher viability than those stored under otherexperimental conditions. Germination was more than 60 per centon a number of defined and undefined media. Rates above 80 percent were obtained on a greenhouse potting mix or its extractor distilled water with filter paper as a support. When maintainedin the dark, seedlings cultured in vitro on one of several definedmedia etiolated and produced atypical, elongated internodeswhich resemble those of vining aroids. Plantlets which developedat their nodes were removed and raised to maturity. Colocasia esculenta (L.) Schott, taro, seed storage, seed germination, seedling proliferation     

The Dry Storage of Citrus Seeds

The survival of seeds of lemon (Citrus limon L.), lime [C. arantifolia(Christm.) Swing.] and sour orange (C. aurantium L.) was examinedunder a wide range of constant moisture contents and temperatures.Seed longevity was increased by decreasing the moisture contentand temperature of the storage environment. Maximum viabilitywas maintained in the combination of storage conditions includingthe lowest moisture content (5 per cent) and lowest temperature(–20 °C) investigated. The practicality of dry storageof citrus seed for genetic conservation is discussed. Citrus limon L., lemon, Citrus aurantifolia (Christm.), Swing, lime, Citrus aurantium L., sour orange, dry storage, moisture content, temperature, seed viability, seed longevity     

Seed Germination and Radicle Growth of a Halophyte, Kalidium caspicum(Chenopodiaceae)

Effects of temperature, light, NaCl and polyethylene glycol(PEG)-6000 on seed germination and radicle growth in a halophyticshrub, Kalidium caspicum(L.) Ung.-Sternb. were investigated.When seeds were incubated in deionized water at constant temperaturesbetween 10 and 30°C, the percentage germination in the darkexceeded 75%; light suppressed seed germination at alternatingtemperatures. Incubating seeds with a hypersaline solution ofNaCl for 30 d had no adverse effect on their germinability.The percentage germination of seeds incubated with a –0.8MPa NaCl solution was 73, 80 and 54% at 10, 20 and 30°C,respectively, but all radicles died before their length exceeded5 mm. In contrast, when seeds were incubated with a –0.8MPa PEG solution at 20°C, 68% of seeds germinated, and 95%of the emerging radicles survived beyond 5 mm. The high sensitivityof small radicles of this species to salinity indicated thatsalt must be removed from the soil surface for seedling establishment.Copyright2000 Annals of Botany Company Chinese desert, radicle growth, germination, halophyte, Kalidium caspicum, salinity     

Soya Bean Seed Growth and Maturation In vitro without Pods

Immature Glycine max (L.) Merrill seeds, initially between 50and 450 mg f. wt, were grown and matured successfully in vitro.Excised seeds were floated in a liquid medium containing 5 percent sucrose, minerals and glutamine in flasks incubated at25 °C under 300 to 350 µE m^–2 s^–1 fluorescentlight. During 16 to 21 d in culture, seeds grew to a matured. wt of 100 to 600 mg per seed at an average rate of 5 to 25mg d. wt per seed d^–1 depending on initial size. Growthrates were maximal during the first 8 to 10 d in vitro but declinedwith loss of green colour in the cotyledons. Seed coats rupturedwith rapid cotyledon expansion during the first 2 d in culture.Embryos were tolerant to desiccation and 80 to 90 per cent germinatedif removed from culture before complete loss of green colour.The growth of excised seeds in vitro exceeded the growth ofseeds in detached pods, but when windows were cut in pods topermit direct exposure of seeds to the medium, seed growth wascomparable. Glycine max (L.) Merrill, soya bean, seed culture, seed growth, seed maturation, germination     

Protein Synthesis During Rehydration, Germination and Seedling Growth of Naturally and Precociously Matured Soybean Seeds (Glycine max)

Soybean seeds [Glycine max (L.) Merr.] synthesize de novo andaccumulate several non-storage, soluble polypeptides duringnatural and precocious seed maturation. These polypeptides havepreviously been coined ‘maturation polypeptides’.The objective of this study was to determine the fate of maturationpolypeptides in naturally and precociously matured soybean seedsduring rehydration, germination, and seedling growth. Developingsoybean seeds harvested 35 d after flowering (mid-development)were precociously matured through controlled dehydration, whereasnaturally matured soybean seeds were harvested directly fromthe plant. Seeds were rehydrated with water for various timesbetween 5 and 120 h. Total soluble proteins and proteins radio-labelledin vivo were extracted from the cotyledons and embryonic axesof precociously and naturally matured and rehydrated seed tissuesand analyzed by one-dimensional PAGE and fluorography. The resultsindicated that three of the maturation polypeptides (21, 31and 128 kDa) that had accumulated in the maturing seeds (maturationpolypeptides) continued to be synthesized during early stagesof seed rehydration and germination (5–30 h after imbibition).However, the progression from seed germination into seedlinggrowth (between 30 and 72 h after imbibition) was marked bythe cessation of synthesis of the maturation polypeptides followedby the hydrolysis of storage polypeptides that had been synthesizedand accumulated during seed development. This implied a drasticredirection in seed metabolism for the precociously maturedseeds as these seeds, if not matured early, would have continuedto synthesize storage protein reserves. Glycine max (L.) Merr, soybean, cotyledons, maturation, germination/seedling growth     

Sinapine Leakage from Non-viable Cabbage Seeds

Seeds leak many compounds during the early phases of germinationand seed viability may be associated with differential leakageof specific compounds. Leakage of a fluorescent compound fromnon-viable cabbage (Brassica oleracca var. capitata L.) wasdocumented and studies were performed to identify the fluorescentcompound. Imbibed samples of both heat-killed (HK) and viablecabbage seeds were submerged in a viscous colloidal gel. After2 h to 4 h, a fluorescent halo was observed under U.V. lightaround the heat-killed seed but not around the viable seed.Viable and HK seeds were imbibed in water for 8 h and the pHof the leachate was adjusted to either 7 or 10. The absorptionspectra of leakage from HK seeds revealed peak values at 322nm and 388 nm at pH 7 and 10, respectively. This pattern wasnot observed from viable seed leakage. Two-dimensional paperchromatography was conducted on the HK seed leachate. Four fluorescentspots were observed after development first with BAW (n-butanol:acetic acid: water, 4: 1: 5 by vol.) followed by 6% acetic acid.One of the fluorescent spots (spot 3) was studied further dueto its observed intensity. Sinapine thiocyanate was preparedfrom rapeseed oilmeal and used as a reference sample. Absorptionspectra of spot 3 and sinapine thiocyanate were similar at bothpH 7 and 10. Spot 3 had identical R_F values using three solventsystems and identical colour reactions in five tests comparedto sinapine thiocyanate. It was concluded that sinapine wasthe major compound responsible for the fluorescent leakage fromHK cabbage seeds. Key words: Leachate, viability, Brassica     

Soya Bean Seed Growth and Maturation by In Vitro Pod Culture

Immature Glycine max (L.) Merr. seeds initially at 50–70mg fresh weight were successfully grown and matured in vitroin detached pods. Surface sterilized pods were floated in aliquid medium containing 5 per cent sucrose, minerals, and glutaminein 125 ml Erlenmeyer flasks and incubated at 25 °C under350–400 µE m^–1 s^–1 white light. Seedswhich were matured in vitro increased tenfold in dry weight,were visually similar to commercial seeds of the same size,were tolerant to desiccation and germinated with normal seedlinggrowth. Excised pods transported dye from the pedicel to thegrowing seed within 120 min. Soya bean pod culture is a usefultechnique to study the influence of single or combinations ofchemical or environmental parameters on regulation of seed growth,seed maturation, and subsequent germination events without theconfounding interactions with the mother plant. Glycine max (L.) Merr., soya bean, pod culture, seed culture, seed growth, seed maturation, germination     

Germination Rate of Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] Seeds Affected by Their Orientation

Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] seeds developwith the adaxial surface continuous with the cone scale. Onseparation, the scale side of the seed is lighter coloured andflatter than the side not previously in contact with the scale.Seeds oriented scale-side-up on a moist surface germinate morerapidly than seeds placed scale-side-down. When seeds were germinatedunder laboratory conditions, the response to orientation variedamong seeds of different geographic origin. Most of the geographicvariation in germination rate occurred when seeds were orientedscale-side-up. Pseudotsuga menziesii (Mirb.) Franco, Douglas-fir germination, orientation of seeds     

Studies on Mature Seeds of Cuscuta pedicellata and C. campestris by Electron Microscopy

The seeds of Cuscuta pedicellata have been investigated by transmissionand scanning electron microscopy. Additional observations havebeen made on seeds of C. campestris by SEM only. The seed coatconsists of an outer single epidermis, two different palisadelayers, and an inner multiparenchyma layer. The outer epidermalwall in C. pedicellata has a thick cuticle and zones rich inpectic substances. The thicker ‘U-shaped’ cell wallsin the outer palisade layer are strengthened by a wall layerof hemicellulose. The inner palisade layer has thick walledcells with a ‘light line’. The inner cell wall ofthe compressed multiparenchyma layer has a thin cuticle. A fairlythick cuticle is positioned directly on the endosperm surface.The aleurone cell walls are different from the remaining endospermwalls. The latter are thick and believed to be of galactomannans.There is a ‘clear’ zone between the plasmalemmaand the cell wall in the aleurone cells. The embryo cells arepacked with lipids and proteins. In Cuscuta campestris mostendosperm has been absorbed during the seed development. Theembryo apex has two minute leaf primordia. The features of theCuscuta seeds are discussed in relation to functional and environmentalconditions. Cuscuta pedicellata, Cuscuta campestris, seed, seed coat, cuticle, cell walls, endosperm, aleurone cells, galactomannan, embryo, TEM, SEM     

Germination and Early Growth of Plants Studied Using Neutron Radiography

Seed swelling, germination, root extension, lateral root initiationand shoot growth were studied in soils of different water contents,using non-destructive, serial neutron radiography. Seeds fromthree varieties of soya beans (Glycine max L.) and one varietyeach of maize (Zea mays L.) and vetch (Vicia sativa L.) wereused. The seeds germinated when they had increased in size bya certain amount, if germination is taken as the time when theradicle first appears. The rate at which roots and shoots extendalso depend on soil water content. Glycine max L., Vicia sativa L., Zea mays L., Soya bean vetch, maize, seed germination, root extension, lateral root initiation, neutron radiography     

Seed development and maturation in Phaseolus vulgaris I. Ability to germinate and to tolerate desiccation

The onset and development of both the ability to germinate andto tolerate rapid enforced desiccation were investigated duringthe development and maturation of seeds of bean (Phaseolus vulgahsL.) at different temperatures and also after different slow-dryingtreatments. The onset of germinability occurred when seeds wereless than half-filled in the absence of both a post-ovule abscissionprogramme and water loss from the seeds. Maximum ability togerminate normally and maximum tolerance to rapid enforced desiccationto 14–16% moisture content did not occur until 2–23d and 6–23 d after mass maturity (end of the seed-fillingperiod), respectively. The slow-drying of immature seeds for7 d ex planta before rapid enforced desiccation increased theability to germinate and stimulated the onset of desicationtolerance. Holding seeds moist for 7 d (during which time moisturecontent declined by <5%) had similar effects, but seed germinationafter rapid enforced desiccation was consistently greater inseeds first dried slowly than held moist. Comparisons betweenseeds less than half-filled dried slowly ex planta and fullseeds undergoing maturation drying in planta showed that a similar(slow) rate of water loss over a 7 d period had a similar effecton the subsequent ability of seeds to tolerate rapid enforceddesiccation. Thus, neither a post-ovule abscission programmenor loss of water were required for the onset of the abilityto germinate in developing bean seeds, but both were requiredfor the development of the ability to germinate and resistanceto solute leakage, when rehydrated, after rapid enforced desiccation. Key words: Bean, Phaseolus vulgaris L., seed germination, seed development, desiccation tolerance     

Seed Maturity and the Effects of Different Drying Conditions on Desiccation Tolerance and Seed Longevity in Foxglove (Digitalis purpurea L.)

The effects of different drying methods on desiccation toleranceand longevity in seeds of foxglove (Digitalis purpurea L.) wereassessed from just prior to mass maturity (when seeds have attainedmaximum dry weight), and at intervals during the post-abscissionphase of development. Tolerance of drying under seed conservationconditions (15% relative humidity, RH, and 15 °C), was acquiredclose to mass maturity at 36 d after flowering (DAF). Increasesin desiccation tolerance were induced when drying was delayedfor 4 d by placing seeds in a near-saturated atmosphere (approx.100% RH), or if seeds were pre-dried for 7 d at either approx.32% or approx. 73% RH. Irrespective of the drying treatment, seed longevity increasedthroughout the sampling period, i.e. beyond the point of massmaturity and throughout the post-abscission phase, up to thepoint of incipient natural dispersal. At each developmentalstage, delayed drying or pre-drying led to an increase in seedlongevity under controlled ageing conditions compared with seedsdried directly under seed conservation conditions. Increasesin longevity were apparent as increases in the estimates forthe intercept of transformed seed survival curves (K_i) and forthe standard deviation of the normal distribution of seed lifespans,and also in the mean time to death of individuals in storage,consistent with a continuation of ripening events. The results are discussed in relation to the assessment of seedlongevity and to current post-harvest drying practices for seedsintended for long-term ex-situ conservation.Copyright 1995,1999 Academic Press Digitalis purpurea L., foxglove, seed development, seed drying, seed longevity