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     

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     

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     

Some Morphological and Chemical Characteristics of Developing Fruits of Raphia hookeri

Ndon, B. A. 1985. Some morphological and chemical characteristicsof developing fruits of Raphia hookeri.—J. exp. Bot. 36:1817–1830. Fruits which were at different stages of development were randomlysampled from different inflorescences of Raphia hookeri palms.The morphological characteristics and chemical (the dry matter,lipid and carbohydrate) contents of the exocarp and seeds weredetermined. The results showed that the seed length, circumferenceand volume were optimal at 24 months after pollination whichindicates that Raphia seeds attained maximum size at that period.The seed endosperm was liquid or semi-liquid between 6–18months after pollination but became solid with a prominent embryoat 24 months. The seed dry matter was low at the early stagesof development but there was a rapid increase in seed dry weightat 18–33 months after pollination. The seeds were physiologicallymatured at 30–33 months after pollination, while the exocarpmatured at 24–30 months after pollination. The Raphia seeds were low in lipid (about 2%) compared to theexocarp which had 30–40% lipid at full maturity. Maximumamount of lipid was accumulated within the exocarp at 36–42months after pollination and this period indicates the timefor harvesting Raphia fruits for maximum oil which is probablythe most economic part of the fruit. The total sugar concentration increased in the exocarp withincrease in maturity. Conversely the concentration of sugarsdecreased within the seeds as the fruit matured. Maximum totalsugar concentration (about 309 mg g^–1 dry fat free sample)was found in the exocarp at 36–42 months after pollination.Mature seeds at 48 months after pollination had about 50 mgof total sugars per g of fat free sample. There was insignificantaccumulation of starch in the exocarp. The mature seeds werelow in starch (5–10% of the dry weight). Key words: Raphia hookeri, development, fruit     

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     

Seed Maturation in Soybeans (Glycine max L. Merr.) is Independent of Seed Mass and of the Parent Plant, Yet is Necessary for Production of Viable Seeds

Freshly harvested seeds of soybean (Glycine max L. Merr.) orseeds shelled, then dried, were non-viable. Seeds dried in intactpods, even when only 17% of normal size, matured into viableseeds and produced healthy plants. These seeds maintained activityof various enzymes but gained little soluble protein while air-dryingin intact pods. There was a qualitative change in seed proteinsassociated with maturation. Seeds matured in intact pods havea greater proportion of protein as slow-moving bands and havecompletely lost one fast-moving band compared with seeds shelledbefore drying. Seed maturation is a distinct phase of seed production,is independent of the parent plant, and can be imposed on theseed at many stages of development.     

Moisture Loss as a Prerequisite for Seedling Growth in Soybean Seeds (Glycine max L. Merr.)

Rosenberg, L. A. and Rinne, R. W. 1986. Moisture loss as a prerequisitefor seedling growth in soybeanseeds (Glycine max L. Merr.).—J.exp. Bot. 37: 1663–1674. As soybean seeds [Glycine max (L.) Merr.] develop, they undergoa change in seed moisture. When excised prematurely from thepod and planted, seeds do not exhibit seedling growth until63 d after flowering (DAF) when the seed moisture has fallenbelow 60%. In contrast, seed germination (radicle protrusion)can occur when seeds as young as 35 DAF (68–79% moisture)are excised, but this germination docs not lead to comparableseedling growth frequencies unless seeds are first given a moistureloss treatment to artificially reduce their moisture below 60%.A moisture loss treatment applied at 35 DAF thus enables seedto undergo the transition from germination (cell expansion)to seedling growth (cell division and expansion) to the extentthat treated immature seed have a vigour index comparable toseeds matured on the plant (100%). The pattern of protein synthesisin vivo was examined in 35 DAF seed using [³⁵S]-methionine incorporation.When moisture loss treatment was applied for 24 h to 35 DAFseeds, seeds synthesized several new polypeptides when comparedwith untreated seeds at the same developmental stage. The sameseed samples showed 0% seedling growth in the absence of moistureloss treatment and 80% seedling growth when the treatment hadbeen applied. Moisture loss from soybean seeds appears to bea prerequisite for the synthesis of new proteins which may bepart of the metabolic process or processes that allow the soybeanseed to undergo the transition from seed germination to seedlinggrowth. Key words: Moisture loss, germination/growth, soybean     

The role of auxin in cell separation in the dehiscence zone of oilseed rape pods

Concentrations of both free and conjugated indole-3-acetic acid(IAA) were studied during development of pod wall, dehiscencezone and seeds of Brassica napus pods. A decrease in auxin contentprior to moisture loss in the pods was observed specificallyin the dehiscence zone, which was correlated with a tissue specificincrease in ß-1,4-glucanase activity. Furthermore,treatment of the pods with the auxin mimic 2-methyl-4-chlorophenoxyaceticacid resulted in a delay of 10 d of ß-1,4-glucanaseactivity and con-comitant cell separation in the dehiscencezone. This indicates that the activity of hydrolytic enzymesinvolved in cell separation in the dehiscence zone is regulatedby auxin activity. Comparison of parthenocarpic pods with seeded pods pointed tothe seeds as the source of IAA. Levels in the dehiscence zoneof these pods were low over the entire sampling period, whilecell separation in the dehiscence zone was delayed by about4 d. These results indicate that a low level of auxins in thedehiscence zone is necessary for dehiscence to take place, butother factors may also be important. Key words: Brassica napus, pod dehiscence, auxin, cellulase     

Localization of 4-Chloroindole-3-acetic Acid in Seeds of Pisum sativum and Its Absence from All Other Organs

4-Chloroindole-3-acetic acid (4-Cl-IAA) was shown by GC-MS analysisto be present in immature and mature seeds of Pisum sativum,but not in any other organs of this plant. Its content was maximalat one week after anthesis and decreased as the seeds matured.Only indole-3-acetic acid (IAA) was detected in the other organsof P. sativum, its content being particularly high in the flowersand young pods during anthesis and the early pod set. (Received January 18, 1988; Accepted April 6, 1988)     

Biochemical Characteristics Associated with the Development of the Desiccation-sensitive Seeds of Machilus thunbergii Sieb. & Zucc.

Biochemical properties, i.e. endogenous abscisic acid, proline,sugars, respiration, adenosine phosphates and adenylate energycharge, and growth and moisture content were measured duringthe development of seeds of Machilus thunbergii. As dry matteraccumulated in the embryo during development, moisture content,ABA, proline, respiration and sugars all declined. At maturity,the dry mass of the seeds failed to attain a plateau beforethe period of natural seed shedding; the axis and cotyledonsreached moisture contents of 58 and 45%, respectively. Dryingof immature seeds at 73% relative humidity and 25 °C for30 d resulted in a complete loss of viability at all developmentalstages tested with the exception of mature seeds that were ableto tolerate a 5% decrease in moisture content before germinationdeclined. ABA was detected in all embryos tested, with a maximum value16.·16 µg g^-1 d. wt about midway through development.Although the presence of ABA induced no tolerance to desiccationof mature seeds, it did coincide with decreased content of waterin the developing seeds and decreased respiration. Desiccationdamage of M. thunbergii seeds occurred when moisture contentwas still high (45%) and this damage was not related to theabsence of oligosaccharides in the mature seeds. We concludethat developing embryos and mature seeds of M. thunbergii haveproperties common to many recalcitrant seeds, with seeds beingsensitive to desiccation at all stages, having a prominent ABApeak, little proline, lacking oligosaccharides, and specifically,little dormancy and a moderate rate of respiration of matureseeds (0·9 µmol O₂ min^-1 g^-1 f. wt). Adenosinetriphosphate content and energy charge decreased from stagefour to stage eight of seed development, then increased againto 103 nmol g^-1 d. wt and 0·73, respectively, in matureseeds. The moderate energy charge observed in mature seeds indicatesthat continuous metabolism is also a characteristic of recalcitrantseeds.Copyright 1995, 1999 Academic Press Machilus thunbergii, seed development, recalcitrant seed, abscisic acid, energy charge     

Primary dormancy in tomato (Lycopersicon esculentum cv. Moneymaker): studies with the sitiens mutant

Intact wild-type tomato (Lycopersicon esculentum cv. Moneymaker)seeds do not complete germination to the same percentage orat the same speed as intact ABA-deficient sitiens (sit^w) mutantseeds when seeds of both genotypes are imbibed on polyethyleneglycol (PEG) solutions of –0.3 to –1.5 MPa osmoticpotential. However, if the thicker testas of wild-type seedsare removed (stripped) from the micropyle without damaging theendosperm, both the percentage and speed of germination at lowexternal water potential are similar to that of sit^w mutantseeds. Removing the micropylar end of the testa from sit^w seedsdid not enhance either the speed or percentage of germinationon PEG solution. Despite similar germination percentage and speed between strippedwild-type seeds and either stripped or intact sit^w seeds underosmotic stress, some differences in seed metabolism are evidentbetween genotypes. The activity of endo-ß-mannanasewas greater in the endosperm of sit^w mutant seeds compared tothe endosperm of wild-type seeds when seeds were exposed toosmotic stress. Although     

Effect of Water Potential and Temperature on the Germination of Four Species of African Savanna Trees

The germination responses of seeds from the African tree speciesColophospermum mopane, Combretum apiculatum, Acacia tortilisand Acacia karroo under varying regimes of temperature and waterstress (induced by incubation in PEG 8000) are reported Withthe exception of Combretum (at –0.14 and –0.29 MPa)and Colophospermum (at –0.29 MPa), incubation in PEG decreasedthe maximum achieved germination percentage (90–100% forall species), but did not extend the germination lag (exceptin Combretum) or affect the time required to reach maximum germinationCombretum and Colophospermum were found to germinate under thewidest range of temperatures and water potentials, for example,as strongly negative as –1.0 MPa at 20 and 30 °C,respectively These seeds also showed greater or equivalent hypocotylelongation in PEG solutions creating potentials of –0.14,–0.29 or –0.51 MPa when compared with seeds germinatedin water, indicating an additional stress adaptation Acaciaspecies showed progressive reduction in germination rates andradicle elongation in response to decreasing water potentialExperiments giving pre-imbibition treatments in water priorto transfer to PEG solutions showed that both Acacia speciesgerminated at approximately 90% if given such pre-treatmentand less than 10% if transferred directly to PEG It is concludedthat the most stress-adapted species studied are Colophospermummopane and Combretum apiculatum, a finding generally correlatedwith the growth habit of these trees Colophospermum mopane, Combretum apiculatum, Acacia tortilis, Acacia karroo, germination, water stress, Zimbabwe     

The Effects of Ethanol on the Germination of Seeds of Japonica and Indica Rice (Oryza sativa L.) under Anaerobic and Aerobic Conditions

An examination was made of the effects of ethanol at 0.2–6.0%(v/v) on the germination, under aerobic conditions, of intactand dehusked seeds of indica rice (cv. Assam IV), which hadbeen harvested 14, 21 and 28 d after anthesis, and of the japonicarice (cv. Sasanishiki), which had been harvested 30 and 60 dafter anthesis. The inhibition of germination caused by dehuskingjaponica rice was overcome by 0.5–5% ethanol, with maximumgermination (frequently 100%) achieved at 3–5% (30 d afteranthesis) or 1–4.5% (60 d after anthesis) ethanol. Furtherincreases in the ethanol concentration reduced germination.The germination of dehusked indica rice was slightly inhibitedat 0.5 and 1% ethanol, whilst the promotion of germination by2% ethanol increased as the seeds matured. At all harvests germinationwas greatest at 3% ethanol, and at 5–6% ethanol germinationfell to 0%. Inhibition, no effect, or minimal stimulation ofthe germination of intact seeds of both japonica and indicarice by ethanol was observed at the concentrations examined.The absence of oxygen stimulated germination of dehusked japonicarice, but this germination was inhibited by ethanol. In contrastethanol had little or no effect on the failure of dehusked indicaseeds to germinate in anaerobic conditions. Thus ethanol treatmentmay help break the strong dormancy of dehusked seeds of indicaand japonica rice. The possible role of ethanol in stimulatinggermination in rice is discussed. Rice; Oryza sativa L.; seed germination; dehusking treatment; ethanol; indica; japonica; oxygen; dormancy; germination inhibition; seed formation     

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.     

The Effect of Desiccation on the Longevity of Seeds of Araucaria hunsteinii and A. cunninghamii

Seeds of Araucaria hunsteinii K. Schum. dried quicker at 29°Cthan at 19°C and quicker with the seed-coat removed thanwhen intact; seeds enclosed in polyethylene bags increased inmoisture content. At 15°C, seeds in a flow of air driedquicker than seeds in a box with silica gel, which in turn driedquicker than seeds in a box with no desiccant. No loss of germinationability occurred on drying fresh seed from 53 to about 32 percent moisture content (fresh weight basis); during further desiccationthe percentage germination was related to percentage moisturecontent in the form of a sigmoid curve, culminating in a completefailure to germinate at approximately 14 per cent moisture content.A consistent relationship was observed for all treatments andthe mean critical moisture content for seed death (failure togerminate) was near 23 per cent. Excised embryos grew on 1 percent agar but died if previously subjected to 14 h of desiccationat 15°C. In contrast, no relationship was found between germination andmoisture content of A. cunninghamii D. Don on desiccation from21 to 7 per cent moisture content. Possible causes for the observeddifference in response to desiccation are discussed and methodsfor seed storage are considered. Araucaria hunsteinii, Klinkii pine, Araucaria cunninghamii, Hoop pine, desiccation, seed longevity, storage of seeds     

Water as a Storage Medium for Elaeis guineensis (pisifera) Seeds under Aseptic Conditions

Over 50 per cent germination has been obtained from Elaeis guineensisform pisifera seeds stored in unaerated sterile distilled waterfor 6 months. The moisture levels of the seeds and excised embryoswere of the same order (20–30 and 60–70 per cent,respectively) as those of fully imbibed fresh seeds. The implicationsof an apparent lower oxygen requirement by seeds stored underwater as against germinating seeds are discussed in the contextof the successful storage. Elaeis guineensis, pisifera, germination, seed dormancy, embryo     

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     

Beneficial Effects of Moist Chilling on the Seeds of Black Spruce (Picea mariana [Mill.] B.S.P.)

Moist chilling (cold stratification) is well-known as a simpleand effective means of overcoming physiological seed dormancyand enhancing germination of many temperate tree and shrub species.However, the apparent activation of intracellular repair mechanismsin non-dormant black spruce (Picea mariana) seeds followingmoist chilling, has not previously been reported. The presentcontribution records the beneficial effects of moist chillingafter accelerated ageing (40°C/98% relative humidity) for3–10 d of black spruce seeds. This treatment was maximallyeffective after 7 d of accelerated ageing, increasing germinationfrom 43 to 61%. Ultrastructural examination of embryo cellsindicated that intracellular activation occurred during moistchilling even when seeds had not been subjected to acceleratedageing, and that the increasing levels of damage accumulatingover the 3–7 d period of ageing were substantially reversedduring subsequent moist chilling. The results of cold stratificationof non-dormant seeds are discussed in terms of the value ofthis practice in nursery seedling production from seeds thatare of less than the highest quality.Copyright 2000 Annals ofBotany Company Accelerated ageing, black spruce, cold stratification, embryonic axes, germination, moist chilling, Picea mariana, repair mechanisms, ultrastructure     

Effects of High Temperatures on the Permeability and Germinability of the Hard Seeds of Rhus javanica L.

The effects of temperature, 40–85 °C, on the permeabilityand germinability of the hard seeds of the pioneer tree Rhusjavanica L. with a fire syndrome were studied. The temperatureeffective for removal of the water-impermeable coat dormancyof the seeds was 55 ± 7·4 °C. With increasingtemperature, shorter exposure became sufficient to render theseeds permeable, but at temperatures above 75 °C, heat impairmentof germinability resulted in less than 60% germinability, evenwith long exposure. The most favourable regimes among thosetested were temperatures of 65–75 °C for durationsof 30–120 min, which frequently occur on denuded groundduring the midday hours of clear spring or summer days. Rhus javanica L., water-impermeable coat dormancy, seed germination, high temperature     

Influence of Irradiated Pollen on Embryo and Endosperm Development in Kiwifruit

Development of seeds following pollination with irradiated pollenwas studied inActinidia deliciosa(kiwifruit) ‘Hayward’.Pollinations were carried out using two different sources ofpollen (‘Tomuri’ and ‘Matua’) irradiatedwith gamma rays at doses of 700 and 900 Gy. Non-irradiated crosseswere used as controls. Pollen irradiation had little effectonin vitropollen germination. Irradiated pollen affected seedset and seed content, and induced the formation of parthenogeneticembryos. In comparison to the control, the embryo growth ratewas slower and the endosperm contained very low amounts of storageproducts. Seed set was significantly reduced following bothdoses of irradiation. Two types of seeds were observed: (1)seeds with endosperm only; and (2) seeds with both embryo andendosperm. The proportion of seeds containing endosperm onlywas almost ten-fold higher than those containing both embryoand endosperm. Embryo production by gamma-irradiated pollenwas genotype- and dose-dependent. The induction of parthenogenesiswas higher following gamma ray doses of 900 Gy than 700 Gy,which suggests the ‘Hertwig Effect’; the best efficiencywas obtained with ‘Tomuri’ pollen. Ploidy levelof parthenogenetic embryos was evaluated by nuclear size (area)with the use of image analysis. There was a large differencein embryo nuclei size between control and parthenogenetic embryos(mean size 90.8 and 49.1 µm², respectively). It is concludedthat parthenogenetic embryos represent trihaploids.Copyright1998 Annals of Botany Company. Actinidia deliciosa, kiwifruit, pollen irradiation, induced parthenogenesis.