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     

Protein Synthesis during Natural and Precocious Soybean Seed (Glycine max [L.] Merr.) Maturation

Protein synthesis was studied during precocious and natural soybean seed (Glycine max [L.] Merr.) maturation. Developing seeds harvested 35 days after flowering were precociously matured through controlled dehydration. Total soluble proteins and proteins labeled with [³⁵S]methionine were extracted from control, developing seeds and from precociously and naturally matured seeds and were analyzed by one-dimensional PAGE and fluorography. The results demonstrated that several polypeptides which were designated “mature polypeptides,” were synthesized de novo during precocious and natural seed maturation. Two of these polypeptides, 31 and 128 kilodalton in mass, also stained intensely with Coomassie blue, suggesting their abundant accumulation during seed maturation. Results from in vitro translation experiments showed that the mRNAs corresponding to these “maturation polypeptides” accumulated during precocious maturation and in naturally matured seeds, but not in seeds freshly harvested 35 days after flowering (control). The role of the “maturation polypeptides” is currently unknown; however, their presence and that of their corresponding mRNAs was coincident with the ability of matured seeds to establish seedling growth. This study has demonstrated that precocious seed maturation treatments may be extremely useful for investigations of metabolic events and molecular control mechanisms affecting soybean seed maturation.     

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     

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.     

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     

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     

Growth Analysis of Soybean Seedlings During the Lifespan of the Cotyledons

Patterns of seedling growth of Glycine max in light and darknesswere compared during the period from germination to cotyledonabscission. Fitted growth curves and the derived functions,relative growth rate, unit leaf rate, leaf area ratio and specificleaf area, were used to assess the relative importance in seedlinggrowth of cotyledon storage reserves, cotyledon photosynthesisand leaf photosynthesis. The cotyledons are of an intermediatetype with a predominant storage and a minimal photosyntheticfunction. Cotyledon reserves support seedling growth until theprimary leaves expand, after which growth depends on leaf photosynthesis. Glycine max (L.) Merr., soybean, cotyledons, growth analysis, seedling development     

Abscisic Acid and Precocious Germination in Soybeans

Immature embryos of soybeans (Glycine max L. Merr.) can be inducedto germinate precociously by depleting the endogenous pool ofabscisic acid (ABA). Washing the embryos or allowing the embryosto dry slowly within or out of detached pods causes a gradualdecline in ABA content. The extent of germination is correlatedwith the length of washing or drying treatments, which in turn,affects the level of endogenous ABA. Providing embryos are threeweeks of age or older, maturation and precocious germinationcan be induced by treating embryos in a way that causes a reductionin embryo ABA content. Drying is not an obligatory requirementfor soybean seed maturation or germination. Key words: ABA, Germination, Embryos     

Dehydration Injury in Germinating Soybean (Glycine max L. Merr.) Seeds

The sensitivity of soybean (Glycine max L. Merr. cv Maple Arrow) seeds to dehydration changed during germination. Seeds were tolerant of dehydration to 10% moisture if dried at 6 hours of imbibition, but were susceptible to dehydration injury if dried at 36 hours of imbibition. Dehydration injury appeared as loss of germination, slower growth rates of isolated axes, hypocotyl and root curling, and altered membrane permeability. Increased electrolyte leakage due to dehydration treatment was observed only from isolated axes but not from cotyledons, suggesting that cotyledons are more tolerant of dehydration. The transition from a dehydration-tolerant to a dehydration-susceptible state coincided with radicle elongation. However, the prevention of cell elongation by osmotic treatment in polyethylene glycol (−6 bars) or imbibition in 20 micrograms per milliliter cycloheximide did not prevent the loss of dehydration tolerance suggesting that neither cell elongation nor cytoplasmic protein synthesis was responsible for the change in sensitivity of soybean seeds to dehydration. Furthermore, the rate of dehydration or rate of rehydration did not alter the response to the dehydration stress.     

Characterization of Solute Efflux from Dehydration Injured Soybean (Glycine max L. Merr) Seeds

Soybean (Glycine max L. Merr) seeds lose their tolerance of dehydration between 6 and 36 hours of imbibition. Soybean axes and cotyledons were excised 6 hours (tolerant of dehydration) and 36 hours (susceptible) after commencing imbibition and subsequently dehydrated to 10% moisture. Kinetics of the efflux of potassium, phosphate, amino acid, sugar, protein, and total electrolytes were compared in the four treatments during rehydration. Only slight differences were observed in the kinetics of solute efflux between the two cotyledon treatments dehydrated at 6 and 36 hours suggesting that the cotyledons may retain their tolerance of dehydration at this stage of germination. Several symptoms of injury were observed in the axes dehydrated at 36 hours. An increase in the initial leakage of solutes during rehydration, as quantified by the y-intercept of the linear regression line for solute efflux between 2 and 8 hours suggests an increased incidence of cell rupture. An increase in the rate of solute efflux (slope of regression line between 2 and 8 hours) from fully rehydrated axes was observed in comparison to axes dehydrated at 6 hours. The Arrhenius activation energy for potassium, phosphate, and amino acid efflux decreased and for protein remained unchanged. Both observations indicate an increase in membrane permeability in dehydration-injured tissue. Increasing the H⁺ concentration of the external solution increased K⁺ efflux from both control and dehydrated/rehydrated samples, increased sugar efflux from axes at 6 hours imbibition but decreased sugar efflux from axes at 36 hours imbibition, indicating changes in membrane properties during germination. The dehydration treatment did not alter the pattern of the pH response of axes dehydrated at 6 or 36 hours but did increase the quantity of potassium and sugar efflux from dehydration injured axes. These results are interpreted as indicating that dehydration of soybean axes at 36 hours of imbibition increased both the incidence of cell rupture during rehydration and altered membrane permeability of the rehydrated tissue.     

Reduced phytic Acid content does not have an adverse effect on germination of soybean seeds

Altering the level of phytic acid phosphorus by nutritional means had no effect on the ability of soybean (Glycine max L. [Merr.], cv `Williams 79') seeds to germinate under laboratory or greenhouse conditions. Dry matter moved out of the cotyledons at similar rates whether the germinating seeds initially contained low (0.19), medium (0.59), or high (1.00 milligram per seed) phytic acid phosphorus. Growth of roots and shoots from 3 to 9 days after planting was similar for seeds containing low and medium levels of phytic acid phosphorus. The medium level of phytic acid resembles that found in field-grown seed, so it is clear that soybean seeds normally contain a phosphorus reserve far above that needed for germination and early seedling growth.     

Protein Modification and Utilization of Starch in Soybean (Glycine max L. Merr.) Seed Maturation

Seeds of soybean (Glycine max L. Merr.) harvested at variousstages of development and allowed to dry in intact pods undergoa maturation process and are viable. Defatted powders of seedharvested 24–66 d after flowering were extracted to yieldbuffer-soluble and alkali-soluble proteins. Imposition of amaturation process increased the level of buffer-soluble proteinsbut had no effect on the disulflde content of these proteins.After undergoing maturation, seeds showed an accumulation ofbuffer-soluble polypeptides in the molecular weight range of43–94 kD. Maturation may be associated with the synthesisof specific polypeptides having a molecular weight of approximately85 kD. Alkali-soluble proteins, which represents the storageproteins, did not show any responses to maturation. Their quantityincreased substantially during seed development and the disulfidelevel was only half that of buffer-soluble proteins, attaininga maximum value of 10.9 mol S per 10⁵ g protein. Matured seedat all harvest dates had a final starch content close to thatof normal seed, 10–20 mg g^–1, and soluble sugarswere maintained at quite high levels, 51–83 mg g^–1.The metabolic program for synthesis and degradation of starchseems quite rigidly followed and is independent of harvest dateor of attachment to the parent plant. Soybean seeds retain considerablesoluble proteins and soluble sugars throughout maturation, andthese collectively may be important in maintaining a desiccationresistant structure.     

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     

The Development of Desiccation-tolerance and Maximum Seed Quality During Seed Maturation in Six Grain Legumes

‘Physiological maturity’, i.e. the time when seedsreach their maximum dry weight during development, occurredwhen maturation drying on the parent plant in the field hadreduced seed moisture content to approximately 60 per cent infaba bean (Vicia faba L.), lentil (Lens culinaris Medic.), chickpea(Cicer arietinum L.), white lupin (Lupinus albus L.), soya bean(Glycine max [L.] Merr.) and pea (Pisum sativum L.) The onsetof desiccation-tolerance, i.e. the ability of seeds to germinatefollowing harvest and rapid artificial drying, coincided withphysiological maturity, except in pea where it occurred a littleearlier at about 70 per cent moisture content. Maximum seedquality as determined by maximum viability, minimum seedlingabnormalities and maximum seedling size occurred in pea, chickpeaand lupin when seeds were harvested for rapid drying at physiologicalmaturity; but for maximum seed quality in the other speciesmaturation drying had to proceed further - to about 45 per centmoisture content in soya bean and to about 30 per cent moisturecontent in lentil and faba bean seed crops. Much of this variationamongst the six species, however, was due to differences inthe variation in maturity within each seed crop. Results forindividual pods showed that peak maturity, i.e. maximum seedquality following harvest and rapid artificial drying, was achievedin all six species once maturation drying had reduced the moisturecontent of the seeds to 45–50 per cent. In pea, faba beanand soya bean there was a substantial decline in viability andan increase in seedling abnormalities when harvest was delayedbeyond the optimal moisture content for harvest.     

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     

Role of Lectins in Plant-Microorganism Interactions: II. Distribution of Soybean Lectin in Tissues of Glycine max (L.) Merr

Three different assay procedures have been used to quantitate the levels of soybean (Glycine max [L.] Merr.) lectin in various tissues of soybean plants. The assays used were a standard hemagglutination assay, a radioimmunoassay, and an isotope dilution assay. Most of the lectin in seeds was found in the cotyledons, but lectin was also detected in the embryo axis and the seed coat. Soybean lectin was present in all of the tissues of young seedlings, but decreased as the plants matured and was not detectable in plants older than 2 to 3 weeks. Soybean lectin isolated from seeds of several soybean varieties were identical when compared by several methods.     

Partitioning and Utilization of Photosynthate Produced at Different Growth Stages after Anthesis in Soybean (Glycine max L. Merr.): Analysis by Long-term 13C-Labelling Experiments

Yamagata, M., Kouchi, H. and Yoneyama, T. 1987. Partitioningand utilization of photosynthate produced at different growthstages after anthesis in soybean (Glycine max L. Merr.): Analysisby long term ¹³C-labelling experiments.—J. exp. Bot. 38:1247–1259. Soybean (Glycine max L. Merr. var. Akishirome) plants were allowedto assimilate ¹³CO₂ with a constant specific activity for 10h at different growth stages (a total of seven times at aboutone week intervals) after anthesis. The plants were harvestedperiodically until the time of full maturity and the partitioningof ¹³C into individual plant parts was investigated with anemphasis on the contribution of carbon assimilated at differentgrowth stages to the seed formation. Carbon assimilated at the middle to late seed-filling stagecontributed most to the seed production; one day contributionaccounted for 3–4% in total carbon of the seed at fullmaturity. Integrated contribution of carbon assimilated afteranthesis was estimated as 96% of the final seed carbon. An approximationbased on the temporal data of the incorporation of labelledcarbon into the seeds indicates that 77% of the final seed carboncame from direct transfer of current photosynthate from sourceleaves, which occurred within a few days after the photosyntheticfixation, while the rest originated from remobilization of carbonreserved mainly in leaves and stems plus petioles. In comparison with the total carbon accumulation in the seeds,protein carbon in the seeds was relatively more dependent onphotosynthate produced during the early period of reproductivegrowth stage, whereas lipid carbon was more dependent on photosynthateproduced during the later reproductive stage. Key words: Photosynthate partitioning, soybean (Glycine max L. Merr.), ¹³CO₂ assimilation, seed formation     

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     

Abscisic Acid Levels in Soybean Reproductive Structures during Development

Abscisic acid (ABA) concentrations and growth rates of developing soybean (Glycine max [L.] Merr. cv. Wye) seeds and pod walls were determined from anthesis to maturation using high pressure liquid chromatographic techniques. Developing soybean seeds contain up to 12,200 ng/g fresh weight of ABA compared to 330 ng/g fresh weight for pod walls. In the developing seeds ABA levels correlated with growth rates, being the highest during the most active growth period of seed enlargement, and then decreasing to less than 10 ng/g fresh weight at maturity. Higher levels of ABA were found to occur in the cotyledons and seed coats than the root-shoot axes at 21 days postanthesis. The time required for excised root-shoot axes to initiate growth in liquid culture decreased as seed development progressed and ABA levels of the seeds declined.     

The Role of Maturation Drying in the Transition from Seed Development to Germination: IV. PROTEIN SYNTHESIS AND ENZYME ACTIVITY CHANGES WITHIN THE COTYLEDONS OF RICINUS COMMUNIS L. SEEDS

Drying of immature seeds of Ricinus communis L. cv. Hale (castorbean) during the desiccation-tolerant phase of development causesthem to germinate upon subsequent rehydration. This desiccation-inducedswitch from development to germination is also mirrored by achange in the pattern of soluble and insoluble protein synthesiswithin the cotyledons of the castor bean. Following rehydrationof seeds prematurely dried at 40 d after pollination (DAP),cotyledonary proteins characteristic of development (e.g. storageproteins) are no longer synthesized; hydrolytic processes resultingin their degradation commence (after 12 h) in a manner similarto that observed following imbibition of the mature seed. Apattern of protein synthesis recognizable as germination/growth-associatedoccurs; premature drying has elicited a redirection in metabolismfrom a developmental to a germinative mode. Desiccation is alsorequired for the induction (within cotyledons of 35 DAP seeds)of enzymes involved in protein reserve breakdown (leucyl ß-naphthylamidase;LeuNAase) and lipid utilization (isocitrate lyase; ICL), anevent intimately associated with the post-germinative (growth)phase of seedling development. Thus, at a desiccation-tolerantstage of development, premature drying results in the suppressionof the developmental metabolic programme and a permanent switching-onof the germination/growth metabolic programme. Key words: Desiccation, metabolism, seed development, seed germination, castor bean, cotyledons