Generative Cell Division and Sperm Cell Association in the Pollen Grain of Sambucus nigra

Generative cell division in tricellular pollen grains of Sambucusnigra L. (Caprifoliaceae) has been examined with light and electronmicroscopy. During division the generative cell is located inthe centre of the pollen grain, near to the nucleus of a surroundingvegetative cell. Conventional mitosis of the generative cellis followed by cytokinesis through centrifugal cell plate formation.Two sister sperm cells remain in spatial contact with each otherand are surrounded, as formerly their progenitor cell was, bythe vegetative cell. From the changes of shape of the generativecell during division and of the sperm cells it may be assumedthat the space between these cells and the vegetative one containsa labile, non-rigid, wall material. No plastids have been observedin the generative cell and its mitochondria appear to be unequallydistributed between the two future sperm cells during division. Sambucus nigra L., generative cell division, pollen, sperm cell association     

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 Chloroplast and Leaf Developmental Mutant, pale cress, Exhibits Light-conditional Severity and Symptoms Characteristic of its ABA Deficiency

The PALE CRESS gene (PAC) is essential for proper chloroplastand leaf development in Arabidopsis thaliana. The ability ofpac mutants to accumulate significantly more chlorophyll whengrown in low light conditions than in high light conditionssuggests that carotenoid deficiency is at least partly responsiblefor premature cessation of chloroplast development. In additionto accumulation of low levels of chlorophyll and carotenoidpigments,pac mutants are abscisic acid (ABA) deficient and havecharacteristics which may be explained by this deficiency. Theseinclude reduced seed viability and, in enclosed growth conditions,increased leaf growth. Plants transformed with an antisensePAC construct often bear viviparous embryos which may be symptomaticof a deficiency in ABA. Since carotenoids are precursors ofABA, a role for PAC in carotenoid biosynthesis is further supported.The nuclear-encoded, chloroplast-localized PAC protein has beenimplicated in the maturation of plastid-encoded mRNAs. Thus,PAC may affect the abundance of one or more chloroplast proteinswhich function in the synthesis or stability of carotenoids.Using thePROLIFERA gene as a marker for cell division, it isshown that cell division profiles in the pac shoot apex aredisrupted. pac leaves are relatively normal in size and shapedespite the light intensity-induced variability of leaf celldefects. Copyright 2000 Annals of Botany Company Abscisic acid, carotenoid, chloroplast development, leaf development, organismal theory, PALE CRESS,PROLIFERA , vivipary     

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     

Cessation of cell elongation in rye coleoptiles is accompanied by a loss of cell-wall plasticity

The mechanism by which endogenous cessation of coleoptile elongationafter emergence of the primary leaf is brought about was investigatedin rye seedlings (Secale cereale L.) that were either grownin darkness or irradiated with continuous white light. In 3-d-oldetiolated (growing) coleoptiles a turgor pressure of 0.59 MPawas measured. In 6-d-old coleoptiles, which had ceased to elongate,cell turgor was 0.51 MPa and thus only 13% lower than in therapidly growing organ. Hence, the driving force for growth (turgor)is largely maintained. Cell-wall plasticity (E_pl) and elasticity(E_Ql were determined with a constant load extensiometer bothin vivo (turgid coleoptile segments) and in vitro (frozen-thawedsamples). Cessation of coleoptile elongation was correlatedwith a 95% reduction in E_pl9 whereas E_Ql was only slightly affected.Extension kinetics were measured with living and frozen-thawedsegments cut from growing and non-growing coleoptiles. The correspondingstress-strain (load-extension) curves indicate that the cellwall of the growing coleoptile behaves like an elastic-plasticmaterial whereas that of the non-growing organ shows the behaviourof an elastic solid. These data demonstate that E_pl representsa true plastic (irreversible) deformation of the cell wall.It is concluded that cessation of coleoptile growth after emergenceof the primary leaf is attributable to a loss of cell-wall plasticity.Hence, a mechanical stiffening of the cell wall and not a lossof turgor pressure may be responsible for the deceleration ofcell elongation in the rye coleoptile. Key words: Extension growth, rye coleoptile, cell-wall extensibility, turgor pressure     

Manganese Nutrition of Lupinus spp. Especially in Relation to Developing Seeds

Patterns of transport and accumulation of manganese were studiedin Lupinus albus L. and Lupinus angustifolius L. in a wide rangeof availability levels in the rooting medium. The recently described‘split seed’ disorder, involving discolouration,splitting, and deformity of seeds, was reproduced in sand cultureusing critically low levels of manganese. The disorder was preventedby maintaining adequate manganese in the medium and its incidencein field and glasshouse was quantitatively related to the managneselevel in seed and fruit phloem sap. The use of phloem sap analysisfor early diagnosis of the disorder is suggested. High levelsof manganese in parent seed is suggested to afford protectionagainst the disorder by improving early vegetative growth ina manganese deficient situation. Direct carry-over of manganesefrom one seed generation to the next was insignificant. Manganese proved to be fully mobile in xylem but only sparinglymobile in phloem from vegetative structures to seed. It wasaccumulated in massive amounts in leaves and fruits when availabilitywas high. Seed manganese content increased 80–100 foldas the level in the rooting medium was increased from 0•1to 500 mg Mn 1^–1. L. albus was superior to L. angustifoliusin accumulating manganese in leaves and pods, and more efficientin translocating the element to its seeds. These differenceswere greatest at low or moderate manganese levels. Xylem intakeby a fruit was small relative to phloem intake when manganeseavailability was low, but became increasingly important as thesupply in the rooting medium was raised.     

Development of Lateral Root Primordia in Vicia faba, Pisum sativum, Zea mays andPhaseolus vulgaris: Rates of Primordium Formation and Cell Doubling Times

Lateral root primordium development has been examined in primaryroots of Vicia faba L., Pisum sativum L., Zea mays L. and Phaseolusvulgaris L. Following their initiation from an estimated minimumnumber of 77–162, 20–57, 17 and 12 cells respectivelyin Vicia, Phaseolus, Pisum and Zea, the primordia rapidly increasedin cell number to emerge as secondary roots about 2.8–3.6days later depending on the species being examined. Cell doublingtimes were estimated directly from cell numbers at differenttimes following primordium inception and were found to increasewith increase in primordium size in each of the species investigated. The number of primordia formed per cm of root growth per daywas greatest in Zea and least in Pisum. A comparison of thedata obtained for Vicia with that in the literature led to theconclusion that although the number of primordia produced percm of root growth was independent of the rate of primary elongation,the number produced per day increased in a linear fashion withincrease in the rate at which the primary lengthened. Vicia faba L, Pisum sativum L, Zea mays L, Phaseolus vulgaris L, broad bean, garden pea, maize, dwarf bean, root primordia, cell division, cell doubling time     

Cellular pathway of photosynthate transport in coats of developing seed of Vicia faba L. and Phaseolus vulgaris L. II. Principal cellular site(s) of efflux

The cells responsible for the photosynthate efflux from coatsof developing seed of Vicia faba L. and Phaseolus vulgaris L.were elucidated using known properties of the efflux mechanism.Sensitivity of sucrose efflux to NEM and high potassium concentrationswas retained by seed-coat halves of Phaseolus following pectinaseremoval of the branch parenchyma cell layer. In contrast, removalof the thin-walled parenchyma transfer cell layer from Viciaseed-coat halves abolished this sensitivity. The membrane-impermeantthiol-binding fluorochrome, qBBr, selectively stained the surfaceof the thin-walled parenchyma transfer cells. This phenomenonwas inhibited by the slowly permeable sul-phydryl agent, PCMBS,indicating that the plasma membranes of these cells are enrichedin sulphydryl groups characteristic of membrance porter proteins.On the basis that carrier-mediated sucrose efflux from seedcoats appears to be proton coupled, the putative plasma membraneH+-ATPase was used as a marker for the cells responsible forcarrier-mediated photosynthate efflux. When seed-coat halveswere exposed briefly at pH 8.5 to the weak acid fluorochrome,SRG, the ground parenchyma and thin-walled parenchyma transfercell layers selectively accumulated the dye. The apparent lowpH environment in the walls of these cells that renders SRGmembrane permeant appeared to be maintained by a VAN-sensitiveproton pump. The findings with SRG were corroborated by thecyto-chemical localization of plasma membrane ATPase activityto the ground parenchyma and thin-walled parenchyma transfercells using precipitation of cerium phosphate. Together, ourobservations provide qualified support for the conclusion thatcarrier-mediated photosynthate efflux from coats of Phaseolusand Vicia seed is primarily restricted to the ground parenchymaand thin-walled parenchyma transfer cell layers, respectively. Key words: Ground parenchyma, Phaseolus vulgaris L., photosynthate efflux, seed coat, transfer cell, Vicia faba L.     

Interrelationship Between Plant Developmental Stage, Plant Growth Rate, Nitrate Utilization and Nitrogen Fixation in Hydroponically Grown Soybean

The growth rate of the indeterminate soybean plant [Glycinemax (L.)Merr.] slows as it proceeds from vegetative phase intoreproductive growth. Yet, the well-nodulated plant acquiresmost of its nitrogen during reproductive growth. Thus, the interrelationshipbetween plant developmental stage and nitrogen fixation wasexamined. It is shown that, regardless of the age of the hydroponicallygrown soybean plant at the time of its inoculation with Bradyrhizobiumjaponicum, the highest rate of nitrogen fixation occurs duringthe pod-filling stage (R5). Nevertheless, maximum total nitrogenfixation is generally achieved when inoculation occurs at thefull-bloom stage (R2). It is shown, however, that flower budsand flowering are not responsible for the enhanced nodulationand nitrogen fixation. Rather, the data suggest that the onsetof rapid nodulation occurs soon after the initiation of thedevelopmentally programmed drop in foliar nitrate reductaseactivity. The ensuing increase in nitrogen fixation providesthe plant with much of its needed nitrogen and hence stimulatesplant mass accumulation during pod-fill. It is suggested thatnitrogen fixation enhances growth of the soybean plant by increasingits net photosynthetic efficiency during reproductive growthand by providing the needed nitrogen at the appropriate timefor maximum seed growth. Key words: Glycine max, nitrate, nitrogen fixation, nodulation     

Membrane State and Pollen Viability

The relationship between germinability and fluorochromasia (FCR)has been studied in pollen of eight genera, Secale, Iris, Carex,Eleocharis, Cytisus, Digitalis, Plantago and Lonicera. The FCRtests two properties of the pollen, (a) the integrity of theplasmalemma of the vegetative cell and (b) the presence of anesterase capable of cleaving the fluorogenic ester, fluoresceindiacetate. In general, the correlations between FCR and germinabilitywere found to be very highly significant. This is interpretedas meaning that the primary determinant of pollen viabilityin short-term storage is the state of the vegetative cell membranes.It is suggested that in the partly dehydrated grain at the timeof dispersal the membranes are largely dissociated and do notform an osmotic barrier, but that normal properties are recoveredduring controlled hydration which would normally take placeon the stigma. According to this view, the decay of apparentviability is related to the progressive loss of the capacityof the vegetative cell membranes to regain a normal structureon rehydration. The genera investigated varied in their longevityin storage in low and high humidity conditions. After low-humiditystorage, most showed some enhancement of germination with 1h exposure to a humid atmosphere before transfer to the germinationmedium. During this ‘conditioning’ period the membranesrecover their fluorescein retentivity in step with the increasein germinability. pollen testing, pollen membranes, pollen fluorochromasia, Secale cereale L., Iris pseudacorus L., Carex ovalis Good., Carex nigra (L.) Reichard, Eleocharis palustris (L.) R.Br., Cytisus battandieri Maire, Plantago lanceolata L., Digitalis purpurea L., Lonicera periclymenum L.     

Control of Seed Growth in Soya Beans [Glycine max (L.) Merrill]

The seed is the primary sink for photosynthate during reproductivegrowth and an understanding of the mechanisms controlling therate of seed growth is necessary to understand completely theyield production process. The growth rate of individual seedsof seven soya bean [Glycine max (L.) Merrill] cultivars withgenetic differences in seed size varied from 10.8 to 3.9 mgseed^–1 day^–1. The growth rates were highly correlatedwith final seed size. The growth rate of cotyledons culturedin a complete nutrient medium was highly correlated with thegrowth rate of seeds developing on the plant and with finalseed size. The number of cells per seed in the cotyledons variedfrom 10.2 to 5.7 x 10⁶ across the seven cultivars. The numberof cells per seed in the cotyledons was significantly correlatedwith final seed size and the seed growth rate both on the plantand in the culture medium. The data suggest that genetic differencesin seed growth rates are controlled by the cotyledons and thenumber of cells in the cotyledons may be the mechanism of control. Glycine max L., soya bean, seed size, growth rate, cell number, sink activity     

Auxospore Structure, Initial Valves and the Development of Populations of Stephanodiscus in Farmoor Reservoir

A population of the planktonic diatom Stephanodiscus growingin Farmoor Reservoir has been investigated with regard to auxosporeformation and the subsequent change in cell size. This is thefirst account of auxospores in Stephanodiscus. The auxosporewall is believed to be organic without any siliceous components.Germination results in two large hemispherical initial valveswhich are rather crudely formed and lack the ring of spinescharacteristic of the vegetative cells which are figured forcomparison. The division of the initial cell to form hemisphericalcells is recorded. These then form a normal vegetative celland a hemispherical cell. The hemisperhical cells acted as markersin the natural population and their longevity was determined.The natural populations did not exhibit a continuous decreasein cell diameter, at times remaining almost constant and atother times decreasing. The origin and ratio of occurrence ofbiconcave, biconvex and concavo-convex cells in the populationis discussed. Stephanodiscus, diatom, auxospore, diatom cell size     

Pigmented Plant Tissues in Culture II. Growth, Development and Decline of Chlorophyllous Tissues

During the phase of exponential growth in chlorophyllous calluscultures derived from Haplopappus gracilis, Hypochaeris radicata,and Acer pseudoplatamus, cells double their number on the average,and also their volume, in about 4.3, 6.6, and 9–2 daysrespectively. The two rates decline subsequently but cell expansioncontinues for a short time after division has ceased. With culturesof Oxalis dispar, however, which have an average cell generationtime of about 10 days, there is first a short exponential phasedominated by division, and this is followed by a series of phasesdominated alternately by either division or expansion. Chlorophyll accumulation does not occur in Haplopappus duringthe exponential phase (chlorophyll a decreases) but there isa slow accumulation of caro-tenoids. The bulk of the pigmentsaccumulate during the declining phase of growth mainly afterdivision has ceased. With Hypochaeris and Acer, on the otherhand, accumulation is most intense during the exponential phase,and few pigments are added later. With Oxalis, most of the accumulationoccurs after the exponential phase; carotenoids accumulate untilthe cessation of growth whereas chlorophylls start to declinebefore this. With all species, pigments decline after the cessationof growth. The loss is small in Haplopappus and the tissuesare still bright green when the medium dries out: Hypochaerisand Oxalis, in contrast, eventually become colourless. The data are discussed in relation to the changes in pigmentcontent that accompany the growth and development of a singlecell in each species.     

Some Aspects of Ontogeny of the Shoot Apices of Solanum melongena L. and Capsicum annuum L.

The ontogeny of the vegetative shoot apices of two importantvegetable plants, Solanum melongena L. and Capsicum annuum L.is described. Each shoot apex is studied at different stagesof seed germination. The relationship between time of germinationand (i) area of vacuoles in the cell, (ii) total area of thecell, (iii) area of the nucleus in the cell, and (iv) ratioof area of vacuoles in the cell to the total area of the cellin each apex is examined. The differentiation of cytohistologicalzonation in both apices is distinct only after one or two leavesare initiated and developed. At a certain stage heterogeneityin staining in the peripheral region of the shoot apex of S.melongena is found. The zonation in both plants differentiatesgradually in histological and cytological features. Vacuolationincreases or decreases in all the cells of the shoot apex orin the cells of a particular region of the shoot apex at differentstages of its ontogeny.     

Changes in Ultrastructure and Abscisic Acid Level, and Response to Applied Gibberellins inTaxus maireiSeeds Treated With Warm and Cold Stratification

Seeds ofTaxus maireiare known for their deep dormancy whichcan only be broken by a procedure involving warm stratificationfollowed by cold stratification. Treatments with alternatingtemperatures of 25/15 or 23/11 °C (12 h light) for 6 monthsfollowed by 5 °C for 3 months were successful in overcomingseed dormancy. After 6 months of warm stratification, cytologicalchanges observed included: enlargement of the embryo; a decreasein the number of lipid bodies; appearance of ER; and increasesin mitochondria, plastids, dictyosomes, vacuoles and microbodiesin the shoot apical meristem. Cold stratification followingthe warm treatment induced cell division, and one or two distinctnucleoli in the shoot apical meristem cells were observed. Bothwarm and cold stratification reduced endogenous ABA concentrationsfrom the original 8888 pg per freshly harvested seed to 392and 536 pg, respectively. Treatment with exogenous gibberellinsafter seeds had been warm-stratified showed that GA₄and GA₇wereeffective at promoting seed germination, but GA₃was not. Theseresults suggest that the strong seed dormancy ofT. maireicouldbe caused by a high ABA content and underdevelopment of theembryos in freshly shed seeds. We conclude that warm stratificationwith alternating temperatures increases the growth of embryosby cell expansion and enlargement and decreases ABA content,but seeds still remain ungerminated. Cold stratification mayinduce the response to GAs and initiate cell division resultingin release from physiological dormancy and subsequent germinationofT. maireiseeds.Copyright 1998 Annals of Botany Company Taxus mairei; ultrastructure; abscisic acid; gibberellin; seed dormancy; stratification; germination.     

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     

Pod Formation and Its Geotropic Orientation in the Peanut, Arachis hypogaea L., in Relation to Light and Mechanical Stimulus

Gynophore elongation, pod formation and pod orientation in thepeanut plant (Arachis hypogaea L.) were studied in relationto the effects of light and dark conditions, mechanical stimulus,and growth substances. It was found that the proembryos controlgynophore elongation, probably by secretion of growth regulatorswhich stimulate cell division in the intercalary meristem locatedproximal to the ovules. The stimulus of pod production causesthe development of the proembryo into a mature embryo simultaneouslywith the growth of pod tissues and the cessation of gynophoreelongation. Darkness was found to be an essential factor forthe induction of pod formation. Pod formation did not occurin any of the treatments performed in the light, including theapplication of different growth substances on the ovary. A mechanicalstimulus is needed, in addition to darkness, for the normalthickening and diageotropic orientation of the pod, caused bya higher growth rate of the basal proximal side of the pod.The two ovules are always located on the upper wall of the diageotropicallyoriented pod (ventral suture). A possible mechanism which causessuch an orientation is discussed.     

Young Reproductive Structures Promote Nitrogen Fixation in Soya Bean

In many legumes the transition from the vegetative to the reproductivephase of development is associated with a marked increase inthe rate of symbiotic nitrogen fixation. In soya bean [Glycinemax (L.) Merr.), the removal of reproductive parts at differentstages of their development showed that the increase in nitrogenfixation rate was primarily due to the presence of flower buds.The increase in the fixation rate of intact reproductive plantswas accompanied by a rapid increase in the weight of noduleson lateral roots and it is suggested that these nodules areresponsible for much of the nitrogen fixation which occurs duringreproductive growth. Maintaining plants in the vegetative stateprovided evidence which suggests that it is the flower budsand not the flowering stimulus which are responsible for theincrease in fixation rate. The marked effects on vegetativegrowth of removing reproductive parts suggests that the mechanisminvolved in the promotion of nitrogen fixation may be hormonal. Glycine max (L.) Merr., soya bean, nitrogen fixation     

The Comportment of the Vegetative Nucleus and Generative Cell in the Pollen and Pollen Tubes of Helleborus foetidus L

It has been reported that in species of Plumbaginaceae, Chenopodiaceae,Cruciferae and Amaryllidaceae a ‘male germ unit’is formed in which the two male gametes remain inter-connected,with one of the pair linked intimately to the vegetative nucleus.In two species the unit has been shown to remain intact in thepollen tube, and some accounts imply that it is polarized inits movement, the vegetative nucleus leading in the tube. Evidence given in this paper indicates that such a unit is unlikelyto be present in Helleborus foetidus L. (Ranunculaceae). Applicationof an optical sectioning technique has shown that at no timeis there a persistent linkage between the generative cell andthe vegetative nucleus in unhydrated, hydrated and germinatingpollen, nor is one present in the early pollen tube. Furthermore,no inter-connections between the two entities were seen in protoplastsfrom living, hydrated and incipiently germinating grains isolatedmechanically in an osmotically balancing medium. Following germination,the vegetative nucleus leaves the grain in advance of the generativecell in most instances, but in the samples examined the generativecell led in about 30 per cent of the tubes. Assembling a polarisedmale germ unit in these circumstances would require (a) theformation of an inter-connection between the vegetative nucleusand the generative cell or one of the gametes derived from itduring passage through the tube, and (b) where the generativecell initially leads in the tube, an exchange in relative positions.It is considered improbable that these conditions could consistentlybe met. Mature, incipiently germinating pollen of H. foetidus releasesa fibrillar component when extruded into suitable media. Websor clusters of fibrils are commonly seen to be associated withboth the vegetative nucleus and the generative cell. The possibilitythat the fibrils are composed of aggregates of microfilamentsis considered. Helleborus foetidus L., pollen germination, generative cell, vegetative nucleus, male germ unit     

Accumulation of Betacyanin in Phytolacca americana Cells and of Anthocyanin in Vitis sp. Cells in Relation to Cell Division in Suspension Cultures

In suspension cultures of Vitis sp., maximal accumulation ofanthocyanin was observed during the stationary phase. Accumulationof anthocyanin occurred in parallel with the cessation of celldivision under conditions such as a reduction of the concentrationof phosphate in the medium, or the presence of aphidicolin,an inhibitor of DNA synthesis. By contrast, in suspension culturesof Phytolacca americana, aphidicolin inhibited the accumulationof betacyanin and cell division. When aphidicolin was removedfrom cells by washing, partially synchronized division of cellswas induced and the accumulation of betacyanin also occurred,in conjunction with cell division. In the absence of phosphatefrom the medium, cell division did not occur and accumulationof betacyanin also ceased. Readdition of phosphate to cellsstarved for phosphate induced both cell division and the accumulationof betacyanin. These results indicate a positive correlationbetween the accumulation of betacyanin and cell division inPhytolacca which contrasts with a negative correlation betweenthe accumulation of anthocyanin and cell division in Vitis. (Received April 17, 1989; Accepted December 23, 1989)