GROWTH, DEVELOPMENT AND RESPIRATION IN THE OVULES OF ZEPHYRANTHES LANCASTERI AT DIFFERENT STAGES OF MATURATION

Changes in the rate of respiration and activity of succinicdehydrogenase in the ovules, embryo and endosperm have beeninvestigated at different stages of seed development in Z. lancasteri.An attempt has been made to find any possible relation betweenphysiological changes and the growth pattern of ovule. The fertilizedovules mature into seeds in 16–19 days from the time ofpollination. The respiratory course and the growth of ovulesfollow identical trends. The curve for oxygen uptake is characterizedby at least two peaks; the first on the sixth or seventh dayis related to the laying down of walls in the coenocytic micropylarchamber of the endosperm and the second on the 11th day precedesthe elongation of cotyledon. In excised endosperm the activityof succinic dehydrogenase is highest during its transformationfrom freenuclear to cellular state, while in the embryo it isso after the attainment of mature size. ¹Present address: AEC Plant Research Laboratory, Michigan StateUniversity, East Lansing, Mich., U.S.A.     

CHANGES IN THE CARBOHYDRATES, PROTEINS AND NUCLEIC ACIDS DURING SEED DEVELOPMENT IN OPIUM POPPY

This investigation is concerned with the major changes in thelevels of carbohydrates, proteins, nucleic acids and some enzymesin the ovules of Papaver somniferum at various stages of seeddevelopment. Of the soluble sugars, fructose and glucose arepresent in large amounts up to the free nuclear stage of theendosperm but decrease rapidly when the latter turns cellular;then sucrose becomes most abundant. At this time the concentrationof insoluble carbohydrates is almost one-seventh that of solubleand the activities of - and ß-amylases are at theirhighest. The nitrogen in the seed is accumulated in two phases,the first coinciding with the development of endosperm and thesecond with the development of embryo. Cell wall formation inthe coenocytic endosperm is accompanied with a marked decreasein the soluble nitrogen. The activity of glutamic-alanine transaminaseincreases concurrently with the increase of protein in the ovules.The maximal RNA content is attained after the ovules have completedtheir increase in size but prior to the deposition of proteinaceousreserves in the endosperm. The amount of DNA increases withthe growth of endosperm but decreases during the maturationof embryo. Both ribonuclease and deoxyribonuclease show twopH optima corresponding to pH 5.5 and 7.5 (ribonuclease) and5.0 and 7.5 (deoxyribonuclease). Their activities seem to becorrelated to the levels of nucleic acids. ¹ Present address: AEC Plant Research Laboratory, Michigan StateUniversity, East Lansing, Mich., U.S.A.     

Endogenous cis,trans-Abscisic Acid and Pea Seed Development: Evidence for a Role in Seed Growth from Changes Induced by Temperature

Measurements were made using GC/MS SIM¹ of the effects of temperatureon cis,trans-ABA levels in developing ovules and embryos oftwo pea genotypes contrasted in seed size. These effects werethen related to differences in the growth of the pods, seeds,embryos, and testae. In both genotypes high temperatures hastenedthe onset and rate of logarithmic and then linear growth, greatlyshortening the duration of pod and seed development but withoutgreatly altering seed size. Cis,trans-ABA was most concentratedxin the ovules immediately after fertilization. It also accumulatedin the embryo, more rapidly in the larger-seeded line, duringseed maturation. The stage when accumulation in the embryo beganwas the same irrespective of temperature. Accumulation ceasedwhen the pods started to desiccate. The effects of differentconstant temperatures on the maximum levels of embryo cis,trans-ABAwere relatively small and confounded in one genotype by variationin ovule abortion and in the other by differences in the stagewhen cis,trans-ABA accumulation ceased. However, when plantswere transferred from 13 °C to 29 °C at two differentstages during seed maturation, further seed growth was greatlyinhibited coincident with a substantial increase in embryo cis-trans-ABA.The results suggested a role for cis,trans-ABA in the controlof cotyledon enlargement during the linear phase of seed growth.     

Hormonal and Structural Aspects of Fruit Development in the Orchid, Epidendrum

Endogenous cytokinin and gibberellin-like activity were measuredby bioassay in developing fruit of the orchid Epidendrum ibaguense.Cytokinins decline during the first 30 d after pollination,then begin to accumulate, with very high levels (1–13µ g zeatin eq. g^–1 dry wt. ) in the mature fruitand seed. The major structural change in developing fruit duringthe first 30 d is the ongoing cell division in the fruit wall.By day 30 most ovules have been fertilized and embryo developmentbegins. The increase in cytokinin activity thus coincides withthe onset of embryo development. Gibberellin levels declinein the fruit throughout development, although high activity(0.9 µ g GA₃ eq. g^–1 dry wt. ) is observed in themature seed. The mature embryo shows no obvious structural differentiationinto embryonic axis and cotyledon and no endosperm develops.     

CHANGES IN THE CONTENT OF GIBBERELLIN-LIKE SUBSTANCES IN RIPENING SEED AND POD OF LUPINUS LUTEUS

The amount of gibberellin-like substances in seed, pod, embryoand "endosperm" of Lupinus luteus in relation to their developmentwas studied by means of the rice seedling test. The amount of gibberellin-like substances per seed increasedremarkably at the first stage of development, when the growthof seed and pod was very slow, and attained its maximum at suchan early stage as the dry weight of seed reached a value ofonly 10 per cent and that of pod 28 per cent of their respectivemaximum levels. Changes in amount of gibberellin-like substancespresent in the pod were very closely related to the changesin growth of pod. Gibberellin-like substances equivalent to0.3 µg gibberellin A₃ per seed and 0.028 µg perpod were found at the 20th day after anthesis, and no activityin mature seed and 0.09 µg per pod at the 50th day. Noactivity was found in the embryo itself at all stages, indicatingthat the gibberellin-like substances in seed occur only in the"endosperm," i.e. in the tissues that will eventually form testain fully ripened stage. (Received December 18, 1962; )     

Seedlessness and Parthenocarpy inPistacia veraL. (Anacardiaceae): Temporal Changes in Patterns of Vascular Transport to Ovules

Pistacia vera‘Kerman’ (pistachio nut) typicallyproduces high numbers of seedless or blank fruits. Patternsof vascular transport into fruits and ovules were studied over3 years by following the movement of disodium fluorescein solutionfrom cut branches into developing fruitlets. Early in the season,vascular conductivity is intact through to the chalazal endof the ovule. Soon afterwards, the percentage of ovules withvascular conductivity through to the chalaza declines, and ina variable fraction of fruits, movement of the fluorochromesolution becomes blocked either at the placenta or in the funiculus.Six to 9 weeks after anthesis there is blockage in 90 (1 year)to 100% (2 years) of fruits. Subsequently, vascular conductivityresumes in 83.3% (3 year mean) of ovules, a percentage thatcorrelates well with the mean percentage of seeded nuts at harvest(77.5%). Ovules from fruits with dysfunctional vascular conductionearly in the season are smaller than those with fully functionalvascular tissue. At the time conductivity declines, a high percentageof those ovules with blocked vascular movement lack endospermand appear to be unfertilized; none of the ovules that retainfull vascular flow lack endosperm. Pollination using gamma-irradiatedpollen (⁶⁰Co, 1.0 kGy) led to a nearly three-fold increase inthe production of blank nuts. The results indicate that fluoresceintransport may be a valuable tool to predict the fate of ovules,and are consistent with the hypothesis that parthenocarpic fruitset may be an important factor in blank nut production in pistachio.Copyright1999 Annals of Botany Company Pistachio,Pistacia veraL., fluorescein, seed set, seedlessness, parthenocarpy, blanking, ovule, funiculus, chalaza, embryo.     

GIBBERELLIN-LIKE SUBSTANCES OCCURRING IN THE SEED OF PHARBITIS NIL CHOIS. AND THEIR CHANGE IN CONTENTS DURING THE SEED DEVELOPMENT

The amount of gibberellin-like substances in the seed of Pharbitisnil increased in parallel with the growth of the seed, and attained20 days after anthesis to a maximum of 0.115 µg gibberellinA₃ eqivalent per seed, when the seed reached its maximum freshweight or four-fifths of its final dry weight. At this maximumlevel, 0.03 µg and 0.1 µg gibberellin A₃ equivalentswere localized in the embryo and in the "endosperm", respectively.Three gibberellin-like substances (Factors I, II and III) wereseparated upon paper chromatography. In view of the changesin amount of the factors with respect to the seed maturation,these factors, especially Factor II, in the "endosperm" andembryo were assumed to participate in the initial or the mainpart of growth of the embryo. Dwarf rice seedling and maizedwarfs 1, 3 and 5 responded to the three factors nearly in thesame way. Pharbitis dwarf, however, responded only to FactorI, but not to Factors II and III. (Received February 21, 1963; )     

The Role of Maturation Drying in the Transition from Seed Development to Germination: VII. EFFECTS OF PARTIAL AND COMPLETE DESICCATION ON ABSCISIC ACID LEVELS AND SENSITIVITY IN RICINUS COMMUNIS L. SEEDS

During mid-development (25–40 d after pollination: DAP)of the castor bean seed the amount of abscisic acid (ABA) increasesin both the endosperm and the embryo, declining substantiallythereafter until there is little present in the mature dry (60DAP) seed. Premature desiccation of the seed at 35 DAP alsoleads to a major decline in ABA within the embryo and endosperm.Partial water loss from the seed at 35 DAP which, like naturaland premature desiccation, leads to subsequent germination uponreturn of the seed to full hydration, causes a much smallerdecline in ABA levels. In contrast, ABA declines substantiallyin the non-dried (hydrated) control at 35 DAP, but the seedsdo not germinate. Hence, a clear negative correlation betweenABA content and germinability is not observed. Both drying,whether natural or imposed prematurely, and partial drying decreasethe sensitivity of the isolated embryo to exogenous ABA by about10-fold. The protein synthetic response of the castor bean embryo exposedto 0.1 mol m^–3 ABA following premature desiccation exhibitssome similarity to the response of the non-dried developingembryo—in both cases the synthesis of some developmentalproteins is enhanced by ABA, and germination is suppressed.Germination of mature seeds is also suppressed by 0.1 mol m^–3ABA, but the same developmental proteins are not synthesized.In the cotyledons of prematurely-desiccated seed, some proteinsare hydrolysed upon imbibition in 0.1 mol m^–3 ABA, a phenomenonthat occurs also in the cotyledons of similarly treated matureembryos, but not in developing non-dried embryos. Hence theembryo exhibits an ‘intermediate’ response uponrehydration in 0.1 mol m^–3 ABA following premature desiccation;viz. some of the responses are developmental and some germinative.Following natural or imposed drying, the isolated embryo becomesrelatively insensitive to 0.01 mol m^–3 ABA: germinationis elicited and post-germinative reserve breakdown occurs inthe radicle and cotyledons. The reduced sensitivity of the embryoto ABA as a consequence of desiccation may be an important factorin eliciting the switch to germination and growth within thewhole seed. Key words: Abscisic acid, desiccation, astor bean endosperm, seed development, germination, protein synthesis, isolated embryos, hormone sensitivity     

The Uptake and Degradation of 1-14C-Abscisic Acid by Apple Seeds during Stratification

The distribution and degradation of I-¹⁴C-ABA in apple seedsduring stratification was investigated. It was found that totalradioactivity of labelled ABA present in the apple seed after4⁸ h of soaking in I-¹⁴C-ABA solution decreased gradually duringthe stratification period. The decrease of radioactivity wasobserved in seed coats and endosperm with the simultaneous considerableincrease in the cotyledon and embryo axis. Decarboxylation ofI-¹⁴C-ABA was demonstrated and it is postulated as a mode ofABA degradation in apple seed.     

Ovule and Embryo Ontogenesis in Bombacopsis glabra (Pasq.) A. Robyns

Ontogeny of the ovule and development of the embryo in Bombacopsisglabra (Pasq.) A. Robyns were examined. The ovule is bitegmic,crassinucellate, and anatropous. The exostome is eccentric relativeto the endostome; stomata occur on the outer integument. Thesingle archesporial cell functions directly as the megasporemother cell. The embryo-sac is bisporic. The organization ofthe nuclei in the mature embryo-sac is normal. The antipodalcells disintegrate soon after formation. Double fertilization takes place; the zygote undergoes a longperiod of dormancy, but the primary endosperm nucleus dividesimmediately to produce first a nuclear-type, later a cellular-typeendosperm. The zygote is of the caryophyllad type. Adventive embryos arise from single cells of the nucellus inthe vicinity of the micropyle, and develop faster than the sexuallyproduced embryo; this leads to anomictic renroduction.     

Metabolism and Translocation of Exogenous Zeatin Riboside in Germinating Seeds and Seedlings of Zea mays

High specific activity [³H]-zeatin riboside (ZR) was suppliedto germinating seed and developing seedlings of Zea mays tostudy its metabolism and translocation The major metabolitesof ZR in endosperm, embryo, and first leaves were adenosine,adenine, and adenine nucleotide When ZR was supplied to theradicle tip a significant proportion of the radioactivity extractedfrom the radicle was identified as zeatin-9-glucoside (Z9G).However, some ZR was also transported to the shoot and vestigialembryo During the initial stage of germination, movement ofzeatin riboside from the embryo to the endosperm was pronouncedbut little movement occurred in the reverse direction Key words: Zea mays cytokinin, zeatin riboside, metabolism, translocation     

Structure and In Vitro Growth of Zygotic Embryos of Taro (Colocasia esculenta var. antiquorum)

Zygotic embryos of taro, Colocasia esculenta var. antiquorumwere examined using both light and scanning electron microscopyand cultured on Linsmaier-Skoog (LS) medium without the additionof growth regulators. Embryos present within mature seed consistof a hypocotyl-root axis and an undeveloped cotyledon and aresurrounded by two major types of endosperm cells, aleurone andstarchy endosperm. Embryos cultured on LS medium developed intomature plants only in the presence of endosperm tissue. Excisedembryos turned green after 2–4 d in culture and reacheda rapid growth period between days 4 and 6. Culture of taroembryos leading to viable plantlet development depends upon(1) removal of the outer and inner integument, and (2) the presenceof endosperm tissue (including an intact aleurone layer). Colocasia esculenta var. antiquorum, Araceae, taro, embryo culture, integument, endosperm     

Import of Sucrose and its Partitioning in the Synthesis of Galactomannan and Raffinose-oligosaccharides in the Developing Guar (Cyamopsis tetragonolobus) Seed

Import of sucrose and its transformation to galactomannan andraffinose-oligosaccharides have been studied in the developingguar seed. The amount of galactomannan gradually increased withthe ageing of the seed. During the entire period of pod development,sucrose constituted the major portion of the free sugars inthe seed (both endosperm and cotyledons) as well as in the podwall. Besides myo-inositol, the free sugars detected in thedeveloping endosperm and cotyledons were glucose, fructose,raffinose and stachyose. Some compounds, possibly glycosides(RG values higher than that of fructose), were also detectedin the endosperm. In the later stages of seed development, therelative proportion of raffinose in the free sugars increased,reaching 50% of the total free sugars in 77-d-old cotyledons.With pod maturity, the activities of soluble acid and boundacid invertases in the pod wall increased manifold with a concomitantdecline in the non-reducing sugar content. These enzymes seemto be involved in the mobilization of sucrose from this fruitingstructure into the seed. An increased synthesis of raffinose-oligosaccharidesboth in the endosperm and cotyledons was associated with highactivities of soluble acid invertase (pH 4.8) and sucrose-UDPglucosyl transferase in these tissues. Feeding uniformly labelled¹⁴C-sugars to the detached intact pods as well as to the isolatedendosperm and cotyledons resulted in labelling of all endogenousfree sugars and galactomannan. The uptake and incorporationinto galactomannan of ¹⁴C was stimulated by Co²⁺, Mn²⁺ and Mg²⁺.Except for mannose, a major proportion of the ¹⁴C from glucose,fructose and sucrose appeared in sucrose in both endosperm andcotyledons indicating a fast reconstitution of sucrose in situ.Based on the present results, a possible mode of transformationof sucrose to galactomannan and raffinose-oligosaccharides hasbeen proposed. Key words: Sucrose, galactomannan, raffinose-oligosaccharides, invertase, sucrose-UDP glucosyl transferase, ¹⁴C-incorporation, guar seed     

THE ROLE OF THE SEED-COATS IN GERMINATION OF PHOTOSENSITIVE LETTUCE SEEDS

1. In order to analyze the role of both the fruit-coat and theendosperm in the germination of Grand Rapids lettuce seed andin the elongation of the embryo, the seeds were subjected toa number of operations. The fruit-coat was found to make onlya minor con tribution to the photosensitivity of the seed. Thefact that 100% germination of these seeds can occur in completedarkness, when the restraining influence of the endosperm isremoved by appropriate cuts, makes clear that it is the integrityof the endosperm layer that lecides the photosensitivity ofthe seed to red and far-red light. Thus it is deduced that theeffect of light is exerted primarily on the mechanical propertiesof the endosperm layer.
2. Neither red nor far-red light wasfound to affect the elongationof the radicle of de-coated seeds,whereas both powerfully affectthe germination of intact seeds.It follows that radicle elongationis not itself the limitingfactor in germination, although ofcourse it follows immediatelyupon the germination process proper.
3. In order to account forthe ability of red light to initiategermination, it is proposedthat the final step in the germinationcontrol process is theproduction of an enzyme whose actionenables the tip of theradicle to penetrate through the coat.This makes it possiblefor the radicle to begin elongating.Although the detectionof such an enzyme in the radicle wasnot successful, the cotyledonsdo produce both a pectinase andan enzyme hydrolyzing carboxymethylcellulose.Furthermore, seedsinjected with a cellulolytic, hemicellulolyticor pectolyticenzyme germinated to nearly the maximum extentin the dark.This gives considerable support to the hypothesis.

¹ Present address: Johnson Foundation for Medical Physics, Universityof Pennsylvania, Philadelphia, Pa., U. S. A. (Received December 5, 1962; )     

Functional Anatomy of the Ovule in Broad Bean (Vicia faba L.): Ultrastructural Seed Development and Nutrient Pathways

Ovules of broad bean (Vicia faba L.) were studied to discloseultrastructural features, which can facilitate nutrient transportto the embryo sac from 10 d after pollination (DAP) to the matureseed. Fertilization occurs during the first 24 h after pollination.The endosperm is a coenocyte, which is eventually consumed bythe embryo. By 10 DAP the inner integument is degraded and theouter integument adjoins the embryo sac boundary. The heart-shapedembryo approaches the embryo sac boundary at two sites, whichhere are named contact zones. Small integument cells in theneighbourhood of the first formed contact zones become separatedby prominent intercellular spaces. A heterogenous scatteringmaterial, probably representing secretion products accumulatesin these spaces. By 14-16 DAP the integument exudate disappears,and the suspensor degenerates. As the contact zones increasein size, wall ingrowths form a bridging network in the narrowspace between the embryo sac boundary and the extra-embryonicpart of the endosperm wall. The epidermal cells of the embryoseparate adjacent to these zones, and develop conspicuous wallingrowths. At 20 DAP vacuoles showing various stages in formationof protein bodies appear in the cells of the embryo.Copyright1994, 1999 Academic Press Vicia faba, broad beans, ovule, seed, nutrient transport     

STUDIES ON RESPIRATION IN DEVELOPING POPPY SEEDS

The excised ovules of Papaver somniferum have been used to investigatethe endogenous uptake of oxygen during their maturation intoseeds. Effect of some activators and inhibitors and the activityof succinic dehydrogenase have been studied. There appears tobe an intimate correlation between respiration and the anatomicalchanges in the ovule. There is an increased oxygen uptake duringor immediately preceding the events of (a) pollination, (b)divisions of the endosperm nuclei, (c) cell wall formation inthe endosperm, and (d) elongation of cotyledons. The activityof succinic dehydrogenase follows a curve essentially similarto that of oxygen uptake. The low intensity of respiration duringcertain period of development appears not to be due to exhaustionof endogenous substrates. Endogenous substrates do, however,appear to be limiting on the days when maximum respiration occurssince added sugar does significantly enhance oxygen uptake.The results with two inhibitors, iodoacetate and fluoroacetate,suggest the operation of glycolysis and tricarboxylic acid cycleduring the earlier phase of seed development when the rate ofgrowth is highest. (Received September 1, 1964; )     

An Analysis of Seed Development in Pisum sativum: VI. ABSCISIC ACID ACCUMULATION

The abscisic acid (ABA) content of wrinkled (rr) pea seed tissueshas been quantified during development using multiple-ion-monitoringcombined gas chromatography-mass spectrometry and a deuteratedinternal standard. The level of ABA in the embryo generallyincreased with increasing cotyledon fresh weight while thatin the testa showed a distinct maximum at the time of maximumendosperm volume and the slowing in the growth of the testa.Pods contained relatively little ABA on a fresh weight basis.The total seed ABA content showed a biphasic distribution, thefirst maximum following the maximum growth rate of the testaand the second that of the embryo. The biphasic distributionof ABA in the pea seed was confirmed using a second pea genotype,near-isogenic to the first except for the r locus, and by theanalysis of individual seeds using a radioimmunoassay for ABA.The first maximum was composed mainly of a testa component andthe second mainly of an embryo component. When plants were grownin different environments, wrinkled seeds were found to containslightly more ABA than round (RR) but this was only significantlate in development. Immature seeds were capable of metabolizing17'-deoxy ABA to ABA, as determined by incorporation of either³H or ²H, and the metabolite was present mainly in the testa.The production of ABA in pea seeds is discussed in relationto the development of the different seed tissues. Key words: Abscisic acid, peas, seed development     

An Analysis of Seed Development in Pisum sativum: IV. COTYLEDON CELL POPULATIONS IN VIVO AND IN VITRO

A method for quantifying changes in the cell population of Pisumsativum cotyledons during development is described. The methodis based on determining the frequency distribution for cellarea following the random sampling of a single-cell suspensionof cotyledon cells. The population profile of these cells changedprogressively and systematically from a single population, similarin size to meristematic cells, found in embryos less than 3.0mg in fresh weight, to a bimodal population in embryos greaterthan 100 mg fresh weight. This method was used to compare embryosof similar size from two genotypes near-isogenic except forgenes at the r locus. No significant differences were foundbetween the cell population profiles of embryos up to 30 mgfresh weight. However, a significant difference was found betweenembryos with fresh weights of 100 mg, the wrinkled (rr) linehaving a higher mean and maximum cell area (2 951 µm²and 9 240 µm² respectively) than the round (RR) line (2591µm²and 6470 µm²respectively). Comparisons were alsomade between cotyledon cell populations from round (RR) embryosgrown in vivo and in vitro. The most obvious differences werethe higher mean and maximum cell size of the large cell populationof in vitro grown embryos which were twice those found in vivo.Embryos grown to either 30 mg or 100 mg fresh weight in vitrohad a much greater proportion of large cells in the populationwith a corresponding reduction in total cotyledon cell number,compared with similar sized embryos grown in vivo. These data suggest that comparisons between different genotypes,or, between cultured and in vivoembryos, based on morphologicalsimilarities between embryos, may be invalid and subject tomisinterpretation. Key words: Pea, seed development, cell population