The Relationship between Embryo and Endosperm in Interspecific Hybrids in PennisetumL. Rich

In the interspecific cross [diploid Pennisetum typhoides X hexaploid(P. typhoides X P. purpureum)] X diploid P. typhoides, embryosand endosperms in the same ovule may respond differently totreatment with the growth substance gibberellic acid, as maydo different genotypes. Size of embryo and size of its endospermare positively correlated in some genotypes but not in others.Large fully developed normal embryos may be associated withnormal, retarded, degenerate or even no endosperm at all. Conversely,large normal endosperms, even in mature seeds, may contain large,small, or no embryos. Fully developed embryos may be viableregardless of whether they had been associated with normal,retarded or degenerate endosperm, or even no endosperm at all. It is concluded that the embryo is not always dependent on theendosperm during its early development and that a degeneratingendosperm need not have a detrimental effect on the developmentof its embryo or on the viability of the latter.     

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 CARBOHYDRATES, PROTEINS AND NUCLEIC ACIDS IN THE OVULES OF ZEPHYRANTHES LANCASTERI AT DIFFERENT STAGES OF MATURATION

The major changes in the levels of soluble and insoluble carbohydrates,nitrogenous compounds and nucleic acids were investigated atdifferent stages of seed development in Zephyranthes lancasteri.The activities of amylases, glutamic-alanine transaminase, ribonucleaseand deoxyribonuclease were also studied. The alcohol-solublenitrogen and carbohydrates attain their maximal levels priorto the elongation ofthe cotyledon. Both of these decrease markedlyduring further maturation of the seed. The accumulation of totalnitrogen in the ovule follows a sigmoid pattern. The glutamic-alaninetransaminase activity appears to be exclusively localized inthe endosperm and is absent from the embryo. The embryo seemsto derive its organic nitrogen fromthe endosperm. The peak inthe level of DNA per ovule is attainedprior to the elongationof the cotyledon while that of RNA is foundsoon after. ¹Present address: AEC Plant Research Laboratory, Michigan StateUniversity, East Lansing, Mich., U.S.A.     

Visualization of Membrane Calcium and Calmodulin in Embryo Sacs in situ and Isolated from Petunia hybrida L. and Nicotiana tabacum L.

We have used chlorotetracycline (CTC) and fluphenazine (FPZ)as fluorescent probes to visualize the distributional patternsof membrane calcium (mCa²⁺) and the Ca²⁺-receptor protein calmodulin(CaM) in various cell types of unfixed living isolated and unisolatedembryo sacs of Petunia hydrida L. and Nicotiana tabacum L. Ourresults indicate that in the young embryo sacs of Petunia, bothsynergids and the central cell sequester relatively higher amountsof mCa²⁺ and CaM than the egg cell and the antipodals. Much of the mCa²⁺ in the synergids is polarized in its distributionin that the mCa²⁺ is higher towards the micropylar end of thesynergids. Interestingly, in the mature embryo sacs of Petuniaonly one of the two synergids and the egg cell proper manifesta higher level of mCa²⁺. In vivo only one of the synergids inthe young as well as in the mature embryo sacs if Nicotianaconsistently show higher mCa²⁺. In the mature embryo sacs of Petunia the level of CaM is almostuniform in all the cell types except that one of the synergidsand the three antipodal cells show a slightly higher level ofCaM. The possible implications of these findings in the late eventsof vectorial orientation of pollen tube tip, pollen-tube-synergidinteractions and sperm delivery mechanism are discussed.Copyright1993, 1999 Academic Press Membrane-Ca²⁺, calmodulin, living embryo sacs, Petunia, Nicotiana, pollen tube-synergid interaction     

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     

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.     

Effects of Growth Regulators and Glutamine on In Vitro Development of Zygotic Embryos of Taro (Colocasia esculenta var. antiquorum)

Zygotic embryos of taro, Colocasia esculenta var. antiquorumcultured on Linsmaier-Skoog (LS) medium without the additionof hormones develop into mature plants only in the presenceof endosperm tissue. Growth is usually evident within the firstweek of culture when embryos swell and become green. Embryosexcised from endosperm and cultured on LS containing 0-01 mg1^–1 naphthaleneacetic acid (NAA), and 0–01 mg 1^–16-dimethylaminopurine (6-DMAP) grow at a rate comparable withcontrols for the first week of culture. During the second week,growth rates are higher than controls primarily because embryosform elongated hypocotyl regions which often produce swollentissues and/or callus. In the presence of 200 mg 1^–1 glutamineand a range of concentrations of 6-dimethylaminopurine, benzyladenine,or NAA, elongation of the hypocotyl axis is inhibited, and acompact callus may develop. Embryos grown on LS containing 200mg 1^–1 glutamine and 2.0 mg 1^–1 2, 4, 5-trichlorophenoxyaceticacid form friable callus which was used to generate short-livedsuspension cultures. Growth Regulators, Glutamine, tygotic embryos, Colocasia esculenta, endosperm     

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     

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     

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; )     

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.     

Histological Observations on Initiation and Morphogenesis in Immature and Mature Embryo Derived Callus of Barley (Hordeum vulgare L.)

Callus was induced from immature and mature embryos of barley(cv. Haruna Nijo) on Murashige and Skoog medium containing 2mg l^-1 2,4-D and 5 mg l^-1 picloram, respectively. Paraffin sections(10 µm thick) were prepared for histology during callusinitiation and plant regeneration. Meristems were regeneratedfrom nodular compact callus (NC) derived from scutellar epidermisin immature embryos, whereas they were regenerated from NC derivedfrom epidermal cells of leaf or coleoptile bases in mature embryos.Regardless of the explant source, regeneration was predominantlythrough organogenesis, although regeneration through somaticembryogenesis infrequently occurred. Thus, the callus inducedfrom immature and mature embryos of barley was regarded as 'nodularcompact' rather than 'embryogenic'.Copyright 1995, 1999 AcademicPress Barley, callus, Hordeum vulgare, histology, immature embryo, mature embryo, regeneration     

Accumulation of Metabolites during Seed Development in Trifolium repens L.

Metabolite deposition during seed development was examined histochemicallyin Trifolium repens by light- and fluorescence microscopy. Allendosperm haustorium at the chalazal pole of the embryo sacand wall protrusions in cell walls of the suspensor and theembryo sac suggest that transfer of metabolites from maternalto offspring tissue takes place primarily at these sites. Thisis further supported by prominent cutinization of the interpolarregion of the embryo sac wall, accumulation of starch in integumentaltissue at the embryo sac poles, and breakdown of interpolarendothelial cells. Decomposition of osteosclereid starch isfollowed by accumulation in the cellular endosperm and subsequentlyin the embryo parallel to endosperm degradation. The starchaccumulates gradually inward from the subepidermal cells ofthe embryo to the stele. Protein bodies are formed in the vacuolesalong the tonoplast, later to be cut off in vesicles releasedinto the cytoplasm. At maturity the embryo is packed with proteinand starch, but without lipid reserves. Phytin is observed inthe protein bodies. The mature embryo is surrounded by a proteinand starch containing aleurone layer which originates from theendosperm.Copyright 1994, 1999 Academic Press White clover, protein, starch, cuticle, embryo sac wall     

Embryo and Endosperm Function and Failure inSolanumSpecies and Hybrids

Although the importance of the endosperm as a food store inmany angiosperm seeds is well known, its significance duringearly embryogenesis has been neglected. In many interspecifichybrids, and in some other situations, embryos do not developfully and abort. It has often been stated that this is causedby the endosperm failing to conduct sufficient nutrients tothe embryo, but seldom has it been suggested that the endospermactively controls most of the early stages of morphogenesisof the embryo. Information gleaned from a broad survey of theliterature, combined with additional evidence presented here,obtained fromSolanum incanumand interspecific hybrids, indicatethat the endosperm is dynamic and very active in regulatingearly embryo development. This requires highly integrated geneticcontrol of rapidly changing metabolism in the endosperm. Ininterspecific hybrids, lack of coordination may cause unbalancedproduction of growth regulating substances by the endospermand hence abortion of the embryo, or even unregulated productionof nucleases and proteases resulting firstly in autolysis ofthe endosperm and then digestion of the embryo. The endospermmay thus serve to detect inappropriate hybridization of speciesor ploidy levels and so prevent waste of resources by producingseeds that would result in sterile hybrids or unthrifty subsequentgenerations. This discriminatory function of the endosperm hasdiminished during evolution and domestication of the crop plantSolanummelongenaL.Copyright 1998 Annals of Botany Company Solanum, embryo morphogenesis, endosperm, hybrid, seed development.     

ASPECTS OF THE LIFE CYCLE, POPULATION DYNAMICS,GROWTH AND SECONDARY PRODUCTION OF THE EDIBLE SNAIL HELIX LUCORUM LINNAEUS, 1758 (GASTROPODA, PULMONATA) IN GREECE

This paper examines the biology and ecology of Helix lucorumL. which lives in mainland Greece, as well as its growth andsecondary production. A demographic study revealed that (a)3 cohorts exist at any time during the year (when adults ofall generations belong to the same cohort) (b) egg-laying andhatching occur during the months of July and August respectively,(c) the most rapid growth takes place during spring. Study ofH. lucorum genitalia in relation to age showed that the snailsaxe sexually mature 3 years after hatching, when the largestdiameter of their shell (D) is equal to or greater than 35 mm Von Bertallanffy's method suggests that Helix lucorum may liveup to 14 years or more in order to reach its possible maximumsize (48.80 mm) The study of relative growth of D in relation to P_s (peristomesurface) of Helix lucorum shows that D grows faster than P_swhen D12.50 mm; juveniles change their growth rate when theirD arrives at 22.05 mm, and growth becomes slower when adultsarrive at 36.27 mm Annual secondary production calculated by the size frequencymethod gave a mean annual density of 3.39 individuals per m²,a mean annual crop (biomass) of 4.04 g-m^–2 and an annualproduction (P) of 5.02 g · m^–2. The annual turnoverratio (P//b) is equal to 1.24 (Received 23 June 1987;     

HORMONAL CONTROL OF ROOT GROWTH I

The growth of the primary root of a single gene dwarf mutantof Zea mays L., dwarf-1, is dependent upon gibberellic acidor one of its derivatives as a growth hormone. Moreover, forthe root to respond to gibberellin treatment both embryonicand mature tissues must contribute growth stimuli. ¹ This study was financed by the National Institute of Health,Grant GM 08706-03.     

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     

Structural Changes of the Grain Associated with Black Region Formation in Pennisetum americanum

The structure and ontogeny of grains of Pennisetum americanumare described with particular reference to the black regionon the abgerminal surface of the grain. Darkly pigmented materialwas deposited in the cells of the chalazal pad at black regiondevelopment. This colour development was associated with thecrushing of the transfer cells of the basal endosperm by theembryo and the cessation of transfer of ¹⁴C-labelled assimilateinto the grain. It is proposed that the growth of the embryointo the basal endosperm transfer cells and the subsequent accumulationof the pigmented material are the mechanisms by which graingrowth is halted.