Compositional changes of douglas fir seeds during germination

Changes in weight, water content, nucleic acids, nucleotides, carbohydrates, lipids, nitrogenous and phosphorus compounds in embryo and gametophyte of Douglas fir seeds (Pseudotsuga menziesii Franco) were studied at 6 stages of germination. Lipids, proteins, and reserve phosphorus compounds in the gametophyte were utilized for the synthesis of carbohydrates, structural components, and soluble compounds in the seedling.     

Rapid Increase in Adenosine 5'-Triphosphate during Early Wheat Embryo Germination

The ATP content of isolated wheat (Triticum aestivum L. var. Polk) embryos increases 5-fold during the first 30 minutes and 10-fold during the first hour of germination to 80% of maximum. The ATP level remains at approximately 800 nanomoles per gram of tissue during the next 15 hours. ADP, AMP, and total adenosine phosphates decrease between 1 and 6.5 hours, while adenylate energy charge increases from 0.6 to 0.8 and remains constant. The rapid increase in ATP during imbibition is consistent with the energy requirement for polyribosome formation and protein synthesis during the first hours of germination. A method for determining nanomole quantities of ATP in tissue extracts by isotopic dilution of γ-³²P-ATP in the hexokinase reaction is outlined.     

獐牙菜的胚胎发生

对獐牙菜大孢子发生、雌配子体形成、受精、胚及胚乳发育过程进行了研究。主要结果如下：子房2心皮,1室,4列胚珠,侧膜胎座;薄珠心,单珠被,倒弯生胚珠。大孢子母细胞减数分裂形成4个大孢子直线形排列,合点端的大孢子具功能,胚囊发育为蓼型。3个反足细胞宿存,每个细胞均多核和异常膨大,反足吸器明显,并在胚乳之外形成染色较深的类似“外胚乳”的结构。珠孔受精,受精作用属于有丝分裂前类型。胚乳发育为核型;胚胎发育为茄型。果实成熟时,种子发育至球形胚阶段。反足细胞在龙胆科一些短命植物中的宿存与分裂具有重要的生殖适应与进化意义。     

Rapid Increase in UDP-Glucose during Early Wheat Embryo Germination

The cellular content of UDP-glucose in isolated wheat (Triticum aestivum L.) embryo increases 8-fold during the first 40 minutes of imbibition. An additional 3-fold increase in the amount of UDP-glucose was observed in the next 5 hours of germination. This communication also describes a unique, quantitative method to achieve a high sensitivity in a direct determination of UDP-glucose with Na [³²P]pyrophosphate and UDP-glucose pyrophosphorylase. The sensitivity of the assay for UDP-glucose is 10 picomoles.     

Imprinting of the MEDEA polycomb gene in the Arabidopsis endosperm.

In flowering plants, two cells are fertilized in the haploid female gametophyte. Egg and sperm nuclei fuse to form the embryo. A second sperm nucleus fuses with the central cell nucleus that replicates to generate the endosperm, which is a tissue that supports embryo development. MEDEA (MEA) encodes an Arabidopsis SET domain Polycomb protein. Inheritance of a maternal loss-of-function mea allele results in embryo abortion and prolonged endosperm production, irrespective of the genotype of the paternal allele. Thus, only the maternal wild-type MEA allele is required for proper embryo and endosperm development. To understand the molecular mechanism responsible for the parent-of-origin effects of mea mutations on seed development, we compared the expression of maternal and paternal MEA alleles in the progeny of crosses between two Arabidopsis ecotypes. Only the maternal MEA mRNA was detected in the endosperm from seeds at the torpedo stage and later. By contrast, expression of both maternal and paternal MEA alleles was observed in the embryo from seeds at the torpedo stage and later, in seedling, leaf, stem, and root. Thus, MEA is an imprinted gene that displays parent-of-origin-dependent monoallelic expression specifically in the endosperm. These results suggest that the embryo abortion observed in mutant mea seeds is due, at least in part, to a defect in endosperm function. Silencing of the paternal MEA allele in the endosperm and the phenotype of mutant mea seeds supports the parental conflict theory for the evolution of imprinting in plants and mammals.     

Starch Degradation Metabolism towards Sucrose Synthesis in Germinating Araucaria araucana Seeds

As starch is the main seed reserve material in both species of Araucaria of South America, A. araucana and A. angustifolia, it is important to understand starch breakdown in both embryo and megagametophyte tissues of Araucaria seeds. Sugar analysis by thin layer chromatography indicates that sucrose is the main sugar produced in both tissues. Enzyme reactions coupled to benzidine oxidation indicate that sucrose is the main sugar moved from the megagametophyte to the growing regions of the embryo via the cotyledons.

Phosphorylase was detected in both embryo and megagametophyte tissues by the formation of [³²P]glucose-1-P and by formation of [¹⁴C] amylopectin from [¹⁴C]glucose-1-P. The enzyme activity increases 5-fold in both embryo and gametophyte to a peak 18 hours after the start of imbibition. Debranching enzyme, α-glucosidase, and hexokinase are also present in both embryonic and megagametophytic tissues.

Branched glucan oligosaccharides accumulate during this time, reaching a maximum 40 hours after imbibition starts, and decline after germination occurs.

The pattern of activity of the enzymes studied in this work suggests that starch degradation is initiated by α-amylase and phosphorylase in the embryo and by phosphorylase mainly in the megagametophyte. Sucrose-P synthase seems to be the enzyme responsible for sucrose synthesis in both tissues.

     

红直獐牙菜的胚胎学

首次报道了红直獐牙菜大小孢子发生,雌雄配子体形成和胚胎发育过程。主要结果如下：花药四室,药壁发育为双子叶型,绒毡层异型起源,接近腺质绒毡层,中层３层,花药壁表皮宿存,细胞柱状伸长,纤维状加厚,药室内壁退化,小孢子母细胞减数分裂为同时型,四分体的排列方式为四面体形;成熟花粉为３－细胞;子房２心皮,１室,１２列胚珠,侧膜胎座,薄珠心,单珠被,倒生胚珠,蓼型胚囊,反足细胞３个,宿存时间短,胚乳发育为核型     

Kin selection and conflict in seed maturation

There are four genetically distinct components in the developing seeds of flowering plants: maternal sporophyte, gametophyte, endosperm, and embryo. Each component can potentially influence the quantity or quality of nutrients provided to the embryo of its seed, thereby reducing the amount available to embryos in other seeds of that plant. The theory of kin selection predicts that each component will be selected to favor its own embryo over the other embryos to the extent that it is more closely related to its own. Under this criterion, an embryo should be selected to try to acquire more nutrients than the endosperm should be selected to provide, the endosperm should try to supply more than the gametophyte should, and the gametophyte more than the parent sporophyte. Evidence for this conflict of interests is found in the higher frequency of endopolyploidy, nutrient-absorbing haustoria, and food storage tissues in the embryo and endosperm than in the gametophyte of maternal tissues.This theory also suggests how the gametophyte, which is the nurse tissue of gymnosperm seeds, was displaced from this role in the flowering plants by an endosperm initiated by a secondary fertilization. “Neoteny” in the pro-angiosperms created conditions in which (1) an endosperm initiated by double fertilization would be more closely related to the embryo than is the gametophyte and (2) the endosperm would be formed early enough to be of significant aid to the embryo.If this theory is correct it (1) requires a different approach to the study of seed morphology and physiology, (2) increases the plausibility of arguments that flowering plants are a polyphyletic group, (3) provides evidence that parents cannot always control the outcome of conflict with their offspring, and (4) forges a conceptual link in our understanding of the evolution of social interactions in plants and animals.     

Antipodal cells persist through fertilization in the female gametophyte of Arabidopsis

Key message

Extended antipodal life-span.

Abstract

The female gametophyte of most flowering plants forms four cell types after cellularization, namely synergid cell, egg cell, central cell and antipodal cell. Of these, only the antipodal cells have no established functions, and it has been proposed that in many plants including Arabidopsis, the antipodal cells undergo programmed cell death during embryo sac maturation and prior to fertilization. Here, we examined the expression of female gametophyte-specific fluorescent reporters in mature embryo sacs of Arabidopsis, and in developing seeds shortly after fertilization. We observed expression of the fluorescence from the reporter genes in the three antipodal cells in the mature stage embryo sac, and continuing through the early syncytial endosperm stages. These observations suggest that rather than undergoing programmed cell death and degenerating at the mature stage of female gametophyte as previously supposed, the antipodal cells in Arabidopsis persist beyond fertilization, even when the other cell types are no longer present. The results support the concept that the Arabidopsis female gametophyte at maturity should be considered to be composed of seven cells and four cell types, rather than the previously prevailing view of four cells and three cell types.     

The Multiple Forms of alpha-Amylase Enzyme of the Araucaria Species of South America: A. araucana (Mol.) Koch and A. angustifolia (Bert.) O. Kutz : A Comparative Study

α-Amylase is one of the major enzymes present in the seeds of both Araucaria species of South America and it initiates starch hydrolysis during germination and early seedling growth. The pattern of the multiple forms of α-amylase of the two Araucaria species was investigated by electrophoresis and isoelectrofocusing of the native enzyme in polyacrylamide gels. The enzyme forms were compared in the embryo and megagametophyte of quiescent seeds and of seeds imbibed for 18, 48, and 90 hours. Specific α-amylase enzyme forms appear and disappear during these imbibition periods showing both similarities and differences between tissues and species. Before imbibition, there are five α-amylase forms identical in both tissues, but different between species. After 18 hours of imbibition, there are two enzyme forms in both tissues of Araucaria araucana seeds, only one form in the embryo of Araucaria angustifolia but two forms in the megagametophyte of this specie. After 48 hours of seed imbibition, most of the enzyme forms present in quiescent seeds reappear. At 90 hours of imbibition different enzyme forms are detected in the embryo with respect to the gametophyte. The changes in form patterns of α-amylase are discussed according to a possible regulation of gene expression by endogenous gibberellins.     

Developmental events in ovules of the ornamental plant Rudbeckia bicolor Nutt

Microscopic observations of R. bicolor ovules showed that tetrasporic embryo sacs of Fritillaria type are formed. In the mature female gametophytes modifications in antipodal cell formation and egg apparatus organization were observed, e.g. morphological resemblance was evident of antipode or synergid to the egg cell. In the central cell cytoplasm of the mature gametophytes the presence of small bodies was a characteristic feature. Development of both embryo and endosperm was observed in ∼73% of ovules at the embryo stage, while retarded or arrested development of the endosperm was found in ∼26% of them. Occasionally, two embryos occurred in the embryo sac. This is the first record of polyembryony in this species. Although hemigamy has been previously described in Rudbeckia bicolor Nutt., in the present investigations mosaic structure of embryos was not detected. Measurements of the C-DNA amount (flow cytometry) revealed embryo nuclei with 2C DNA content only, and endosperm nuclei with 5C DNA content in the mature seeds. No peak corresponding to 1C nuclei was detected in the histograms obtained from the nuclear preparation of seeds or seedling parts. These results suggest that hemigamy is not an obligatory phenomenon in R. bicolor. The mean 2C DNA value was determined as 14.51 pg (the first estimation for this species).     

Adenylate energy pool and energy charge in maturing rape seeds

A study of energy state and chemical composition of pod walls and seeds of maturing rape (Brassica napus L.) was conducted on two varieties, Victor and Gorczanski. Total adenosine phosphates, ATP, and adenylate energy charge increased with increasing cell number and cellular synthesis during the early stages, remained high at maximum dry weight accumulation and maximum substrate influx time, and decreased with ripening. A temporal control of energy supply and ATP concentration is evident in developing tissues with determined functions; whereas the association of a high energy charge and active cellular biosynthesis occurs only in tissues with a stabilized cell number.     

Polyembryony increases embryo and seedling mortality but also enhances seed individual survival in Handroanthus species (Bignoniaceae)

Polyembryony seems to be advantageous to mother plants in detriment of their siblings which face competition since the beginning of seed development. This competition may limit the turnover of embryos into seedlings and their survival ability. We analysed polyembryony frequency and embryo to seedling turnover in three Handroanthus species with sporophytic apomixis. We tested if the embryo number per seed affected seed and embryo morphometry, seedling survival ability and seed individual survival (i.e. survival of at least one seedling per seed). The number of embryos per seed was compared with seedling number at different developmental stages. All 14 populations showed high frequencies of polyembryonic seeds (21–91%). As the number of embryos per seed increased (up to eight embryos/seed), there was a reduction of mean embryo mass, area, seedling length, individual seedling survival ability, and embryo to seedling turnover. There was also an increase in embryo morphological anomalies. However, enhanced seed individual survival was also observed. Thus, the high frequency of polyembryonic seeds and the increase in seed individual survival support the idea that polyembryony represents an alternative reproductive mechanism which can favours these species.     

Seed Development in Podocarpus henkelii: an Ultrastructural and Biochemical Study

Biochemical and fine-structural studies were undertaken on theembryo and female gametophyte of seeds of Podocarpus henkeliiduring the 24-week period of post-fertilization growth and development. Although the d. wt of the embryo and female gametophyte showeda steady increase during development, the levels of sugars,amino acids, proteins and lipids differed between seed partsand showed changes during the four sampling intervals. Onlystarch was seen to increase steadily throughout development.Lipid levels were high in the mature embryo and free amino acidsshowed a steady increase until the seeds were shed. At thisstage seeds were fully hydrated, possessed abundant reservesand all organelles appeared fully functional. This was interpretedas part of the development strategy of neotonous (recalcitrant)seeds, namely, the maintenance of full metabolic competencefor continued growth in the absence of development arrest whichfollows drying in orthodox seeds. Podocarpus henkelii, yellow wood, embryo and endosperm growth, neotonous seed, recalcitrance     

Calcium fluxes in hydrozoan embryos depend, in part, on exocytosis and fluid phase endocytosis.

In the hydrozoan Phialidium gregarium, the constitutive calcium influx of cleavage stage embryos in sea water is 1.96 +/- 0.75 x 10(-15) moles/embryo/minute. Treating embryos with 227 mM KCl in seawater briefly increases the calcium influx more than 100-fold, to 3.9 x 10(-13) mol/embryo/min. About 62% of the KCl-induced calcium influx is due to calcium flowing through voltage-sensitive calcium channels. This causes a marked intracellular calcium transient and secretion of intracellular vesicles. The other component (approximately 38%) of the calcium influx occurs via fluid phase endocytosis of the extracellular medium (detected using extracellular 3H-sucrose). KCl-treatment of 45Ca loaded embryos induces a 45Ca efflux which can reach peak fractional rates of 0.98/min, during which 55-75% (mean 66%) of the total 45Ca is lost. The KCl-induced calcium efflux is due, in part, to secretion because loaded 3H-sucrose is effluxed simultaneously. This pathway may be important for the calcium efflux necessary for long-term calcium homeostasis in cells.     

Terpene composition of Pinus pinaster seedlings and plants

-The production of essential oils in Pinus pinaster increases with the age of the seedlings, being higher in plants grown under continuous illumination. In the seedlings, nearly all the terpene is α- and β-pinene, the relative proportions of which are completely reversed between the 6th and 10th days of growth, regardless of the illumination period. Another reversion takes place after 60–65 days, the proportion found in the adult pine being very constant with α-pinene as the main component. The amount of oil in seedlings from high resin yielding parent trees was 4-fold higher than that in seedlings of normal (wild type) seeds. This finding is interesting because it can be employed for the preselection at the early stage of seedling of seeds to be used in forestry to obtain plantations of a high resin yield.     

Far Red and White Light-promoted Utilization of Calcium by Seedlings of Phaseolus vulgaris L

The cotyledons and embryo axes of seeds of Phaseolus vulgaris L. cv. Pinto contained 16% of the total calcium in the seed. The remaining 84% was in the testas. There was no evidence that calcium in testas was used in seedling growth or that calcium was leached from seedlings during growth.     

Distributional changes of 32P-labeled acid-soluble phosphates and phospholipids among subcellular fractions during early vertebrate embryonic development

Early developing embryos of the toad Bufo arenarum Hensel were employed to study the content and in vivo labeling with ³²P of the acid-soluble phosphates and phospholipids at the subcellular level. The radionuclide was administered to the female toad along with the pituitary extract used to induce the ovulation.Most of the total phospholipids (68%) and proteins (84%) are confined to the yolk platelet fractions. Up to the heart beat stage (130 h of development) there are no significant changes detectable in protein and phospholipid content.The total P content in trichloroacetic acid-soluble fraction was distributed mainly between postmitochondrial supernatant (58%) and yolk platelet fraction (37%) in the unfertilized oocyte. As development proceeds an increase was observed in the former and a decrease in the latter. The acid-solube phosphates in the mitochondrial fraction only amount to 4% of the total embryo throughout the examined stages.The unfertilized oocyte contains about 98% of acid-soluble phosphates labeled with ³²P in the postmitochondrial supernatant and as development proceeds a striking decrease was found to occur while the radioactivity in the acid-soluble phosphates of mitochondrial and yolk platelet fractions increases significantly during the studied stages. About 11.5% of the lost radioactivity from the acid-soluble phosphates was found to be used to label the phospholipids.     

Late Embryogeny of Fokienia

The young embryo of Fokienia became massive and columnar in the middle of July in Chekiang, China. The polarity of the embryo has already been evident at this time. The cell in the free apex of the columnar embryo are smaller, while in the opposite end the cells continuous to the suspensor are larger in size and irregular in arrangement. Then a group of root initials appears at the middle part of the arc formed by the arrangement of the cells about 10 cells deep from the free apex. All kinds of tissues and organs of the embryo were differentiated in the first week of August. And the root initials become evident. Finally the root initials give rise to the procambium and the embryonic cortex upward and the root cap downward. There are about 20 layers of cells of the procambium in width and only about 10 layers of cells in cotyledonal procambium strand in the mature embryo. The cells of the embryonic cortex are continuous to those of the pericolumn of the root cap. The embryonic epidermis is absent in root cap. The embryo became fundamentally mature about the end of September. The hypocoty and cotyledons are well developed and each constitutes about 40% of the total length of the mature embryo. The root cap is rather weak, only about 10% of the total length. And the rest is the degenerated suspensors. The pith and secretory cells are absent in the mature embryo. The cotyledonal number of the embryo is 2. In mature ovule, there are 15 layers of nucellar cells in width in micropylar part but only 4—5 layers around the rest of the female gametophyte. The megaspore membrane is about 3.6μin thickness. When the young embryo is in the columnar stage, the nuclei of the female gametophytic cells are dividing and forming polynucleate cells. Thus, each cell usually has 2—4 nuclei. In this case, the cells of female gametophyte are large and isodiametric and about 60—120 μ in diameter. But the cells in the outer layer of the female gametophyte are rather small and they are usually uninuclear, rarely binuclear. The present article also deals with the starch distribution during the late embryogeny.     

Allocation of proteolytic activity in the seedling of Cynara cardunculus L. in the initial growth stages

Cynara cardunculus L. seeds were germinated in vitro under environmentally controlled conditions. Seeds showed a 60% germination rate, and three growth stages were established based on the seedling mean relative growth rate (RGR). Root, stem and cotyledons were compared in these stages with respect to the emergence of total proteases and cardosin activity and its allocation in the seedling. In growth stage I (1st-5th post-germinative days), seedlings grew very slowly. Total proteases and cardosins were already active at the onset of seedlings in the stem. Total soluble protein remained constant in cardoon seedlings during stage I, and the content of all free amino acids (aa) but proline (Pro) was equally allocated on the 1st post-germinative day. In growth stage II (5th-10th post-germinative days), seedlings grew intensively and exhibited fully developed cotyledons. A pronounced increase in the content of all free aa up to the middle of growth stage II in both stems and roots was observed. In addition, the allocation of the total proteolytic activity and cardosins followed a gradient from the root to the seedling shoot. However, the whole seedling soluble protein remained constant up to the 7th day in and tended to peak on the 10th post-germinative day, being allocated mainly to the seedling stem. In growth stage III (10th-15th post-germinative days), cardoon seedlings exhibited the lowest mean RGR and the highest R/S growth ratio. An intensive degradation of total soluble protein present in the whole seedling except for cotyledons (ca. 5-fold) was observed. Nevertheless, in growth stage III, both the gradients exhibited by total proteases and cardosins activities between the root and the seedling shoot were enhanced, as were contents of all aa except Pro, exhibiting the highest levels in cotyledons on the 15th post-germinative day.