The phenology of sexual reproduction inGinkgo biloba: Ecological and evolutionary implications

This study examines how the latitude of cultivation ofGinkgo biloba affects the timing of all phases of its sexual reproductive cycle, from pollination through germination. Seeds produced by trees growing in warm-temperate climates germinate earlier in the year than seeds produced in cold-temperate climates, and they have a longer period of time available for seedling establishment. The embryos ofG. biloba seeds possess a temperature-dependent developmental-delay mechanism that allows seeds to survive winter by preventing premature germination in the fall. This and other cold-climate adaptations appear to have evolved within the genusGinkgo during the early Cretaceous, when the Northern Hemisphere was undergoing dramatic cooling after a long period of stable, warm conditions.Ginkgo biloba seeds possess an odoriferous sarcotesta that attracts mammalian scavengers in Asia-most notably members of the Carnivora—presumably by mimicking the smell of carrion. Seeds cleaned of their sarcotesta germinated faster and at higher percentages than those with their sarcotesta intact, suggesting that animal dispersal plays an important role in promoting seedling establishment. During the Cretaceous, potential dispersal agents included mammals, birds, and carnivorous dinosaurs.     

Inhibitors of endogenous proteinases in Scots pine seeds: Fractionation and activity changes during germination

Resting seeds of Scots pine (Pinus sylvestris L.) contain inhibitors which inhibit the proteinase activity present in germinating seeds but have no effect on trypsin or chymotrypsin. When a crude inhibitor preparation was chromatographed on Sephadex G-75, the inhibitor activity separated into four peaks with elution volumes corresponding to the molecular weights 24,000, 14,600, 14,000, and 9000. Each of the inhibitors affected both the hydrolysis of haemoglobin at pH 3.7 and the hydrolysis of casein at pH 5.4 and 7.0 by proteinase extracts prepared from “germinating” endosperms. These results suggest that one major proteinase was possibly acting in all the assays. In resting seeds inhibitor activity was present in both the embryo and the endosperm, the activity (per mg dry weight) in the embryo being about 2-fold that in the endosperm. In the endosperms of germinating seeds the inhibitor activity per seed decreased at about the same rate as total N and dry weight. In the seedlings the activity per seedling remained approximately constant. The patterns of the activity changes suggest that the inhibitors do not control the breakdown of storage proteins; a more probable function is the protection of other cellular components from the high proteinase activities required for the rapid proteolysis during germination.     

Rapporti Fra Endosperma ed Embrione Nella Germinazione di Pinus Pinea L.: Nota i: Effetti del Grado di Maturità del Seme

Abstract

Endosperm embryo interrelationships during germination of PINUS PINEA L. 1rst note. — Seeds and excised embryos have been cultured in vitro in the dark. Experiments with ripe seeds and embryos have been paralleled with exper. with unripe ones (6 months before natural seed dispersal). The results are as follows:

Both ripe and unripe seeds germinate in the dark into green seedlings.

In both types of seedlings the rootlet is the only part which rapidly increases in length, actually growing, but the growth speed of the root is greater in ripe seeds than in the unripe ones.

Both ripe and unripe excised embryos grow slowly, with greenles cotyledons and dwarf roots, showing a very slight geotropic curve.

The root stops growing more sharply in ripe embryos than in unripe ones.

The hypothesis is raised that the rootlet reaches its full growing power, under the endosperm control, and that this control encreases more and more, as the seed approaches its full maturity.     

Rapporti Fra Endosperma ed Embrione Nella Germinazione di Pinus Pinea L.

Abstract

Endosperm embryo interrelationships during germination of PINUS PINEA L. (2nd Note). — Experiments have been performed with unripe fresh seeds paralleled with unripe stored seeds (50 days of storage).

The effect of storage is very similar to the effect of further permanence on the tree.

It is confirmed the importance of the endosperm in the germination process already put in evidence in the 1rst note.

The relationships existing between endosperm and embryo during the germination of the seed become more and more strict as the seed ripens, so that in a ripe seed the embryo develops in a normal seedling only in connection of its own endosperm.

In an unripe seed the embryo is uncompletely controlled by its endosperm.

Some remarks have been made about the degree of maturity of the different parts of the embryo.     

Peptide transport by germinating barley embryos

Glycylsarcosine, a dipeptide which is resistant to peptidase activity, was accumulated intact against a concentration gradient by germinating embryos of Hordeum vulgare L., var. Maris Otter, Winter. This is the first clear evidence for the presence of a dipeptide transport system involved in the movement of protein reserves across the scutellum from the endosperm to the embryo during germination.Abbreviations gly-sar glycylsarcosine - TCA trichloroacetic acid     

A view of plant dehydrins using antibodies specific to the carboxy terminal peptide

Dehydrins are characterized by the consensus KIKEKLPG amino acid sequence found near the carboxy terminus, and usually repeated from one to many times within the protein. A synthetic peptide containing this consensus sequence was used to produce specific antibodies that recognize dehydrins in a wide range of plants. This range covered two families of monocots, viz. Gramineae (Hordeum vulgare L., Triticum aestivum L., Zea mays L., Oryza sativa L.) and Liliaceae (Allium sativa L.), and five families of dicots, Malvaceae (Gossypium hirsutum L.), Solanaceae (Lycopersicon esculentum L.), Brassicaceae (Raphanus sativus L.), Fabaceae (Vigna unguiculata L.), and Cucurbitaceae (Cucumis sativus L.). Two families of gymnosperms, Pinaceae (Pinus edulis Engelm.) and Ginkgoaceae (Ginkgo biloba L.), were also included. For several plants in which dehydrin cDNA and genomic clones have previously been characterized, it now appears that the dehydrin family of proteins is larger, and the regulation of dehydrin expression much more complex, than earlier studies have shown.     

Ginkgo biloba Expresses Calreticulin,the Major Calcium-Binding Reticuloplasmin in Eukaryotic Cells

Calreticulin, the main Ca²⁺ binding protein in the endoplasmic reticulum of eukaryotic cells, was characterized in Ginkgo biloba L. pollen and seeds. Electrophoretic analysis of the partly purified extracts showed the presence of two protein bands of 57 and 50kDa apparent molecular masses, which strongly cross-reacted with antibodies against plant calreticulins. Amino acid sequence comparison with other plant and animal calreticulins revealed a much higher similarity of the N-terminus of Ginkgo calreticulins with the homologue from angiosperms rather than with that from mammals. The finding of calreticulin in Ginkgo is indicative of the conservation also in gymnosperms of Ca²⁺ homeostatic mechanisms, which seem to rely on the same molecular components as all eukaryotic cells.     

Permian ginkgophyte fossils from the Dolomites resemble extant <Emphasis Type="Italic">O-ha-tsuki</Emphasis> aberrant leaf-like fructifications of <Emphasis Type="Italic">Ginkgo biloba</Emphasis> L

Background  

Structural elucidation and analysis of fructifications of plants is fundamental for understanding their evolution. In case of Ginkgo biloba, attention was drawn by Fujii in 1896 to aberrant fructifications of Ginkgo biloba whose seeds are attached to leaves, called O-ha-tsuki in Japan. This well-known phenomenon was now interpreted by Fujii as being homologous to ancestral sporophylls. The common fructification of Ginkgo biloba consists of 1-2 (rarely more) ovules on a dichotomously divided stalk, the ovules on top of short stalklets, with collars supporting the ovules. There is essentially no disagreement that either the whole stalk with its stalklets, collars and ovules is homologous to a sporophyll, or, alternatively, just one stalklet, collar and ovule each correspond to a sporophyll. For the transition of an ancestral sporophyll resembling extant O-ha-tsuki aberrant leaves into the common fructification with stalklet/collar/ovule, evolutionary reduction of the leaf lamina of such ancestral sporophylls has to be assumed. Furthermore, such ancestral sporophylls would be expected in the fossil record of ginkgophytes.     

<Superscript>1</Superscript>H NMR-based metabolomics combined with HPLC-PDA-MS-SPE-NMR for investigation of standardized <Emphasis Type="Italic">Ginkgo biloba</Emphasis> preparations

Commercial preparations of Ginkgo biloba are very complex mixtures prepared from raw leaf extracts by a series of extraction and prepurification steps. The pharmacological activity is attributed to a number of flavonoid glycosides and unique terpene trilactones (TTLs), with largely uncharacterized pharmacological profiles on targets involved in neurological disorders. It is therefore important to complement existing targeted analytical methods for analysis of Ginkgo biloba preparations with alternative technology platforms for their comprehensive and global characterization. In this work, ¹H NMR-based metabolomics and hyphenation of high-performance liquid chromatography, photo-diode array detection, mass spectrometry, solid-phase extraction, and nuclear magnetic resonance spectroscopy (HPLC-PDA-MS-SPE-NMR) were used for investigation of 16 commercially available preparations of Ginkgo biloba. The standardized extracts originated from Denmark, Italy, Sweden, and United Kingdom, and the results show that ¹H NMR spectra allow simultaneous assessment of the content as well as identity of flavonoid glycosides and TTLs based on a very simple sample-preparation procedure consisting of extraction, evaporation and reconstitution in acetone-d ₆. Unexpected or unwanted extract constituents were also easily identified in the ¹H NMR spectra, which contrasts traditional methods that depend on UV absorption or MS ionizability and usually require availability of reference standards. Automated integration of ¹H NMR spectral segments (buckets or bins of 0.02 ppm width) provides relative distribution plots of TTLs based on their H-12 resonances. The present study shows that ¹H NMR-based metabolomics is an attractive method for non-selective and comprehensive analysis of Ginkgo extracts.     

The Effect of Arsenate and Other Inhibitors on Early Events during the Germination of Lettuce Seeds (Lactuca sativa L.)

The effect of arsenate, arsenite, 2,4-dinitrophenol, and anaerobiosis on early events in seed germination was investigated using both intact and punched seeds of lettuce (Lactuca sativa L.). It was found that punching the seed removes penetration barriers to the entrance of inhibitors without an undue loss of germination or light responses. The kinetics of the action of germination inhibitors were established by 2-hour pulse experiments. Arsenate and 2, 4-dinitrophenol have very different kinetics. The inhibition of germination in punched seeds by arsenate given in conjunction with phosphate compared with the lack of inhibition of arsenate plus phosphate on the growing seedling, suggest a distinct metabolic change in the germinating embryo at some time between the onset of germination and subsequent seedling growth.     

Effect of disturbed photoperiod on the surface structures of ripening Scots pine (Pinus sylvestris L.) seeds

 The role of disturbed photoperiod on the developing surface layers of ripening Scots pine (Pinus sylvestris L.) seeds was studied from August 14 to October 30. The embryo sizes of both the light-treated and the control seeds were examined by the X-ray method prior to germination tests. The anatomical details of resin-embedded seeds were examined by fluorescence and light microscopy. The timing of the ripening process of the surface structures was described and documented. The greatest anatomical changes in the ripening seeds occurred in the sarcotesta and in the nucellar layers. Maturation of the surface structures was essentially slower than could have been interpreted by the size of the embryo and responded clearly to the disturbance of photoperiod. Accumulation of phenolic substances and degeneration of cells, particularly at the chalazal region, advanced faster in the light-treated than in the control seeds up to mid-September. The ripening effect of the altered photoperiod diminished, however, after mid-September. This was also confirmed by the brown colour of the seed coat getting darker only in the control seeds at the end of the test period. Consequently, fully ripe structures were first found about a fortnight earlier in the control than in the light-treated seeds. The coincidental advancement of the anatomical potential examined by the X-ray method supported the role of the surface structures on the anatomical maturity of pine seeds and the timing of cone collecting. Received: 26 January 1998 / Accepted: 30 March 1998     

773. GINKGO BILOBA – Connections with people and art across a thousand years

We review the representation of Ginkgo biloba L., a quintessentially Chinese plant, in the art of those Western botanists who first encountered it in China and Japan. We also show how several later representations of Ginkgo, in both the West and the East, were inspired and made possible by specimens of Ginkgo in cultivation.     

In vitro Propagation of Ginkgo biloba by Using Various Bud Cultures

The purpose of this research is micropropagation of Ginkgo biloba L. Apical and nodal meristems removed from plantlets or apical buds from a tree were used as explants. Meristems produced an extensive callus and single or rare multiple shoots on Murashige and Skoog medium with different growth regulators and endosperm extract (En) obtained from mature seeds of the same species. For successful root production it was necessary to transfer the shoots to a rooting medium with En. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparative study of storage compound breakdown in germinating seeds of three lupine species

Storage lipid and protein breakdown in germinating seeds of yellow (Lupinus luteus L.), white (L. albus L.), and Andean lupine (L. mutabilis Sweet) and regulatory function of sucrose were investigated. Less oil bodies were detected in organs of yellow lupine seeds, whereas the highest content of oil bodies was noticed in the Andean lupine seeds. Mature, air-dried yellow, white and Andean lupine seeds do not contain starch. Starch grains appear the earliest in white lupine seeds during imbibition. Sucrose deficiency in tissues enhances breakdown of storage lipid, protein and temporary starch in cotyledons. In sucrose starved embryo axes of all investigated lupine species, an increased level of vacuolization was noted. Interconnections between catabolism of storage protein and storage lipid in germinating lupine seeds were identified by applying ¹⁴C-acetate. To assess the importance of key processes in storage lipid breakdown NaF (inhibitor of glycolysis and gluconeogenesis), KCN, NaN₃ and SHAM (inhibitors of mitochondrial electron transport chain) and MSO (inhibitor of glutamine synthetase) were used. Radioactivity coming from ¹⁴C-acetate was released as ¹⁴CO₂ but mostly was incorporated into ethanol-soluble fraction of embryo axes and cotyledons. Respiratory inhibitors caused a significant decrease in ¹⁴CO₂ and ethanol fractions in all three lupine species studied. MSO stimulated release of ¹⁴CO₂ and radioactivity of ethanol fractions in yellow lupine organs fed with sucrose, but in Andean lupine MSO enhanced the production of ¹⁴CO₂ and radioactivity of ethanol fractions both in organs fed and not fed with sucrose. Different strategies of storage compound breakdown are proposed, depending on relative proportion in storage protein and lipid content in lupine seeds.     

Osservazioni Preliminari sui Sistemi Degradanti dell'RNA nei semi di Ricino

Abstract

Preliminary observations on the enzymatic degradation of RNA in castor bean seeds. — Cocucci, Maggio, Monroy and Marrè have shown the decrease of RNA content during ripening in castor bean seeds, and its increase during germination. Furthermore, these Authors have demonstrated that in the dry ripe seeds the ribosomes are undetectable, and that they increase rapidly during germination. Two peaks of ribosomes are easily detected upon ultracentrifugal analysis in germinating seeds (Cocucci and Sturani). These observations were the basis for our investigations of the enzymes of RNA metabolism in castor bean seeds. This paper deals with our preliminary observations on RNA degrading enzymes in these tissues. We have been able to measure RNase activity, phosphodiesterase, 3′-,5′- and 2′-nucleotidases in castor bean seeds at different stages of development. RNase activity (measured in crude extracts) changes little during the ripening process, its rate corresponding to 40–50 μMoles of nucleotides liberated from RNA per hour and per gram of fresh weight. In the dry seeds, RNase activity is 30–40 μMoles of nucleotides/h.g.f.w., and it increases to about 60–70 μMoles/h/g.f.w. after 72 hours of germination.

Phosphodiesterase activity is about 4–5 μMoles/h.g.f.w.

The following rates have been found in seeds almost completely ripe seeds for 3′-, 5′- and 2′-nucleotidase activities, respectively 45–50 μMoles/h.g.f.w.; 6–7 μMoles/h.g.f.w.; 8 μMoles/h.g.f.w.; ATP-ase activity was of about 80–100 μMoles of phosphate liberated /h.g.f.w. - The high activity of 3′-nucleotidase, of the same order of that of RNase, suggests that these two enzymes are responsible for degradation of RNA to nucleosides and inorganic phosphate. Further investigations are being carried on to define the biochemical properties of castor bean RN-ase.     

Sull'attivazione Funzionale dei Semi Germinanti. Nota v. l'attività Nicotinamide Adenina Dinucleotide Ridotto: - e Nicotinamide Adenina Dinucleotide Fosfato Ridotto: 2,6 - Diclorofenolo Indofenolo Ossidoreduttasica

Abstract

On the functional activation of germinating seeds. Note V. NADH₂:— and NADPH₂: DIP oxidoreductase activity. - NADH ₂:— and NADPH ₂ : DIP oxidoreductase activities are ascertained, in Pisum sativum and Zea mays, in the acetone precipitates obtained from extracts of dry and germinating seeds.

Such activities are always present and increase during the first days of germination.

In the mays, where in the embryo the reduction of the DIP is particularly high, at the point that it could be measurable on less than 1 mg of material, the NADPH ₂ : DIP oxidoreductase activity prevails. In the pea, the NADH ₂ : DIP oxidoreductase activity prevails.

Endosperm of mays and cotyledons and embryonic axis of pea have NADH ₂ : DIP and NADPH ₂ : DIP oxidoreductase activities which, referred to the dry weight, are of the same order of extent and about thirty-fifty times inferior to those of the embryo of mays.     

Effetto di Inibitori Della Sintesi Proteica Sull'aumento di Attività Enzimatiche e sul Risveglio del Metabolismo Nell'Endosperma di Semi di Ricino in Germinazione

Abstract

Effects of inhibitors of protein synthesis on the development of metabolic activity in the endosperm during the germination of castor bean seeds. — The effect of chloramphenicol, streptomycin and actinomycin-C on the increase of the activities of glyceroaldehyde-phosphate dehydrogenase, aldolase, glucose-6-phosphate dehydrogenase, fructose 1–6 diphosphate-1-phosphatase, phosphomonoesterase, in the endosperm of germinating castor bean seeds was investigated.

In all cases, the protein synthesis inhibitors depressed the activation of the enzymes tested: in particular, actinomycin (50 μg/ml) completely suppressed the increase of the activities.

The development of the rate of oxygen uptake and the conversion of fats to sugars was strongly affected by the inhibitors.

These data suggest that the increase of the activities of several enzymes in the germinating endosperm is dependent on enzyme synthesis rather than on the conversion from the inactive to the active form of the enzymes.     

Development of microsatellite markers in ginkgo (Ginkgo biloba L.)

Ginkgo biloba L. is one of the oldest unevolved tree species on Earth. We isolated five polymorphic microsatellite loci from G. biloba using a dual‐suppression polymerase chain reaction technique. These loci provided microsatellite markers with high polymorphism ranging from three to 13 alleles per locus. The observed and expected heterozygosities ranged from 0.667 to 0.952 and from 0.640 to 0.897, respectively. The markers will contribute to research on the conservation, genetic diversity and mating patterns of G. biloba.     

Role for plant peroxiredoxin in cadmium chelation

Abstract

Studies on heavy metal underline the role of thiols in plants and attribute tolerance to metal binding. The thiol and peroxiredoxins (Prx) contents and guaiacol peroxidase (GPOX) activity were analyzed in the cotyledons and embryo of pea (Pisum sativum L.) germinating seeds exposed to toxic Cd concentration. The 2-cysteine peroxiredoxin (2Cys-Prx) level as well as the non-protein thiol (–SHNP) pool increased in both tissues treated with Cd compared to the control. An oxidized dimer of 2Cys-Prx was resolved in the presence of Cd ions. The obtained results suggest that Prx constitute a main key target in Cd toxicity. Despite of the decrease in GPOX activity due to the generation of an intracellular oxidative stress, a protective action via increasing Prx expression on thiols is possible to improve the redox status.     

Is Ginkgo biloba (Ginkgoaceae) really an oviparous plant?

Germination of Ginkgo biloba seeds with intact and removed sarcotesta was compared to test the role of the seed coat in germination biology. The presence of an intact sarcotesta significantly reduced total germination percentage when compared to seeds with the sarcotesta removed. Some seeds were also cold stratified. This treatment was not necessary for germination, but it did improve total germination percentage. The seeds were collected during the period of natural abscission. Contrary to the accepted literature, we found that Ginkgo seeds contain well-developed embryos at the time of dispersal. These data demonstrate that the seed coat contributes to winter dormancy of G. biloba, and that the phenology of this species is less primitive than popularly believed.