Attività Respiratoria Livelli Enzimatici e Tenore in Acqua Durante la Maturazione del Seme di Ricino

Abstract

Respiratory activity, enzyme levels and water contents in the ripening castor beans seed endosperm. — In the endosperm of the developing castor bean seed oxygen uptake, water contents and the « in vitro » measurable activity of various enzymes parallely drop during the terminal strages of ripening. The present investigations shows that also the capacity of water uptake decreases (and that of water loss increases) during ripening.

When developing seeds at a stage close to ripeness are removed from the fruits and incubated under a condition of easy water availability, both respiration and enzyme activities rapidly rise; while this is not observed for seeds removed from the fruit at an earlier stage of development.

These results are interpreted as indicating that the dehydration of the seed during ripening is both a consequence and a cause of the inactivation of enzyme systems.     

Sull'evoluzione Dei Mitocondri Nell'endosperma di Semi di Ricino in Germinazione

Abstract

On the behavior of mitochondria in the castor bean seed endosperm during the early phases of germination. — In the endosperm of the castor bean seed the oxidative activity and the protein nitrogen contents of the mitochondrial fraction markedly increase during the first period of germination (Beevers and coworkers). The activation of the mitochondrial system is paralleled by a similar increase of the activity of several soluble enzymes; the latter process is severely depressed by protein synthesis inhibitors (Cornaggia, Aberghina).

The present research is aimed to understand at what extent phenomena of activation and/or, respectively, of « ex novo » synthesis are responsible of the increase of mitochondrial activity. The following aspects of the mitochondrial behavior during the early period of germination were investigated:

a) Changes in the activity of cytochrome oxydase, malate dehydrogenase and of the succinate-citochrome reductase system.

b) Changes in the morphology of mitochondria and other particulated cell structures, as revealed by electron microscopy.

In the mitochondrial preparation all of the three enzymatic activities investigated were found to increase rapidly during the first days of germination. The increase during the first 24 hours was almost as large when measured as specific activity (activity per mg protein in the mitochondrial fraction) than when measured on an absolute (i.e. per seed) basis; moreover, it was not significantly inhibited by puromycin or by actinomycin. The increase of the three activities during the following period of germination (second-third day) was accompanied by an increase of the protein nitrogen (per seed) in the mitochondrial fraction, and was consistently depressed by the protein synthesis inhibitors.

In the mitochondrial preparation all of the three enzymatic activities investigated were found to increase rapidly during the first days of germination. The increase during the first 24 hours was almost as large when measured as specific activity (activity per mg protein in the mitochondrial fraction) than when measured on an absolute (i.e. per seed) basis; moreover, it was not significantly inhibited by puromycin or by actinomycin. The increase of the three activities during the following period of germination (second-third day) was accompanied by an increase of the protein nitrogen (per seed) in the mitochondrial fraction, and was consistently depressed by the protein synthesis inhibitors.

These results, integrated with those of other investigations on the same material are in agreement with the hypothesis that the activation of metabolism in the endosperm during germination depends in a very early phase mainly on the transition of enzyme systems from an inactive to an active state; while in a second phase synthesis « ex novo » of enzymes and cell structures predominates.     

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.     

Effetto della Presenza di Zuccheri Sull'attivita Esochinasica in Cotiledoni Germinanti di Ricino

Abstract

The effects of sugars on the development of hexokinase and fructokinase activities in isolated cotyledons from germinating castor bean seeds. — The possibility of an inductive effect of hexose concentration on the rate of synthesis of enzymes involved in the phosphorylation of sugars has been investigated. Cotyledons were removed from castor bean seeds germinated 48 h at 27 °C in the dark, and incubated 12 h in water or in 0,05–0,1 M glucose or fructose. The activities of hexokinase and of fructokinase (determined spectrophotometrically in the soluble fraction from cell free extracts) was found to increase, upon incubation, at a rate more than 100% higher for the cotyledons incubated in the presence of sugars than for those in water. The results suggest some specificity of the effect of fructose on fructokinase and of glucose on hexokinase. « Insoluble » hexokinase was not affected by the sugars. Protein synthesis inhibitors such as actinomycin D and puromycin inhibited any increase of kinase activities in the isolated cotyledons.     

Variazioni di Metaboliti Nel Seme di Ricino in Fase di Maturazione

Abstract

Changes of glycolytic substrates level during ripening of the castor bean seed. — The changes of the concentration of carbohydrates and of the main glycolytic substrates in the castor bean seed during the ripening phase were investigated. The following results were obtained:

The level of unphosphorylated sugars and of acid hydrolysable polysaccharides remains almost unchanged, with a tendency to a rise during the ripening phase. The slight increase of these compounds, together with the transition of the R. Q. from high to low values, might be interpreted as an indication of a shift of the seed from the a metabolism of fat synthesis to one of conversion of lipids into sugars, such as is observed in the germinating castor bean seeds.

Hexose monophosphate level sharply decreases during the last period of maturation. However, the level of these substrates does not fall so low as to suggest a severe limitation for the pentose-P pathway activity.

Fructose diphosphate, DOAP, GAP, 3 PGA, 2 PGA, PEP and pyruvate levels consistently increase during the ripening process. This indicates that the drop of oxygen uptake observed in this phase cannot be due to a lack of glycolytic substrates. On the other hand, the ratios between some substrates are shifted, during ripening, from values close to the theoretical equilibrium constants to quite different values. This finding, when correlated with the one of the strong decrease of the glycolytic flow, strongly suggests a severe inactivation of the glycolyting enzymes during ripening.

The increase of pyruvate in tissues showing a decreasing respiratory activity indicates a fall of the oxidative capacity of mitochondria. This might be due to a lack of ADP, or other high energy bond acceptor, following a block of synthetic processes. However, no decrease of ADP level, and an increase of the ADP/ATP ratio during ripening is observed, Among the alternative hypothesis: a) lack or excess of oxalacetate; b) increase of concentration of some Krebs cycle inhibitor; c) inactivation of mitochondrial enzymes, the latter is thought most probable, in view of the finding of a sharp decrease of some other enzyme activities during ripening, of the above mentioned interpretation of the shift of the ratios between glycolytic substrates, and of the very low level of mitochondrial activity in preparation from the mature castor bean seed. These results when correlated with those from parallel investigations on the biochemistry of castor bean seed maturation and germination suggest, as a working hypothesis, that the respiratory metabolic inactivation accompanyng seed repening is due to a general block of the metabolism of ribonucleic acid and thus protein synthesis.     

Osservazioni Sull'effetto Inibente di Elevate Pressioni Osmotiche su Diversi Tipi di Crescita

Abstract

On the inhibition of seeds germination and of growth by cell enlargement by the osmotic pressure of the medium. — The mechanism of inhibition by osmotic pressure (O.P.) of the medium on growth and respiration of germinating wheat, castor bean and lettuce sèeds and of etiolated pea internode segments was investigated.

The following results were obtained:

1 - External osmotic pressure (up to 0.3 M) of various substances such as mannitol, urea, glucose, NaCl, was shown to inhibit the germination and growth of lettuce, wheat and castor bean seeds.

2 - a) A remarkable decrease of the development of respiration during the first 48 h of germination was demonstrated in embryos of wheat seeds germinated and maintained in mannitol solutions at concentration from 0,2 to 0,3 M.

b) A slight but reproducible inhibition of óxygen uptake by O.P. was also observed in embryos isolated from wheat seeds germinated in water for 24 and 34 h and transported respectively in water or into 0,2 M mannitol solutions.

This is interpreted as indicating that high external O.P. inhibits both the respiratory metabolism and the development with time of enzyme systems supporting respiration.

3 - Mannitol solutions (0,2–0,3 M) inhibited completely growth by cell enlargement in pea internode sections, while they did not at all affect oxygen uptake and protein synthesis ( ¹⁴ C - leucine incorporation). The stimulatory effect of auxin on pea elongation was almost completely suppressed by mannitol, whereas the hormone stimulation of respiration remained unchanged.

These data are interpreted as indicating that in tissues, presenting an advanced differentiation, high external O.P. inhibits growth by a direct physico-chemical mechanism; while the inhibitory effect in embrional tissues seems to comprehend, besides this direct effect, a complicated metabolic component, apparently influencing protein synthesis.     

Primi Processi di Germinazione in Pinus Pinea L. var. Fragilis Du Hamel

Abstract

Early stages of germination in PINUS PINEA L. var. FRAGILIS Du Hamel. — The main stages of the hydration process preceeding germination and accompaning root elongation have been observed in Pinus pinea L. seeds, by means of vital colours (Congo red, neutral red, acid fuchsine).

The results are as follows:

a) water penetrates easily through the outer shell of the seed reaching its deepest layer which is less permeable to water. Two or three days were required in our experience for water could overcome this barrier.

b) The inner shell (known as « soft shell ») is almost water-proof and seems to draw water towards the micropilar pole of the seed, so that the first region of the seed which sucks up water is the micropile.

c) Through the micropile water enters the seed and imbibes the column, the pericolumn and the endosperm cells.

The endosperm swells with water until the seed shell blows up, because of the inside pressure. At this time water freely penetrates the seed everywhere.

In natural conditions we may infer that the first tissues which take contact with the soil water are the column and pericolumn. As a certain amount of time is required for penetration of water as far as the column (two or three days in experience conditions) germination starts only after a given amount of water is available in the soil for a certain period of time.

When seed hydration is performed the embryo root starts elongating and gets out of the seed.

The behaviour of the column, the pericolumn and the root cap during the early stages of germination are dexcribed.     

Nuove Prospettive Nello Studio dei Fattori che Controllano la Germinazione Dei Semi

Abstract

New perspectives in the study of factors which control seed germination. — Seedlings of Triticum durum, cv. « Cappelli », coming from unirradiated embryos grafted on to irradiated endosperms (EM(u)/EN(i) of presoaked seeds (in distilled water for 24 h. at 20[ddot]C.) (treatment: X-rays, doses, 2,4,6,8,10 and 20 Kr), grow more than seedlings of control EM(u)/EN(u) (dose 0) (fig. 1). To have this reaction, it is necessary that the used seeds be after-ripe; at the various stages of seed ripening, be ginning from the milk stage, the phenomenon is not present.

On this basis, the author has thought that a natural inhibitor occurs in the after-ripe endosperm of « Cappelli », which is neutralized or destroyed by X-rays.

As an experimental demonstration, some trials have been made of growing wheat seedlings in Petri dish, on moistened (distilled water) filter paper, together with excised embryos or isolated endosperms (fig. 2): the after-ripe endosperm is able to depress the seedling growth (fig. 3). In the same experimental conditions, X-rays, dose 6 Kr, neutralize the inhibition effect given by the endosperm. (fig. 4).

A completely different situation occurs in wheat seed, during its ripening: endosperm is inactive, embryo produces inhibition effects on the seedling growth, which, also in this case, are reduced by X-rays.

These phenomena, put in relation with dormancy in Triticum durum, cv. « Cappelli », which is a relative dormancy, having its maximum at the milk stage, have led the author to the general conclusion that, during dormancy, a germination inhibitor occurs in the embryo of wheat seed; when dormancy is finished, the inhibitor appears in the endosperm, in a situation which becomes stable and definitive.     

Characterization of asparaginyl endopeptidase activity in endosperm of developing and germinating castor oil seeds

A spectrophotometric assay was devised to characterize the asparaginyl (Asn) endopeptidase activity from the endosperm of castor oil seeds. (Ricinus communis L. var. Baker 296). The assay measures the release of p-nitroaniline from the hydrolysis of benzoyl-l-Asn-p-nitroanilide. Assay sensitivity was improved through diazotization of the reaction product with N(]-napthy])-ethylenediamine dihydrochloride: diazotized p-nitroaniline was determined spectrophotometrically at 548 nm (?₅₄₈= 1.64 × 10^?1M^?1 cm^?2). By using this assay. Asn endopeptidase activity was detected in endosperm extracts of developing, mature and germinating castor seeds. Comparison of the Asn endopeptidase activities of developing and germinating castor endosperms revealed that they: 1) have identical pH-activity profiles with optimal activity occuring at pH 5.4: 2) are heat-labile proteins displaying comparable thermal stability profiles, and 3) are activated and inhibited by dithiothreitol and thiol modifying reagents, respectively. Thus, the Asn endopeptidases of developing and germinating castor seeds are very similar, if not identical, cysteine proteases. The most significant increase in the activity of endosperm Asn endopeptidase occurs during the full coryledon to maturation stage of seed development, this period coincides with the most active phase of reserve protein accumulation by ripening castor oil seeds. Asn endopeptidase activity of fully mature (dry) castor seeds was about 2-fold lower than that of muturation stage ripening castor oil seed. Asn endopeptidase activity showed a slight reduction over the inicial 2-day period following seed imbibition, and then rapidly decreased over the next several days of germination. The results are compatible with the proposal that Asn endopeptidase functions both to process storage preproteins following their import into protein bodies of developing seeds, as well as to participate in the mobilization of storage proteins during the early phase of seed germination.     

Inattivazione dei Mitocondri Negli Endospermi di Ricino Durante la Maturazione

Abstract

inactivation of mitochondria in the castor bean seed endosperm during ripening. — The present research deals with the behaviour of mitochondria from the endosperm of castor bean seeds, during the last phases of seed maturation. The activities of citochrome oxidase, of malate dehydrogenase, of the succinate-citochrome c reductase system, and the phosphorylating activity, were chosen as tests of the state of mitochondria.

The results obtained show an increase of the first two activities up to the moment when some ovular tissues are still present, and, successively, a more or less rapid inactivation of the three enzymes investigated, which fall to extremely low values in the dry seed. Also the phosphorylating capacity, high during the first phases, drops quickly as the seed approches to dormancy.

A certain amount of citochrome oxidase is revealed in the supernatant from 20000xg centrifugation made to prepare the mitochondrial fraction; its activity gradually increases as the seed advances to ripeness. A further fractionation of the activity not sedimenting at 20000xg reveals that approximately one half of it sediments when centrifugated for 1 hour at 50000xg, while the other half remains in the supernatant. The particles sedimenting between 20000 and 50000xg show very little, if any, phosphorylating capacity (with succinate).

It is suggested that the gradual inactivation of mitochondrial efficiency during the ripening phase is due to a degradation of mithochondrial structures.     

Azione della luce sulla crescita allo stadio di plantula di un mutante clorofilliano di Triticum durum Desf

Abstract

ACTION OF LIGHT ON SEEDLING GROWTH IN A YELLOW-GREEN MUTANT OF TRITICUM DURUM DESF. — Dry grains of Triticum durum cv. « Cappelli » and its radiation-induced yellow-green mutant were sown in sand in four environment, different for light conditions (daylight, artificial light, ultraviolet rays, darkness), having all the same temperature (23°C.). Seven days after sowing, the following data were registered: a) growth (in cm.) of the shoot and of the longest root; b) fresh weight (in gr.) of shoots and roots; c) water content of shoots and roots.

The following main results were obtained:

1. the yellow-green mutant always shows less growth and less weight increase than normal « Cappelli », with the exception of the seedlings grown at daylight, which have shorter shoots and longer roots than the normal « Cappelli » but the same weight:

2. artificial light, besides depressing the growth of the roots of the yellow-green mutant, which becomes green under these conditions, induces a remarkable decrease in the water content of the roots of the two wheats.

     

A cysteine endopeptidase with a C-terminal KDEL motif isolated from castor bean endosperm is a marker enzyme for the ricinosome, a putative lytic compartment

A papain-type cysteine endopeptidase with a molecular mass of 35 kDa for the mature enzyme, was purified from germinating castor bean (Ricinus communis L.) endosperm by virtue of its capacity to process the glyoxysomal malate dehydrogenase precursor protein to the mature subunit in vitro (C. Gietl et al., 1997, Plant Physiol 113: 863–871). The cDNA clones from endosperm of germinating seedlings and from developing seeds were isolated and sequence analysis revealed that a very similar or identical peptidase is synthesised in both tissues. Sequencing established a presequence for co-translational targeting into the endoplasmic reticulum, an N-terminal propeptide and a C-terminal KDEL motif for the castor bean cysteine endopeptidase precursor. The 45-kDa pro-enzyme stably present in isolated organelles was enzymatically active. Immunocytochemistry with antibodies raised against the purified cysteine endopeptidase revealed highly specific labelling of ricinosomes, organelles which co-purify with glyoxysomes from germinating Ricinus endosperm. The cysteine endopeptidase from castor bean endosperm, which represents a senescing tissue, is homologous to cysteine endopeptidases from other senescing tissues such as the cotyledons of germinating mung bean (Vigna mungo) and vetch (Vicia sativa), the seed pods of maturing French bean (Phaseolus vulgaris) and the flowers of daylily (Hemerocallis sp.). Received: 20 December 1997 / Accepted: 18 March 1998     

Insediamento e Diffusione di Buddleia Davidii Franch

Abstract

«Buddleya davidii» in «Valtellina» (North Italy), its settling and diffusion. — Vegetational areas where «Buddleya davidii» is naturalised in Valtellina are described and the subsequent phases are followed through 18 years.     

Inibizione Da Saccarosio Della Demolizione Delle Riserve Lipidiche in Cucurbita Maxima

Abstract

Inhibition by sucrose of utilization of storage lipids in germinating seeds of «Cucurbita maxima». – Previous work demonstrated that sugars inhibit almost completely the respiratory utilization of lipid in isolated cotyledons from germinating «Ricinus communis» seeds as well as in a yeast, «Rhodotorula gracilis». This research was carried out with the aim to further characterize this phenomenon; the material used are the cotyledons and the embryonic axes of «Cucurbita maxima» var. «mammouth», where the lipid contents markedly decrease during the first 72 hours of germination. By incubating the embrional axes, isolated from 48 hours germinating seeds in 0.1M sucrose it was observed: a) a rise of the R.Q. from 0,65 to 1 which is not due to alchoolic fermentation since no degradation of sucrose to ethanol was detected, b) a marked decrease or a complete inhibition of the utilization of fatty acids. On the contrary, no significant increase of R. Q. occurs, when the cotyledons are incubated in presence of 0.1M sucrose. The metabolic utilization of the fatty acids in «Cucurbita maxima» has been studied by using acetate-2-C¹⁴. Labelled acetate was fed after 30 and 90 minutes of incubation of the embryonic axes in water and, respectively in 0.1M sucrose. The radioactivity in the C0₂ and in total fatty acids fractions was measured 30 minutes after feeding the labelled acetate. It was found that: in the presence of sucrose: a) the incorporation of labelled acetate in fatty acids was increased by about 4396 after 30′ and by 10% after 90′. b) the dissimilation of acetate to CO₂ was decreased by about 5096. These results indicate that sucrose replaces lipids as respiratory material. Experiments with labelled acetate are in agreement with the hypotesis that the inhibition of lipid catabolism by sugars (or some derivative) is due chiefly an enhancement of lipid synthesis. At the same time there is probable an inhibition of fatty acids oxidation.     

Changes in stability of isocitrate dehydrogenase (NADP+) during germination of castor bean seeds

In crude extract of castor bean endosperm, isocitrate dehydrogenase (NADP⁺) (EC 1.1.1.42) was stable at 57°C at the beginning of seed germination as well as in maturing and dry seeds. The enzyme gradually became less thermostable as germination proceeded and became unstable after 4 days. Extract from 5-day-old endosperm reduced the thermostability of the thermostable enzyme. The destabilizing factor accumulated in the endosperm as germination progressed and was identified as ricinoleate. Ricinoleate destabilized the purified enzyme which was stabilized by isocitrate and Mg²⁺, but ricinoleate did not affect the activity of NADP⁺-isocitrate dehydrogenase itself. Stearate, oleate, palmitate and myristate were similar to ricinoleate in their effect on the thermostability of the enzyme. The thermolabile enzyme in the crude extract of 5-day-old endosperm was readily inactivated by trypsin and in low concentrations of buffer. The thermostable enzyme in the crude extract of 2-day-old endosperm was not affected by these treatments. The thermostable enzyme treated with ricinoleate showed the same instabilities as the thermolabile enzyme. The role of ricinoleate in ther germinating castor bean endosperm is discussed.     

Evoluzione Del Metabolismo Ossidativo Dei Semi di Ricino Durante la Maturazione

Abstract

Changes of respiratory metabolism in developing castor bean seeds. — The respiratory metabolism and the effect of the removal of the teguments on gas exchanges in castor bean seeds at various stages of development has been investigated. Maximal values of oxygen uptake rate were found in the period of fat accumulation. From this period on, oxidative activity steadly decreased to become, in the mature seeds, almost undetectable.

In all stages of maturation, the removal of the teguments induced a decrease of the respiratory rate of the seeds. This finding indicated that the respiratory rate of the internal tissues is not directly limited by a scarce availability of oxygen caused by a low permeability of the teguments to gases.

The value of the respiratory quotient (R.Q.) was found close to 1 in the first stages, during the growing of the endosperm while in the following period of fat accumulation it rised to 1.6, and falled again, just before the abscission of the seed, to 0.7. This fall of the R.Q. suggests that the oxidative activity of these seeds in the last stages of maturation is supported either by the oxidation of substrates more reduced than carbohydrates (probabily fats), or, partially, by the conversion of fats to sugars.     

Studio Comparativo della Struttura e del Ciclo Biologico di Due Diatomee di Acqua Dolce: Navicula Minima Grun. E Navicula Seminulum Grun.: III. Esame Dei Caratteri di Navicula Minima Grun. E di Navicula Seminulum Grun

Abstract

Comparative study of the structure and of the biological cycle of two fresh water diatoms: « Navicula minima » Grun. and « Navicula seminulum » Grun. III. Examination of the characteristics of « N. minima » Grun. and « N. seminulum » Grun. — After studing separately the Navicula minima Grun. and the Navicula seminulum Grun. the various characteristics of the two have been compared and contrasted for purpose of classification. The form and dimentions of the frustules, the density and the structure of the streaks, the structure of the walls, the life circle, aspects of their growths, and some anomalies, most especially, as regards the streaks were examined. It was found the among such numerous characteristics, only one, that is, the presence of the streaks formed by double rows of pores in N. seminulum rather than of a simple file as in N. minima makes one thinks that, in spite of strong similarities existing between these two Naviculae, they are to be regarded as belonging to e different systematic classes. However, their actual positions in taxonomy May-June be defined by further research into the structures and the variabilities of the frustules of other species of Naviculae similar to those examined.     

Aspetti Enzimatici Della Maturazione e Germinazione dei Semi di Ricino

Abstract

Enzyme levels during ripening and germination of castor bean seeds. — During the development of the endosperm of castor bean seeds two distinct phases can be recognized: pre-maturation and germination. The former is characterized, metabolically, by the rapid conversion of carbohydrates into lipids, and storage proteins. The latter is characterized by the reconversion of these storage materials into sugars. Both these processes are dependent upon the activity of the glycolytic pathway; for this reason the behaviour of some enzymes of this pathway and, in general, of the carbohydrate metabolism have been studied during the two phases. The changes (during the evolution of the seeds) of the following enzymes have been studied:

Gl-6-P-dehydrogenase, 6-P-gluconate dehydrogenase, P-glucomutase, Hexokinase Hexoseisomerase, Aldolase, alcaline and acid Phosphatase, Pyrophosphatase and ATP-ase.

All these activities have been measured in the 20.000 × g supernatant fraction of cell homogenates.

The results show that all the enzymes activities measured increase rapidly during the period of accumulation of storage materials. In the following period all of these activities decrease until the stage of ripeness of the seed. During the first few days of germination the activities increase again rapidly. A particular behaviour is the one of Fr-1-6-P-phosphatase (the enzyme cleaving the phosphate bond in C ¹ position). This enzyme reaches during germination a level much higher than the maximal observed during the ripening process. This might be an important fact correlated with the inversion of the glycolytic reactions during germination.     

Contributo alla Conoscenza dell'idratazione e della Pressione Osmotica Nelle Specie di Tre Associazioni Forestali delle Marche: I° - L'idratazione

Abstract

Researches on the water content and osmotic pressure in plants of three forest associations in the Marche. I° - The water content. — The main forest associations of the Umbro-Marchigiani Appennines are the Orneto-Quercetum ilicis, the Quercetum pubescentis (s. l.) and the Fagetum. In each of these associations a typical station has been chosen where the most important species have been followed from March to November 1963 in order to determine the seasonal variations of their water content (« T.I. »)expressed as percentage of fresh weight.

The annual curves show that the water content reaches very high levels in the early stages of plant development while it falls to lower levels later in the season when the leaves are full grown. The water content of each species follows a different course and can be considered a specific character. Taking into account the maximum and minimum levels of water content of the three groups of species a « hydric spectrum » has been worked out for the three associations.

The water content of each species has also been calculated as percentage of dry weight (imbibition percentage: « G.I. »).     

Higher-plant cofactor-independent phosphoglyceromutase: purification,molecular characterization and expression

Cofactor-independent phosphoglyceromutase (PGM) was purified to homogeneity from developing castor seed endosperm. Immunological characterization using monospecific antisera raised against this protein indicates that the enzyme is located in the cytosol and that there is no immunologically related polypeptide in the leucoplast from this tissue. Isolation and sequence determination of full-length cDNA clones for castor and tobacco PGM demonstrate that the protein is highly conserved in these plants and is closely related to the maize enzyme. A comparison of the amino acid sequence of peptides derived from Neurospora crassa PGM with the cofactor-independent enzyme from higher plants demonstrated that they are related and may have diverged from a common ancestral gene. The previously proposed relationship between higher-plant PGM and alkaline phosphatases is not supported by sequence analysis of the castor and tobacco enzymes. Expression of the single castor cytosolic PGM gene correlates well with other cytosolic glycolytic genes in developing and germinating castor seeds, and with the appearance of enzyme activity and PGM polypeptides in these tissues.Institute of Cell and Molecular Biology