Ricerche Sull'infrastruttura del Coleottile di Avena

Abstract

Researches on ultrastructure of Avena coleoptile. 3. The sieve elements. — A study on the ultrastructural organization of the mature sieve elements of Avena coleoptile has been carried out. Data suggest that functional phloem tubes are alive and remain alive until they are working. Judging on morphological basis, the metabolic activity of sieve elements should be of peculiar type and low in comparison to that of the companion cells. In fact the cytoplasm is located in a narrow parietal strand, mitochondria, Golgi apparatus and endoplasmic reticulum are present, but they appear very modified; plastids and nucleus are absent. The cytoplasm is bounded externally by a normal plasmalemma, whilst the vacuole has no visible limits: a tonoplast is, therefore not identifiable.

The strands connecting the superimposed sieve elements with one another through the sieve plate result to be made by a double membrane system very similar to the endoplasmic reticulum, which we believe to realize cytoplasmic continuity between phloem tubes.

The data reported are more favorable to the existence in the sieve tubes of an active mechanism of translocation of organic solutes than a passive mass-flow.

The collaboration of companion cells in the translocation mechanism has been discussed.     

Ultrastruttura della cellula apicale di Halopteris scoparia (L.) Sauvageau (Phaeophyceae,Sphacelariales)

Abstract

Ultrastructure of the apical cell of Halopteris scoparia (L.) Sauvageau (Phaeophyceae, Sphacelariales). - The ultrastructure of resting apical cells of Halopteris scoparia (L.) Sauvageau from material collected in December is described. The cytoplasm is higly vacuolated with lipids, poliphenolic substances and polisaccharides occurring inside the vacuoles (the classic « physodes »).

Two cell organelles are prominently active at this stage: conspicuosly hypertrophic dictyosomes and the budding endoplasmic reticulum. Both light and electron microscope observations show that the cell wall has an outer stratification and inner discontinuous thickenings, the constituent material of which is uniformerly dispersed.

The above observations point out that the apical cell of Halopteris scoparia at this stage of its life cycle is engaged in an elaboration of vacuolar and parietal substances which will be therefore readly available at the outset of the growing season.     

Fissazione di CO2 da Parte di Entomocecidi di Cuscuta Australis

Abstract

Fixation of carbon dioxide by galls of CUSCUTA AUSTRALIS. — The mechanism of the carbon dioxide fixation by Cuscuta is a controversial biochemical topic. Light induced reactions are involved in MacLeod's opinion while others as Ciferri and Poma believe that mainly a dark-fixation occurs.

In this study use is made of the galls, caused by the insect Smicronyx on Cuscuta australis, that appear more green coloured than the normal tissues.

Equal weights of excised galls were kept both in light and in darkness in contact with C¹⁴O₂for different incubation times, and the magnitude of the fixation was compared under these two conditions by measuring radioactivity of both soluble and acid hydrolysed fractions.

After short exposures to the tracer the fixation in light greatly exceeds that in dark (25–19/1) while with more prolonged exposures the ratio sharply decreases (to about 5/1): these figures can be interpreted with the assumption that a strong light-induced fixation superimposes itself to a low but definite dark-fixation activity.

The ratio does not change if radioactivity is measured in the hydrolysed fractions.

These results are of course referred to as being largerly preliminar and requiring further and more extensive studies.     

Osser Vazioni Anatomiche su Foglie di Olivo Sane ed Attaccate da Spilocaea Oleagina (Cast.) Hugh.

Abstract

Anatomical observations on sound leaves and by SPILOCAEA OLEAGINA (Cast.) Hug infected leaves in Olive-tree. — The author has studied the anatomical characters comparatively between two cv. of Olive tree growing in Messina district (Sicily): cv. Ogliarola, receptive to Spilocaea oleagina (Cast.) Hug, and cv. Leccino not receptive.

Anatomical differences of the leaf as a whole and particularly of the «cuticular membrane » have been observed between sound leaves and attacked leaves of cv. Ogliarola, and sound leaves of cv. Leccino.

The main conclusions drawn from this study are the following ones:

The mesophyll is thicker in cv. Ogliarola and thinner in cv. Leccino.

The cuticular membrane and its 4 layers are thicker in Ogliarola leaves than in Leccino's ones.

Ogliarola leaves attacked by the fungus show a cuticolar membrane thicker than the sound leaves of the same cultivar.

Hiperplastic and hipertrophic tissues have been observed in Ogliarola leaves attacked by Spilocaea, expecially epidermis and palisade.

The anatomical causes of resistence and receptivity are prospected.     

Frammenti di Quereeto a Roverella e di Macchia Mediterranea a Vivara (Isole Flegree)

Abstract

Fragments of oak-crop with QUERCUS PUBESCENS willd. and of mediterranean maquis in the isle of vivara (Flegrean isles). — In these note the types of arboreous and shrubby vegetation pointed out in the islet of Vivara are passed in rapid review. Particular references to the types of vegetation arranged in the Quercion ilicis are given.

A short account is reserved to the presence of Quercus pubescens Willd. and to the importance of these species concerning the construction of the fragments of oak-crop and of mediterranean high maquis that are present to the edges of tableland and on the sides of the smallest Flegrean isles.

It is a question of unusual aspects of vegetation for a microinsular environment. Therefore they are worthy of study before they are destroyed or still more altered.     

Fenomeni di Inattivazione e di Riattivazione Enzimatica in Endospermi di semi di Ricino

Abstract

Inactivation and riactivation of enzymes in endosperms of castor bean seeds. — On the basis of previous results, the possibility has been investigated of the reversible interconversion of active and inactive form of enzymes in castor bean seeds, during their development.

The results described here indicate that:

1. the activity of some glycolytic enzymes increases greatly (81% and 400% increase of, respectively, Gl-6-P-dehydrogenase and aldolase) upon incubation of dry seeds for few hours at 4 °C.

2. The decrease of enzyme activity upon dehydration of seeds and the increase during the subsequent imbibition can be shown reproducibly.

3. This same observation is made for oxygen uptake.

These results are interpreted to indicate the reversible inactivation of enzymes caused by dehydration of seeds.     

ANATOMICAL FEATURES OF METAPHLOEM IN STEMS OF SABAL,COCOS AND TWO OTHER PALMS

Sieve tubes in metaphloem of palm stems function throughout the life of the plant and merit close investigation. A stem of Sabal palmetto estimated to be 50 years old was sampled extensively. Variation in length of sieve-tube elements throughout this stem was measured and is discussed. In the metaphloem of individual vascular bundles companion cells are not sharply differentiated from other phloem parenchyma cells. Definitive callose deposits and slime are normally absent from mature sieve tubes, even in fixed material. Otherwise no conspicuous structural features which might account for the longevity of sieve tubes can be discerned. Occlusion of phloem strands after leaf fall is initially by callose deposition on sieve plates followed immediately by tylosoid formation. Similar sampling of Cocos nucifera, Washingtonia robusta and to a lesser extent Archontophoenix alexandrae confirmed these results except for quantitative differences.     

Translokation und biochemisches Verhalten von D- und L-Phenylalanin bei Vicia faba

Summary D,L-Phenylalanine (phe) applied to the first primary leaf of a young Vicia faba plant moves into the sieve tubes and appears in the honey dew of aphids feeding on the third primary leaf. Ethanol extracts of the treated leaf contain labeled phe and an acidic compound which could be identified as N-malonyl-D-phe.D-phe-l-¹⁴C was obtained pure by enzymatic decarboxylation of the L-isomer of commercially available D,L-phe-l-¹⁴C, using an enzyme from red algae (Hartmann, 1967). The uptake of the D-isomer of phe by the sieve tubes is independent of the age of the treated leaf. L-phe applied to a young leaf is completely incorporated into protein; L-phe taken up by an older leaf is translocated in considerable amounts.Pulse-labeling with the two isomers shows that D-phe entering the sieve tube system is quickly removed to the parenchyma where it is acylated with malonic acid to the phloem immobile N-malonyl-D-phe. L-phe does not react with malonic acid at all. It is translocated to the centers of protein synthesis.

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Herrn Prof. Dr. Maximilian Steiner zum 65. Geburtstag gewidmet.     

Ricerche Sulla Cultura Sommersa di Cellule Singole di Piante Superiori

Abstract

Research on submerged culture of single cells of higher plants. — The author describes a method which allows to obtain submerged cultures of single cells of Phaseolus vulgaris and Nicotiana tabacum. The medium composition in macroelements in the culture on agar appears to effect to a great extent the ability of tissues to dissociate into single cells in the subsequent liquid culture. In this respect Heller's solution results to be more suitable than Gautheret's and Hildebrandt and Ri-ker's.

Cells are grown at 24 [ddot]C in 300 ml flasks containing 60 ml of broth on a rotary shaker at 220 rpm.

To prevent contaminations some antibacterial agents were added to cultures of Phaseolus vulgaris. Among these Penicillin and Neomycin were not tossic at 20 and 5 ppm concentrations respectively.

The presence of septa, which are observed also in largely vacuolate cells, seems to confirm the ability of single cells to divide.

The optimum 2,4-D concentration for growth decreases from 6 × 10^-8 to 6 × 10^-8 during successive liquid cultures, each of them being inoculated with on amount of the previous one. This fact, showing the adaptation of liquid cultures to decreasing concentrations of the growth hormone, is in agreement with previous observations in solid cultures by several authors.     

Azione di Nucleosidi e Relative Basi su Alcuni Aspetti Fisiologici di Rhizobium Leguminosarum Frank

Abstract

On the action of some nucleosides and related bases on some physiologiacal aspects of RHIZOBIUM LEGUMINOSARUM Frank. — Research was carried out on the action of some nucleosides and related bases on some physiological aspects of Rhizobium leguminosarum Frank. The compounds tested were: adenine, adenosine, cytosine, guanine, guanosine, hypoxantine, inosine, thymine, thymidine, uracil, uridine.

Pyrimidinic nucleosides, apart from the disappearance of ribose, behave similarly to their bases.

The bases and nucleosides tested stimulate changes in the respiration of R. leguminosarum. Remarkably different values are observed in the endogenous respiration when compared to the control, and a remarkable delay in the beginning of respiration in the case of cells precultivated in presence of cytidine, uridine and thymidine. A number of these cells were later found to have an anaerobic metabolism, at least in the starting period. These findings may be explained admitting that the regulatory system of Rhizobium, with regard to oxygen uptake, are dependent on the presence of certain compunds, including some nucleosides, which the microorganism is in contact with within the nodule.

Apart from cytidine sulphate, there is a fundamental stimulatory effect on the growth of R. leguminosarum. After 24 hours incubation, purinic bases and their nucleosides are not detectable in cultural media. Thymine and cytosine, however, are still present in the medium, or have been transforned into uracil; the latter undergoes no transformation in the medium     

Structurally,phloem unloading in the maize leaf cannot be symplastic

Developing longitudinal vascular bundles of the leaf blades of maize (Zea mays L. cv. W273) were examined with the transmission electron microscope to determine the frequency of plasmodesmata between the sieve tubes and their neighboring cells. Of particular interest were the protophloem sieve tubes, the first sieve tubes to mature in importing (all large and some intermediate) bundles. The protophloem sieve tubes, most of which lack companion cells, intergrade structurally with the thin-walled metaphloem sieve tubes. Both the protophloem sieve tubes and the thin-walled metaphloem sieve tubes and their companion cells (the sieve tube-companion cell complexes) are virtually isolated symplastically from the rest of the leaf, precluding a symplastic mechanism of phloem unloading in the leaf blade of maize.     

Sul trasporto dei glucidi nelle piante superiori: I. — Aspetti biochimici

Abstract

CARBOHYDRATE TRANSLOCATION IN HIGHER PLANTS. I. - BIOCHEMICAL ASPECTS. — The concentration of soluble sugars and of hexose phosphates and the activity of several enzymes involved in hexose activation and polysaccaride synthesis have been investigated, separately, in the phloematic tissue and in the medullar parenchyma of Cucurbita Pepo internodes.

In the phloematic tissue (including sieve tubes, companion cells and phloematic parenchyma) the concentration of free hexoses appeared of about 50% lower, and that of glucose-6-P and of sucrose of about 100% higher then in the medullar parenchyma. Consistent amounts of raffinose were found only in the phloematic tissue. Paper chromatograms of the sieve tube exudate showed the presence of raffinose and sucrose in a ratio close to unity, and no appreciable amounts of free hexoses.

Determination of enzyme activity on preparations obtained from homogenates from the two types of tissue by repeated ammonium sulfate precipitation showed in the phloematic tissue a high activity of the enzymes hexokinase, UDP-kinase, UDPG-pyrophosphorylase and inorganic pyrophosphatase. The presence in the same tissue of galactosekinase, UDP-Gal-pyrophosphorylase and UDPG-epimerase was also ascertained.

On a protein basis, the activity of UDPG-pyrophosphorylase, inorganic pyrophosphatase and hexokinase appared about 3 times higher in the phloematic tissue than in the parenchyma; while this difference between the two tissues was not so marked for phosphofructokinase, and very small for other enzymes such as ATP-ase and phosphomono-esterase.

These results suggest that the very high activity, in the phloem cells neighbouring the sieve tubes, of the enzyme system catalyzing oligopolysaccaride synthesis could be an important component of the mechanism involved in the accumulation of oligopolysaccarides in the sieve tubes, and thus in sugar translocation. A scheme is proposed according to which the ATP and UTP energy would be utilized by the phloem cells to reach and to maintein a concentration of soluble sugars consistently higher than that prevailing in the contiguous tissues.     

Ricerche Comparate Sulla Morfologia e Sulla Fisiologia di Larix e di Picea

Abstract

Comparative researches on morphology and physiology of PICEA and LARIX. The chlorophyll content of seeds and seedlings during germinations in darkness. — Very different are the chlorophyll contents of the seeds and the seedlings of Picea excelsa and Larix decidua grown in darkness.

In Picea chlorophyll is abundantly synthetized in cotyledons in the passage from seed to seedling stages. In Larix however this synthesis is very poor. These differences are more evident after the outgrowth of cotyledons from the primary endosperm and its exhaustion.

If seedlings of both species are exposed to 3.000 lux of light intensity for 24 hours, after development in darkness, one can observe much stronger chlorophyll synthesis in Larix cotyledons than icea.

These figures quanti atively express the different light-dipendence of chlorophyll synthesis in these two plants and offer a more adherent interpretation of the different ecological behaviours that are caracteristic of these two plants.     

A QUANTITATIVE STUDY OF SIEVE-TUBE DIFFERENTIATION IN VEGETATIVE SHOOT APICES OF COLEUS

Quantitative methods, with round-the-clock collecting of large samples in successive years, have uncovered several new phenomena of sieve-tube differentiation in young leaves of Coleus vegetative shoots. In small leaves (1–350 μ), there are no sieve tubes in the leaf itself, but they differentiate acropetally in the two traces to each leaf. Regression lines fitted to the data for leaf length vs. most distal position of sieve tubes in the traces support the view that differentiation is steady and acropetal, but they also reveal that differentiation in the traces falls steadily farther behind elongation of the leaf. Leaves more than 500 μ long have sieve tubes close to their tips. An intensive search of leaves of intermediate lengths revealed an isolated locus of sieve-tube differentiation. These relationships were reproducible year after year. Every plant with discontinuous strands of sieve tubes in the second leaf pair had discontinuous xylem in the third. This isolated locus was not seen before, probably because of small samples and daytime collections; most of our cases were from night collections.     

Osservazioni sulla microflora fungina di ceppaie di abete bianco

Abstract

Mycoflora of Silver fir (Abies alba Mill.) stumps. - This work is a first contribution to the knowledge of the mycoflora colonizing Silver fir (Abies alba Mill.) stumps in Italy.

Autochthonous fungal species were isolated from the wood of A. alba stumps from three different sites in the Tuscan Apennines.

Frequency values for the isolated species and taxonomic groups were calculated.

Deuteromycetes (57–82.5%) occurred more frequently than basidiomycetes (5–36%), phycomycetes (0.7–11.6%) and ascomycetes (0–1%). The most common genera were found to be: Aleurodiscus, Penicillium, Phellinus, Phialophora, Scytalidium, Sistotrema and Trichoderma.

An effective method of isolation from wood is also described.     

Microautoradiographic studies of phloem loading and transport in the leaf of Zea mays L.

Microautoradiographs showed that [¹⁴C]sucrose taken up in the xylem of small and intermediate (longitudinal) vascular bundles of Zea mays leaf strips was quickly accumulated by vascular parenchyma cells abutting the vessels. The first sieve tubes to exhibit ¹⁴C-labeling during the [¹⁴C]sucrose experiments were thick-walled sieve tubes contiguous to the more heavily labeled vascular parenchyma cells. (These two cell types typically have numerous plasmodesmatal connections.) With increasing [¹⁴C]sucrose feeding periods, greater proportions of thick- and thin-walled sieve tubes became labeled, but few of the labeled thin-walled sieve tubes were associated with labeled companion cells. (Only the thin-walled sieve tubes are associated with companion cells.) When portions of leaf strips were exposed to ¹⁴CO₂ for 5 min, the vascular parenchyma cells-regardless of their location in relation to the vessels or sieve tubes-were the most consistently labeled cells of small and intermediate bundles, and label (¹⁴C-photosynthate) appeared in a greater proportion of thin-walled sieve tubes than thick-walled sieve tubes. After a 5-min chase with ¹²CO₂, the thin-walled sieve tubes were more heavily labeled than any other cell type of the leaf. After a 10-min chase with ¹²CO₂, the thin-walled sieve tubes were even more heavily labeled. The companion cells generally were less heavily labeled than their associated thin-walled sieve tubes. Although all of the thick-walled sieve tubes were labeled in portions of leaf strips fed ¹⁴CO₂ for 5 min and given a 10-min ¹²CO₂ chase, only five of 72 vascular bundles below the ¹⁴CO₂-exposed portions contained labeled thick-walled sieve tubes. Moreover, the few labeled thick-walledsieve tubes of the transport region always abutted ¹⁴C-labeled vascular parenchyma cells. The results of this study indicate that (1) the vascular parenchyma cells are able to retrieve at least sucrose from the vessels and transfer it to the thick-walled sieve tubes, (2) the thick-walled sieve tubes are not involved in long-distance transport, and (3) the thin-walled sieve tubes are capable themselves of accumulating sucrose and photosynthates from the apoplast, without the companion cells serving as intermediary cells.     

Plasmodesmatal distribution and frequency in vascular bundles and contiguous tissues of the leaf ofThemeda triandra

Small and intermediate vascular bundles and contiguous tissues of the leaf blade ofThemeda triandra var.imberbis (Retz.) A. Camus were examined with transmission and scanning electron microscopes to determine the distribution and frequency of plasmodesmata between various cell types. Plasmodesmata are most abundant at the mesophyll/bundle-sheath cell and bundle-sheath/vascular parenchyma cell interfaces, and their numbers decrease with increasing proximity to both thick- and thin-walled sieve tubes. Among cells of the vascular bundles, the greatest frequency of plasmodesmata occurs between vascular parenchyma cells, followed by that of plasmodesmata between vascular parenchyma cells and companion cells, and then by the pore-plasmodesmata connections between companion cells and thin-walled sieve tubes (sieve tube-companion cell complexes). The sieve tube-companion cell complexes of theT. triandra leaf are not isolated symplastically from the rest of the leaf and, in this respect, differ from their counterparts in theZea mays leaf. However, the thick-walled sieve tubes, like their counterparts inZea mays, lack companion cells and are symplastically connected with vascular parenchyma cells that about the xylem.Abbreviations SEM scanning electron microscope - TEM transmission electron microscope     

Alcuni Aspetti Citologici Dell'Endosperma di Phaseolus Coccineus L.

Abstract

Some cytological aspects of Phaseolus coccineus L. endosperm. — Cytological observations were made on the endosperm of Phaseolus coccineus, in a limited stage of development of the seed, using different dyes and labelling with ³H-thymidine and ³H-uridine. The results seem to indicate that, contrary to suspensor cells, in the endosperm of Phaseolus, at least in the considered stage, extra DNA synthesis is not present. Chromosomes in these nuclei, however, undergo several endoreduplication cycles. Nucleoli may be either one or more in a cell, and show a characteristic structure. They are often eccentric in the nucleus and extrusions of nucleolar content in the nucleoplasm or cytoplasm are seen. An apparent alternate activity of RNA synthesis in the nucleoli of the suspensor and in those of endosperm cells is discussed.     

Microtuboli in Cellule Parenchimatiche ed Epidermiche di Coleottile di Avena

Abstract

Microtubules in parenchyma and epidermis cells of avena coleoptile. — The fine structure of differentiating parenchyma and epidermis cells of the oat coleoptile, fixed in glutaraldehyde-osmium tetroxide, or glutaraldehyde-potassium permanganate, was investigated. Tubular structures have been observed aligned in the peripheral cytoplasm, between the cell wall and the plasma membrane, embedded in the cell wall and inside the tonoplast in the vacuoles.

The nature and function of these structures are yet unknown. Microtubular structures, localized beneath and above the plasma membrane, have been associated to the wall development; the function of the microtubules observed in vacuoles results, anyhow, of far problematic interpretation.     

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.