Assorbimento di saccarosio in ipocotili di ravanello (Raphanus sativus L.)

Abstract

In questo lavoro abbiamo determinato le caratteristiche di entrata del saccarosio in funzione della sua concentrazione esterna, incubando i pezzi in saccarosio triziato, in condizioni normali e di stress osmotico. Nelle parti apicali l'assorbimento di saccarosio esogeno è lineare in presenza di basse concentrazioni mentre non lo è ad alte concentrazioni. Per quanto riguarda le parti basali la linearità si mantiene per tutte le concentrazioni; inoltre nelle parti apicali l'assorbimento è maggiore che nelle basali e la differenza aumenta all' aumentare delle concentrazioni di saccarosio esterno. Abbiamo poi studiato l'assunzione di saccarosio con osmotici diversi in cui fosse presente anche saccarosio marcato. I risultati ottenuti dimostrano che le parti apicali stressate prendono meno saccarosio dei controlli, mentre nelle basi stressate l'assorbimento, pur essendo inferiore a quello degli apici, è superiore rispetto ai controlli. Le differenze nel trasporto del saccarosio negli apici e nelle basi potrebbero essere spiegate dal fatto che a parità di peso nelle zone apicali ci sono più cellule di dimensioni inferiori che non hanno completato la distensione cellulare e che essendo in attiva crescita sono metabolicamente più attive. Nelle zone basali è invece presente un maggiore numero di cellule morte (xilema completamente differenziato). Questi due fattori potrebbero determinare nelle parti apicali un maggiore assorbimento con prevalenza del trasporto attivo e nelle parti basali un assorbimento inferiore con prevalenza del trasporto per diffusione.     

Nucleolar Organizer ultrastructure in Allium cepa

The Nuoleolar Organizer Region (NOR) was studied in Allium cepa. The cells analyzed were: meristematic root tip cells, cells from the layers of the anther (exothecium, intermediate layer cells, endothecium and tapetum) and meiocytes from preleptotene to diptotene. Conventional electron microscope techniques and serial section were used in the study. Ultracytochemical tests, using preferential techniques for nucleic acids were also applied. The results show that: a) Although the NOR may have a very variable form in the different cells analyzed, in each case its fine structure is similar, being constituted of chromatin differentiated in high and low density zones. b) In each case, its cytochemical characteristics are similar; highly positively to EDTA (preferential stain for RNP) being observed in the low density zones. As scattered chromatin these would constitute the active regions of the NOR. c) In certain cases a clear relationship is seen between NOR volume and the size of the nucleolus. d) The chromatin associated to the NOR always appears as very dense zones. e) The NOR-Nucleolus relationships are established exclusively with the latter's fibrillar pars. f) The nucleolar segregation phenomenon takes place sometimes as a result of a NOR contraction and in other cases depends on the position of the NOR in relation to the nucleolus. g) In preleptotene, the NOR shows its characteristic morphology and is positive to EDTA, while from zygotene to diplotene the structure of the NOR presents no differences from the rest of the chromatin and only the lateral elements of the synaptonemal complex or its remnants are positive to EDTA. h) The synaptonemal complex runs through the NOR with no differentiation and on occasions its lateral elements show associations with the nucleolus.     

Synaptonemal-like complexes in Allium cepa Microspores

The paper describes the synaptonemal-like complexes found in the course of microsporogenesis in Allium cepa, during the young microspore stage. In longitudinal sections these structures are morphologically more or less like the synaptonemal complexes, which appear during the pachytene stage of meiosis, and consist of two regions resembling the lateral elements, separated from one another by a space measuring about 1000 Å, in which a central element is occasionally observed. When observed in transverse or oblique sections, their shape indicates that they are tubular structures with a central axis; they are generally observed as independent units inside the nucleus, sometimes associated with the chromatin masses.By using the uranyl-EDTA-lead staining method, which picks out the RNA the synaptonemal-like complexes, show up at the level of their lateral elements and a similar result can be achieved by using the alcoholic PTA technique, which is believed to be a selective staining method for histones. We suppose that the synaptonemal-like complexes consist, at least partially, of RNP material.     

Sister chromatid exchanges in Allium cepa

Differential staining of sister chromatids with Giemsa after BrdU incorporation into DNA was performed in Allium cepa L. chromosomes. A treatment solution containing 10^–7 M FdU, 10^–4 M BrdU and 10^–6 M Urd was found to ensure BrdU incorporation without affecting cell cycle duration. After several procedures before staining the slides with Giemsa had been tested, treatment with the fluorochrome compound 33258 Hoechst, exposure to UV light and heating at 55° C in 0.5×SSC, were found to be essential for good differentiation. The distribution of SCEs per chromosome agrees with the expected Poisson distribution. The mean value of SCEs per chromosome occurring when cells were exposed to the treatment solution for two consecutive rounds of replication (=5.5) was double the mean value observed when cells were exposed to the same treatment for only one round of replication (=2.8). SCEs were found to occur more frequently in those chromosome regions corresponding neither to C-bands nor to late replicating DNA-rich regions. Finally, the occurrence of SCEs involving less than the width of a chromatid is discussed.     

Nuclear matrix network in Allium cepa

The presence of a nuclear matrix network in animal cells has been claimed by many workers in the last decade. The purpose of our study was to see whether a similar structure could be identified in the cells of higher plants. Our work revealed the presence of a fibrillar nonchromatinic network in Allium cepa nuclei. This could be impregnated with AgNO₃ in intact cells as well as in isolated matrices at the light microscope level. It was seen to be associated with the chromosomes from early to late prophase and also in telophase. Ultrastructurally a fibrillar network comparable to that reported earlier from animal cells was observed. This network remained associated with metaphase and anaphase chromosomes and could be digested with pepsin. Biochemical estimation of the isolated matrix revealed it to be made up mainly of proteins (more than 90%), traces of DNA (less than 1%) and a small quantity of RNA. SDS PAGE showed three polypeptide bands in the molecular weight range of 55,000–63,000 daltons.     

BAC FISH analysis in Allium cepa.

Onion (Allium cepa L.; 1C=15,000 Mb) is an agriculturally important plant. The genome of onion has been extensively studied at the conventional cytogenetic level, but molecular analyses have lagged behind due to its large genome size. To overcome this bottleneck, a partial bacterial artificial chromosome (BAC) library of onion was constructed. The average insert size of the BAC library was about 100 kb. A total of 48,000 clones, corresponding to 0.32 genome equivalent, were obtained. Fluorescent in situ hybridization (FISH) screening resulted in identification of BAC clones localized on centromeric, telomeric, or several limited interstitial chromosomal regions, although most of the clones hybridized with entire chromosomes. The partial BAC library proved to be a useful resource for molecular cytogenetic studies of onion, and should be useful for further mapping and sequencing studies of important genes of this plant. BAC FISH screening is a powerful method for identification of molecular cytogenetic markers in large-genome plants.     

Uno studio citofotometrico delle proteine durante la mitosi nel meristema radicale di Allium cepa L.

Abstract

A cytophotometric study of proteins during mitosis in root tip meristematic cells of «Allium cepa L.». — Spectrophotometric analyses of the amount of Fast-green stainable proteins at different pH values (8,1; 6; 4; 2) have been accomplished in Allium cepa root tip meristematic cells during the four phases of mitosis. The results seem to indicate that: a) the highest absorption is detectable in correspondence of metaphase at each of the four pH values; b) the transition from prophase to metaphase is characterized by an increase of both proteins reacting at pH 8,1 (histones) and between pH 6 and pH 8,1 (neutral proteins); c) the transition from metaphase to anaphase is characterized by a loss of histones and of proteins reacting between pH 4 and pH 6 (acid proteins); d) the transition from anaphase to telophase is characterized by a loss of neutral proteins. The data are discussed in relation to the problem of chromosome coiling.     

Azione della Cinetina sulla Degradazione della Clorofilla in Foglie di Orzo Irradiate

Abstract

The effects of kinetin on chlorophyll breakdown in irradiated barley leaves. — Kinetin is shown to inhibit the breakdown of chlorophyll in isolated barley leaves. Moreover the kinetin is shown to inhibit, even if with lower effect, the chlorophyll breakdown in irradiated barley leaves. This possible correlation of these observations, with kinetin promoted protein synthesis is suggested.     

Costanza numerica delle cellule di purkinje del cervelletto dell'uomo in individui di varia eta'

Zusammenfassung Es wurde an mikrotomischen Schnitten des Kleinhirns die Zahl der Purkinjeschen Zellen pro mm Länge bei 10 Menschen von verschiedenem Alter (von 6–88 Jahren) berechnet; für jeden Fall wurden etwa 16000–18000 Zellen berechnet. Im Gegensatz zu den Ergebnissen von Ellis habe ich keine Abnahme der Zellenzahl mit fortschreitendem Alter beobachtet; die Variationen der Zellenzahl waren unbedeutend und standen jedenfalls keineswegs in Beziehung zu dem Alter; bei 6jährigen Kindern waren die erhaltenen Werte etwas höher, d. h. waren die Zellen dichter; dies ist erklärlich, wenn man bedenkt, daß bei Kindern trotz der geringeren Größe des Kleinhirns die Zahl der Neuronen ebenso groß ist als beim Erwachsenen.     

Actin in the preprophase band of Allium cepa

F-actin has been identified in the preprophase band of Allium cepa. Cells attached to subbed slides were obtained from formaldehyde-fixed root tips digested in EGTA and Cellulysin. The air-dried cells were extracted in Triton X-100, treated with rhodamine-phalloidin, rinsed briefly in PBS, and viewed in the fluorescence microscope. Interphase cells contain a network of actin fibers that extends into all areas of the cytoplasm. During preprophase, the network is replaced by a band of fibers aligned in the position of the preprophase band. Colocalization of F-actin with rhodamine-phalloidin and microtubules with tubulin immunocytochemistry confirms that the two bands are coincident. The actin appears to comprise a thin layer of fibers next to the plasmalemma. Like the microtubule preprophase band, the actin band narrows as preprophase progresses and disappears by midprophase. Fluorescent actin bands are not seen in fixed cells pretreated with excess unlabeled phalloidin before staining. They are also absent in roots exposed to cytochalasins B and D before fixation, but preprophase band microtubules at all stages of aggregation are still present. Colchicine treatment leads to the loss of both preprophase band microtubules and actin. The possible function of preprophase band actin is discussed.     

Localizzazione intracellulare della clorogenico ossidasi in tessuti di tubero di patata in fase di attivazione

Abstract

INTRACELLULAR LOCALIZATION OF CHLOROGENIC ACID OXIDASE IN AGED POTATO TUBER DISCS. — The localization in the cells of chlorogenic acid oxidase has been investigated in potato tubers and discs of potato tuber. It has been ascertained that the rise of activity per gram of tissue, after preparation of discs, is not due to bacterial or fungal growth. The activity is widely distributed among cell fractions. Some activity is found in mitochondria, while most of the activity is distributed among soluble fraction and a « microsomal » fraction sedimented by centrifugation in the range 15.000–50.000 x g. This fraction appears to contain mitochondrial fragments, fragments of the endoplasmic reticulum and ribosomes. When tuber discs are aged, the rise of chlorogenic acid oxidase activity is much larger in the soluble than in particulate fraction.     

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.     

Extraction and identification of different prostaglandins in Allium cepa

Green onions (Allium cepa) were homogenized in a blender and extracted by normal extraction methods except that diethyl ether was used as the first extracting solvent. Different analytical procedures were used for the identification of the prostaglandins separated. TLC was applied using silica gel 60 F254 plates and a mixture of benzene, dioxane and acetic acid (20:10:1) as eluent, and the Rf values were compared with those of authentic samples. GC analysis on an SE 30 packed column and FID was applied; relative retention times of the onion extract components were measured and matched with authentic prostaglandin samples using cholesterol as an internal standard. GC-MS analyses using the same conditions adopted for GC analysis were conducted on a Finnigan MAT 112S instrument. Four peaks were identified. The prostaglandins identified were F1 alpha, E1, B1 and A2.     

Synaptonemal complex spreading in Allium cepa and A. fistulosum

The general features and fine structure of homologous chromosome alignment and pairing have been investigated in two species of Allium (A. fistulosum and A. cepa), which have similar karyotypes but very different patterns of chiasma distribution. Although there is no support for the occurrence of a general pre-meiotic alignment of homologous chromosomes, both species show some alignment of homologues as an immediate prelude to synaptonemal complex (SC) formation. In both species pairing usually commences at sub-terminal sites and is succeeded by numerous separate intercalary initiations of pairing in interstitial and distal regions and then in proximal regions. The last parts to pair, in both species, are pericentromeric and telomeric regions. There is, therefore, no evident relationship between the sequence of pairing and chiasma distribution in these species. Regularly alternating convergences and divergences of aligned axial cores (ACs), termed multiple association sites, are frequently observed. It is proposed that these represent potential pairing initiation sites and from observations on their spatial distribution it is argued that they may be evenly distributed through most of the genome. Small spherical or ellipsoid nodules are found at association sites and between closely aligned ACs which persist in the SC segments present during zygotene, but most of them disappear abruptly at the end of zygotene. These are termed zygotene nodules (ZN) and it is proposed that they are involved in matching corresponding sites on homologous chromosomes as well as possibly having a recombinational role. Their composition, structure, mode of action and relationship to pachytene recombination nodules are at present unknown.