Flora Palinologica Italiana,Sezione Aeropalinologica: S 205 -Pinus pinea L. (Pinaceae)

Summary According to the program Palynological Italian Flora, Aeropalynological section, the pollen morphological card ofPinus pinea L. is presented. The study is carried out on pollen coming from three Italian localities and regards fresh and acetolyzed pollen. For each sample, measurements are carried out on 30 fresh pollen grains in glicerol jelly with fuchsin and on 30 acetolyzed pollen grains in water/glicerol (1/1); general observations regard 1000 fresh and 1000 acetolyzed pollen grains/sample. Some observations on the main differences between fresh and acetolyzed pollen are mentioned.

---

Riassunto Nell'ambito della Flora Palinologica Italiana, Sezione Aeropalinologica, è presentata la scheda morfopalinologica diPinus pinea L. nella versione su polline fresco e polline acetolizzato, su tre campioni di diversa provenienza. Vengono notate le principali differenze tra polline fresco e polline acetolizzato.

     

Comparative studies of Feulgen hydrolysis for DNA

Summary We compared the Feulgen hydrolysis curves (37° C, 5 M HCl) of human blood lymphocytes fixed by the following four methods: 96% ethanol, 60 min at 20° C; ethanol-acetone, 11, 120 min at 4° C; ethanolglacial acetic acid mixture (31), containing 2% of formaldehyde (EAF), 90 min at 20° C; and ethanol-glacial acetic acid (31), 60 min followed by 5% chrome alum solution for 360 min at 20° C. The best results were obtained with EAF-fixation, with respect to the highest amount of DNA-Schiff complex at the peak point of the curve, the longest plateau region and the smallest scattering of DNA-Schiff amount values along the plateau. With other types of fixation the addition of polyethylene glycol (PEG) 6,000 to the hydrolysis solution resulted in modification of the shape of the hydrolysis curve so that it became nearly similar to the EAF-curve. The effect of PEG₆₀₀₀ on the EAF-curve was minimal. Slides fixed by ethanol were used for a comparison of polyethylene glycols with m.w. 1,500, 6,000 and 20,000. The longest plateau was obtained with PEG₆₀₀₀. The only effect of PEG₁₅₀₀ was a dramatic increase of DNA-Schiff amount at the peak point. PEG₂₀₀₀₀ had no significant effect on the shape of the hydrolysis curve. The results are discussed in terms of Kjellstrand's chain with a stable structure model of Feulgen hydrolysis.     

Iron sulfur proteins of the purple sulfur bacterium Ectothiorhodospira shaposhnikovii

In addition to several cytochromes three iron sulfur proteins were detected in mixotrophically grown cells of Ectothiorhodospira shaposhnikovii, a member of the Chromatiaceae. They were identified as a bacterial ferredoxin and two high potential iron sulfur proteins (HIPIPs). The two HIPIPs were purified and characterized. They were named according to their differing retention times on a DEAE-cellulose column using a continuous NaCl gradient: early and late HIPIP. The HIPIPs contain 4 mol of non-heme iron and 4 mol of acid labile sulfur per mol protein. Under the conditions of purification the early HIPIP (E _{m, 7}+270 mV) was present in a semi-reduced state. Using ion-exchange chromatography the early HIPIP could be split into a reduced green-brown (pI=3.7) and an oxidized red-brown (pI=3.9) fraction. The late HIPIP (pI=3.8) showed a midpoint potential of only+155 mV, the lowest redox potential of a HIPIP described so far.Non-common abbreviations HIPIP high potential iron sulfur protein - MOPS 3(N-morpholino)propane sulfonate - SDS sodium dodecylsulfate     

Circadian changes in protein-synthesis rate and protein phosphorylation in cell-free extracts of Gonyaulax polyedra

The polysomal pattern of the dinoflagellate Gonyaulax polyedra, cultured under constant conditions, demonstrates a circadian rhythm. The relative amount of polysomes increases during the phase corresponding to the previous night period (=subjective night phase) when the rate of protein synthesis reaches its maximum (Cornelius et al., 1985, Planta 160, 365–370). Cell-free extracts were isolated at different circadian phase. The rate of protein synthesis in the extracts changed rhythmically in the same manner as the rate of protein synthesis in vivo. Substances in the postribosomal supernatants influenced the protein-synthesis rate of the cell-free system, depending on the phase when they were isolated: night factors stimulated protein synthesis in day extracts whereas day factors inhibited protein synthesis in night extracts. These effects were abolished by heating the postribosomal supernatant. In-vitro phosphorylation in parallel probes showed changes in the pattern of phosphorylated proteins. Phosphorylation of one of the proteins (95 kDa) was decreased after addition of night factor(s) and increased after addition of day factor(s). Cyclic-AMP enhanced the rates of protein synthesis and phosphorylation in the day extracts.Abbreviations cAMP cyclic-AMP - CT circadian time - D (N) subjective day (night)phase     

Lipid profile of rat myelin subfractions

Myelin from adult rat brains was separated on a discontinuous sucrose gradient into three subfractions. Analysis of light, heavy and membrane fraction lipid classes was performed by HPTLC and densitometry while fatty acid composition was determinated by GLC. The more interesting results observed are: i) the membrane fraction resembles in its lipid and fatty acid composition other cell membranes (particulary oligodentrocytes); ii) light and heavy myelin are quite similar between them but the former has a higher content of sphingomyelin, a lower hydroxy/nohhydroxy cerebrosides ratio and a lower content of monoenoic fatty acids than the heavy subfraction. The results obtained could explain the different structures observed in each myelin subfraction since fatty acid composition, hydroxy fatty acids, sphingomyelin and cholesterol play a key role in the stability and structure of membranes.     

Analysis of rho mutability in Saccharomyces cerevisiae

Summary Two additional types of nuclear determinants involved in the control of spontaneous mutability of rho in S. cerevisiae have been identified: mmc and the pet-ts 1, 2, 10, 52 and 53 genes.These genes in their mutated recessive form increase at various extents the number of respiratory deficient cytoplasmic petite mutants accumulated.The gene mmc does not affect the respiratory activity and is not temperature-dependent whereas the pet-ts genes determine at the non permissive temperature a respiratory deficient phenotypes even if they affect the mutability of rho at the permissve and at the non permissive temperature.The data here reported suggest that a replicative complex exists for the mitochondrial DNA.It is in the purpose of this paper to deal with the relative contrition that mmc and pet-ts gene products have in ensuring the fidelity of this replicative complex.     

Psychological changes associated with self-regulatory treatments of irritable bowel syndrome

The psychological side effects of self-regulatory treatment (a combination of relaxation, thermal biofeedback, and cognitive therapy) for irritable bowel syndrome (IBS) were compared among 20 successfully treated patients, 12 unsuccessfully treated patients, and 9 patients who merely monitored symptoms for 12 weeks. Pretreatment and posttreatment scores on the Beck Depression Inventory, State-Trait Anxiety Inventory, and Psychosomatic Symptom Checklist were examined. Successfully treated patients had significant (p<.01) reductions on all measures and significantly greater reductions on depression and state anxiety than the symptom monitoring group. Interestingly, the failures also showed a significant (p=.027) reduction in trait anxiety and no significant increases on other measures.     

Abbau der 4-Chlorbenzoesäure durch eineArthrobacter-Species

Zusammenfassung EineArthrobacter-Species, die 4-Chlorobenzoesäure als einzige Kohlenstoffquelle verwerten kann, gibt beim Wachstum auf dieser Verbindung 4-Hydroxybenzoesäure und Protocatechusäure ins Medium ab. Der weiter Abbau des aromatischen Ringes erfolgt durch meta-Spaltung. Beim Wachstum derArthrobacter-Species auf Benzoesäure trit im Medium cis,cis-Muconsäure auf. In diesem Fallewird also der ortho-Weg eingeschlagen. Die Enzyme für beide Abbauwege sind induzierbar.

---

Degradation of 4-Chlorobenzoic acid by anArthrobacter species

AnArthrobacter sp. growing on 4-Chlorobenzoic acid as its sole source of carbon excretes 4-hydroxybenzoic acid and protocatechuic acid into the culture medium. Protocatechuic acid is further attacked by meta-cleavage. During growth of theArthrobacter sp. on benzoic acid cis-cis muconic acid can be isolated from the medium, suggesting the involvement of the ortho-cleavage pathway. The enzymes both for the meta- and the ortho-cleavage pathway are inducible.

Dem Andenken an Professor Bernhauer gewidmet     

Regeneration of fertile plants from protoplasts of different Arabidopsis thaliana genotypes

Summary A protocol for obtaining regenerated fertile plants from mesophyll protoplasts of Arabidopsis thaliana is reported. Protoplasts were isolated from leaves of 21-to 28-day-old Arabidopsis plants grown in a controlled environment. Sustained divisions were achieved when protoplasts were embedded in beads formed by 1.4% sodium alginate in the presence of 50mM CaCl₂ in 0.4 mannitol, which was then exchanged againts modified B5 medium. About 0.4%–0.6% of the protoplasts developed into colonies of which 80%–90% formed shoots and subsequently regenerated to fertile plants. Seeds harvested from more than 200 independently regenerated plants were sown and germination frequencies of more than 95% were obtained. Furthermore, the F₁ plants did not show any evidence of somaclonal variation on visual inspection. This protocol was originally developed for Arabidopsis thaliana Columbia; however it was shown to be applicable also for the genotypes Wassilewskija, Landsberg erecta and Estland though with differing efficiencies.Abbreviations FDA fluorescein diacetate - CM culture medium - SRM shoot regeneration medium - SEM shoot elongation medium - RM rooting medium - PE plating effciency - NAA -naphthaleneacetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - BAP 6-benzylaminopurine - Kin kinetin - 2-iP 2-isopentenyladenine - GA₃ gibberetic acid     

Meiotic analysis ofElymus caucasicus,E. longearistatus,and their interspecific hybrids with twenty-threeElymus species (Triticeae,Poaceae)

Interspecific hybridizations were carried out between the two tetraploidsElymus caucasicus andE. longearistatus, and 23 tetraploids and hexaploids ofElymus containing SH, SY, SYH, and SYW genomes and representing various geographical regions. Meiotic pairing was studied in the two target species and their hybrids. It is concluded from this study that (i) interspecific hybridization is fairly easy to perform although strong reproductive barriers exist between the species; (ii)Elymus caucasicus andE. longearistatus are allotetraploids, and share the diverged SY genomes; (iii) the divergence of SY genomes is correlated with the geographic distance between theElymus spp. studied.     

Retention of lead within the digestive vacuole inTetrahymena

Summary The ciliateTetrahymena pyriformis was exposed to lead acetate. Cell proliferation in the presence of 0.1% lead salt (with or without EDTA) equaled, after a variable lag period, that of the control cells. The lead (550 ppm) forms a fluffy precipitate with the organic growth medium; this was in part prevented by addition of EDTA. The cells primarily ingested the fluffy precipitate whereby they became exposed to large amounts of lead. Within the digestive vacuole, the fluffy precipitate became converted into refractile structures (3 m in diameter) which were egested and accumulated at the bottom of the culture flask. The lead content of these defecation balls was higher than that of the fluffy precipitate. In addition to the lead-containing vacuoles, the cells contained small, refractile granules. The apparent, high tolerance ofTetrahymena towards lead is believed to be due in part to the low ionic concentration of lead under the present conditions and in part to a detoxication mechanism consisting of retention of lead within the digestive vacuoles and perhaps of accumulation of lead within the small, refractile granules.     

Mutants of Arabidopsis thaliana with altered phototropism

Thirty five strains of Arabidopsis thaliana (L.) Heynh. have been identified with altered phototropic responses to 450-nm light. Four of these mutants have been more thoroughly characterized. Strain JK224 shows normal gravitropism and second positive phototropism. However, while the amplitude for first positive phototropism is the same as that in the wild-type, the threshold and fluence for the maximum response in first positive phototropism are shifted to higher fluence by a factor of 20–30. This mutant may represent an alteration in the photoreceptor pigment for phototropism. Strain JK218 exhibits no curvature to light at any fluence from 1 mol·m^-2 to 2700 mol·m^-2, but shows normal gravitropism. Strain JK345 shows no first positive phototropism, and reduced gravitropism and second positive phototropism. Strain JK229 shows no measurable first positive phototropism, but normal gravitropism and second positive phototropism. Based on these data, it is suggested that: 1. gravitropism and phototropism contain at least one common element; 2. first positive and second positive phototropism contain at least one common element; and 3. first positive phototropism can be substantially altered without any apparent alteration of second positive phototropism.Abbreviation WT wild-type     

„Kolbenbildungen“ der Muskelfasern des menschlichen Kehlkopfs

Zusammenfassung Fasern der menschlichen Kehlkopfmuskulatur weisen seitliche Aussackungen auf, die unter dem Sarkolemm in lockerer Anordnung Kerne, Lysosomen, Lipofuscingranula und Mitochondrien mit oft konzentrisch angeordneten Cristae enthalten. Sie sind mit den lichtmikroskopisch bekannten End- bzw. Seitenkolben identisch. Diese Aussackungen werden von den gefäßnahen subsarkolemmalen Mitochondrienanhäufungen eines bestimmten, auch in der übrigen Skelettmuskulatur weitverbreiteten Muskelfasertyps abgeleitet. Es wurden keine Verbindungen mit Nervenfasern gefunden. Im Gegensatz zu Goerttler können wir diese Bildungen deshalb nicht als Mechanorezeptoren deuten. Die Endkolben werden als sekantial getroffene Seitenkolben aufgefaßt. Da die Aussackungen der Fasern auch bei gesunden jungen Männern in der Kehlkopfmuskulatur regelmäßig auftreten, handelt es sich hier nicht um eine krankhafte Bildung, wenn auch die Faser im Bereich der Aussackungen einen degenerativen Prozeß durchzumachen scheint.

---

Summary Fibers of human larynx muscles show lateral dilatations of the sarcolemma containing loosely disposed mitochondria, lysosomes and eventually one or more nuclei. The cristae of the mitochondria often are arranged concentrically. Corresponding fiber dilatations have already been described by light microscopists as side- resp. end-bulbs.Our electron-microscopical observations lead to the conclusion that the dilatations are derived from accumulations of mitochondria beneath the sarcolemma which in similar form — but smaller — are characteristic of one type of the different muscle fibers, frequently to be found in other muscles of the body. Both subsarcolemma accumulations of mitochondria are close to the muscle capillaries.The fiber dilatations have no connections with nerve axons, and therefore cannot be interpreted as mechanoreceptors (Goerttler, 1960). The end-bulbs are supposed to be secantially (or obliquely) cut side-bulbs. We do not consider them pathologic since they are regularly found in fibers of healthy young men. On the other hand, they may represent a circumscribed degenerative alteration the origin of which remains to be explored.

     

Mitochondrial genetic damage induced in yeast by a photoactivated furocoumarin in combination with ethidium bromide or ultraviolet light

Summary Ethidium bromide (EB) and ultraviolet light (UV) in combination are known to produce a synergistic induction of petite mutants in yeast. Two other agents were combined with EB, 3-Carbethoxypsoralene (3 CPs) activated by 365 nm light or rays. EB in combination with 3 CPs also resulted in an enhanced production of petite mutants. After the photoaddition of 3 CPs in exponential phase cells, recovery of the petite mutation during dark liquid holding was inhibited by the presence of EB producing an enhanced number of petite mutants. The behavior of mitochondrial antibiotic resistance markers after individual and combined treatments with EB and 3 CPs indicates a random loss of markers after EB and a preferential loss of a certain region for the 3 CPs photoaddition. The combination of the two agents leads to an additivity of total drug marker losses rather than a synergistic loss. The combination of EB with rays produced no enhancement in petite induction. A combination of UV and 3 CPs showed a synergistic interaction for petite induction. These results indicate that the three agents, EB, UV and 3 CPs photoaddition may share a common repair step for mitochondrial lesions.     

Further cytogenetic analyses of the Croatian triploid shallot “Ljutika” (Allium cepa var.viviparum, Alliaceae) and its comparison with the Indian triploid “Pran”

Feulgen and silver-stained karyotypes and meiosis of two triploid viviparous onion forms (Allium cepa var.viviparum), the Croatian Ljutika and the Indian Pran, were comparatively analyzed. The results of chromosome measurements show that Ljutika and Pran are karyologically not identical, although significant similarities were found in the morphology of their chromosomes. Five geographically distant clones of Ljutika showed good agreement in the number and gross morphology of the chromosomes and in the number and position of NORs and interphase nucleoli. Heterotrivalents were predominant in meiosis of Ljutika but a relatively high frequency of higher multivalents together with univalents and bivalents were also observed. The relationship between Ljutika and Pran and their possible origin are discussed.     

Silver staining the chromosome scaffold

Cytological silver-staining procedures reveal the presence of a core running along the chromatid axes of isolated HeLa mitotic chromosomes. In this communication we examine the relationship between this core and the nonhistone chromosome scaffolding, isolated and characterized in previous publications from this laboratory. When chromosomes on coverslips were subjected to the steps used for scaffold isolation in vitro and subsequently stained with silver, the characteristic core staining was unaffected. Control experiments suggested that the core does not contain large amounts of DNA. When scaffolds were isolated in vitro, centrifuged onto electron microscope grids, and stained with silver, they were found to stain selectively under conditions where specific core staining was observed in intact chromosomes. These results suggest that the nonhistone scaffolding is the principal target of the silver stain in chromosomes.     

Vibrations in the orb web of the spider Nephila clavipes: cues for discrimination and orientation

Transmission of natural and artificial vibrations in webs of Nephila clavipes was examined using laser Doppler vibrometry to determine how this spider discriminates and localizes stimuli. 1. Vibration signals of four entrapped insect species peaked at different frequencies from 5–30 Hz, but their spectra overlapped considerably. Peak amplitudes spanned 50 dB. 2. Transmission of longitudinal vibrations along individual radii was attenuated over ca. 12 cm by 4.0 ± 2.7 dB; attenuation values for transverse and lateral vibrations were 22.2 ± 4.6 dB and 26.2 ± 4.3 dB, respectively. Some transmission spectra characteristics may be explained by resonances of the spider and threads. 3. Radial thread transmission increased by 2.2–5.8 dB after cutting the connecting auxiliary spirals, demonstrating that vibrations leak from stimulated radii via these threads. Auxiliary spirals provide structural support to Nephila webs at the expense of degraded directional transmission. 4. Upon single-point stimulation, vibrations measured around the web hub and at the spider's tarsi revealed 2-D vibration amplitude gradients of 20–30 dB indicating the stimulus direction. In contrast, measured vibration propagation velocities of 70–1500 m/s resulted in time-of-arrival differences at the spiders tarsi of < 1.5 ms, which may be too brief for stimulus direction determination.Abbreviations A area - C propagation velocity - E Young's modulus - LDV laser Doppler vibrometer/-metry - r.h. relative humidity - T tension - space constant - radius of gyration - density - 2-D two-dimensional - 3-D three-dimensional     

Somatic embryogenesis in quite a direct way in cultures of mesophyll protoplasts of Brassica napus L.

Somatic embryos and plantlets have been obtained in quite a direct way from mesophyll protoplasts isolated from androgenetic plants of two cultivars (Loras and Tower) of Brassica napus. The procedure consists of four steps. Proembryos were induced in a medium supplemented with a cytokinin and an auxin at comparatively high concentrations. They developed to mature embryos when the auxin was reduced or omitted.Abbreviations ABA abscisic acid - BAP 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - NAA 1-naphthaleneacetic acid Dedicated to Prof. Dr. Wilhelm Halbsguth on the occasion of his 70th birthday.On leave from the Institute of Genetic, Academia Sinica, Beijing, China     

Production of a novel syrup containing neofructo-oligosaccharides by the cells of Penicillium citrinum

A novel syrup containing neofructo-oligosaccharides was produced from sucrose (Brix 70) by whole cells of Penicillium citrinum. The efficiency of fructo-oligosaccharides production was more than 55% and those of the main carbohydrate components, 1-kestose (Fruf 21Fruf 21 Glc), nystose (Fruf 21Fruf 21 Fruf 21 Glc) and neokestose (Fruf 26 Glc12 Fruf), were 22, 14 and 11%, respectively.     

Methodological problems in evolutionary biology VIII. Biology and culture

Biology cannot accommodate all aspects of culture. Aspects of culture that a biological approach can take into account can be covered by the biological categories of phenotype and environment. There is no need to treat culture as a separate category. Attempts to elaborate biological explanations of cultural variation will meet with success only if biologists expand theories of development, and integrate them in evolutionary biology. The alternative — elaborating the idea of so-called cultural inheritance — makes little sense from a biological point of view.