Changes in Endogenous Growth Kegulators in Loblolly Pine Seeds During the Process of Stratification and Germination

The seeds of loblolly pine (Pirns taeda L.) were cold-stratified for 0, 14, 28, 42, and 56 days. Endogenous growth regulators were extracted from these seeds, and also from the germinating and the 28-day warm stratified seeds. Partially purified extracts were separated on chromatographic paper. The chromatograms were cut into 10 equal segments, and these were tested for biological activity using three different btoassays. The results indicated that the unstratified seeds and those stratified up to a period of 28 days contained very little or no growth promoter (GA-like substances), and a relatively high concentration of an inhibitor (presumably abscisic acid), Following 42-day stratification, the promoter concentration gradually increased while the inhibitor level fell almost to zero. A high level of promoter but no inhibitor was detected in germinating seeds. No auxin-like activity was noted in the unstratified seeds. This activity slowly increased up to a period of 28 days and remained at this level for the subsequent stratification periods. However, the activity greatly increased in the germinating seeds. Very little changes in the levels of growth regulators were noted in warm-stratified seeds as compared to the unstrati-fied controls.     

Significance of a low oxygen layer for a Daphnia population in Lake Yunoko,Japan

Population dynamics and vertical migration of Daphnia longispina in Lake Yunoko were studied. The Daphnia population was small in spring and early summer, probably because of high predation pressure by fish. The population grew in midsummer, when thermal stratification developed and the dissolved oxygen became very low in the deeper layer of the hypolimnion. In this season, adults of D. longispina concentrated in the daytime near the lake bottom, where fish were absent because of the anoxic conditions, but ascended at night to the upper layer of the hypolimnion, where food was most abundant. The low oxygen layer near the bottom kept out the predators and protected Daphnia from predation, and consequently contributed to the built-up of its population. However, the low oxygen layer was unfavorable for reproduction of Daphnia, as reflected in the low egg ratio and high percentage of males in the population. The population decreased in the fall, when thermal stratification disappeared and predation pressure seemed to increase.     

Studies on the limnology and primary production of a tropical lake

Summary Limnology of Vihar lake has been studied for one year with reference to primary production. There is an unstable thermal stratification. Complete circulation is brought about by the wind. The thermal stratification does not follow the formulation ofBirge. There was a tendency for the D.O. to increase during late November 1964 to early January 1965. During the six weeks it was possible that a complete mixing is brought about. Carbon dioxide was more in the bottom water than in the surface water. The surface waters were found in general to be less alkaline in cooler months than in warmer months. The fluctuation in the alkalinity may be due to the photosynthetic activity of the algae. There is always a pH stratification, however, it was found to be unstable. The productivity test revealed that the compensation depth during the test period was 30. There was a little relationship between the phosphate and chlorophyll. However, the fluctuations in chlorophyll, pH, and D.O. seem to be interrelated. The nitrates were absent in the surface waters on almost all occasions. The fluctuation in the total iron was not striking. The sulphates were totally absent.Published with the permission of the Director, Central Public Health Engineering Research Institute, Nagpur, India.     

Placentas from spontaneously hypertensive rats and control strain Wistar-Kyoto rats

Placentas from spontaneously hypertensive rats (SHR) were compared to those of control strain Wistar-Kyoto rats (WKY) at 15, 18 and 20 days of gestation using light microscopic techniques. Placental lesions similar to those in pregnant hypertensive women were absent in both strains; however, other abnormalities were noted. Hemorrhage at the lateral edges of the decidua basalis appeared to be more extensive in the SHR than WKY at 15 days. At the same time, bloody vaginal discharges were noted in 18% of the SHR. Leukocytic encapsulation of 20-day placentas with viable fetuses was noted in two SHR dams but not in any WKY. It is thought that these differences may be related to the high maternal blood pressure in the SHR or to hormonal imbalance associated with the stress response in the SHR due to frequent monitoring of blood pressure.     

Pelagic Diatom Populations in Lentic Freshwater Microcosms

Three experiments, involving simultaneous monitoring of selected biological and chemical parameters in 50 l laboratory microcosms and the epilimnia of their parent reservoirs, were carried out from the autumn of 1978 to the winter of 1980. Experiments lasted 8-13 weeks, their aim being to ascertain degree of similarity between laboratory and field systems. Microcosm dynamics, specifically diatom dynamics, most closely paralleled that found in reservoirs during late spring and early summer, a time of thermal stratification. During winter months when thermal stratification was absent or less pronounced, microcosm diatom populations diverged significantly from reservoir populations within 24 days. It is inferred that microcosm design and operating conditions have a major bearing on microcosm usefulness for environmental assessment.     

Germination ecology of the clonal herb Knautia arvensis: Regeneration strategy and geographic variation

Abstract. We investigated germination responses and seed recruitment in the clonal grassland herb Knautia arvensis (Dipsacaceae) throughout its distributional range in Norway. Four predicted relationships between germination responses and field regeneration behaviour were tested using phytotron experiments and experimental and observational field studies. Seedlings appeared in all experimental microsites in the field, corroborating phytotron predictions that gap‐ or depth‐sensing strategies should be absent in the species. Seasonal timing patterns were predicted from a cold stratification response in the phytotron, but these were not supported in the field. The relationship between dormancy, germinability during storage, and seed carry‐over in the field largely conformed to expectations. Seeds from four different geographical regions responded differently to temperature and cold stratification and storage. Dormancy and seed carry‐over was higher in seeds from a coastal population, where winters are relatively mild and the probability of repeated freeze‐thaw events is high, than in populations from mountain and inland areas, where winters are colder. This is discussed against two alternative hypotheses about the relationship between climate and dormancy in seasonal climates.     

Proteins and polyamines during dormancy breaking of European beech (<Emphasis Type="Italic">Fagus sylvatica</Emphasis> L.) seeds

The studies were carried out on Fagus sylvatica seeds during stratification and their germination. After imbibition beechnuts were subjected to cold (3 °C — temperature which breaks dormancy) or warm (15 °C — temperature unable to break dormancy) stratification and alternatively were treated with polyamine synthesis inhibitors: canavanine and DFMO (difluoromethylornithine). After cold stratification in embryo axes we found (using 2-D electrophoresis) about 150 new proteins absent in dry seeds. Exogenous spermidine increased the protein synthesis, percent of germinated seeds and accelerated breaking of dormancy. In contrast, canavanine and DFMO decreased dynamic of protein synthesis, quantity of proteins probably synthesised de novo, and percent of germinated seeds. The maximum of polyamine content in embryo axes during cold stratification preceded such the maximum during warm stratification. Irrespective of the influence of PAs and inhibitors of PA synthesis, the comparison of electrophoregrams and autoradiograms showed that different groups synthesised de novo appeared after different periods of cold stratification. Probably the part of this protein is associated with Fagus sylvatica seeds dormancy breaking.     

Reproductive traits in Nubian dairy goats

For 12 months, ten Nubian doe goats were examined daily for oestrus and blood was obtained twice weekly for serum progesterone (P) analysis. Before the onset of regular ovulatory cycles in the autumn, brief increases in serum P were noted. Ovulation without oestrus was associated with the onset of the breeding season in five of 13 instances. Ovarian activity as assessed by corpus luteum (CL) function began in either September or October and ceased in either January or February in nonpregnant does. CL function was absent postpartum until the beginning of the next breeding season. An instance of male pseudohermaphroditism and mucometra is noted.     

Population dynamics of oxiclinal species in lake Arcas-2 (Spain)

Oxiclinal rotifer species show high concentrations just above the oxic-anoxic interface in the hypolimnion of some lakes. The stratification of their populations is best shown by sampling at close depth intervals and quantifying their densities by the Utermöhl technique. With this technique we were able to count males which otherwise pass through filters and more accurately count egg production. We evaluated female, male and egg numbers of the two main oxiclinal species of lake Arcas-2: Filinia hofmanni and Anuraeopsis fissa, during two annual cycles (1990–91). F. hofmanni was an exclusive oxiclinal species. It had an exponential growth phase at the onset of stratification giving a distinct spring peak. The population then maintained a high density during summer, but was almost absent the rest of the year. This cycle is repeated annually but population density can vary among years, depending on winter-spring circulation. Sexuality was always observed when the animal was present in the samples, with a maximum of males and resting eggs at the peak of the population. Resting eggs were always inside females. The annual cycle of A. fissa is displaced with respect to that of F. hofmanni: A. fissa attained greatest densities during summer, until the autumn overturn. Mixis in A. fissa was restricted to the end of the stratification period. Moreover, A. fissa occured throughout the vertical profile and secondarily occupied the oxicline.     

Seasonal and annual variability in the phytoplankton community of the Raunefjord,west coast of Norway from 2001–2006

Changes in phytoplankton community composition potentially affect the entire marine food web. Because of seasonal cycles and inter-annual variations in species composition, long-term monitoring, covering many sequential years, is required to establish a baseline study and to reveal long-term trends. The current study describes the phytoplankton biomass variations and species composition in relation to hydrographic and meteorological conditions in the Raunefjord, western Norway, over a 6-year period from 2001 to 2006. The extent of inflow or upwelling in the fjord varied from year to year and resulted in pronounced differences in water column stability. The annual phytoplankton community succession showed some repeated seasonal patterns, but also high variability between years. Two to four diatom blooms were observed per year, and the spring blooms occurring before water column stratification in March were dominated by Skeletonema marinoi and Chaetoceros socialis, and other Chaetoceros and Thalassiosira spp. Blooms of the haptophytes Phaeocystis pouchetii and Emiliania huxleyi were irregular and in some years totally absent. Although E. huxleyi was present all year round it appeared in bloom concentrations only in 2003, when the summer was warm and the water column characterized by high surface temperatures and pronounced stratification. The annual average abundance of both diatoms and flagellates increased during the six years. Despite the high variation from year to year, our investigation provides valuable knowledge about annual phytoplankton community patterns in the region, and can be used as a reference to detect possible future changes.     

Intratidal fluctuations in stratification within a mangrove estuary

The dependence of salinity stratification on the vertical structure of the longitudinal salinity gradient is investigated by solving the equations of longitudinal momentum and salt conservation. The results are used to interpret measurements made throughout 31 tidal cycles in the Merbok Estuary, Malaysia, as part of an ecological study of a tropical mangrove estuary. The solutions show that a substantial part of the salinity stratification was caused by vertical shear in the currents coupled with advection of the longitudinal salinity gradient. This mechanism led to maximum stratification around low water and minimum stratification around high water. Observed intratidal variations in stratification at neap tides greatly exceeded modelled values when a depth-independent, longitudinal salinity gradient was assumed. Simulations made using a longitudinal salinity gradient of simple depth-dependent, power-law form implied that this difference arose mainly because the near-surface, longitudinal salinity gradient in the field was significantly larger than that near the bed during neap tides and at times of high run-off. The longitudinal, dispersive transport of salt, fresh water and solutes due to vertical shear is shown to be important during periods of stratification at neap tides and during high run-off. The relevance of stratification to mangrove estuaries is discussed.     

Lipid composition of cultured human keratinocytes in relation to their differentiation

The present study was undertaken to explore the possibility of the use of cultured human keratinocytes for the study of changes in lipid composition in relation to epidermal differentiation. In a submerged culture system, in which the stratification is incomplete, no significant differences have been found between the lipid composition of cells grown either at low calcium concentration (0.06 mM) (at which the keratinocyte differentiation is markedly retarded) or at normal calcium concentration (1.6 mM) (at which some differentiation takes place). Under these conditions the amount of phospholipids and sterols was high and that of ceramides was low. Furthermore, the acylglucosylceramides (AGC) and acylceramides (AC), the latter one known to be involved in water barrier function, were found to be absent. Contrary to this, both AGC and AC were found to be present in significant amounts in an air-exposed model using de-epidermized dermis (DED) as a substrate (in which, as judged from morphologic criteria, the extent of keratinocyte stratification is similar to that seen under the in vivo conditions). Fatty acid analysis revealed significantly lower content of 18:2 and higher content of 16:1 and 18:1 acids with all culture conditions used, as compared to the parent epidermis. This is probably a result of fatty acid levels and composition in fetal calf serum (which was used in the present study) that differ markedly from the in vivo situation. The 20:4 content was similar to that in the epidermis only in cells cultured under the submerged conditions, during which they have been found (Isseroff et al. 1987. J. Lipid Res. 28: 1342-1349) to be able to convert 18:2 to 20:4. In DED cultures, however, the 20:4 content was markedly lower. Under all culture conditions used, the triglyceride content was higher as compared to the non-cultured epidermis. The high content of triglycerides and the fatty acid composition of the various lipid fractions showed a resemblance with what is found in the epidermis in essential fatty acid-deficient animals. This resemblance was confirmed by electron micrographs which revealed the presence of some partially or completely empty lamellar bodies. The results of the present study suggest that the air-exposed culture model, in which the keratinocytes show a high extent of stratification, could be of great value in the study of epidermal lipid metabolism. However, further alterations in culture conditions are necessary to more closely approximate the lipid composition of noncultured epidermis.     

Abnormal patterns of embryogenesis in Dirofilaria immitis treated with ivermectin

The percentage composition and spatial distribution of embryogenic stages in the uteri of female Dirofilaria immitis were examined at various times after treatment with a microfilaricidal dose of ivermectin and compared to nontreated parasites. Worms sampled 42 days post-treatment (PT) exhibited an increased proportion of stretched microfilariae in the distal portion of the uterus. A decreased proportion of developed embryos was noted in the mid body region of worms sampled 42 days PT, and these forms were completely absent from the proximal area of the uterus. Relative numbers and spatial distribution of other stages remained virtually identical to controls. Radical changes in the composition and spatial distribution of embryogenic forms were noted in the uteri of a single worm sampled 80 days PT. Unlike nontreated parasites and worms sampled 42 days PT, stretched microfilariae constituted the predominant form in the distal uterus of this worm, and these stages were found in decreasing numbers throughout the proximal segments. Also, the intermediate embryogenic stages were either rare or absent.     

Limnological studies of and primary production in temple pond ecosystems

Three temple ponds with permanent blooms of blue green algae were highly productive. They all showed high alkalinity, hardness, electrical conductivity and pH. Organic carbon and nitrogen were highest in Sarvatheertham pond—60 to 79.6 mg./l. C and 4.10 to 7.60 mg./l. N. In Tamaraikulam it was 16.5 to 20.3 mg. C/l. and 1.03 to 1.32 mg. N/l. In Sarvatheertham, the gross production ranged from 2.85 to 20.72 g. O₂/m.²/d. Self shading by blanket algae of blue greens reduced productivity in Sarvatheertham, where a persistent thermal and biochemical stratification was noted. Very high organic carbon and nitrogen contents were noted in Sarvatheertham pond. The dry weight of plankton in this pond ranged from 430 to 900 mg./l. Productivity computed from diurnal changes in alkalinity and dissolved oxygen also revealed a high rate in Ayyankulam, Tamaraikulam and Sarvatheertham in descending order. Very wide fluctuations in pH, both diurnally and depth-wise, were recorded in Sarvatheertham and to a lesser extent in the other two ponds. Photosynthetic efficiency was 4.03% in Ayyankulam, 2.09% in Tamaraikulam and 1.56% in Sarvatheertham. By the diurnal oxygen curve method, a gross primary production of 97.5 g. O₂/m.²/d was recorded in Ayyankulam.     

Diversity of the mitochondrial DNA cytochrome b gene of the field mouse Apodemus agrarius Pallas, 1771 in the south of the Russian Far East

We studied the variability of a fragment of the cytochrome b gene and the corresponding amino acid sequence of the field mouse Apodemus agrarius (Pallas, 1771) in the South of Primorskii krai (Far East of Russia). A significant polymorphism of these traits was detected. High indices of molecular diversity were obtained. The topology of the phylogenetic tree and the median networks suggest that the population of field mice in the south of Primorskii krai originated from three maternal lines. It was noted that clustering of haplotypes on a territorial basis is absent, and the field mouse population in southern Primorskii krai can be considered as one of the key points in maintaining a high genetic diversity of the species.     

Morphogenesis and proliferation in mono- and organotypic co-cultures of primary human periodontal ligament fibroblasts and alveolar bone cells

Cells of the periodontal ligament and the alveolar bone lie in close vicinity in the periodontium. The goal of this study was to create an in vitro model to facilitate the study of the morphogenesis and proliferation of these two cell types under more in-vivo-like conditions. This was accomplished by the generation of organotypic co-cultures of primary human periodontal ligament fibroblasts (PDL) and alveolar bone cells (BC) and matched mono-cultures after 1, 2 and 3 weeks. Indirect immunofluorescence (IIF) for vimentin indicated that PDL cells exhibited sustained stratification only in the presence of BC cells, suggesting an important role for BCs in maintaining the stratification of PDL cells. In mono-cultures, only BC cells showed progressing stratification. They also displayed the most pronounced contraction of the cell culture matrix. Moreover, Ki-67 antigen detection by IIF revealed that these features coincided with cell proliferation localized on the matrix surface at the onset of cell stratification. These findings suggest that, in addition to proliferation, a further prerequisite for stratification may be cell migration. Furthermore, the maintained cell stratification, proliferation, and compartmentalization noted for PDL cells in organotypic co-cultures and BCs in mono-cultures can only be observed in a three-dimensional culture system. Thus, our system represents a novel experimental tool to further elucidate the underlying mechanisms of the growth and differentiation of PDL and bone tissue.     

Changes in mRNA populations during loblolly pine (Pinus taeda) seed stratification, germination and post-germinative growth

Changes in the patterns of gene expression were examined during loblolly pine ( Pinus taeda L.) seed stratification, germination, and post-germinative growth. In both the megagametophyte and the embryo, DNA contents remained relatively constant at all stages examined. RNA contents, however, increased in both tissues following seed germination, particularly in the embryo where a 7-fold increase in the RNA content was observed 5 days after germination. Poly(A)⁺ RNA, extracted from megagametophytes and embryos, was translated in vitro in a rabbit reticulocyte lysate cell-free system. Analysis of [³⁵S]-methionine-labelled translation products by two-dimensional electrophoresis/fluorography indicated that there were changes in the populations of mRNAs during all developmental stages examined. In both the megagametophyte and the embryo several distinct mRNA populations, including one constitutively present at all stages examined, were identified. One mRNA population, present in the mature seed, decreased during seed stratification. Another population, not present in the mature seed, rose during the period of stratification that coincided with an increase in seed germinability. A third population, which appeared during seed germination, increased steadily during post-germinative growth. Besides these similarities, specific differences between megagametophyte and embryo were noted. For example, one mRNA population, which was present in the megagametophyte of the mature seed and remained constant during the stratification period, disappeared immediately following seed germination. In the embryo, one set of messages was germination specific. In total, these results show that mRNA populations change in a temporal fashion that is consistent with the patterns of de novo protein synthesis known to occur in loblolly pine during the same developmental periods.     

Changes in protein synthesis during stratification and dormancy release in embryos of sugar maple (Acer saccharum)

Protein synthesis in dormant embryos of sugar maple ( Acer saccharum ) was investigated in seeds stratified at 4°C or incubated at 15°C. Seeds stratified at 4°C germinated after 27 days; seeds incubated at 15°C failed to germinate. Stratification increased the embryo's capacity for protein synthesis by day 11 as measured by in vivo incorporation of [³⁵S]-methionine into purified protein. At 4°C protein synthesis in the embryonic axis rose in a linear fashion prior to germination, whereas in cotyledons it increased until day 20 and then declined. Analysis of radiolabelled proteins by two-dimensional gel electrophoresis revealed that the levels of specific proteins were altered by temperature, primarily in the cotyledons. Several proteins were expressed in the cotyledons at 15°C but were absent in unstratified embryos and in embryos stratified at 4°C. That is, the expression of these proteins was repressed during stratification and release from dormancy. Levels of other proteins in the cotyledons declined at 4°C during stratification. We suggest that one or more of these proteins may be associated with the inhibition of growth of the embryonic axis imposed by the cotyledons.