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[Adaptation strategies of seed germination and seedling growth to sand dune environment].

Sand dune plants possess many adaptation strategies to withstand sand environment, e. g., some desert plant seeds are dormant when matured, which helps them to escape from unfeasible environment conditions and be preserved as seed bank. The seed germination and seedling establishment of psammophytes need moderate sand burial, while excessive burial will inhibit seed germination and seedling emergence. Seeds without germination in deeper sand are in enforced dormancy, and form soil seed bank. Sand dune plant seedlings could tolerant finite sand burial by increasing the number of nodes per culm and elongating internodes. When the seedlings are partially buried, they could survive through the maintenance of photosynthesis organism. Once sand burial exceed the threshold of the plant, seedlings growth will be restrained and the growth ability even permanently lost. Other factors such as salt spray, insect herbivory, and lack of soil nutrients also affect seed germination and seedling establishment. The precipitation in desert and sand land is unpredictable and irregular. Sand erosion leads roots be exposed to the air and dehydrated to die. However, seedlings of some desert plant have the ability to tolerate desiccation for a period of time after germination. Once there is rain, the seedlings will rivive.

Adaptation, Physiological↗

Maize DNA polymerase 2 is a phosphoprotein with increasing activity during germination.

DNA replication is a late event during maize germination and DNA polymerase activity increases as germination proceeds. A replicative alpha-type DNA polymerase has been purified from maize seeds (DNA polymerase 2) and has been shown to be a multisubunit complex [Coello, P., Rodríguez, R., García, E. & Vázquez-Ramos, J. M. (1992) Plant Mol. Biol. 20, 1159-1168; Coello, P., García, E. & Vázquez-Ramos, J. M. (1994) Can. J. Botany 72, 818-822]. DNA polymerase 2 activity increased several fold during maize germination, with no apparent change in either the amount of holoenzyme or in any of the individual subunits. However, the level of phosphorylation of the 90-, 70-, 55- and 45-kDa polypeptides changed in a cyclic fashion with their highest levels occurring at 11-14 h and 45-48 h of germination. Phosphate incorporated into the different polypeptides in the 11-14-h period remained stable for at least the next 10 h (to 24 h of germination), the period of maximal enzyme activity. However, DNA polymerase 2 processivity was very similar in freshly prepared 3-h and 24-h enzymes, and no evidence was found that polymerase activity was modified by in vitro phosphorylation. The significance of these results is discussed.

DNA Polymerase II↗

Bcl-x rather than Bcl-2 mediates CD40-dependent centrocyte survival in the germinal center.

Both rapid B-cell proliferation and programmed cell death (PCD) occur during the differentiation and selection of B cells within the germinal center. To help elucidate the role of Bcl-x in B-cell antigen selection and PCD within the germinal center, we examined its expression in defined B-cell populations and by immunochemistry of tonsil tissue. Purified B-cell fractions enriched for centrocytes express high amounts of Bcl-x and relatively low amounts of Bcl-2, whereas fractions enriched for centroblasts lack significant levels of both proteins. Consistent with this observation, immunocytochemistry localized Bcl-x within cells scattered throughout the germinal center. Stimulation of tonsil B cells with either CD40 or Staphylococcus aureus Cowan increase bcl-x mRNA and protein levels. Treatment of a cell line with a germinal center phenotype (RAMOS) or the tonsillar B-cell centroblast fraction with CD40 rapidly increased Bcl-x levels and partially rescued B cells from PCD. These data suggest that Bcl-x rather than Bcl-2 may rescue centrocytes during selection in the germinal center.

Apoptosis↗

Effect of germination on the protein content and on the level of specific activity of lipoxygenase-1 in seedlings of three soybean cultivars.

The effect of germination on the protein content and in the level of specific activity of lipoxygenase-1 on seedlings of Santa Rosa, FT-2 and Davis soybean cultivars were studied. Soybean seeds of the three cultivars were germinated from 0 to 72 hours in a seed germinator at 30 degrees C temperature, 100% relative humidity, and absence of light. The seeds were then frozen in liquid nitrogen, freeze-dried and milled. The lipoxygenase-1 was extracted with 0.02M thris-HCI buffer at pH = 8.2 and 13% saccharose for 30 minutes at 4 degrees C without shaking. The highest levels of specific activity of lipoxygenase-1 per milligram of flour (dry basis) in the Santa Rosa, FT-2 and Davis cultivars were observed between 18 and 36, 18 and 24, and 0 and 12 hours, respectively. The polinomial quadratic model best fits the regression analysis of the variable levels of specific lipoxygenase activity for cultivar Davis (R2 = 0.9197). Although for cultivars Santa Rosa (R2 = 0.9041)., and FT-2 (R2 = 0.7486) the 4th degree polynomial model has shown the best fit. The protein content in the three cultivars studied reached the maximum values at 48h germination. The germination induced a relative increase of the protein content but caused a reduction of the level of specific lipoxygenase-1 activity.

Germination↗

Oligoclonal development of B cells bearing discrete Ig chains in chicken single germinal centers.

Chicken single germinal centers enable us to analyze the postbursal diversifications of B cells due to their easy isolation. Germinal center formation has peaked by day 7 of primary responses and begins to wane 14 days after immunization. To detail the kinetics of Ig mutation and selection, we analyzed Ig light chain sequences recovered from single germinal centers at 7 and 11 days postimmunization with an artificial Ag. Our observations show that multiple, Ag-activated B cells migrating into single germinal centers are diversified by gene conversion in the very early phase of the germinal center reaction and are subsequently subjected to point mutations and selection for oligoclonality.

Amino Acid Sequence↗

[Antiphysiological and nutritional factor changes in sorghum (Sorghum bicolor (L.) Moench) seeds during germination].

The purpose of this work was to evaluate the effects of germination on the nutritional quality of two commercial varieties of low tannin content Sorghum: brown sorghum without testa (ICSY-CM89513) and white sorghum (ISIAP Dorado). After 24 hours of germination the condensed tannin concentration (catechin equivalent) was reduced 60% and 40% for brown and white sorghum respectively. However, tannin levels increased up 100% at 96 h germination. Phytic acid concentration decreased about 90% in 96 hours for both varieties. The lysine concentration increased up 110% (72 h germination) and 129% (48 h) for white and brown sorghum respectively. The thiamine, niacin and riboflavin contents increased 73, 200 and 353% respectively for brown sorghum in 72 h and 15, 44 and 93% respectively for white sorghum in 48 h. The "in vitro" enzymatic digestibility was increased 39.3% (72 h) for brown sorghum and 100% for white sorghum. The albumin concentration increased 80% and 74% (72 h) for brown and white sorghum respectively. The Calculated Protein Efficiency Ratio indicated nutritional improvements with germination. The sprouting was a practical and simple process for providing better nutritional properties in sorghum seeds to be used as human food.

Amino Acids↗

Inhibition of germination gene expression by Viviparous-1 and ABA during maize kernel development.

Two maize glyoxysomal genes expressed during germination, malate synthase (MS) and isocitrate lyase (ICL), were used to characterize the regulatory roles of the Viviparous-1 (Vp1) regulatory gene and abscisic aicd (ABA) in the induction of embryo quiescence during kernel development. In wild-type maize embryo, MS and ICL transcripts were first detected at 2 (MS) or 3 (ICL) days after germination (DAG), peaked at 5 DAG, and decreased thereafter. By reverse transcriptase-polymerase chain reaction (RT-PCR), the germination-specific genes were amplified in both ABA-insensitive (vp1) and ABA-deficient (vp7 and vp10) mutant embryos at 26 and 33 days after pollination (DAP), but not in wild-type embryos. The repression of these germination-specific genes thus requires the Vp1 gene product and normal levels of ABA to induce embryo quiescence during kernel development. This suggests that a genetic regulatory system exists to prevent vivipary in developing maize embryos. The involvement of the Vp1 gene product and ABA in repressing germination-specific genes complements their previously defined roles in the induction of seed-specific genes such as C1.

Abscisic Acid↗

Ovarian germinal epithelium and folliculogenesis in the common snook, Centropomus undecimalis (Teleostei: centropomidae).

The ovarian germinal epithelium in the common snook, Centropomus undecimalis, is described. It consists of epithelial and prefollicle cells that surround germ cells, either oogonia or oocytes, respectively. The germinal epithelium borders a body cavity, the ovarian lumen, and is supported by a basement membrane that also separates the epithelial compartment of the ovarian lamellae from the stromal compartment. During folliculogenesis, the epithelial cells, whose cytoplasmic processes encompass meiotic oocytes, transform into prefollicle cells, which become follicle cells at the completion of folliculogenesis. The follicle is a derivative of the germinal epithelium and is composed of the oocyte and surrounding follicle cells. It is separated from the encompassing theca by a basement membrane. The cells that form the theca interna are derived from prethecal cells within the extravascular space of the ovarian stroma. The theca externa differentiates from undifferentiated cells within the stromal compartment of the ovary, from within the extravascular space. The theca interna and the theca externa are not considered to be part of the follicle and are derived from a different ovarian compartment than the follicle. Meiosis commences while oocytes are still within the germinal epithelium and proceeds as far as arrested diplotene of the first meiotic prophase. The primary growth phase of oocyte development also begins while oocytes are still within the germinal epithelium or attached to it in a cell nest. The definitions used herein are consistent between sexes and with the mammalian literature.

Animals↗

Nodular lymphoma: an ultrastructural study of its relationship to germinal centers and a correlation of light and electron microscopic findings.

This study describes the ultrastructural findings in 16 cases of nodular lymphoma, and compares these with normal germinal centers in order to determine the possible germinal center origin of the lymphomas. Long branching desmosome-associated dendritic reticulum cells, characteristic of germinal centers, were found in all 16 cases of nodular lymphoma. Desmosomes were observed only between dendritic cells and not between lymphoid cells, and they were seen in only 1 of 7 cases of diffuse lymphoma. Cells comprising the nodular lymphomas share cytologic features, such as nuclear blebs and marked nuclear indentations, with germinal center cells. It appears that nodular lymphomas have a close anatomical relationship to germinal centers, although their actual origin from these sites is not established by this study. They are composed of lymphoid cells; we have no evidence to confirm the view that nodular lymphomas are derived from dendritic cells. Light and electron microscopic findings have been compared in order to establish the nature of large "histiocyte-like" cells in the nodular lymphomas. Many of these cells appear to represent transformed lymphocytes rather than histiocytes. They are more numerous in so-called mixed "histiocytic" lymphocytic lymphomas than in poorly differentiated lymphocytic lymphomas, which otherwise are ultrastructurally similar. This study casts doubt upon the existence of a mixed lymphocytic and histiocytic lymphoma within the spectrum of the nodular lymphomas, and indicates that a cytologic continuum exists between cells interpreted as poorly differentiated lymphocytes and those heretofore identified as histiocytes.

Adult↗

Migration of macrophages from the marginal zone to germinal centers in the spleen of mice.

Histological observations of the mouse spleen were carried out at different times after intravenous carbon injection. Large carbon-laden macrophages appeared in great numbers in the marginal zone soon after injection. They came together favorably around the germinal centers. Possible migration of these cells toward the germinal centers diffusely from the periphery of the white pulp or through the periarterial lymphoid sheath was suggested. These macrophages entered the germinal centers on a large scale and clustered for a long period--at least 180 days. Since the same type of macrophages were observed persistently in the marginal zone, it was thought that some of them might arise from the blood stream. Possible migration of these cells from the marginal zone toward the germinal centers was also persistently observed. A second type of much smaller carbon-laden macrophages was seen in the white pulp. However, they never showed any favorable localization in the germinal centers as did large carbon-laden macrophages.

Animals↗

Alterations in cerebellar germinal cell division induced by graft-versus-host disease.

A systemic immunological syndrome, graft-versus-host disease (GVHD), which does not cause inflammation or cell death in the cerebellum, is shown to retard granule cell production by decreasing the rate of DNA synthesis (S phase) and prolonging mitosis (M), at metaphase. The rate of cell production in diseased animals at postnatal day 14, quantitated by analysis of the rate of labeling of DNA with 3H-thymidine (3H-Tdr), revealed decreased ability to synthesize new DNA. The number of cells taking up 3H-Tdr label per mm2, as detected by autoradiography, was similar in 14-day-old GVHD and control tissue as was the area of the germinal matrix zone and the number of mitotically active germinal cells per mm2 in sagittal sections near the midline. However, because the total volume of the cerebellum was less, the total number of mitotically active cells in the whole cerebellum of 11-, 14-, and 17-day-old diseased animals was less than in littermate controls. Furthermore, DNA synthesis per mitotically active germinal cell was less in diseased animals at each age examined. The mitotic index was unaffected until late in the disease (day 17), suggesting that a prolongation of the cell cycle was responsible for this GVHD-induced decrease in DNA synthesis. Consistent with a prolongation of the cell cycle was the finding that the mitotic figures in 14-day-old GVHD cerebella were mostly metaphase figures, whereas those in control cerebella were, as predicted, mostly prophase. Prolongation of the cerebellar cell cycle in 11- and 14-day-old diseased animals may explain the dramatic decrease in the mitotic index, the thickness of the germinal matrix zone, and the number of germinal cells at postnatal day 17.

Animals↗

Tracing B cell development in human germinal centres by molecular analysis of single cells picked from histological sections.

Germinal centres are areas of intense B lymphocyte proliferation inside primary B cell follicles in spleen and lymph nodes. Rearranged V genes from single human B cells, isolated from histological sections of two such structures by micromanipulation, were amplified and sequenced. Cells from the follicular mantle were clonally diverse and largely expressed germline V genes. Germinal centres were dominated by a few large B cell clones dispersed throughout these structures and exhibiting intraclonal diversity by ongoing somatic hypermutation. Pronounced counterselection of replacement mutations seen in one of the germinal centres may indicate a late phase of the germinal centre reaction. A polyclonal population of activated B cells expressing unmutated antibodies in the dark zone of the other germinal centre may represent the initial founder cells.

Adult↗

Activation and expression of proteins during synchronous germination of aerial spores of Streptomyces granaticolor.

Synchronously germinating aerial spores of Streptomyces granaticolor were used to study protein activation and expression during the transition from dormant to metabolically active vegetative forms. The first phase of protein activation is associated with the solubility of proteins. Three major chaperones, DnaK, Trigger factor, and GroEL, were identified in spores. Enhancement in rate of protein synthesis during germination was accompanied by the association of TF and DnaK with ribosomes. During germination, the chaperones TF, GroEL, and DnaK undergo reversible phosphorylation. GroEL was phosphorylated on both Ser and Thr, whereas phosphorylation of DnaK and TF was detected on Thr only. A proteomic approach was used to gain more information on protein expression during germination on two types of media differing in the ability of cells to produce antibiotic granaticin. To obtain an overview of the metabolic activity of germinating spores, glycolytic enzymes, enzymes of citric acid cycle, metabolism of amino acids and nucleic acids, and components of the protein synthesis system were identified and analyzed using the proteomic database. The results were deposited on the SWICZ proteomic server and are accessible on http://proteom.biomed.cas.cz.

Chaperonin 60↗

Early events of Bacillus anthracis germination identified by time-course quantitative proteomics.

Germination of Bacillus anthracis spores involves rehydration of the spore interior and rapid degradation of several of the protective layers, including the spore coat. Here, we examine the temporal changes that occur during B. anthracis spore germination using an isobaric tagging system. Over the course of 17 min from the onset of germination, the levels of at least 19 spore proteins significantly decrease. Included are acid-soluble proteins, several known and predicted coat proteins, and proteins of unknown function. Over half of these proteins are small (less than 100 amino acids) and would have been undetectable by conventional gel-based analysis. We also identified 20 proteins, whose levels modestly increased at the later time points when metabolism has likely resumed. Taken together, our data show that isobaric labeling of complex mixtures is particularly effective for temporal studies. Furthermore, we describe a rigorous statistical approach to define relevant changes that takes into account the nature of data obtained from multidimensional protein identification technology coupled with the use of isobaric tags. This study provides an expanded list of the proteins that may be involved in germination of the B. anthracis spore and their relative levels during germination.

Amino Acid Sequence↗

Involvement of actin and polarisome in morphological change during spore germination of Saccharomyces cerevisiae.

We studied the morphological changes of Saccharomyces cerevisiae ascospores during germination. Initiation of germination is followed by polarization of actin patches, maintaining their localization to the site of cell surface growth. Loss of polarisome components, Spa2p, Pea2p, Bud6p or Bni1p, results in depolarization of actin patches. Green fluorescent protein-fused polarisome components exhibit the polarized localization, implying that polarisome is involved in the polarized outgrowth during germination. At the late stage of germination, we found that actin patches temporally depolarize before bud emergence. The observation that loss of Cla4p extends the polarized growth period suggests that Cla4p is involved in the actin-depolization step. Actin polarization in the initial stage is accelerated by overexpression of Ras2p, whereas hyperpolarization is continuously observed by overexpression of Rho1p. Thus, yeast spore germination is a morphological event that is regulated by a number of factors implicated in mitotic bud morphogenesis.

Actins↗

Glucose-induced pathways for actin tyrosine dephosphorylation during Dictyostelium spore germination.

In the presence of germination signals, dormant spores of Dictyostelium discoideum rapidly germinate to start a new life cycle. Previously we have shown that half of the actin molecules in spores are maintained in a tyrosine-phosphorylated state, and a decline of the actin phosphorylation levels is a prerequisite for spore swelling. In this study, we have established d-glucose as a trigger molecule for the actin dephosphorylation. Present in a nutrient germination medium, d-glucose both may act as a trigger molecule and/or may serve as a substrate within a pathway for actin dephosphorylation depending upon spore age. However, the glucose-induced actin dephosphorylation was insufficient for spores to swell. Other factors in the nutrient medium were required for complete germination of young spores aged 1 to 5 days. In contrast, dispersion in nonnutrient buffer was necessary and sufficient for a decline of actin phosphorylation levels and even the emergence of amoebae in older spores (6 days and beyond). Moreover, the dephosphorylation pathway in the older spores was independent of energy production. We propose that the diversification of the actin dephosphorylation pathway may enable spores to increase their probability of germination upon spore aging.

Actins↗

Microsporidian intrasporal sugars and their role in germination.

The hypothesis that spores of terrestrial and aquatic microsporidia differ in their utilization of sugars was tested by evaluating the sugars in germinated and ungerminated spores of several species in each category. The aquatic species tested were Vavraia culicis, Edhazardia aedis, and Nosema algerae and the terrestrial species were Vairimorpha necatrix, Nosema disstriae, Nosema apis, Vairimorpha lymantriae, and Nosema spp. from Spodoptera exigua and Plutella xylostella. The percentage germination varied between species, ranging between 40 and 92%. Total sugars (anthrone reactive) and reducing sugars (Nelson's test) remained unchanged through germination in the three terrestrial species tested; however, reducing sugars increased significantly in the aquatic species. High-performance liquid chromatography and gas chromatography revealed a preponderance of trehalose in all species and large quantities of sorbitol in all species except N. algerae and E. aedis. Other sugars were present in some species in much lower concentrations. After germination no changes in sugar content were observed in terrestrial species; however, all aquatic species lost trehalose with a concomitant increase in fructose and/or glucose concentrations. Increased osmotic potential from breakdown of trehalose has been postulated to induce germination of the aquatic species, but another explanation must be found for the terrestrial species.

Animals↗

Pathogenicity of Paecilomyces fumosoroseus isolates to diamondback moth, Plutella xylostella: correlation with spore size, germination speed, and attachment to cuticle.

Infectivity to larvae of the diamondback moth, Plutella xylostella, was compared among eight Paecilomyces fumosoroseus isolates. Isolate infectivity was assessed for correlation with spore length and germination speed. Four isolates applied to P. xylostella cuticle were also compared for number of spores remaining on the cuticle after washing and for percentage germination after 36 h. Infection of larvae inoculated with the different isolates at an average dosage of 4000 conidia/cm2 ranged from 2 to 47%. The correlation of infectivity with spore length and germination speed in broth was highly significant. Fewer spores of the least infective isolate, ARSEF 1576, attached to larval cuticle compared to spores of the more infective isolates ARSEF 3682, 4461, and 4482 (P < 0.05). After 36 h on larval cuticle, the percentage of spores germinated for isolates 1576 and 3682 was 3 and 95%, respectively. Spores of isolate 1576 were smaller, germinated more slowly, and attached to cuticle in smaller numbers than spores of the more infective isolates. Further research will expand our understanding of the mechanisms of virulence among isolates of P. fumosoroseus.

Animals↗