银杏外种皮提取物对病原菌﹙Cylindrocladium colhounii﹚的抑制作用

采用菌丝生长抑制法,研究了银杏外种皮3种不同提取物对病原菌（Cylindrocladium colhounii）的抑制作用并测定了这些提取物的MIC/MBC。结果表明,在相同实验天数内,抑菌效果（最小抑菌率）由高到低依次为银杏外种皮乙醇提取物（37.4%）、石油醚提取物（23.7%）和新鲜外种皮汁液（18.4%）。当提取物的浓度提升到一定程度时,三者的抑菌率均可达到100%;三种提取物的MIC/MBC分别为：86.25/86.25,172.5/276,293.25/345 mg·mL^-1。可见银杏外种皮具有明显抑制该病原菌生长的作用,且抑菌成分在乙醇中的溶解度较大。这为进一步研究该抑菌成分乃至生物农药的开发奠定了基础。     

Leaching of Ions, Organic Molecules, and Enzymes from Seeds of Peanut (Arachis hypogea L.) Imbibing without Testas or with Intact Testas

Reducing sugars, phosphates, potassium ions, total electricallyconducting material, proteins, and phenolics leached from twovarieties of peanut (Arachis hypogea L. ) seeds during 24 himbibition in distilled water. The same substances leached fromthe seeds when testas were removed before imbibition. The quantitiesof substances leached from seeds without testas after 24 h,except protein and phenolics, were much greater than from seedswith intact testas. Time courses of leaching of sugars, phosphates,potassium ions, and of total electrically conducting materialshowed fastest leaching rates in the first hour of imbibition.In the first 4 h of imbibition more sugars leached from intactseeds than from seeds without testas. Peroxidase was presentin leachates from intact seeds but was undetectable in leachatesfrom seeds without testas. Hydrogen peroxide-reducing activitywas detected in 24 h leachates from intact seeds and from seedswithout testas. The activity was partly inhibited by 1 mM ascorbate.It was concluded that catalase was present in the leachates. The peanut testas were thin. Their presence around the embryodid not reduce the rate of imbibition as compared with thatof seeds without testas. Thus the greater leaching of some substancesin the absence of the testa could not be ascribed to the absenceof a physical barrier to water uptake.     

Cytokinin Activity in Lupinus albus L: IV. Distribution in Seeds

Endogenous levels of cytokinin activity were examined in Lupinus albus L. seed at intervals of 2 weeks after anthesis using the soybean callus bioassay. High levels of cytokinin activity per gram seed material were present in the seeds at 2, 4, and 6 weeks after anthesis. The cytokinin activity per gram seed material was low at 8 and 10 weeks after anthesis. Cytokinin activity associated with each seed was greatest at 6 weeks after anthesis. The majority of the activity in the seeds at 4, 6, and 8 weeks after anthesis was in the endosperm. Cytokinin activity was also detected in the testas and embryos at 4, 6, 8, and 10 weeks, and the suspensors at 4 weeks. Column chromatography of extracts of the different seed fractions on Sephadex LH-20 indicated that the cytokinins present coeluted with zeatin, zeatin riboside, and the glucoside cytokinins. It is suggested that cytokinins are accumulated in the seeds and are stored in the endosperm mainly in the form of ribosides and glucosides of zeatin. The reduction in cytokinin activity in the seed coincides with the reduction in endosperm volume and embryo growth and suggests that these compounds are utilized during the course of seed maturation.     

The effects of salinity,and calcium and potassium variations on the growth of two estuarine red algae

The effects of varying salinity and calcium and potassium concentrations on the growth of two species of estuarine Rhodophyta, Bostrychia radicans Montagne and Caloglossa lepricurii (Montagne) J. Agardh, were examined in unialgal culture. Inocula of settled tetraspores on glass coverslips were incubated in six concentrations of potassium (0. 100, 200, 300, 400 and 500 mg/l) at five salinities (0, 5, 15, 25, and 35%) in the presence of calcium. Growth responses of each alga were determined from the average cell number of 75 tetrasporelings after 4 days in Ott's synthetic sea-water medium. The concentrations of dissolved potassium and calcium in sea water along a salinity gradient in an estuary were determined with atomic absorption spectrophotometry. Four-day-old tetrasporeling of Bostrychia and Caloglossa demonstrated growth over wide ranges of potassium concentrations with growth maxima at 400 500 and 200 400 mg/l, at the optimal salinities for growth of 25 and 15%, respectively. These studies indicate also that the presence of calcium is essential for appreciable growth of both species at each salinity and the effects of variations in potassium are dependent upon the presence of calcium. The abundance of both species in the Mullica River estuary. New Jersey, appears to be a response to a total osmotic effect along the salinity gradient because sufficient levels of the major cations are present. However, the upper limit of both species towards the head of the Mullica River estuary may be determined by combinations of low salinity and low levels of dissolved calcium and potassium rather than by low salinity alone.     

Isolation and characterization of biofilm-forming bacteria and associated extracellular polymeric substances from oral cavity

The current work deals with the studies on characterization of two biofilm-forming bacteria isolated from the oral cavity. The major constituent of biofilm other than bacterial cells is the extracellular polymeric substance (EPS) matrix, which is secreted by the bacterial cells themselves. Physical properties of biofilms such as attachment, mechanical strength, antibiotic resistance can be attributed to EPS matrix. Molecular phylogeny confirmed these two isolates as Pseudomonas aeruginosa and Bacillus subtilis. It was observed that cell attachment in both the strains was maximal when xylose was used as the sole carbon source. The EPS characterization result indicated the presence of a macromolecular complex constituting of carbohydrate, protein, lipids and nucleic acids. Test for biofilm formation in the presence of metal salts of iron and zinc showed moderate to high inhibition of biofilm formation. However, calcium, iron and copper have been found to enhance biofilm growth significantly. There was more than 50 % increase in biofilm growth by P. aeruginosa with an increase in calcium concentration up to 80 ppm (Two tailed t-test P?<?0.05), whereas ≥ 15 % increase in biofilm growth by B. subtilis was observed in the presence of 80 ppm of calcium. However, variations were significant (Two way ANOVA, P?<?0.01) between different metals in different concentrations. In this study, attempts have been made to examine the effect of different carbon sources and physiological conditions on biofilm growth.     

Calcium-Induced Alteration of Cellular Morphology Affecting the Resistance of Lactobacillus acidophilus to Freezing

Examination of factors affecting the resistance of Lactobacillus acidophilus NCFM culture concentrates to freeze injury induced during frozen storage at -20°C revealed that calcium supplementation of the growth medium contributed to the storage stability of cells prepared in static culture. Culture concentrates of L. acidophilus NCFM were prepared from cells propagated in MRS broth or MRS broth supplemented with 0.1% calcium carbonate, calcium chloride, or calcium phosphate. After 28 days of frozen storage at -20°C, concentrated cells (3.2 × 10⁹ colony-forming units per ml) prepared from MRS broth cultures showed an 84% reduction in viable cells. Of the remaining viable cells, 88% were sublethally injured and unable to form colonies on MRS agar supplemented with 0.15% bile. Cells prepared in calcium-supplemented MRS broths demonstrated more resistance to frozen storage. Viability and injury losses in the frozen concentrates were limited to 10 to 39% and 3 to 23%, respectively. It was observed that calcium supplementation of MRS medium resulted in a morphological transition of L. acidophilus NCFM from filamentous to bacilloid rods, and the bacilloid cells were more resistant to freezing and storage at conventional freezer temperatures. The results suggest that the morphology of the L. acidophilus cell may be an important consideration in the preparation of freeze-stable culture concentrates.     

Calcium malate overproduction by Penicillium viticola 152 using the medium containing corn steep liquor

In this study, after screening of eight fungal strains for their ability to produce calcium malate, it was found that Penicillium viticola 152 isolated from marine algae among them could produce the highest titer of calcium malate. At the same time, it was found that corn steep liquor (CSL) could stimulate calcium malate production and 0.5 % (v/v) CSL was the most suitable for calcium malate production. Under the optimal conditions, a titer of calcium malate in the supernatant was 132 g/l at flask level. During a 10-l fermentation, a titer of 168 g/l, a yield of 1.28 g/g of glucose, and a productivity of 1.75 g/l/h were reached within 96 h of the fermentation, and 93.4 % of the sugar was used for calcium malate production and cell growth, demonstrating that the titer, yield, and productivity of calcium malate by this fungal strain were very high and the fermentation period was very short. After analysis of the partially purified product with high-performance liquid chromatography, it was found that the main product was calcium malate. The results demonstrated that P. viticola 152 obtained in this study was the most suitable for developing a novel one-step fermentation process for calcium malate production from glucose on a large scale.     

Detection of infection by Pseudomonas phaseolicola (Burkh.) Dowson in white-seeded dwarf bean seed stocks

Seeds of white-seeded varieties of dwarf beans infected with Pseudomonas phaseolicola may show fluorescent areas in the testa under ultra-violet light. Fluorescence is not visible in seed with dark-coloured testas and is masked by coloured seed dressings. Bacterial infection is not the only cause of fluorescence in testas and the type and colour of the fluorescence cannot be used to diagnose infection. Of 542 fluorescing seeds examined 112 were infected by the pathogen, and only six infected seeds were found in 495 non-fluorescing seeds from the same samples. It is estimated that in two seed stocks used for these experiments 64 and 68 % of infected seeds were of the fluorescent type. A 34 lb sample of seedstock has to be taken for examination to detect, with 95 % confidence, ten infected seeds in 1 cwt. The selection of all fluorescing seeds from such large samples effectively concentrates the infected seed into a sample small enough to be tested by bacteriological methods. Test seeds are examined bacteriologically by enrichment-culture in nutrient broth. Ps. phaseolicola is identified in these cultures from the diagnostic symptoms of halo-blight disease induced in bean seedlings inoculated with aliquots of the cultures.     

The biology of Peronospora viciae on pea: factors affecting the susceptibility of plants to local infection and systemic colonization

Peronospora viciae (Berk.) Casp. penetrated leaf disks of Pisum sativum L. through the cuticle. Resistance of pea plants and of individual leaves to infection by P. viciae increased with age, but decreased again at senescence. Resistance was shown by a restriction in fungal growth and sporulation and by a chlorotic reaction in the leaves. Systemic invasion followed infection of meristematic tissue, and was induced by inoculation into the apical bud of young plants, or on to the epicotyl or hypocotyl, but not roots of germinating seedlings. Most plants whose growth was retarded showed an increased resistance to systemic infection. Pods were infected externally by sporangia, rather than by mycelial growth through the peduncle and pedicel. Oospores and mycelium were found in the testas of some seeds, but seeds from infected pods did not give rise to infected seedlings.     

Surface reactivity and in vitro biological evaluation of sol gel derived silver/calcium silicophosphate bioactive glass

Ag ions are known for their antibacterial effects. Ag containing silicate glasses have been extended to create bioactive glasses that exhibit inhibitory effects on bacterial growth using different techniques. In this work, calcium and calcium/silver silicophosphate glasses were synthesized from the sol-gel process and their physicochemical and in vitro biological properties were studied and compared. The effect of silver concentration on in vitro bioactivity and antibacterial properties of the glasses was investigated. Ag₂O was substituted for CaO in the glass formula up to 2 mol% and in vitro bioactivity of the samples was evaluated by soaking them in simulated body fluid followed by structural characterization using XRD, FTIR and SEM techniques. The results showed that both glasses favored precipitation of the calcium phosphate layer when they were soaked in simulated body fluid; however, the morphology of apatite crystals changed for the 2% mol silver containing sample. Substitution of 2% mol Ag₂O for CaO seemed to slightly stimulate the rate of precipitation. The in vitro biodegradation rate of the silver/ calcium silicophosphate glasses was lower than that of the silver-free one (control). Also, the antibacterial properties of the samples indicated that these effects were improved by increasing silver concentration in bioactive glass composition.     

Cuburbitacins from the cruciferae

The distribution of cucurbitacins in seeds and growing plants of Iberis umbellata L. has been investigated. The testas of 'Giant Pink', 'Purple', 'Crimson' and 'Iceberg' (Dutch) strains of Iberis umbellata contain Cucurbitacin B with traces of D and E. Seeds of 'Iceberg' (Danish) strain were exceptional in containing cucurbitacins E and I but no B, and this strain was also distinguished by larger redder seeds and by testas which were not readily separable into two layers. Cucurbitacins are rare in other crucifers but they are present in I. gibraltarica L., I. sempervirens L. and Lepidium sativum L.     

Inhibitors of polygalacturonase and laccase of Botrytis cinerea and their application to the control of this fungus

EDTA, calcium chloride, and two siderophores (rhodotorulic acid produced by Rhodotorula glutinis BNM 0524, and enterochelin from the bacterium Rahnella aquatilis BNM 0523) were evaluated as possible inhibitors of polygalacturonase (PG) and laccase (LC) from Botrytis cinerea. The aim was to apply them to the control of this pathogen, taking into account the fact that these enzymes are related to the invasion and installation of the fungus in the host. Two B. cinerea Pers.:Fr strains (BNM 0527 and BNM 0528) were used. Enzyme activities were measured in the supernatant of 7-day-old cultures. EDTA, calcium chloride, rhodotorulic acid, or enterochelin were added in the reaction mixture. Laccase activity from two strains was more affected by enterochelin (70-80% inhibition) than by the other compounds, while polygalacturonase was more inhibited (45% inhibition) by calcium chloride. The inhibitors were added to the growth medium and after 7 days of culture, the activities of the enzymes were measured in the supernatants. The production of PG and LC in both strains was lower when enterochelin or calcium chloride was added. In the third step, when the inhibitors were tested on apple, all them provided both effects, preventive and curative, against infections caused by B. cinerea, with EDTA and rhodotorulic acid exhibiting more preventive effects while calcium chloride and enterochelin provided more control of pre-existing infections (curative effect), coinciding with their ability to inhibit the production of polygalacturonase and laccase.     

Use of extracts from oyster shell and soil for cultivation of Spirulina maxima

Calcium ion and trace metals play important roles in various metabolisms of photosynthetic organisms. In this study, simple methods were developed to extract calcium ion and micronutrients from oyster shell and common soil, and the prepared extracts were tested as a replacement of the corresponding chemicals that are essential for growth of microalgae. The oyster shell and soil were treated with 0.1 M sodium hydroxide or with 10 % hydrogen peroxide, respectively. The potential application of these natural sources to cultivation was investigated with Spirulina maxima. When compared to standard Zarrouk medium, the Spirulina maxima cultivated in a modified Zarrouk media with elements from oyster shell and soil extract exhibited increases in biomass, chlorophyll, and phycocyanin by 17, 16, and 64 %, respectively. These results indicate that the extracts of oyster shell and soil provide sufficient amounts of calcium and trace metals for successful cultivation of Spirulina maxima.     

Similarity between Cytokinin and Blue Light Inhibition of Cucumber Hypocotyl Elongation

The cytokinin benzyladenine inhibited endogenous hypocotyl elongation in intact etiolated seedlings of cucumber (Cucumis sativus L.). In hypocotyl segments, the inhibitory effect of benzyladenine on growth was clearly detectable in the presence of indoleacetic acid. Fusicoccin-induced elongation was unaffected by the presence of cytokinin. The effect of cytokinin on elongation of the segments was determined by measuring changes in fresh weight, a linear function of extension growth. The effect of benzyladenine on hypocotyl growth was at least as large in segments prepared from red-light-grown seedlings as in those from seedlings grown in total darkness. A comparison was made between the inhibitory effects of cytokinin and blue light. The use of the calcium chelator ethyleneglycol-bis(β-aminoethyl ether)-N, N′-tetraacetic acid indicated that calcium ions are required for manifestation of benzyladenine-induced inhibition.     

Effects of Metals on Streptomyces coelicolor Growth and Actinorhodin Production

Actinorhodin production by Streptomyces coelicolor was used as a model system to study the effects of metals on growth and polyketide synthesis in a streptomycete. Numerous metals were tested in cultures grown in liquid media. Mercury and cadmium were highly toxic, and copper, nickel, and lead were less so, but all tended to inhibit both growth and antibiotic synthesis to a similar extent. Unexpectedly, manganese, cobalt, zinc, and, to a lesser extent, chromium caused complex effects that in general resulted in some enhancement of growth yield but a reduction in antibiotic titers. These complex effects meant that cobalt, manganese, and zinc had lower 50% inhibitory concentrations for antibiotic yields compared with those for biomass. The physiologically active divalent cations calcium and magnesium were also tested. Calcium at high concentrations was particularly effective in reducing antibiotic titers and enhancing growth yields. By adding calcium at different phases of growth, it could be demonstrated that it was most effective in reducing the antibiotic yield when added during the early growth phase. Addition during the antibiotic-producing phase resulted in little reduction of final actinorhodin titers.     

Role of cell wall bound calcium in Neurospora crassa

Cell wall bound calcium constitutes a significant fraction (25%) of total mycelia calcium in Neurospora crassa. Wall bound calcium increases as a function of growth and calcium concentration, while cell wall bound calcium decreases in Ca-free medium. Removal of wall bound calcium causes its rapid replacement from intracellular pool, inhibited by verapamil, nifedipine, concanamycin A, and wortmanin in a vacuolar mutant (Vma-5), but is unaffected by trifluoropyrazine, and calmidizoluim in a calcineurin mutant (Cnb-1) of N. crassa. Ca²⁺ removal from surface with EGTA resulted in leakage of periplasmic enzymes invertase and alkaline phosphatase. Scanning and transmission electron microscopy revealed gross abnormalities represented by giant vacuoles. Toxic metal ions bound to wall fraction by displacing calcium. Our data underline the physiological importance of wall bound calcium in N. crassa.     

Significance of various nitrogen sources for calcium deficient seedlings ofCucurbita pepo L.

The aim of the present investigation was to examine whether some of the growth defects of calcium-deficient pumpkin (Cucurbita pepo L.) seedlings are due to an incapability of the plants to assimilate nitrate nitrogen, such being the mechanism by which according to some of the previous authors growth defects in calcium deficient plants may be produced. The results of the present study, however, demonstrate that the accessibility of various possible products of nitrate reduction is not the limiting factor responsible for growth symptoms of calcium deficiency.     

Osmotic stabilizer-coupled suppression of NDR defects is dependent on the calcium-calcineurin signaling cascade in Aspergillus nidulans

Establishment and maintenance of cell polarity are coordinated by signaling pathways such as NDR (nuclear Dbf2-related) protein-kinase signaling and calcium signaling pathway. The NDR family of kinase is structurally related to the human myotonic dystrophy kinase, which, when impaired, confers a disease that involves changes in cytoarchitecture and ion homeostasis. CotA kinase, a member of the NDR protein kinase family, forms a complex with MobB to regulate cell polarized growth in Aspergillus nidulans. Our previous study demonstrated that mobB/cotA defects could be suppressed by the osmotic stress in the presence of external calcium. In this study, via the genetic and molecular approach, we further demonstrated that Ca²⁺-permeable stretch-activated nonselective cation channel-MidA, calcium/calmodulin-dependent protein phosphatase catalatic subunit-CnaA and external calcium, but not voltage-gated calcium channel homolog-CchA, were required for the osmotic stabilizer-coupled suppression. The up-regulation of calcium/calcineurin signaling pathway induced by osmotic stress might be the reason for bypassing the requirements of NDR kinase complex, which is otherwise necessary for polar morphogenesis. Our results suggest that calcium-calcineurin signaling pathway coordinates with MobB/CotA kinase complex in regulating polarity growth via maintaining cellular calcium homeostasis. However, CchA may act differently as other components in calcium signaling pathway in Aspergillus nidulans. These findings provide an excellent opportunity to identify the potential pathway linking NDR protein-kinase network to calcium signaling pathway.     

Formate Oxidation-Driven Calcium Carbonate Precipitation by Methylocystis parvus OBBP

Microbially induced carbonate precipitation (MICP) applied in the construction industry poses several disadvantages such as ammonia release to the air and nitric acid production. An alternative MICP from calcium formate by Methylocystis parvus OBBP is presented here to overcome these disadvantages. To induce calcium carbonate precipitation, M. parvus was incubated at different calcium formate concentrations and starting culture densities. Up to 91.4% ± 1.6% of the initial calcium was precipitated in the methane-amended cultures compared to 35.1% ± 11.9% when methane was not added. Because the bacteria could only utilize methane for growth, higher culture densities and subsequently calcium removals were exhibited in the cultures when methane was added. A higher calcium carbonate precipitate yield was obtained when higher culture densities were used but not necessarily when more calcium formate was added. This was mainly due to salt inhibition of the bacterial activity at a high calcium formate concentration. A maximum 0.67 ± 0.03 g of CaCO₃ g of Ca(CHOOH)₂⁻¹ calcium carbonate precipitate yield was obtained when a culture of 10⁹ cells ml⁻¹ and 5 g of calcium formate liter⁻¹ were used. Compared to the current strategy employing biogenic urea degradation as the basis for MICP, our approach presents significant improvements in the environmental sustainability of the application in the construction industry.     

Preparation of Refractile Spores of Clostridium thermosaccharolyticum Involves a Solventogenic Phase

Conversion of vegetative cells of Clostridium thermosaccharolyticum to refractile endospores was achieved by sequential transfer and dilution at each generation, with a final dilution into a sporulation medium that contained xylan supplemented with excess calcium. The subsequent growth was synchronous and resulted in elongated, solventogenic cells that were then shifted to 35°C to permit further differentiation without cell division. The synchronized cells grown in xylan medium supplemented with Ca gluconate produced total solvents that reached 9.63% (vol/vol). One hundred percent of these elongated solventogenic cells (4.84 × 10⁹ cells per ml) entered the sporangial stage and continued to differentiate into refractile spores. Only cells sequentially transferred and diluted at a critical time of the growth cycle are synchronized, induced to elongate (≥fourfold), become highly solventogenic in the presence of excess calcium, and are converted to a homogeneous population of refractile spores.