Comparison between pressure-volume and dewpoint-hygrometry techniques for determining the water relations characteristics of grass and legume leaves

Summary The water relations characteristics of three grass species (Panicum maximum var. trichoglume, Cenchrus ciliaris, Heteropogon contortus), and a legume (Macroptilium atropurpureum) grown in the field were measured using both a modified pressure/volume technique with pressure bomb measurements on single leaves and a dewpoint hygrometry technique applied to fresh and to frozen and thawed leaf discs.The two techniques agreed well in the estimates of osmotic potential at full turgor and the water potential at zero turgor. However, for parameters such as the relative water content at zero turgor, bound water and bulk modulus of elasticity there was a poor correlation between the estimates from the two methods. The pressure/volume technique gave less variable results and is more convenient for field use than the hygrometry technique. The determination of the modulus of elasticity from various functions relating pressure potential to relative water content is discussed.     

Location of satellite DNAs in the chromosomes of the kangaroo rat (Dipodomys ordii)

The location of satellite DNA sequences in metaphase chromosomes has been studied in the kangaroo rat by the in situ hybridization technique, staining techniques and phase contrast microscopy. The HS- satellite DNA is located at the kinetochores of all but three chromosome pairs. The HD satellite is located predominantly in the short arms of the chromosomes containing HS- and in the kinetochores of chromosome pairs that lack HS-. The regions that contain the satellite DNA sequences can also be identified by the Giemsa staining technique, and can be visualized with phase contrast microscopy or following Feulgen staining of fixed chromosome preparations.     

The adenosine triphosphate content of cysts of Globodera pallida and G. rostochiensis as a possible quantitative estimate of field populations

A rapid method is described for estimating the number of juveniles of Globodera pallida and G. rostochiensis in cysts collected from field soils. The technique uses a photometer to measure the adenosine triphosphate (ATP) content of the cysts by bioluminescence. Results for ten to twelve soil samples of 100 g taken from each of four fields showed that the number of juveniles that emerged when the samples were placed in potato root diffusate was significantly related to ATP content of a second series of cyst samples from these fields. Cysts from one field gave a higher ATP content per hatched juvenile than was recorded for the remaining three fields. There are several, untested, explanations of this discrepancy, but some evidence suggests that the hatching test and not the ATP method may have given an unreliable estimate of the viable egg content of cysts from this field. Further work may result in an enhanced precision and increased rapidity in estimating field populations of cyst nematodes by this technique.     

Histochemical localization of β-glycosidases in roots ofZea mays

Summary A simultaneous coupling azo dye technique using 6-bromo-2-naphthyl-D-glucoside as substrate has been developed for the localization of-glucosidase in hand sections of plant root tissue. This technique is also applicable to-galactosidase localization. Using corn root hairs and epidermal cells as an experimental system, the localization obtained in simultaneous coupling has been compared with that obtained in the older post-coupling method, and in various control sections. It is concluded that the localization obtained in root hairs in the post-coupling method is an artifact produced by diffusion of the product before coupling to substantive non-enzymatic sites in the cytoplasm.     

Histochemical lectin affinity technique by means of FITC-labeled serum protein fractions

Summary A two-step affinity technique is described for light microscopic demonstration of the Concanavalin A, Agaricus bisporus lectin and Ricinus communis lectin binding sites by means of various FITC-labeled human and rabbit serum protein fractions. Experiments for the visualization of the Lens culinaris lectin and the Pisum sativum lectin binding sites gaves negative results. The technique consist of two reaction steps which involve the incubation of tissue sections in the lectins followed by the visualization of receptor-bound lectins with FITC-labeled serum protein fractions basing on their carbohydrate content. The specificity of the technique could be demonstrated by the addition of the hapten or by incubation in the FITC-labeled serum protein fractions only. In contrast to the direct or indirect staining methods only very small amounts of purified lectins are necessary.     

Determination of low boron concentrations in nutrient solution

To study boron (B) deficiency and toxicity in plants in flowing solution culture, it is necessary to establish a wide range of solution B concentrations. The ability of inductively coupled plasma atomic emisson spectrophotometry (ICPAES) to determine solution B concentrations ranging from 0.15 M to 925 M was investigated. The reliablity of B concentration determination in nutrient solutions containing <10 M B is poor. A technique, involving sorption by a B-specific resin (Amberlite-743), was established to concentrate the B present in low B nutrient solutions and to enable reliable measurement using ICPAES following elution from the resin. Acceptable, reproducible recoveries of B from low B solutions, containing known concentrations of B, were obtained using this technique. The technique enables the imposition, monitoring and maintenance of solution B concentrations well below the direct detection limits of ICPAES and colorimetric procedures. ei]L V Kochian     

Simple and rapid method for the isolation of Aspergillus niger mutants with enhanced cellulase and xylanase activity

Summary An agar plate isolation technique was developed for screening the mutants of Aspergillus niger. Fungal growth in solid media containing carboxymethylcellulose(CMC) was selected within 2–3 days. The mutants selected by the new technique showed a remarkable enhancement of CMCase, FPase, -glucosidase, xylanase, and -xylosidase activities. These results clearly show that the new isolation technique is a practical method for the selection of better strains.     

Quantitative histochemistry of oxalate in vesiculated hairs of Atriplex halimus L.

Summary A quantitative microchemical technique was devised to detect 2 to 3 g oxalate in plant materials. This technique was used to measure the molarity of oxalate in the giant vesiculated hairs covering leaves of Atriplex halimus L. Oxalate concentration in the hairs was found to be approximately 1.4 molar. This technique is much simpler and more sensitive than previously reported methods.     

Détection par immunofluorescence des cellules corticotropes et mélanotropes dans l'hypophyse de la grenouille,Rana temporaria L., au cours du développément

Summary During the embryonic and larval developmental stages of the frog, Rana temporaria L, anti- 1–24, 17–39 corticotropine, and MSH antibodies were used to define, with immunofluorescence technique, the appearence of corticotropic and melanotropic cells.A very small number of fluorescent corticotropic cells appears for the first time during the embryonic stage (10 mm), just before the differentiation of the pars intermedia. The cells are small, their large nucleus is surrounded by a fine rim of fluorescent cytoplasm.During premetamorphic stage, the anti-ACTH antibodies (anti- 1–24 and anti- 17–39 corticotropine) reveal more fluorescent cells in the whole pars distalis. The pars intermedia cells can also be visualized by both antisera.At the end of prometamorphosis and during climax the corticotropic cells show a more precise localization. As in adult frog pars distalis, they are concentrated in the rostral half of the lobe. With the same technique anti- and MSH antibodies reveal only the cells of the pars intermedia. No other cell type of the pars distalis reacts with these antibodies. This technique has the advantage to show that the ACTH and the MSH cells appear very early during the embryonic life.

     

Isolation of auxotrophic mutants of the methylotrophic yeastCandida boidinii and determination of its ploidy

Auxotrophic mutations in the methylotrophic yeast strainCandida boidinii 11Bh were induced by different mutagens and their combinations (nitrosoguanidine, UV light, HNO₂+UV). Majority of the mutants obtained carried defects in histidine, arginine, proline and/or adenine biosynthetic pathways. His^- mutants were distributed into four complementation groups using the protoplasts fusion technique. Ploidy determination ofCandida boidinii 11Bh was performed by measuring its DNA content and by following its survival after chemical mutagens treatment. The DNA content of this strain was found to be similar to that of aSaccharomyces cerevisiae diploid strain. Also the kinetics of survival ofCandida boidinii cells indicate thatCandida boidinii 11Bh is a diploid.     

Immobilization of biocatalysts in thermogels using the resonance nozzle for rapid drop formation and an organic solvent for gelling

Summary The resonance nozzle immobilization technique was tested with thermo-hardening hydrogels (agar, gellan and -carrageenan) and compared with the conventional needle technique. After nozzling the warm gel solution, the formed droplets were caught in an ice-cold two-phase system consisting of an organic solvent (n-butylacetate, hexane or nonane) as top phase, in which gelling occurred, and aqueous medium as bottom phase. The cells used were yeast cells (Saccharomyces cerevisiae), bacterial cells (Mycobacterium aurum L1), plant cells (Tagetes minuta) and insect cells (Spodoptera frugiperda). The retention of the respiration activity was used as a criterion for the suitability of the conditions applied. The relatively polar solvent n-butylacetate with the low log P value gave the poorest activity retention, which is in agreement with experimentally validated theory. For all gels, the resonance nozzle technique proved to give satisfactory results, although the retention of respiration activity was generally lower than obtained with the needle technique.     

Location and quantification of metal ions in enzymes combining polyacrylamide gel electrophoresis and particle-induced X-ray emission

A method is presented to identify and determine the relative amounts of protein-bound metal ionsin situ. Proteins or their subunits are directly scanned by a collimated proton beam of 3 MeV energy, and the characteristic X-rays produced are detected. The determination of Fe content of an iron-sulfur protein (HiPiP), as well as the Fe and Ni analysis of the hydrogenese fromThiocapsa roseopersina, have shown the feasibility of this technique.     

Preparation of Single Cell Detritus from <Emphasis Type="Italic">Laminaria saccharina</Emphasis> as a Hatchery Diet for Bivalve Mollusks

A high-yield technique is described for the elaboration of single cell detritus (SCD) from Laminaria saccharina, based on the sequential action of C1H, enzymes (endoglucanases and cellulases) and 2 bacteria showing a high degree of cellobiotic, proteolytic, and alginolytic activity (CECT 5255 and CECT 5256). Over 85% of dried particles of L. saccharina were transformed into a suspension of free cell and bacterial and detrital particles after 24 hours of bacterial activity with this technique. These particles were less than 20 m in diameter, constituting a suitable diet for bivalve mollusks. After 72 hours 99% of the total particulate volume consisted of particles less than 20 m in diameter. Tests of hatchery diets for the seed of clam Ruditapes decussatus revealed increases of 54% and 68% for live weight and length, respectively, when SCD from L. saccharina was used as the sole dietary component compared with a live phytoplankton diet. However, SCD from L. saccharina is not a suitable food for the larvae of R. decussatus.     

Änderungen der Ultrastrukturen in den Grünalgen Ankistrodesmus braunii und Chlorella fusca var. rubescens bei Stickstoffmangel

Summary Ultrastructural changes as well as changes in pigment and lipid content induced by nitrogen deficiency in the green algae Ankistrodesmus braunii and Chlorella fusca var. rubescens have been studied. Electron micrographs (freezeetching technique) and analyses show that the content of chlorophylls and the number of thylakoids decrease under these conditions synchronously with the increase of secondary carotenoids (astaxanthin-like type) and lipid vacuoles. However, there is no change, as compared with the green control, in the number and structure of the plastoglobuli localized in the chloroplasts. After addition of nitrogen, synthesis of the thylakoids and chlorophylls begins. The lipid vacuoles and secondary carotenoids disappear gradually. These extraplastidic lipid vacuoles do not correspond with the intraplastidic plastoglobuli (lipid globuli) found in higher plants and in Acetabularia.     

纤维型植物南荻的生物学及其应用研究进展

南荻(Triarrhena lutarioriparia L.Liu)是我国特有高产纤维素型多年生草本植物,其独特的生态适应性、广泛的用途及其生产开发潜力已引起相关行业的高度关注。概述了南荻的资源分布、生物学特性、繁育栽培及应用的研究现状,并对其纤维素资源的应用进行了展望。     

Patch-clamp study on membrane properties and transmitter activated currents of rabbit area postrema neurons

Using the patch-clamp technique in combination with sliced tissue preparation the membrane properties of newborn rabbit area postrema neurons were investigated. The neurons responded upon depolarization with a fast Na ⁺-current followed by an inactivating and non-inactivating K ⁺-current. GABA-activated currents were investigated resulting in a large Cl^--conductance, indicating the expression of GABA_A-receptors. The expression of glutamate receptor mRNA was studied by in situ hybridization and electrophysiological measurements of these receptors by means of the patch-clamp technique. As a main result it was found that ionotropic glutamate receptors in the area postrema are composed of flop variants of the GluA-, GluB- and GluC-subunits.Abbreviations AP area postrema - GABA -aminobutyric acid - Glu glutamate - I–V current-voltage - SFO subfornical organ     

Hair Trace Element Levels in Han and Indigenous Hualien Inhabitants in Taiwan

The boron content was determined in 42 different foods consumed in Istanbul, Turkey. Eleven species of fruit, ten species of vegetable, eight species of food of animal origin, four species of grain, two species of nuts, two species of legume, and five other kinds of foods were included to this study. They were analyzed by two methods: Inductively coupled plasma mass spectrometry (ICP-MS) technique and carminic acid assay, and the results of two methods were also compared. Boron concentration in foods ranged between 0.06–37.2 mg/kg. Nuts had the highest boron content while foods of animal origin had the lowest. A strong correlation was found between the results of the carminic acid assay and the ICP-MS technique (p = 0.0001, Pearson correlation coefficient: r = 0.956). Bland Altman analysis also supported this correlation. ICP-MS is one of the most common, reliable, and powerful method for boron determination. The results of our study show that spectrophotometric carminic acid assay can provide similar results to ICP-MS, and the boron content in food materials can be also determined by spectrophotometric method.

     

Legionella pneumophila virulence conserved after multiple single-colony passage on agar

Multiple passage ofLegionella pneumophila on either supplemented Mueller-Hinton or charcoal yeast extract agar by the conventional batch passing technique results in loss of virulence. In this studyL. pneumophila virulence was maintained after multiple passage on buffered charcoal yeast extract (BCYE ) agar by a single-colony transfer technique. Virulence was determined by assessing the growth ofL. pneumophila for thioglycolate-induced susceptible A/J mouse macrophages in vitro and infectivity for susceptible A/J mice in vivo. Passage of the virulentL. pneumophila, 30 times on BCYE agar by the single-colony transfer procedure still resulted in virulence, as compared with the nonpassaged parental bacteria, both in vitro and in vivo. Lethality for susceptible, A/J mice by systemic infection was comparable for the 30th colony-passaged bacteria and the parentalL. pneumophila. These results show that theL. pneumophila phenotype associated with intracellular growth in macrophages or infectivity for susceptible mice is stable following passage by the single-colony transfer procedure on BCYE agar.     

Correlation between potassium and phosphorus content and their nonuniform distribution in Acanthamoeba castellanii

Summary Biologically important elements: K, Na, Mg, Ca, Cl, P, and S were analyzed in Acanthamoeba castellanii. A higher potassium content, as compared with other cations, was detected. Total content of the cation-forming elements: K, Na, Mg, and Ca was ca. 360 mmoles/kg dry weight of the cells. Phosphorus content was estimated as 492 mmoles/kg dry weight. Content of chlorine, a basic cellular anion, was 173 mmoles/kg dry weight. The low level of chlorine appears not the be sufficient to balance all the cations in Acanthamoeba.Distribution of potassium in Acanthamoeba cells was nonuniform and similar to that of phosphorus as shown by X-ray microanalysis technique. Quantitative correlation between phosphorus and potassium as well as the similar distribution of these elements suggests that in Acanthamoeba phosphorus is an essential anion which, being nonuniformly distributed in the cell, determines also a nonuniform distribution of potassium.     

In vitro production of perithecia ofDiaporthe helianthi

In trials with various agar media and autoclaved stem pieces of weeds and cultivated plants, perithecia ofDiaporthe helianthi Muntañola-Cvetkoviet al., developed only on autoclaved stem pieces. The simple, useful culture technique for producing perithecia is fully described.