Localization of the Functional p34cdc2 Homolog of Maize in Root Tip and Stomatal Complex Cells: Association with Predicted Division Sites

We have used an antibody against the functional homolog of the cdc2 kinase of maize to localize the p34cdc2 protein within dividing cells of the root apex and the stomatal complex of leaf epidermis. The microtubule cytoskeletal structure of plant cells was visualized concomitantly with a monoclonal antibody specific for [alpha]-tubulin. We found that the cdc2 protein is localized mainly to the nucleus in plant cells at interphase and early prophase. This finding contrasts markedly with the predominantly cytoplasmic staining obtained using antibody to the PSTAIRE motif, which is common to cdc2 and numerous cdc2-like proteins. In a subpopulation of root cells at early prophase, the p34cdc2 protein is also distributed in a band bisecting the nucleus. Double labeling with the maize p34cdc2Zm antibody and tubulin antibody revealed that this band colocalizes with the preprophase band (PPB) of microtubules, which predicts the future division site. Root cells in which microtubules had been disrupted with oryzalin did not contain this band of p34cdc2 protein, suggesting that formation of the microtubule PPB is necessary for localization of the p34cdc2 kinase to the plane of the PPB. The p34cdc2 protein is also localized to the nucleus and PPB in cells that give rise to the stomatal complex, including those cells preparing for the highly asymmetrical divisions that produce subsidiary cells. Association of the p34cdc2 protein with the PPB suggests that the cdc2 kinase has a role in establishing the division site of plant cells and, therefore, a role in plant morphogenesis.     

Fine-scale mapping of physicochemical and microbial landscapes of the coral skeleton

The coral skeleton harbours a diverse community of bacteria and microeukaryotes exposed to light, O₂ and pH gradients, but how such physicochemical gradients affect the coral skeleton microbiome remains unclear. In this study, we employed chemical imaging of O₂ and pH, hyperspectral reflectance imaging and spatially resolved taxonomic and inferred functional microbiome characterization to explore links between the skeleton microenvironment and microbiome in the reef-building corals Porites lutea and Paragoniastrea benhami. The physicochemical environment was more stable in the deep skeleton, and the diversity and evenness of the bacterial community increased with skeletal depth, suggesting that the microbiome was stratified along the physicochemical gradients. The bulk of the coral skeleton was in a low O₂ habitat, whereas pH varied from pH 6–9 with depth. Physicochemical gradients of O₂ and pH of the coral skeleton explained the β-diversity of the bacterial communities, and skeletal layers that showed O₂ peaks had a higher relative abundance of endolithic algae, reflecting a link between the abiotic environment and the microbiome composition. Our study links the physicochemical, microbial and functional landscapes of the coral skeleton and provides new insights into the involvement of skeletal microbes in the coral holobiont metabolism.     

Development of predictive models of laboratory animal growth using artificial neural networks

Traditional regression analysis of body weight growth curvesencounters problems .when the data are extremely variable. Whiletransformations are often employed to meet the criteria of theanalysis, some transformations are inadequate for normalizingthe data. Regression analysis also requires presuppositionsregarding the model to be fit and the techniques to be usedin the analysis. An alternative approach using artificial neuralnetworks is presented which may be suitable for developing predictivemodels of growth. Neural networks are simulators of the processesthat occur in the biological brain during the learning process.They are trained on the data, developing the necessary algorithmswithin their internal architecture, and produce a predictivemodel based on the learned facts. A dataset of Sprague–Dawleyrat (Rattus norvegicus) weights is analyzed by both traditionalregression analysis and neural network training. Predictionsof body weight are made from both models. While both methodsproduce models that adequately predict the body weights, theneural network model is superior in that it combines accuracyand precision, being less influenced by longitudinal variabilityin the data. Thus, the neural network provides another toolfor researchers to analyze growth curve data.     

Intracellular distribution of the hydrophobic triterpene,bryonolic acid,in cultured cells of Luffa cylindrica L.

Summary Intracellular localization of bryonolic acid, an antiallergic pentacyclic triterpene, in cultured cells of Luffa cylindrica was investigated with reference to the sites of its biosynthesis and accumulation. The results of cell fractionation showed that bryonolic acid was mostly located in the cell wall fraction. The addition of FC-43 emulsion to the culture medium was found to cause the release of bryonolic acid from the cell wall into the medium without affecting cell growth and bryonolic acid production. Under this culture condition, ¹⁴C-labeled sodium acetate administered to the cells was rapidly incorporated into bryonolic acid which was then excreted into the medium within 10 min after administration. Electron microscopic observations suggested that spherical vesicles (ca 0.1 m in diameter) derived from the rough endoplasmic reticulum may be associated with the biosynthesis and excretion of this compound into the cell wall. Furthermore, the activity of 2,3-oxidosqualene cyclase, a key enzyme involved in the biosynthesis of bryonolic acid, was detected in the microsomal fraction containing the endoplasmic reticulum.Abbreviations BA bryonolic acid - ER endoplasmic reticulum - LS Linsmaier-Skoog - NAA 1-naphthaleneacetic acid - MES 2-(N-morpholino)ethanesulfonic acid - PVPP polyvinyl polypyrrolidone     

New branched Porolithon species (Corallinales,Rhodophyta) from the Great Barrier Reef,Coral Sea,and Lord Howe Island

Porolithon is one of the most ecologically important genera of tropical and subtropical crustose (non-geniculate) coralline algae growing abundantly along the shallow margins of coral reefs and functioning to cement reef frameworks. Thalli of branched, fruticose Porolithon specimens from the Indo-Pacific Ocean traditionally have been called P. gardineri, while massive, columnar forms have been called P. craspedium. Sequence comparisons of the rbcL gene both from type specimens of P. gardineri and P. craspedium and from field-collected specimens demonstrate that neither species is present in east Australia and instead resolve into four unique genetic lineages. Porolithon howensis sp. nov. forms columnar protuberances and loosely attached margins and occurs predominantly at Lord Howe Island; P. lobulatum sp. nov. has fruticose to clavate forms and free margins that are lobed and occurs in the Coral Sea and on the Great Barrier Reef (GBR); P. parvulum sp. nov. has short (<2 cm), unbranched protuberances and attached margins and is restricted to the central and southern GBR; and P. pinnaculum sp. nov. has a mountain-like, columnar morphology and occurs on oceanic Coral Sea reefs. A rbcL gene sequence of the isotype of P. castellum demonstrates it is a different species from other columnar species. In addition to the diagnostic rbcL and psbA marker sequences, the four new species may be distinguished by a combination of features including thallus growth form, margin shape (attached or unattached), and medullary system (coaxial or plumose). Porolithon species, because of their ecological importance and sensitivity to ocean acidification, need urgent documentation of their taxonomic diversity.     

Flower senescence in daylily (Hemerocallis)

This paper reviews recent developments in the use of molecular probes for analyzing the genetic makeup of somatic hybrids. Successful application of somatic hybridization to the interspecific transfer of traits encoded in the nucleus is still having limited success. A major difficulty is hybrid infertility, particularly in hybrids between sexually incompatible species. The formation of asymmetric hybrids is being explored as an approach for improving hybrid fertility. Evaluation of the degree of chromosome elimination and chromosome stability and instability in asymmetric hybrids is difficult when the traditional approaches of chromosome counting and isozyme analysis are used. Two new approaches are resolving this difficulty. The use of species-specific repetitive DNA probes in dot blotting and in situ hybridization to chromosomes is providing quantitative data on chromosome elimination and allows detection of translocations. Use of restriction fragment length polymorphism (RFLP) probes for analysis of hybrids between genetically mapped species makes it possible to account for the presence or absence of individual chromosomes and chromosomes arms. Wider use of such molecular probes should greatly improve our understanding of the genetics of both symmetric and asymmetric somatic hybrids and may lead to new strategies for the effective interspecific transfer of nucleus-encoded traits by protoplast fusion.     

Screening of Korean herbal medicines used to improve cognitive function for anti-cholinesterase activity.

Methanolic extracts of seven herbs (Acorus calamus, Acorus gramineus, Bupleurm facaltum, Dioscorea batatas, Epimedium koreanum, Poria cocos and Zizyphi jujuba) used in traditional Korean medicine for improvement of memory and cognition in old age were tested for cholinesterase inhibitory properties using the Ellman colorimetric method. Significant inhibition of the enzyme at 200 microg/ml was observed for extracts from A. calamus and E. koreanum. The possible bases for the reputation of these and the other herbs tested are discussed in the light of previous investigations into their chemistry and biological activity.     

Cell separation from high cell density broths of Alcaligenes eutrophus by using a coagulant

Summary Alcaligenes eutrophus was successfully recovered from high cell density broths by pre-treatment with polyaluminium hydroxide chloride silicate as a coagulant at 36–90 mg Al/l. The optimum pH range for cell coagulation was 10–12. Subsequent centrifugation (45×g) and filtration (pore size 0.5 mm) gave a cell recovery of higher than 90%. The energy demand for cell recovery with the coagulant was only 3–11% of that without it.