<Emphasis Type="Italic">Capsicum</Emphasis> Use in Cambodia: The Continental Region of Southeast Asia Is Not Related to the Dispersal Route of <Emphasis Type="Italic">C. frutescens</Emphasis> in the Ryukyu Islands

Capsicum Use in Cambodia: The Continental Region of Southeast Asia Is Not Related to the Dispersal Route of C. frutescens in the Ryukyu Islands. The local nomenclature and use of Capsicum by Khmer and other ethnic groups in Cambodia and the distribution of the diagnostic ShDH-B isozyme pattern of C. frutescens were studied. People in Cambodia use Capsicum in various ways, not only as a condiment but also as a vegetable, a medicine, and a colorant, and in popular beliefs, agricultural rituals, taboos, and rice malt. The findings showed that the ShDH-B phenotype may not have occurred as a mutation in Asia but in the Americas and then was introduced to Asia. Also, the ShDH-B phenotype is distributed in the insular regions of Southeast and East Asia and Oceania, but seems not to be distributed in the continental region of Southeast Asia. One possible hypothesis is that C. frutescens possessing the ShDH-B phenotype was introduced directly from the Americas via Oceania to the Philippines, and it thereafter dispersed into the insular regions.     

Potential use of the astaxanthin-producing microalga, <Emphasis Type="Italic">Monoraphidium</Emphasis> sp. GK12, as a functional aquafeed for prawns

We determined the nutritional profile of Monoraphidium sp. GK12, a newly isolated astaxanthin (AXA)-producing microalga, and investigated its potential use as a functional aquafeed by evaluating its effect on prawn pigmentation. GK12 contained high levels of pantothenic acid. The β-carotene content of GK12 was higher than that of Haematococcus, a well-studied AXA producer, and was similar to that of Spirulina. GK12 also had a high content of unsaturated fatty acids, of which linolenic acid (C18:3 n−3) was the most plentiful. A GK12-containing feed resulted in significant pigmentation of the prawns, comparable to that of prawns fed on synthetic AXA or Haematococcus. A GK12-containing feed also increased the survival rate of the prawns. Therefore, in addition to improving cultivation methods for Haematococcus, further research is needed into the use of GK12 as an alternative AXA source and as an ingredient of functional aquafeed for farmed fish.     

<Emphasis Type="Italic">In vitro</Emphasis> conservation of European bryophytes

The use of in vitro techniques for conservation has been rising steadily since their inclusion in The Convention on Biological Diversity and The Global Strategy for Plant Conservation. Unfortunately, bryophytes are often overlooked in conservation initiatives, but they are important in a number of large-scale ecosystem processes, i.e. nutrient, water and carbon cycling. There is a long history of the use of tissue culture in cultivating bryophytes, and many species respond well to in vitro techniques. For 6 yr (2000–2006), The Royal Botanic Gardens, Kew and the UK statutory conservation agencies supported a project for the ex situ conservation of bryophytes. Living and cryopreserved collections of UK threatened species were successfully established and the cryopreserved collection continues to be maintained. Other in vitro conservation collections are maintained over Europe, at botanic gardens, museums and by individual university researchers, but there is no coherent European collection of bryophytes for conservation, or standardisation of techniques. A major issue for many in vitro collections is the maintenance of within species genetic diversity. Such diversity is considered to be important, as it is the basis by which populations of species can adapt to new conditions and evolve. We are proposing to establish a European network for in vitro conservation of bryophytes. We envisage that this will include living collections, cryopreserved collections and spore collections. Conservation of genetic diversity would be a priority and the collections would provide a valuable resource for conservation initiatives and support research into rare and threatened species.     

Introgression molecular analysis of a leaf rust resistance gene from <Emphasis Type="Italic">Coffea liberica</Emphasis> into <Emphasis Type="Italic">C. arabica</Emphasis> L.

Leaf rust caused by the fungus Hemileia vastatrix is the most devastating disease of arabica coffee (Coffea arabica). Therefore, developing leaf rust-resistant varieties has been a breeding objective of the highest priority in many countries. The purpose of the present work was to gain insight into the mechanism of introgression into C. arabica of a leaf rust resistance gene from C. liberica (i.e. S_H3 resistance factor) and to identify associated molecular markers. An F₂ progeny (i.e. 101 individuals) derived from a cross between Matari, an arabica accession and liberica-introgressed line S.288, was evaluated for resistance against three different races of H. vastatrix. The progeny segregated for the S_H3 gene in a 3:1 ratio, as expected for a single dominant gene. Amplified fragment length polymorphism analysis of a population subset using 80 different primer combinations revealed that at least half of the total polymorphism observed in the population is associated with introgression of C. liberica chromosome fragments. Furthermore, 15 primer combinations generating candidate marker bands associated with the S_H3 resistance gene were used to analyse the whole F₂ population. A total of 34 marker bands originating from S.288 and attributable to introgression were scored. None exhibited segregation distortion. Linkage analysis revealed only three distinct introgressed fragments corresponding to a total length of 52.8 cM. Twenty-one markers were strongly associated (LOD score >14) with the S_H3 gene and were grouped together in a single linkage group of 6.3 cM. The results are discussed in relation to the efficient use of genetic resources in arabica breeding.     

<Emphasis Type="Italic">Pseudomonas seleniipraecipitatus</Emphasis> sp. nov.: A Selenite Reducing γ-<Emphasis Type="Italic">Proteobacteria</Emphasis> Isolated from Soil

Microbial fuel cells (MFCs) are a technology that provides electrical energy from the microbial oxidation of organic compounds. Most MFCs use oxygen as the oxidant in the cathode chamber. This study examined the formation in culture of an unidentified bacterial oxidant and investigated the performance of this oxidant in a two-chambered MFC with a proton exchange membrane and an uncoated carbon cathode. DNA, FAME profile and characterization studies identified the microorganism that produced the oxidant as Burkholderia cenocepacia. The oxidant was produced by log phase cells, oxidized the dye 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS), had a mass below 1 kD, was heat stable (121°C) and was soluble in ethanol. In a MFC with a 1000 Ω load and ABTS as a mediator, the oxidizer increased cell voltage 11 times higher than atmospheric oxygen and 2.9 times higher than that observed with ferricyanide in the cathode chamber. No increase in cell voltage was observed when no mediator was present. Organisms that produce and release oxidizers into the media may prove useful as bio-cathodes by improving the electrical output of MFCs.     

<Emphasis Type="Italic">Bacillus anthracis</Emphasis>, <Emphasis Type="Italic">Francisella tularensis</Emphasis> and <Emphasis Type="Italic">Yersinia pestis</Emphasis>. The most important bacterial warfare agents — <Emphasis Type="Italic">review</Emphasis>

There are three most important bacterial causative agents of serious infections that could be misused for warfare purposes: Bacillus anthracis (the causative agent of anthrax) is the most frequently mentioned one; however, Fracisella tularensis (causing tularemia) and Yersinia pestis (the causative agent of plague) are further bacterial agents enlisted by Centers for Disease Control and Prevention into the category A of potential biological weapons. This review intends to summarize basic information about these bacterial agents. Military aspects of their pathogenesis and the detection techniques suitable for field use are discussed.     

Global diversity of two widespread,colony‐forming diatoms of the marine plankton,Chaetoceros socialis (syn. C. radians) and Chaetoceros gelidus sp. nov.

Marine phytoplankton samples containing diatoms of the Chaetoceros socialis group were collected from Thailand, China, Denmark, and Greenland, and cells were isolated into culture for light and electron microscopy and DNA sequencing of D1–D3 of the LSU rDNA. Species of this lineage are characterized by three short and one long setae extending from each cell, the long setae from several cells joining into a common center to form large colonies, which are sometimes visible with the naked eye. Phylogenetic analyses including sequences from other parts of the world revealed segregation into three groups. Most sequences fell into two large clades, one comprising material from cold waters, whereas the other contained material from warmer waters. Strain CCMP 172 from the Strait of Georgia, Washington State, USA, formed a separate group. The warm‐water species included Chinese and Thai material and therefore probably also material from the type locality of C. socialis, Hong Kong. It is characterized by all setae being covered by spines and the setae extending from the valve at some distance from the margin. In the resting spores, both valves are ornamented with spines. The cold‐water material is characterized by three spiny and one mostly smooth long setae, and the setae extend from the valve near the margin. Both valves of the resting spore are smooth. This material is described as C. gelidus sp. nov. C. radians, described from the Baltic in 1894, is considered a synonym of C. socialis. CCMP172 is in many ways intermediate and probably constitutes a separate species. The published evidence on this globally distributed and sometimes bloom‐forming group of species indicates higher species diversity than presently thought.     

Life in soil by the actinorhizal root nodule endophyte <Emphasis Type="BoldItalic">Frankia</Emphasis>. A review

Frankia is a genus of soil actinomycetes famous for its ability to form N₂-fixing root nodule symbioses with actinorhizal plants. Although Frankia strains display a high diversity in terms of ecological niches in soil, current knowledge about Frankia is dominated by its life as an endophyte in root nodules. Increased use of molecular methods has refined and expanded insights into endophyte-host specificities and Frankia phylogeny. This review has focus on Frankia as a soil organism, including its part of microbial consortia, and how to study Frankia in soil. We highlight the use of nodulation tests and molecular methods to reveal population size and genetic diversity of Frankia in soil and discuss how autoregulation of nodulation and interactions with other soil microorganisms may influence the results. A comprehensive record of published interactions between Frankia and other soil microbes is summarized.     

Optimization of the Water-Insoluble Procedures for <Emphasis Type="Italic">USP</Emphasis> General Chapter <Emphasis Type="Italic">Residual Solvents</Emphasis> <467>

The water-insoluble procedures in US Pharmacopeia (USP) General Chapter Residual Solvents <467>, which are based on European Pharmacopoeia procedures, were optimized and modified before their inclusion in the chapter to improve their scope, performance, and ruggedness. The optimized procedures use a static headspace introduction system with a gas chromatograph equipped with a flame ionization detector. This article describes some of the key changes made to the USP published procedures, including use of dimethyl sulfoxide (DMSO) or dimethylformamide (DMF) as the solvent, addition of 5 mL of water and 1 mL of sample (dissolved in DMSO or DMF) to the headspace vial, use of a 3:1 GC split ratio, and use of new matrix-matched system suitability solutions. These procedures were verified with two different active pharmaceutical ingredients—hydroxyzine pamoate and prednisone. In the investigation, the more polar material (hydroxyzine pamoate) showed greater recoveries for the optimized procedures when prepared in DMSO. The less polar material (prednisone) typically had greater recoveries in DMF for the optimized procedures. During experimentation, insights into sample preparation, additional types of headspace instrumentation, solvent purity, and other parameters were also gained.     

Stable chloroplast transformation in cabbage (<Emphasis Type="Italic">Brassica oleracea </Emphasis>L. var. <Emphasis Type="Italic">capitata </Emphasis>L.) by particle bombardment

The objectives of this research were first to isolate plastid gene sequences from cabbage (Brassica oleracea L. var. capitata L.), and to establish the chloroplast transformation technology of Brassica. A universal transformation vector (pASCC201) for Brassica chloroplast was constructed with trnV–rrn16S (left) and trnI–trnA–rrn23S (right) of the IR_A region as a recombination site for the transformed gene. In transforming plasmid pASCC201, a chimeric aadA gene was cloned between the rrn16S and rrn23S plastid gene borders. Expression of aadA confers resistance to spectinomycin and streptomycin antibiotics. The uidA gene was also inserted into the pASCC201 and transferred into the leaf cells of cabbage via particle gun mediated transformation. Regenerated plantlets were selected by 200 mg/l spectinomycin and streptomycin. After antibiotic selection, the regeneration percentage of the two cabbage cultivars was about 2.7–3.3%. The results of PCR testing and Southern blot analysis confirmed that the uidA and aadA genes were present in the chloroplast genome via homologously recombined. Northern blot hybridizations, immunoblotting and GUS histochemical assays indicated that the uidA gene were stable integrated into the chloroplast genome. Foreign protein was accumulated at 3.2–5.2% of the total soluble protein in transgenic mature leaves. These results suggest that the expression of a variety of foreign genes in the chloroplast genome will be a powerful tool for use in future studies.     

Substrate-borne Vibrations as a Component of Intraspecific Communication in the Groundhopper <Emphasis Type="Italic">Tetrix ceperoi</Emphasis>

Substrate-borne vibrational signals used in communication of Tetrix ceperoi (Orthoptera: Tetrigidae) are described for the first time. Among all studied Tetrix species, T. ceperoi use the richest repertoire of vibrational signals. These signals consist of isolated pulses or pulses united into rhythmically repeated or irregularly repeated groups and are used in rivalry, pre-copulation, and copulation behaviour (1st, 3rd, and 4th structural types of signals), and are also used by single males or females (2nd type of signal). A 5th type of vibrational signal is produced by wing tremulation, which was unknown Tetrigidae. Based on the results of this study, the vibrational signalisation seems to play an important role in species-specific mate recognition of T. ceperoi. The significance of individual signals is discussed based on results of manipulative contact experiments.     

Documented Utility and Biocultural Value of <Emphasis Type="Italic">Aloe</Emphasis> L. (Asphodelaceae): A Review

Documented Utility and Biocultural Value of Aloe L. (Asphodelaceae): A Review. The genus Aloe L. (Asphodelaceae) comprises 548 accepted species, of which at least one-third are documented as having some utilitarian value. The group is of conservation concern due to habitat loss and being extensively collected from the wild for horticulture and natural products. Cultural value is increasingly important in the effective conservation of biodiversity. The present study evaluated the biocultural value of the known uses of Aloe, excluding the domesticated and commercially cultivated A. vera. Over 1,400 use records representing 173 species were collated from the literature and through personal observation; this paper presents a synopsis of uses in each of 11 use categories. Medicinal uses of Aloe were described by 74% of the use records, followed by social and environmental uses (both 5%). Species yielding natural products, notably A. ferox and A. perryi, were most frequently cited in the literature. Consensus ratios indicate that the most valued uses of Aloe are in medicine and pest control against arthropods and other invertebrates.     

Einsatz von V. lecanii als biologisches Schädlingsbekämpfungsmittel gegen den Bohnenrostpilz U. appendiculatus var. appendiculatus im Feld und im Gewächshaus

The use of Verticillium lecanii as a biological control agent against the bean rust fungus Uromyces appendiculatus var. appendiculatus in the field and in the glasshouse The deuteromycete V. lecanii parasites uredo- and teliospores of the bean-rust-fungus U. appendiculatus var. appendiculatus. We investigated the conditions for the use of the hyperparasite as biological control agent in the field and in glasshouses. The growth rate of the hyperparasite was 0,3 cm per day at 25 °C. Under suitable conditions in the lab (25 °C, 100 % r. h.) it took about 20 days to invade 100 % of uredospores and 65 % of teliospores. We failed to prevent the spread of bean-rust-fungus spores in the field, but we succeeded in the glasshouse by 68 %, compared to the untreated controls, using the hyperparasite V. lecanii as biological control agent.     

Use of <Emphasis Type="Italic">Capsicum frutescens</Emphasis> L. by the Indigenous Peoples of Taiwan and the Batanes Islands

Use of Capsicum frutescens L. by the Indigenous Peoples of Taiwan and the Batanes Islands. The local nomenclature, use, and distribution of C. frutescens among indigenous peoples in Taiwan and the Batanes Islands were studied. Among Taiwanese indigenous peoples, the distribution, frequency of use, and importance of C. frutescens were found to increase with decreasing latitude, which appears to have affected the local names of Capsicum and C. frutescens. The local name for Capsicum in the Batanes Islands—“sili”—is used by several indigenous peoples in Taiwan, suggesting that Capsicum was brought from the south to the north. Indigenous peoples in Taiwan and the Batanes Islands used C. frutescens fruits as condiments, medicines, ornaments, or for ritual uses; also, they used its leaves for soup. A complex of both green and yellowish-green types possessing ShDH-B was introduced from Indonesia into the Batanes Islands and Taiwan, and later only the type with yellowish-green immature fruit was introduced to the Ryukyu Islands under the bottleneck effect.     

Polyhydroxyalkanoates in Gram-positive bacteria: insights from the genera <Emphasis Type="Italic">Bacillus</Emphasis> and <Emphasis Type="Italic">Streptomyces</Emphasis>

Gram-positive bacteria, notably Bacillus and Streptomyces, have been used extensively in industry. However, these microorganisms have not yet been exploited for the production of the biodegradable polymers, polyhydroxyalkanoates (PHAs). Although PHAs have many potential applications, the cost of production means that medical applications are currently the main area of use. Gram-negative bacteria, currently the only commercial source of PHAs, have lipopolysaccharides (LPS) which co-purify with the PHAs and cause immunogenic reactions. On the other hand, Gram- positive bacteria lack LPS, a positive feature which justifies intensive investigation into their production of PHAs. This review summarizes currently available knowledge on PHA production by Gram- positive bacteria especially Bacillus and Streptomyces. We hope that this will form the basis of further research into developing either or both as a source of PHAs for medical applications.     

Effect of larval brain extracts derived from three swallowtail butterflies,Papilio helenus L., P. machaon L. and P. memnon L., on seasonal morph development of P. xuthus L. (Lepidoptera: Papilionidae)

Adults of the three papilionid butterflies, Papilio helenus L., Papilio machaon L. and Papilio memnon L., exhibit seasonal diphenism comprising spring and summer morphs. To elucidate the physiological mechanism underlying seasonal morph development in papilionid butterflies, we investigated whether a cerebral factor showing summer‐morph‐producing hormone (SMPH) activity is present in the brain of three Papilio species using an assay system with chilled male short‐day pupae of P. xuthus L. When 2% NaCl extracts derived from 20 larval brains of the three species were injected into abdomens of chilled male short‐day pupae of P. xuthus, all recipients destined to develop into spring‐morph adults developed into summer‐ and intermediate‐morph adults. On the other hand, all recipients injected with distilled water as a control developed into spring‐morph adults. These results indicate that a cerebral factor showing SMPH activity is present in the larval brain of the three Papilio species. Additionally, all recipients injected with 2% NaCl extracts derived from 20 adult brains of Bombyx mori L. also developed into summer‐ and intermediate‐morph adults. The results revealed that SMPH or a cerebral factor showing SMPH activity is widely distributed among lepidopteran insects.     

Impact of biological control agents on fusaric acid secreted from <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">gladioli</Emphasis> (Massey) Snyder and Hansen in <Emphasis Type="Italic">Gladiolus grandiflorus</Emphasis> corms

Fusaric acid (FA) (5-n-butylpuridine 2-carboxyl acid), a highly toxic secondary metabolite produced by Fusarium oxysporum strains, plays a significant role in disease development. The abilities of three F. oxysporum f. sp. gladioli (Massey) Snyder and Hansen isolates (G010; 649-91; and 160-57) to produce FA in infected Gladiolus corm tissues was evaluated in vitro in relation to the presence of two biological control agents, Trichoderma harzianum T22, and Aneurinobacillus migulanus. Pathogenicity tests were used to differentiate between the abilities of the F. oxysporum strains to secrete FA. FA was identified using LC/MS and quantified using HPLC. Isolate G010 was significantly more virulent (P < 0.01) on Gladiolus grandiflorus corms; it secretes 1.8 μM FA/g fresh weight corm into inoculated Gladiolus. Moreover, G010 was the only isolate that produced FA among the three examined isolates. There was a correlation between the corm lesion area and the FA secretion ability of F. oxysporum f. sp. gladioli (P < 0.001; r ² = 0.96). No FA was detected in PDA cultures of F.oxysporum f. sp. gladioli isolates. The presence of T. harzianum T22 appeared to prevent FA secretion into the corms. In the presence of A. migulanus, however, the amount of FA secreted into the corm tissues increased. These results support the use of T. harzianum as an effective biological control agent against F. oxysporum f. sp. gladioli.     

Improved Nile Red staining of <Emphasis Type="Italic">Nannochloropsis</Emphasis> sp.

High-throughput screening of microalgae for use as a potential feedstock for biodiesel requires a reliable method for the rapid detection of intracellular neutral lipid content. In this study, we report a modified and improved Nile Red (NR) fluorescence staining procedure for use as a rapid and sensitive screening tool to estimate levels of intracellular neutral lipid in the picopleustonic microalgae, Nannochloropsis sp. Addition of either glycerol or dimethyl sulfoxide (DMSO) into microalgae cultures greatly enhances lipid staining efficiency and increases the fluorescence intensity of stained cells. The optimized procedure requires glycerol and DMSO at the concentration of 0.1 and 0.165 g mL⁻¹, respectively, for peak fluorescence in a live culture of Nannochloropsis sp. Incubation for 5 min for glycerol-NR staining and 10 min for DMSO-NR staining at room temperature, in darkness, is used for the NR concentration of 0.3 and 0.7 μg mL⁻¹ for glycerol and DMSO, respectively. For the selection of lipid-rich cells of Nannochloropsis sp. using flow cytometric cell sorting, the glycerol-NR procedure is recommended as glycerol, unlike DMSO, does not inhibit subsequent growth of sorted cells.     

Testing for Homogeneity in Meta‐Analysis I. The One‐Parameter Case: Standardized Mean Difference

Summary Meta‐analysis seeks to combine the results of several experiments in order to improve the accuracy of decisions. It is common to use a test for homogeneity to determine if the results of the several experiments are sufficiently similar to warrant their combination into an overall result. Cochran’s Q statistic is frequently used for this homogeneity test. It is often assumed that Q follows a chi‐square distribution under the null hypothesis of homogeneity, but it has long been known that this asymptotic distribution for Q is not accurate for moderate sample sizes. Here, we present an expansion for the mean of Q under the null hypothesis that is valid when the effect and the weight for each study depend on a single parameter, but for which neither normality nor independence of the effect and weight estimators is needed. This expansion represents an order O(1/n) correction to the usual chi‐square moment in the one‐parameter case. We apply the result to the homogeneity test for meta‐analyses in which the effects are measured by the standardized mean difference (Cohen’s d‐statistic). In this situation, we recommend approximating the null distribution of Q by a chi‐square distribution with fractional degrees of freedom that are estimated from the data using our expansion for the mean of Q. The resulting homogeneity test is substantially more accurate than the currently used test. We provide a program available at the Paper Information link at the Biometrics website http://www.biometrics.tibs.org for making the necessary calculations.     

Comparison of seed and seedling functional traits in native Helianthus species and the crop H. annuus (sunflower)

• Seed functional traits of native Helianthus species contribute towards ecosystem services but limitations to their use in managed programmes exist. Many perennial Helianthus possess seed dormancy. The ability for germination to occur under different temperature and drought conditions, as well as the capacity of germinated seeds to convert into normal seedlings is rarely considered. Our aim was to identify and quantify these constraints through functional trait analyses.
• In five seed lots of native Helianthus (four perennial and one annual) and five genotypes of sunflower (H. annuus) for comparison, dormancy, thermal and hydro thresholds and times, morphology, mass, oil content and conversion into normal seedlings were quantified. The influence of the seed collection site environment on these traits was also explored.
• Seed dormancy of the perennial species was overcome by scarification followed by germination in 5 mm GA₃. Thermal and hydro‐time analyses revealed slower germination for the native seed lots (>1350 °Ch) in comparison to the sunflower genotypes (<829.9 °Ch). However, native seed lots had a higher capacity to convert into normal seedlings at high temperatures and low water potentials than sunflower genotypes. For the native seed lots, the average monthly temperature of the collection site was negatively correlated with thermal time.
• Variability in seed functional traits of native Helianthus and greater capacity for germinated seeds to convert into normal seedlings suggests they are better equipped to cope with high temperature and drought scenarios than sunflower. Effective dormancy alleviation is required to facilitate the use of native Helianthus species.