High-frequency<Emphasis Type="Italic">in vitro</Emphasis> flowering in six species of<Emphasis Type="Italic">ceropegia</Emphasis>

High-frequencyin vitro flowering is reported here fromin vitro regenerated shoots ofin vitro-raised seedlings of rare and endemicCeropegia lawii, Ceropegia maccannii, Ceropegia oculata, andCeropegia sahyadrica, as well as the widely distributedCeropegia bulbosa var.bulbosa andCeropegia hirsuta. In our first set of experiments, the MS medium contained 87 mM sucrose and was supplemented with varying concentrations of BAP (4.4 to 26.6 μM). For the second set of trials, varying concentrations of sucrose (87 to 233 mM) were tested in MS media containing a constant 4.4 p.M BAP. Sub-cultured apical as well as axillary buds flowered with similar frequencies after 30 d of incubation. For all six species, the highest percentage of flowering shoots was obtained with either 26.6 μM BAP or 175 mM sucrose. Although smaller in size, theirin vitro flowers were morphologically comparable within wVo-derived flowers. Variations among species were noted for the number of flower buds per shoot and the percentage of flower formation. Because all six species showed similar responses in both experiments, we can suggest that this protocol is applicable across the wide range ofCeropegia species.     

Coumarins in Eremocitrus glauca; Rutaceae; chemotaxonomy; coumarins; artifact; MS fragmentation.

2′,3′-Dihydroxydihydrosuberosin its acetonide derivative and 7-hydroxycoumarin has been isolated from the extracts of Eremocitrus glauca. The s     

Ndst1, a heparan sulfate modification enzyme, regulates neuroectodermal patterning by enhancing Wnt signaling in Xenopus

Neural tissue is derived from three precursor regions: neural plate, neural crest, and preplacodal ectoderm. These regions are determined by morphogen-mediated signaling. Morphogen distribution is generally regulated by binding to an extracellular matrix component, heparan sulfate (HS) proteoglycan. HS is modified by many enzymes, such as N-deacetyl sulfotransferase 1 (Ndst1), which is highly expressed in early development. However, functions of HS modifications in ectodermal patterning are largely unknown. In this study, we analyzed the role of Ndst1 using Xenopus embryos. We found that ndst1 was expressed in anterior neural plate and the trigeminal region at the neurula stage. ndst1 overexpression expanded the neural crest (NC) region, whereas translational inhibition reduced not only the trigeminal region, but also the adjacent NC region, especially the anterior part. At a later stage, ndst1 knocked-down embryos showed defects in cranial ganglion formation. We also found that Ndst1 activates Wnt signaling pathway at the neurula stage. Taken together, our results suggest that N-sulfonated HS accumulates Wnt ligand and activates Wnt signaling in ndst1-expressing cells, but that it inhibits signaling in non-ndst1-expressing cells, leading to proper neuroectodermal patterning.     

Effect of abscisic acid and proline on <Emphasis Type="Italic">in vitro</Emphasis> flowering in <Emphasis Type="Italic">Vigna aconitifolia</Emphasis>

An experiment was taken up to find out possibilities of manipulating the in vitro flowering in moth bean. Abscisic acid (ABA) and proline both alone and in combination influenced days to flower induction, number of flowers per plant, number of pods per plant and seeds per pod. Frequency of flowering plants approached 100 % at 1 and 3 μM ABA and 800 μM proline. The range of flowering period (3 to 23.6 d) has also been influenced by various treatments.     

Codon-Anticodon Binding in tRNA<Superscript>phe</Superscript>

THE anticodon loop of tRNA^phe of baker's yeast has the sequence (5′ to 3′) AY A A MeG U MeC. The unusual base Y, adjacent to the anticodon (AA MeG), is the only nucleotide in this tRNA which fluoresces at room temperature and because it absorbs to the red of all other bases, the excitation energy is localized on it exclusively. (7-Methyl guanine is another base in tRNA^phe which fluoresces in these conditions but its emission is so weak that it can only be observed in tRNA^phe from which Y has been excised.) The fluorescence spectrum undergoes a small blue shift in the presence of the complementary codon¹ and we report now the use of this shift to determine the association constants for this binding at several temperatures. The results suggest a simple thermodynamic model for the codon recognition step during protein synthesis.     

Survey of Helminth Parasites in Populations of <Emphasis Type="Italic">Alouatta palliata mexicana</Emphasis> and <Emphasis Type="Italic">A. pigra</Emphasis> in Continuous and in Fragmented Habitat in Southern Mexico

The 2 howler species that occur in southern Mexico, Alouatta palliata mexicana and Alouatta pigra are endangered, mainly as a result of habitat loss and fragmentation from human activity. Little is known about the gastrointestinal parasite communities affecting their populations, and lack of baseline information for populations of howler species in continuous forest habitats, makes evaluations of gastrointestinal parasite prevalence in populations in fragmented landscapes difficult. We report the results of a one-time broad survey of gastrointestinal parasites in fecal samples of individuals from several demographically stable populations of Alouatta palliata mexicana and A. pigra existing in continuous and/or protected forests. We further report similar data for populations of both species in human-fragmented landscapes. We detected 6 parasites for each howler monkey species, but only 3 of them (Trematode I, Controrchis biliophilus, Trypanoxyuris sp.) were common to both species. While parasitic prevalence in populations of both howler species was, in general, higher in the fragmented habitat than in continuous and/or protected forests. The difference is only marginally significant in Alouatta pigra. Some parasites (Coccidia and Strongylid) only appeared in populations in fragmented landscapes. Preliminary data suggest that adult males tended to have higher parasite prevalence values than those of adult females in both howler species. Parasite prevalence is associated to average group size, but not to population density in Alouatta pigra.     

Species specificity in settling-plant selection by foundress ant queens in <Emphasis Type="Italic">Macaranga</Emphasis>–<Emphasis Type="Italic">Crematogaster</Emphasis> myrmecophytism in a Bornean dipterocarp forest

Previous studies have demonstrated that the obligate myrmecophytism between Macaranga ant-plants and Crematogaster plant-ants is highly species specific, although multiple Macaranga species can coexist in a microhabitat. However, the species specificity has been described based on the study of trees with established plant-ant colonies. We studied how the process of settling into the partner Macaranga seedlings by single foundress Crematogaster queens contributes to species specificity. By sampling seedlings of three sympatric Macaranga myrmecophytes species in the field, we tested two hypotheses. The first is that foundresses correctly select their specific partner plant species when they settle into seedlings. The second hypothesis is that the seasons in which seedlings available for settling by foundresses appear are segregated among the Macaranga species, and the seasons in which foundress queens settle are synchronized to the appearance of seedlings of specific partner species; thus species specificity is consequently generated. Our results support the former hypothesis but not the latter: we always observed foundresses settling species-specific host plants, and seedlings suitable for settling were always available in each Macaranga species. Electronic Publication     

Occurrence of <Emphasis Type="Italic">Chlamydiaceae</Emphasis> in non-symptomatic free-living raptors in Spain

Few studies have investigated the role of raptors as natural reservoirs of Chlamydiaceae spp. and the preferred anatomical sites where these bacteria can be detected in non-symptomatic wild birds. We investigated the occurrence of Chlamydiaceae in 54 non-symptomatic adult free-living birds belonging to 14 species sampled upon reception in a raptor rehabilitation centre in Spain, and ten juvenile birds from five species born and reared in the centre for subsequent release into the wild. Swabs from conjunctivae, choanae and cloacae were taken to detect Chlamydiaceae DNA by a family-specific polymerase chain reaction (PCR) and a nested genus- and species-specific PCR. Chlamydiaceae DNA was detected in adult raptors belonging to 12 species (85.7%), mainly in conjunctival (40.6%) and, to a lesser extent, choanal (17.2%) swabs, but never in cloacal samples. Neither the genus nor the species of Chlamydiaceae could be confirmed by the nested PCR assay. Our results suggest that most of the raptor species investigated, especially the Common Kestrel (Falco tinnunculus) and Eurasian Griffon Vulture (Gyps fulvus), can be natural reservoirs of Chlamydiaceae spp. in the wild. Failure to identify the organisms at genus and species level might have been due to the poor quality and low concentration of DNA in the samples or to the presence of hitherto unclassified Chlamydiaceae species.     

Hormonal control of the outgrowth of axillary buds in <Emphasis Type="Italic">Alstroemeria</Emphasis> cultured <Emphasis Type="Italic">in vitro</Emphasis>

We study apical dominance in Alstroemeria, a plant with an architecture very different from the model species used in research on apical dominance. The standard explant was a rhizome with a tip and two vertically growing shoots from which the larger part had been excised leaving ca. 1 cm stem. The axillary buds that resumed growth were located at this 1-cm stem just above the rhizome. They were released by removal of the rhizome tip and the shoot tips. Replacement of excised tips by lanolin with indole-3-butyric acid (IBA) restored apical dominance. The auxin transport inhibitors 2,3,5-triiodobenzoic acid (TIBA) and N-1-napthylphthalamic acid (NPA) reduced apical dominance. 6-Benzylaminopurine (BAP) enhanced axillary bud outgrowth but the highest concentrations (> 9 μM) caused fasciation. Thidiazuron (TDZ) did not show improvement relative to BAP. Even though the architecture of Alstroemeria and the model species are very different, their hormonal mechanisms in apical dominance are for the greater part very similar.     

Identification and transcriptional analysis of genes involved in <Emphasis Type="Italic">Bacillus cereus</Emphasis>-induced systemic resistance in <Emphasis Type="Italic">Lilium</Emphasis>

Bacillus cereus C1L was demonstrated to induce systemic resistance in Lilium formosanum against leaf blight caused by Botrytis elliptica. Suppression subtractive hybridization library of L. formosanum triggered by B. cereus C1L were screened and 3 differentially expressed genes were identified. Based on sequence analysis, these genes encoding putative glycine-rich protein, metallothionein-like protein, and PsbR protein of photosystem 2, were designated LfGRP1, LfMT1, and LfPsbR, respectively. The results of Northern blot analysis showed that expressions of LfGRP1, LfMT1 and LfPsbR increased in response to B. elliptica infection. On the other hand, expression of LfMT1 increased but expressions of LfGRP1 and LfPsbR decreased when the rhizosphere of L. formosanum was drenched with suspension of B. cereus C1L with or without subsequent challenge with B. elliptica on lily leaves. Similar expression profiles of homologues of LfGRP1, LfMT1, and LfPsbR (named LsGRP1, LsMT1, and LsPsbR, respectively) were presented in Lilium oriental hybrid Star Gazer. In addition, application of the photosynthetic inhibitor, 3-(3,4-dichlorophenyl)-1,1-dimethylurea, on the leaves reduced disease severity and expressions of LsGRP1 and LsPsbR just as that in response to B. cereus C1L treatment.     

Recovery of photosynthetic capacity in <Emphasis Type="Italic">Vaccinium vitis</Emphasis>-<Emphasis Type="Italic">idaea</Emphasis> during mild spells in winter

Recent studies suggest that evergreen plants may maintain their photosynthetic capacity through the winter. Since mild winters are predicted to be more frequent in the future, the metabolic activity of plants is also likely to increase. The aim of the present study was to assess how various environmental factors, such as temperature, photoperiod and preceding frost, affect the recovery of photosynthesis during a mild spell in winter. The recovery of photosynthesis was studied in a series of growth chamber experiments where the overwintering of lingonberry (Vaccinium vitis-idaea) was interrupted by an intermittent warm spell of 1 week during different phases of winter. Rapid activation was observed in all the experiments during the first 3–4 days. No obvious effects of the phase of winter or photoperiod on the recovery of photosynthesis were observed, but a severe freezing treatment prior to the warm spell retarded the recovery significantly. Once recovered, however, lingonberry was able to maintain high rates of photosynthesis even at near-freezing temperatures, which prevail in their natural sub-nivean environment. The apparent quantum yield of photosynthesis remained high through the winter for lingonberry. This may prove advantageous for evergreen dwarf shrubs which overwinter in dim environments under snow.     

Light respiration in <Emphasis Type="Italic">Chlorella sorokiniana</Emphasis>

Respiration and photosynthesis are two important processes in microalgal growth that occur simultaneously in the light. To know the rates of both processes, at least one of them has to be measured. To be able to measure the rate of light respiration of Chlorella sorokiniana, the measurement of oxygen uptake must be fast, preferably in the order of minutes. We measured the immediate post-illumination respiratory O₂ uptake rate (OUR) in situ, using fiber-optic oxygen microsensors, and a small and simple extension of the cultivation system. This method enables rapid and frequent measurements without disturbing the cultivation and growth of the microalgae. Two batch experiments were performed with C. sorokiniana in a short light-path photobioreactor, and the OUR was measured at different time points. The net oxygen production rate (net OPR) was measured online. Adding the OUR and net OPR gives the gross oxygen production rate (gross OPR), which is a measure for the oxygen evolution by photosynthesis. The gross OPR was 35–40% higher than the net OPR for both experiments. The respiration rate is known to be related to the growth rate, and it is suggested that faster algal growth leads to a higher energy (ATP) requirement, and as such, respiratory activity increases. This hypothesis is supported by our results, as the specific OUR is highest in the beginning of the batch culture when the specific growth rate is highest. In addition, the specific OUR decreases toward the end of the experiments until it reaches a stable value of around 0.3 mmol O₂ h⁻¹ g⁻¹. This value for the specific OUR is equal to the maintenance requirement of C. sorokiniana as determined in an independent study of (Zijffers et al. 2010 (in press)). This suggests that respiration could fulfill the maintenance requirements of the microalgal cells.     

Quorum sensing regulation in <Emphasis Type="Italic">Pseudomonas</Emphasis>

Expression of many bacterial genes is regulated in a cell density-dependent manner via small signal molecules known as autoinducers; this type of regulation is termed quorum sensing (QS). The QS systems that employ N-acyl-homoserine lactones (HSLs) are best un derstood in Gram-negative bacteria. QS regulates expression of various genes, including the genes responsible for the production of virulence factors, synthesis of exoenzymes and antibiotics, antagonistic properties of bacteria, etc. The QS systems of the genus Pseudomonas are linked to other global regulatory networks of the cell, and their functions are controlled by numerous additional regulatory factors. Such regulators and the QS systems together form an intricate multifactorial cascade regulatory network. The review considers the QS systems of several Pseudomonas species, their interaction with other regulatory systems, and their roles in the regulation of cell processes.     

Photosynthetic characteristics in <Emphasis Type="Italic">Oryza</Emphasis> species

Photosynthetic rate (P _N), SPAD value, specific leaf area (SLA), flag leaf area (FLA), and nitrogen content (LN) of genus Oryza were investigated and their correlation was analyzed to assess some of the main photosynthetic traits among different species in the genus Oryza. The results revealed wide variation in these traits. The species O. rufipogon and O. australiensis exhibited maximum photosynthetic rate. Comparison of different types of genomes (diploid: 2n=2x=24; tetraploid: 2n=4x=48) and growth habit (shade- or sun-grown) showed the species of diploid (with genome symbol EE; 2n=2x=24) genomes, with perennial and sun-grown species, had high apparent photosynthesis compared to others. The species with BB/BBCC, shade-grown and the tetraploids showed high SPAD value, and the flag leaf in sun-grown species and diploids were thicker (low SLA) compared with others. However, no significant difference could be noticed among the different types of genomes. Higher leaf area was noticed among the species of CC/CCDD genome, perennial shade-grown species and tetraploids than in others. The variety IR 36 exhibited highest leaf nitrogen concentration. Correlation analysis showed a strong relationship between P _N and leaf nitrogen concentration while no marked relationships were observed among other characteristics. It implies that the species with thick and small leaves with high nitrogen concentration and high photosynthesis evolved better than others. O. rufipogon, with the same genome as O. sativa, could be one of the wild rice resources for elite crop improvement.