Nuclear localization of the Arabidopsis blue light receptor cryptochrome 2

The cryptochrome blue light photoreceptor family of Arabidopsis thaliana consists of two members, CRY1 and CRY2 (PHH1). CRY2 contains a putative nuclear localization signal (NLS) within its C-terminal region. We examined whether CRY2 is localized in the nucleus and whether the C-terminal region of CRY2 is involved in nuclear targeting. Total cellular and nuclear protein extracts from Arabidopsis were subjected to immunoblot analysis with CRY2-specific antibodies. Strong CRY2 signals were obtained in the nuclear fraction. Fusion proteins consisting of the green fluorescent protein (GFP) and different fragments of CRY2 were expressed in parsley protoplasts and the localization of the fusion proteins was determined by fluorescence and confocal laser scanning microscopy. GFP-fusions containing the entire CRY2 protein or its C-terminal region were found exclusively in the nucleus. We conclude from these results that CRY2 is localized in the nucleus and that nuclear localization is mediated by the C-terminal region of CRY2.     

The Arabidopsis blue light receptor cryptochrome 2 is a nuclear protein regulated by a blue light-dependent post-transcriptional mechanism

Cryptochrome 2 is a flavin-type blue light receptor mediating floral induction in response to photoperiod and a blue light-induced hypocotyl growth inhibition. cry2 is required for the elevated expression of the flowering-time gene CO in response to long-day photoperiods, but the molecular mechanism underlying the function of cry2 is not clear. The carboxyl domain of cry2 bears a basic bipartite nuclear localization signal, and the cry2 protein was co-fractionated with the nucleus. Analysis of transgenic plants expressing a fusion protein of CRY2 and the reporter enzyme GUS (GUS-CRY2) indicated that the GUS-CRY2 fusion protein accumulated in the nucleus of transgenic plants grown in dark or light. The C-terminal domain of cry2 that contains the basic bipartite nuclear localization signal was sufficient to confer nuclear localization of the fusion protein. Phenotypic analysis of transgenic plants expressing the fusion protein GUS-CRY2 demonstrated that GUS-CRY2 acts as a functional photoreceptor in vivo, mediating the blue light-induced inhibition of hypocotyl elongation. These results strongly suggest that cry2 is a nuclear protein. Although no obvious light regulation was found for the nuclear compartmentation of GUS-CRY2 fusion protein, the abundance of GUS-CRY2 was regulated by blue light in a way similar to that of cry2.     

The taxonomy and affinities of Papaver gorgoneum from the Cape Verde Islands

A new subspecies of Papaver gorgoneum , ssp. theresias is described from the Cape Verde Islands, West Africa. A detailed comparison of the 2 subspecies is made and the ecology, the affinities of the species in the genus and the origin are discussed. The chromosome numbers of both subspecies (2n = 38) are reported for the first time and a note on the compatibility system is made.     

Hematozoa in endemic subspecies of common kestrel in the Cape Verde Islands

We examined 130 Common Kestrel (Falco tinnunculus) representing two endemic subspecies and nine resident island populations on the Cape Verde archipelago between 1996 and 1999 to study diversity, prevalence, and intensity of hematozoa. Hematozoan diversity was very low; we detected only Plasmodium fallax, a species that is rarely found in Falconoformes, and, possibly, Haemoproteus brachiatus. Moreover, prevalence of Plasmodium fallax was low (1.5%) with a mean intensity of infection of 0.05 protozoa/10(-3) erythrocytes. Only one bird (0.8%) was infected with a gametocyte that was most likely Haemoproteus brachiatus; the intensity in this infected bird was 1.5 protozoa/10(-3) erythrocytes. A single parasite or two parasites were observed in blood smears in four additional birds, but identification to genus was not possible. This is the first record of blood parasites in birds on the Cape Verde Archipelago. The low prevalence of these parasites might be because of arid and less-favorable conditions for the pathogen's vectors. The sedentary nature and high level of isolation of the island kestrel populations are also factors that could decrease the probability of infection.     

The apid cuckoo bees of the Cape Verde Islands (Hymenoptera, Apidae)

The apid cuckoo bees of the Cape Verde Islands (Republic of Cape Verde) are reviewed and five species recognized, representing two genera. The ammobatine genus Chiasmognathus Engel (Nomadinae: Ammobatini), a specialized lineage of cleptoparasites of nomioidine bees is recorded for the first time. Chiasmognathus batelkaisp. n. is distinguished from mainland African and Asian species. The genus Thyreus Panzer (Apinae: Melectini) is represented by four species - Thyreus denoliisp. n., Thyreus batelkaisp. n., Thyreus schwarzisp. n., and Thyreus aistleitnerisp. n. Previous records of Thyreus scutellaris (Fabricius) from the islands were based on misidentifications.     

Resistance to antibiotics in humans and animals on the Cape Verde Islands

From the Cape Verde Islands no studies are known about an efficient treatment with antibiotics and chemotherapeutics. Because of this human samples and samples of domestic animals were collected, the different bacterias were isolated and the antibiograms were analyzed by agar-diffusions-test. The human samples consisted of wound-, eye-, anal-, vaginal- and throat-smears, the animal samples consisted of jaw-, anal- and wound-smears. the aim of these investigations was to make efficient treatment with antibiotics and chemotherapeutics possible. The best efficacy had Cefotaxim, Gentamicin and Trimethoprim + Sulfonamid. Very poor were Penicillin G, Aminopenicillin and Cephalexin.     

The role of cryptochrome 2 in flowering in Arabidopsis

We have investigated the genetic interactions between cry2 and the various flowering pathways in relation to the regulation of flowering by photoperiod and vernalization. For this, we combined three alleles of CRY2, the wild-type CRY2-Landsberg erecta (Ler), a cry2 loss-of-function null allele, and the gain-of-function CRY2-Cape Verde Islands (Cvi), with mutants representing the various photoreceptors and flowering pathways. The analysis of CRY2 alleles combined with photoreceptor mutants showed that CRY2-Cvi could compensate the loss of phyA and cry1, also indicating that cry2 does not require functional phyA or cry1. The analysis of mutants of the photoperiod pathway showed epistasis of co and gi to the CRY2 alleles, indicating that cry2 needs the product of CO and GI genes to promote flowering. All double mutants of this pathway showed a photoperiod response very much reduced compared with Ler. In contrast, mutations in the autonomous pathway genes were additive to the CRY2 alleles, partially overcoming the effects of CRY2-Cvi and restoring day length responsiveness. The three CRY2 alleles were day length sensitive when combined with FRI-Sf2 and/or FLC-Sf2 genes, which could be reverted when the delay of flowering caused by FRI-Sf2 and FLC-Sf2 alleles was removed by vernalization. In addition, we looked at the expression of FLC and CRY2 genes and showed that CRY2 is negatively regulated by FLC. These results indicate an interaction between the photoperiod and the FLC-dependent pathways upstream to the common downstream targets of both pathways, SOC1 and FT.     

Notes on the genus Thamnobryum in Macaronesia (excluding the Cape Verde Islands)

Abstract

The genus Thamnobryum is reviewed from Macaronesia (excludingthe Cape Verde Islands). In this area there are three species, T. alopecurum (Hedw.) Gang., T. maderense (Kindb.) Hedenäs, comb. nov. and T. fernandesii C. Sérgio. T. fernandesii is only known from Madeira; the other two also occur on the Canary Islands and the Azores. Thamnobryum canariense (Ren. & Card.) Long is transferred to Isothecium, as I. atlanticum Hedenäs, comb. et nom. nov.     

Quantitative trait loci controlling aluminium tolerance in two accessions of Arabidopsis thaliana (Landsberg erecta and Cape Verde Islands)

Quantitative trait loci (QTL) analysis of aluminium (Al) tolerance was performed using Ler/Cvi recombinant inbred (RI) lines of Arabidopsis thaliana. Relative root length (RRL) (root length with 4 µm Al/root length with no Al at pH 5.0) on day 5 was used as the Al tolerance index for QTL analysis. Al tolerance judged by RRL was well correlated to tolerance judged by other indexes, including accumulation of callose, reactive oxygen species in the root apex and growth performance on acid soil containing a large amount of exchangeable Al. Using data sets with an h_b² of 0.91, two QTLs were detected at the top of chromosome 1 and bottom of chromosome 3. These QTLs explained 40 and 16% of the phenotypic variation of Al tolerance, respectively, and the positive effect of the Cvi allele. The QTL on chromosome 1 overlapped with a major QTL in another recombinant inbred population, and is possibly related to malate excretion. A complete pair-wise search revealed 11 sets of epistatic interacting loci pairs, which accounted for the transgressive segregation among the RI population. Several epistatic interactions shared the same chromosomal region, indicating the possible involvement of regulatory proteins in Al tolerance in Arabidopsis.     

Revision of the genus Diplotaxis (Brassicaceae) in the Cape Verde Islands, W Africa

The genus Diplotaxis (Brassicaceae) in the Cape Verde Islands, W Africa, is revised. Nine taxa are accepted, of which five are described as new: D. antoniensis sp. nov., D. glauca, D. gorgadensis sp. nov., D. gorgadensis ssp. brochmannii ssp. nov., D. gracilis, D. hirta, D. sundingii sp. nov., D. varia sp. nov., and D. vogelii . All species are suffruticose, yellow-flowered perennials and belong to sect. Catocarpum . The species are endemic to the Cape Verde Islands, but show morphological and karyolog-ical affinity to the N African and Mediterranean D. harra s. lat. Analyses of morphological variation in 90 populations (400 plants) revealed a complex pattern, in particular in vegetative characters, most likely evolved by parallel ecogeographical differentiation in different islands. The chromosome number is 2n = 26 (n = 13) in the five taxa investigated. Self-pollination experiments indicate that the species are self-incompatible and outcrossing. Experimental F₁ hybrids with full seed set after open pollination were obtained in 30 interspecific combinations, and the taxa are considered fully interfertile. The taxa are thus isolated mainly by geographical and partly by ecological barriers and have evolved by vicariant evolution (most islands) and adaptive radiation (one island).     

XXIV.—Further Notes on the Ornithology of the Cape Verde Islands.

The following observations are the result of a second visit to the Cape Verde Islands in October 1897. Before giving them, however, I should like to withdraw two suppositions made in my previous paper ( suprà , pp. 74–118).     

On the drinking water situation on the Cape Verde Islands (Island of Santiago)

On the Cape Verde Islands drinking water investigations were made. The drinking water was tested in chemical and bacteriological respects as well as on amebas and rotaviruses. The isolated germs were investigated on their reaction against several antibiotics and chemotherapeutics by means of the agar diffusion test. The changing of drinking water quality from water-supply-places to the households was investigated as well. According to the fact, that only 10.6% of 207 public water supplies investigated show drinking water quality after ONORM, and that nearly 30% and 80.2% of them contained E. coli and coliforme germs respectively, the urgency of extensive sanitations of water supplies on the Cape Verde Islands is evident.     

Population structure and conservation implications for the loggerhead sea turtle of the Cape Verde Islands

The Cape Verde Islands harbour the second largest nesting aggregation of the globally endangered loggerhead sea turtle in the Atlantic. To characterize the unknown genetic structure, connectivity, and demographic history of this population, we sequenced a segment of the mitochondrial (mt) DNA control region (380 bp, n = 186) and genotyped 12 microsatellite loci (n = 128) in females nesting at three islands of Cape Verde. No genetic differentiation in either haplotype or allele frequencies was found among the islands (mtDNA F _ST = 0.001, P > 0.02; nDNA F _ST = 0.001, P > 0.126). However, population pairwise comparisons of the mtDNA data revealed significant differences between Cape Verde and all previously sequenced Atlantic and Mediterranean rookeries (F _ST = 0.745; P < 0.000). Results of a mixed stock analysis of mtDNA data from 10 published oceanic feeding grounds showed that feeding grounds of the Madeira, Azores, and the Canary Islands, in the Atlantic Ocean, and Gimnesies, Pitiüses, and Andalusia, in the Mediterranean sea, are feeding grounds used by turtles born in Cape Verde, but that about 43% (±19%) of Cape Verde juveniles disperse to unknown areas. In a subset of samples (n = 145) we evaluated the utility of a longer segment (~760 bp) amplified by recently designed mtDNA control region primers for assessing the genetic structure of Atlantic loggerhead turtles. The analysis of the longer fragment revealed more variants overall than in the shorter segments. The genetic data presented here are likely to improve assignment and population genetic analyses, with significant conservation and research applications.     

Internal and External Influences on Near-Surface Microbial Community Structure in the Vicinity of the Cape Verde Islands

Microbial community structure in the subtropical north-east Atlantic Ocean was compared between 2 years and variation attributed to environmental variables. Surface seawater communities were analysed by flow cytometry and fluorescence in situ hybridisation. Probes specific to Alphaproteobacteria, Cyanobacteria, Gammaproteobacteria and Bacteroidetes identified 67–100% of cells. Due to natural variation in the study region due to the occurrence of major currents and islands, data could not be pooled but were instead divided between distinct water masses. Community structure did not differ greatly around the Cape Verde Islands between sampling periods but varied substantially in the open ocean, suggesting different environmental perturbations favour specific bacterial groups. Wind speed varied significantly between years, with moderate to strong breeze in winter 2008 and gales in winter 2006 (8.9 ± 0.2 ms⁻¹ and 16.0 ± 0.4 ms⁻¹, respectively). Enhanced wind-driven turbulence was associated with domination by the SAR11 clade of Alphaproteobacteria, which were present at 2.4-fold in the abundance of Prochlorococcus (41.8 ± 1.6% cells, compared to 17.7 ± 7.1%). Conversely, the calmer conditions of 2008 seemed to favour Prochlorococcus (40.0 ± 1.2% cells). Prochlorococcus high-light adapted clade HLI were only numerous during wind-driven turbulence, whereas oligotrophic-adapted clade HLII dominated under calm conditions. Bacteroidetes were most prominent in turbulent conditions (9.5 ± 1.3% cells as opposed to 4.7 ± 0.3%), as were Synechococcus. In 2008, a considerable dust deposition event occurred in the region, which may have led to the substantial Gammaproteobacteria population (22.5 ± 4.0% cells compared to 4.6 ± 0.6% in 2006). Wind-driven turbulence may have a significant impact on microbial community structure in the surface ocean. Therefore, community change following dust storm events may be linked to associated wind in addition to dust-derived nutrients.     

The signaling state of Arabidopsis cryptochrome 2 contains flavin semiquinone

Cryptochrome (Cry) photoreceptors share high sequence and structural similarity with DNA repair enzyme DNA-photolyase and carry the same flavin cofactor. Accordingly, DNA-photolyase was considered a model system for the light activation process of cryptochromes. In line with this view were recent spectroscopic studies on cryptochromes of the CryDASH subfamily that showed photoreduction of the flavin adenine dinucleotide (FAD) cofactor to its fully reduced form. However, CryDASH members were recently shown to have photolyase activity for cyclobutane pyrimidine dimers in single-stranded DNA, which is absent for other members of the cryptochrome/photolyase family. Thus, CryDASH may have functions different from cryptochromes. The photocycle of other members of the cryptochrome family, such as Arabidopsis Cry1 and Cry2, which lack DNA repair activity but control photomorphogenesis and flowering time, remained elusive. Here we have shown that Arabidopsis Cry2 undergoes a photocycle in which semireduced flavin (FADH(.)) accumulates upon blue light irradiation. Green light irradiation of Cry2 causes a change in the equilibrium of flavin oxidation states and attenuates Cry2-controlled responses such as flowering. These results demonstrate that the active form of Cry2 contains FADH(.) (whereas catalytically active photolyase requires fully reduced flavin (FADH(-))) and suggest that cryptochromes could represent photoreceptors using flavin redox states for signaling differently from DNA-photolyase for photorepair.     

Parallel ecoclinal evolution and taxonomy of F'rankenia (Frankeniaceae) in the Cape Verde Islands, W Africa

One species, the Macaronesian and N African Frankenia ericifolia , with three subspecies is recognized in the Cape Verde Islands: ssp. ericifolia (a coastal, xerophytic ecotype), ssp. caboverdeana ssp. nov. (a coastal, mesophytic ecotype), and ssp. montana ssp. nov. (a montane, hygrophytic ecotype). Subspecies caboverdeana and ssp. montana are endemic to the Cape Verde Islands. In addition, ssp. latifolia comb. et stat. nov. is tentatively recognized as endemic to the Canary Islands. Reproductive data suggest that the Capeverdean plants are self-compatible but mainly outcrossing. Mor-phometric analyses of field-collected material and progeny families cultivated under uniform conditions demonstrate that there is genetically based, clinal morphological variation connecting ssp. ericifolia and ssp. caboverdeana in several coastal areas that vary in humidity conditions because of varying exposures to the trade wind. Clinal variation was observed in leaf-, calyx-, petal-, and seed characters, probably caused by one or a few genetic factors that control organ size and are subjected to differential selection with respect to drought tolerance. The variation in progeny families derived from morphologically intermediate parents was no higher than the variation in progeny families derived from parents belonging to the "pure" subspecies. It is thus likely that the clinal variation is caused by primary evolution along continuous humidity gradients rather than hybridization. The existence of similar ecoclines in different islands and the potentially simple genetic basis of such clinality suggest that ssp. caboverdeana has evolved several times in mesic environments from xerophytic, ericifolia-like ancestors.     

An integrative taxonomic revision of the Tarentola geckos (Squamata,Phyllodactylidae) of the Cape Verde Islands

Recent phylogeographical analyses using mitochondrial DNA (mtDNA) sequences indicate that the Tarentola geckos from the Cape Verde archipelago originated from a propagule that dispersed from the Canary Islands approximately 7.7 Mya and that underwent a fast evolutionary radiation. Molecular analyses carried out to date clearly show some incongruences with the current taxonomy of Tarentola from the Cape Verde Islands, with some species being paraphyletic or polyphyletic, and several independently evolving lineages needing formal taxonomic recognition. The aim of this study was to clarify the systematics of this group to unravel its taxonomy by applying an integrative approach based on information from three independent sources: mtDNA, nuclear genes, and morphology. As a result of this taxonomic revision, two novel species for the islands of S. Nicolau and Fogo are described and eight subspecies are upgraded to species level. Moreover, an identification key for the genus Tarentola from the Cape Verde archipelago is presented. This study reconciles taxonomy and phylogeny in this group, provides a better understanding of diversity patterns, new insights on evolutionary hypotheses, and supports the basic framework for the future management and conservation of this unique reptile radiation. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 164 , 328–360.