Demographic response by a small epiphytic orchid

? Premise of the study: Biotic changes are an inevitable consequence of climate change. Epiphytes may be more susceptible to changes in climate variation, but data regarding responses to climate variability under field conditions are limited. We evaluated whether the abundance of demographic stages in the epiphytic orchid Lepanthes rupestris at the Luquillo Experimental Forest in Puerto Rico was associated with short-term changes in climate variation over an 8-yr period. ? Methods: We used cross-correlation analyses to evaluate associations between the abundance of seedlings, juveniles, adults, and fruits per subpopulation, population growth, colonization and extinction rates in L. rupestris with variables related to precipitation and temperature, with and without lag- responses. ? Key results: We detected significant negative correlations between the average number of seedlings and the number of dry days, between the average number of fruits and minimum average temperature with a 6-mo response lag, and between the average number of adults and the maximum temperature with a 1-yr response lag. Neither population growth rate nor probability of colonization and extinction were directly related to climatic variation between 2000 and 2007. ? Conclusions: Associations between climatic variables and demographic stages could have negative implications for this orchid within the context of expected drying trends in the Caribbean. Results argue for the establishment of long-term monitoring studies of orchid populations, because only long-term studies would provide the appropriate temporal scale to detect and predict climate change effects and adaptive management of orchid populations.     

Genetic analysis of the O-antigen gene clusters of Yersinia pseudotuberculosis O:6 and O:7

Among the 21 O-polysaccharide (OPS) O-antigen-based serotypes described for Yersinia pseudotuberculosis, those of O:6 and O:7 are unusual in that both contain colitose (4-keto-3,6-dideoxy-d-mannose or 4-keto-3,6-dideoxy-l-xylo-hexose), which has not otherwise been reported for this species, and the O:6 OPS also contains yersiniose A (4-C[(R)-1-hydroxyethyl]-3,6-dideoxy-d-xylo-hexose), another unusual dideoxyhexose sugar. In Y. pseudotuberculosis, the genes for OPS synthesis generally cluster together between the hemH and gsk loci. Here, we present the sequences of the OPS gene clusters of Y. pseudotuberculosis O:6 and O:7, and the location of the genes required for synthesis of these OPSs, except that there is still ambiguity regarding allocation of some of the glycosyltransferase functions. The O:6 and O:7 gene clusters have much in common with each other, but differ substantially from the group of 13 gene clusters already sequenced, which share several features and sequence similarities. We also present a possible sequence of events for the derivation of the O:6 and O:7 gene clusters from the most closely related set of 13 sequenced previously.     

Knockdown of dystrophin Dp71 impairs PC12 cells cycle: localization in the spindle and cytokinesis structures implies a role for Dp71 in cell division

The function of dystrophin Dp71 in neuronal cells remains to be established. Previously, we revealed the involvement of this protein in both nerve growth factor (NGF)-induced neuronal differentiation and cell adhesion by isolation and characterization of PC12 neuronal cells with depleted levels of Dp71. In this work, a novel phenotype of Dp71-knockdown cells was characterized, which is their delayed growth rate. Cell cycle analyses revealed an altered behavior of Dp71-depleted cells, which consists of a delay in G0/G1 transition and an increase in apoptosis during nocodazole-induced mitotic arrest. Dp71 associates with lamin B1 and β-dystroglycan, proteins involved in aspects of the cell division cycle; therefore, we compared the distribution of Dp71 with that of lamin B1 and β-dystroglycan in PC12 cells at mitosis and cytokinesis by means of immunofluorescence and confocal microscopy analysis. All of these three proteins exhibited a similar immunostaining pattern, localized at mitotic spindle, cleavage furrow, and midbody. It is noteworthy that a drastic decreased staining in mitotic spindle, cleavage furrow, and midbody was observed for both lamin B1 and β-dystroglycan in Dp71-depleted cells. Furthermore, we demonstrated the interaction of Dp71 with lamin B1 in PC12 cells by immunoprecipitation and pull-down assays, and importantly, we revealed that knockdown of Dp71 expression caused a marked reduction in lamin B1 levels and altered localization of the nuclear envelope protein emerin. Our data indicate that Dp71 is a component of the mitotic spindle and cytokinesis multi-protein apparatuses that might modulate the cell division cycle by affecting lamin B1 and β-dystroglycan levels.     

Altered expression of chitin synthetase activity and biochemical changes in the cell wall in a developmental mutant ofPhycomyces

The Phycomyces developmental mutant S356 elaborates spores which show a much poorer viability and a higher affinity for Calcofluor White than the wild-type spores. Protease-activated extracts of the mutant spores showed higher levels of chitin synthetase activity than the parental strain-derived spores. High levels of enzyme activity in the mutant extracts, but not in the corresponding wild-type extracts, could be detected in the absence of an exogenous protease. The high basal active chitin synthetase is not the result of activation by endogeneous proteases during cell breakage since protease inhibitors did not reduce, but rather increased, the activity levels. The analysis of cell wall composition in the mutant spores revealed significant changes in the proportion of uronic acids and protein but not in chitin. The mutant phenotype is discussed in relation to the developmental stage at which the alterations connected with cell wall metabolism occurred.     

Disobedient epiphytes: colonization and extinction rates in a metapopulation of Lepanthes rupestris (Orchidaceae) contradict theoretical predictions based on patch connectivity

The metapopulation concept is widely established in population biology. It predicts that the likelihood of colonization of an empty patch is positively correlated with its connectivity, because colonizers from occupied patches will be more likely to reach an empty patch if they are closer to it. Another prediction is that the likelihood of extinction of an occupied patch will be negatively correlated with its connectivity to other patches, as the occupied patch can be ‘reinforced’ by immigrants from patches that are close by. We tested these predictions using an extensive data set for an epiphytic orchid, Lepanthes rupestris from Puerto Rico. Our data did not support the first prediction, but we found that the likelihood of extinction is negatively correlated with patch connectivity. We hypothesize that this might be because most orchid seeds are wind dispersed and seeds that do not fall immediately below the mother plant are uniformly distributed after a steep leptokurtic distribution. We predict that taxa with similar seed and gene flow characteristics should show similar patterns in the association between colonization/extinction rates and patch connectivity. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 598–606.     

Specific genetic markers for detecting subtypes of dengue virus serotype-2 in isolates from the states of Oaxaca and Veracruz,Mexico

Background  

Dengue (DEN) is an infectious disease caused by the DEN virus (DENV), which belongs to the Flavivirus genus in the family Flaviviridae. It has a (+) sense RNA genome and is mainly transmitted to humans by the vector mosquito Aedes aegypti. Dengue fever (DF) and dengue hemorrhagic fever (DHF) are caused by one of four closely related virus serotypes (DENV-1, DENV-2, DENV-3 and DENV-4). Epidemiological and evolutionary studies have indicated that host and viral factors are involved in determining disease outcome and have proved the importance of viral genotype in causing severe epidemics. Host immune status and mosquito vectorial capacity are also important influences on the severity of infection. Therefore, an understanding of the relationship between virus variants with altered amino acids and high pathogenicity will provide more information on the molecular epidemiology of DEN. Accordingly, knowledge of the DENV serotypes and genotypes circulating in the latest DEN outbreaks around the world, including Mexico, will contribute to understanding DEN infections.     

Nuclear and nuclear envelope localization of dystrophin Dp71 and dystrophin-associated proteins (DAPs) in the C2C12 muscle cells: DAPs nuclear localization is modulated during myogenesis

Dystrophin and dystrophin-associated proteins (DAPs) form a complex around the sarcolemma, which gives stability to the sarcolemma and leads signal transduction. Recently, the nuclear presence of dystrophin Dp71 and DAPs has been revealed in different non-muscle cell types, opening the possibility that these proteins could also be present in the nucleus of muscle cells. In this study, we analyzed by Immunofluorescence assays and Immunoblotting analysis of cell fractions the subcellular localization of Dp71 and DAPs in the C(2)C(12) muscle cell line. We demonstrated the presence of Dp71, alpha-sarcoglycan, alpha-dystrobrevin, beta-dystroglycan and alpha-syntrophin not only in plasma membrane but also in the nucleus of muscle cells. In addition, we found by Immunoprecipitation assays that these proteins form a nuclear complex. Interestingly, myogenesis modulates the presence and/or relative abundance of DAPs in the plasma membrane and nucleus as well as the composition of the nuclear complex. Finally, we demonstrated the presence of Dp71, alpha-sarcoglycan, beta-dystroglycan, alpha-dystrobrevin and alpha-syntrophin in the C(2)C(12) nuclear envelope fraction. Interestingly, alpha-sarcoglycan and beta-dystroglycan proteins showed enrichment in the nuclear envelope, compared with the nuclear fraction, suggesting that they could function as inner nuclear membrane proteins underlying the secondary association of Dp71 and the remaining DAPs to the nuclear envelope. Nuclear envelope localization of Dp71 and DAPs might be involved in the nuclear envelope-associated functions, such as nuclear structure and modulation of nuclear processes.