Molecular analysis of genomic changes in synthetic autotetraploid Phlox drummondii Hook

Polyploidization is a major force that has shaped the evolution of flowering plants. Newly‐generated polyploids have been used to investigate the changes that occur in genomes immediately after polyploidization. The present research programme created colchitetraploids in Phlox drummondii and followed them through different generations (C₀, C₁, C₂, and C₃) to determine genomic changes. Genomic changes that followed autopolyploidization were investigated using amplified fragment length polymorphism (AFLP) and methylation‐sensitive amplified polymorphism (MSAP) technologies. The markers for which variation was evident were characterized by cloning and sequencing. Dot‐blotting was used to confirm increased or decreased abundance of the fragments for which variation was noted. We used bisulfite sequencing to determine changes in the levels of DNA methylation in the vicinities of the fragments that show variation. Our data indicate that these areas were more methylated in C₂ and C₃ colchitetraploids than in C₀ and C₁ colchitetraploids. The overall levels of methylation polymorphism found in C₂ and C₃ colchitetraploids were 1.0% and 1.65%, respectively. Changes were observed in both coding and noncoding regions. Some MSAP fragments that showed variation were similar to retrotransposons. Bisulfite sequencing indicated that CHH sites accounted for all the observed variation, whereas CG and CHG did not show any variation. The present study suggests that the genomic changes that accompany polyploidization in synthetic autopolyploids resemble those in allopolyploids. However, the extent of variation is lower in synthetic autopolyploids than in allopolyploids. These changes may help in the stabilization of the genomes of neo‐autopolyploids. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 591–605.     

Ranging and foraging of Himalayan grey langurs (Semnopithecus ajax) in Machiara National Park, Pakistan

Grey langurs (Semnopithecus spp.) occupy a variety of habitats, ranging from lowland forests and semi-desert to alpine forests. Little is known about their foraging and ranging in alpine forests, which appear to contain less food than lowland forests. We conducted a 1-year study of Himalayan grey langurs (Semnopithecus ajax) in Machiara National Park, Pakistan, where they occur at relatively high altitudes (range 2000–4733 m). We followed three groups of different sizes and compositions and examined the effects of ecological and social factors on ranging and feeding. The home-range sizes of a small bisexual group (SBG), a large bisexual group (LBG), and an all-male group (AMG) were 2.35 ± 0.92 (mean ± SD; average of four seasons), 3.28 ± 0.55, and 3.52 ± 1.00 km², respectively, and were largest in winter for all groups. The daily path lengths of the SBG, LBG, and AMG were 1.23 ± 0.28 (mean ± SD; average of four seasons), 1.75 ± 0.34, and 1.84 ± 0.70 km, respectively; that of the LBG was longer in winter, while that of the AMG was shorter in summer. Both the home-range size and daily path length of the AMG were larger than those of the other groups, even after partialling out the effect of group size differences. The mean altitude used by the langurs and the proportion of animals seen feeding did not differ among seasons or group types. As the mean temperature increased, the altitude used by langurs significantly increased for the SBG and LBG, but not for the AMG. On the other hand, as the temperature increased, the home-range sizes significantly decreased for the SBG and AMG, but not for the LBG. Rainfall did not show any correlation with ranging or feeding in any of the groups. Our results suggested that grey langurs in Machiara National Park employ a high-cost, high-return foraging strategy in winter, and that the ranging of the AMG also reflects its reproductive strategy.     

Sequence analysis of the internal transcribed spacer 2 (ITS2) from Philornis seguyi (García, 1952) and Philornis torquans (Nielsen, 1913) (Diptera: Muscidae)

Philornis Meinert, 1890 (Diptera: Muscidae) is a genus of Neotropical dipterans that parasitise birds. The currently used external morphological characters to distinguish between species within this genus present some limitations. We used the second internal transcribed spacer region (ITS2) of the rRNA gene as a molecular marker to differentiate adult specimens of Philornis identified morphologically as Philornis torquans and Philornis seguyi from different localities. Specimens identified as P. seguyi from Magdalena (Buenos Aires Province) showed an ITS2 sequence different from that for P. torquans, whereas all other specimens of P. seguyi had sequences identical to those for P. torquans. These findings do not necessarily confirm that specimens from Magdalena indeed belong to P. seguyi, nor that P. seguyi is a valid species. Instead, they alert us about the potential for species misidentification when using morphological characters alone. The use of molecular approaches to aid the identification of Philornis spp. will shed light on the systematics of this group. P. torquans is reported for the first time in Mendoza Province and Uruguay.     

Soil volume and carbon storage shifts in drained and afforested wetlands of the Paraná River Delta

Wetland ecosystems have a high carbon storage potential as a result of high primary productivity and low decomposition rates dictated by water saturation. In the herbaceous wetlands of the Paraná River Delta, drainage and afforestation with poplars represents one of the dominant land uses. We explored the effects of these interventions on the volume and carbon storage of the young sedimentary soils of the region. At three sites we identified paired stands occupying similar landscape positions and soil types but subject to natural flooding and covered by natural herbaceous communities or drainage and flood control by dikes and covered by poplar plantations established 12, 17 and 19 years ago. Soil sampling at these sites revealed a reduction of the litter compartment (?86 %) and decreasing volume and porosity of its underlying mineral layer (0–10 cm in the wetland reduced to 0–4 cm in the plantation). Our comparisons of carbon storage accounted for these volumetric shifts by using accumulated mineral mass rather than depth as a reference, showing that tree plantations gained in the mineral soil (22 Mg C ha^?1) almost as much as what they lost in the litter. These gains were particularly large at intermediate depths (4–43 cm in the plantations) were soil porosity remained unaffected and C was raised by 64 % explained by (1) the pulse of inputs from overlaying litter and organic layers subject to rapid decomposition and mobilization after drainage and (2) root colonization, since tree plantations had 75 % of their fine root biomass at these intermediate soil depths, whereas roots in the wetlands did not explore the mineral soil profile and were completely confined to the organic layer. A neutral C balance following wetland drainage and afforestation resulted from the opposing effects of aeration, favoring decomposition in the organic layer, root colonization and organic matter stabilization, favoring its accumulation in the mineral soil.     

RVBs Are Required for Assembling a Functional TIP60 Complex

RVB1/RVB2 (RuvBL1/RuvBL2 or pontin/reptin) are enigmatic AAA⁺ ATPase proteins that are present in multiple cellular complexes. Although they have been implicated in many cellular functions, the exact molecular function of RVB proteins in the various complexes is not clear. TIP60 complex (TIP60.com) is a tumor suppressor chromatin-remodeling complex containing RVB proteins. RVBs are required for the lysine acetyltransferase activity of TIP60.com but not for that of the pure recombinant TIP60 polypeptide. Here we describe two molecular functions of RVBs in TIP60.com. First, RVBs negate the repression of catalytic activity of TIP60 by another protein in TIP60.com, p400. RVBs competitively displace the SNF2 domain of p400 from the TIP60 polypeptide. In addition RVBs are also required for heat stability of TIP60.com by a p400-independent pathway. RVB1 and RVB2 are redundant with each other for these functions and do not require their ATPase activities. Thus, RVB proteins act as molecular adaptors that can substitute for one another to facilitate the optimal assembly, heat stability, and function of the TIP60 complex.     

Manifestations of native topology in the denatured state ensemble of Rhodopseudomonas palustris cytochrome c'

To provide insight into the role of local sequence in the nonrandom coil behavior of the denatured state, we have extended our measurements of histidine-heme loop formation equilibria for cytochrome c' to 6 M guanidine hydrochloride. We observe that there is some reduction in the scatter about the best fit line of loop stability versus loop size data in 6 M versus 3 M guanidine hydrochloride, but the scatter is not eliminated. The scaling exponent, ν(3), of 2.5 ± 0.2 is also similar to that found previously in 3 M guanidine hydrochloride (2.6 ± 0.3). Rates of histidine-heme loop breakage in the denatured state of cytochrome c' show that some histidine-heme loops are significantly more persistent than others at both 3 and 6 M guanidine hydrochloride. Rates of histidine-heme loop formation more closely approximate random coil behavior. This observation indicates that heterogeneity in the denatured state ensemble results mainly from contact persistence. When mapped onto the structure of cytochrome c', the histidine-heme loops with slow breakage rates coincide with chain reversals between helices 1 and 2 and between helices 2 and 3. Molecular dynamics simulations of the unfolding of cytochrome c' at 498 K show that these reverse turns persist in the unfolded state. Thus, these portions of the primary structure of cytochrome c' set up the topology of cytochrome c' in the denatured state, predisposing the protein to fold efficiently to its native structure.     

EDA-Containing Fibronectin Increases Proliferation of Embryonic Stem Cells

Embryonic stem cells (ESC) need a set of specific factors to be propagated. They can also grow in conditioned medium (CM) derived from a bovine granulosa cell line BGC (BGC-CM), a medium that not only preserves their main features but also increases ESC´s proliferation rate. The mitogenic properties of this medium were previously reported, ascribing this effect to an alternative spliced generated fibronectin isoform that contains the extra domain A (FN EDA⁺). Here, we investigated if the FN EDA⁺ isoform increased proliferation of mouse and human ES cells. We analyzed cell proliferation using conditioned media produced by different mouse embryonic fibroblast (MEF) lines genetically engineered to express FN constitutively including or excluding the EDA domain (FN EDA^-), and in media supplemented with recombinant peptides containing or not the EDA. We found that the presence of EDA in the medium increased mouse and human ESC’s proliferation rate. Here we showed for the first time that this FN isoform enhances ESC’s proliferation. These findings suggest a possible conserved behavior for regulation of ES cells proliferation by this FN isoform and could contribute to improve their culturing conditions both for research and cell therapy.     

Link between MnSOD Ala16Val (rs4880) polymorphism and asthma risk is insignificant from sequential meta-analysis

The mitochondrial manganese superoxide dismutase (MnSOD) enzyme protects lungs against oxidative stress by neutralizing the free radical superoxide produced in the respiratory function. This has relevance to asthma. Therefore, it is of interest to describe the potential effect of MnSOD Ala16Val genetic polymorphism to asthma risk. Known data in this context is inconclusive in nature. The possible link between MnSOD Ala16Val polymorphism and asthma is explored using sequence meta-analysis. Data from the pooled analysis of MnSOD Ala16Val polymorphism using five genetic models i.e., allelic (Val vs. Ala: p=0.846; OR=1.033, 95% CI=0.742 to 1.440) is discussed. Homozygous (Val Val vs. Ala Ala: p=0.517; OR=1.307, 95% CI=0.582 to 2.932) and heterozygous (Val Ala vs. Ala Ala: p=0.307; OR=1.138, 95% CI=0.888 to 1.459) data using the described models are documented. Data from the dominant model (Val Val + Val Ala vs. Ala Ala: p=0.301; OR=1.289, 95% CI=0.797 to 2.085) and the recessive model (Val Val vs. Val Ala + Ala Ala: p=0.761; OR=0.924, 95% CI=0.555 to 1.538) analyses for several ethnic subgroups in this context is reported.