Systematics within Gyps vultures: a clade at risk

Background  

Populations of the Oriental White-backed Vulture (Gyps bengalensis) have declined by over 95% within the past decade. This decline is largely due to incidental consumption of the non-steroidal anti-inflammatory veterinary pharmaceutical diclofenac, commonly used to treat domestic livestock. The conservation status of other Gyps vultures in southern Asia is also of immediate concern, given the lack of knowledge regarding status of their populations and the continuing existence of taxonomic uncertainties. In this study, we assess phylogenetic relationships for all recognized species and the majority of subspecies within the genus Gyps. The continuing veterinary use of diclofenac is an unknown but potential risk to related species with similar feeding habits to Gyps bengalensis. Therefore, an accurate assessment of the phylogenetic relationships among Gyps vultures should aid in their conservation by clarifying taxonomic uncertainties, and enabling inference of their respective relatedness to susceptible G. bengalensis.     

Precursor supply for insect juvenile hormone III biosynthesis in a cockroach

The biosynthesis of the sesquiterpenoid juvenile hormone III (JH III) was studied using corpora allata of the cockroach Diploptera punctata incubated in vitro and a radiochemical assay for the hormone produced. The influence of several exogenous precursors such as glucose, trehalose, acetate, amino acids, and mevalonate on JH synthetic rates was studied. Glucose or trehalose were needed for an optimal rate of JH synthesis. Highest rates were achieved at trehalose concentrations below the normal hemolymph levels (35-40 mM). About one-third of the glucose utilized for the biosynthesis of JH III was metabolized through a pentose pathway, but acetyl-CoA derived from glucose was significantly diluted by acetyl-CoA from other sources. Amino acids provided both a source of carbon for JH III synthesis and a source of energy that allowed JH III synthesis from acetate and stimulated JH III synthesis from glucose. Acetate was a poor substrate, because it could not support JH III synthesis in long term incubations. The incorporation of exogenous mevalonate into JH III was dependent on the physiological state of the glands, but there was a significant dilution with endogenous mevalonate. This dilution reflected in part the poor penetration of mevalonate into the corpora allata cells, because JH synthesis in mevinolin-treated cells was not fully rescued by mevalonate.     

Prenyl proteins in eukaryotic cells: a new type of membrane anchor

Recent studies have indicated that eukaryotic cells contain proteins that are post-translationally modified by long-chain, thioether-linked prenyl groups. These proteins include yeast mating factors, ras proteins and nuclear lamins. The modification occurs on a cysteine residue near the C terminus and appears to initiate a set of additional protein modification reactions that promote attachment of the proteins to specific membranes.     

A barrier to diffusion in pseudoplasmodia of Dictyostelium discoideum

Several enzymatic activities have been reported to be preferentially localized in the anterior portions of pseudoplasmodia of Dictyostelium discoideum. Since anterior cells are responsible for formation of the stalk during fruiting body construction, it has been suggested that accumulation of these enzymes may direct the cells toward stalk differentiation. However, the evidence for enzyme localization has come only from histochemical studies. We have assayed for succinoxidase and several dehydrogenase activities in cell free extracts of isolated anterior and posterior fragment and found no significant differences in specific activities, although by histochemical techniques each is apparently localized in the anterior cells. The preferential staining appears to be a consequence of a barrier to diffusion that is more effective at the back than at the front in limiting entry of the histochemical chromogen into pseudoplasmodia. The barrier appears to be the glycoprotein surface sheath that surrounds the pseudoplasmodium. The consequences of a barrier to diffusion of compounds into and out of pseudoplasmodia are discussed in relation to a mechanism that could give cells information concerning their position in this organism.     

Human lamin B contains a farnesylated cysteine residue

We recently showed that HeLa cell lamin B is modified by a mevalonic acid derivative. Here we identified the modified amino acid, determined its mode of linkage to the mevalonic acid derivative, and established the derivative's structure. A cysteine residue is modified because experiments with lamin B that had been biosynthetically labeled with [3H]mevalonic acid or [35S]cysteine and then extensively digested with proteases yielded 3H- or 35S-labeled products that co-chromatographed in five successive systems. A thioether linkage rather than a thioester linkage is involved because the mevalonic acid derivative could be released from the 3H-labeled products in a pentane-extractable form by treatment with Raney nickel but not with methanolic KOH. The derivative is a farnesyl moiety because the Raney nickel-released material was identified as 2,6,10-trimethyl-2,6,10-dodecatriene by a combination of gas chromatography and mass spectrometry. The thioether-modified cysteine residue appears to be located near the carboxyl end of lamin B because treatment of 3H-labeled lamin B with cyanogen bromide yielded a single labeled polypeptide that mapped toward this end of the cDNA-inferred sequence of human lamin B.     

Elevated CO2 alters anatomy,physiology, growth,and reproduction of red mangrove (Rhizophora mangle L.)

Mangroves, woody halophytes restricted to protected tropical coasts, form some of the most productive ecosystems in the world, but their capacity to act as a carbon source or sink under climate change is unknown. Their ability to adjust growth or to function as potential carbon sinks under conditions of rising atmospheric CO₂ during global change may affect global carbon cycling, but as yet has not been investigated experimentally. Halophyte responses to CO₂ doubling may be constrained by the need to use carbon conservatively under water-limited conditions, but data are lacking to issue general predictions. We describe the growth, architecture, biomass allocation, anatomy, and photosynthetic physiology of the predominant neotropical mangrove tree, Rhizophora mangle L., grown solitarily in ambient (350 ll^–1) and double-ambient (700 ll^–1) CO₂ concentrations for over 1 year. Mangrove seedlings exhibited significantly increased biomass, total stem length, branching activity, and total leaf area in elevated CO₂. Enhanced total plant biomass under high CO₂ was associated with higher root:shoot ratios, relative growth rates, and net assimilation rates, but few allometric shifts were attributable to CO₂ treatment independent of plant size. Maximal photosynthetic rates were enhanced among high-CO₂ plants while stomatal conductances were lower, but the magnitude of the treatment difference declined over time, and high-CO₂ seedlings showed a lower P_max at 700 ll^–1 CO₂ than low-CO₂ plants transferred to 700 ll^–1 CO₂: possible evidence of downregulation. The relative thicknesses of leaf cell layers were not affected by treatment. Stomatal density decreased as epidermal cells enlarged in elevated CO₂. Foliar chlorophyll, nitrogen, and sodium concentrations were lower in high CO₂. Mangroves grown in high CO₂ were reproductive after only 1 year of growth (fully 2 years before they typically reproduce in the field), produced aerial roots, and showed extensive lignification of the main stem; hence, elevated CO₂ appeared to accelerate maturation as well as growth. Data from this long-term study suggest that certain mangrove growth characters will change flexibly as atmospheric CO₂ increases, and accord with responses previously shown in Rhizophora apiculata. Such results must be integrated with data from sea-level rise studies to yield predictions of mangrove performance under changing climate.     

Patterns of divergence during evolution of alpha 1-proteinase inhibitors in mammals

alpha 1-Proteinase inhibitor (alpha 1-PI), a member of the serine proteinase inhibitor superfamily, has a primary role in controlling neutrophil elastase activity within the mammalian circulation. Several studies have indicated that the reactive center region of alpha 1-PI, the amino acid sequence of which is critical to recognition of and binding to target proteinases, is highly divergent within and among species. This appears to be a consequence of accelerated rates of evolution that may have been driven by positive Darwinian selection. In order to examine this and other features of alpha 1-PI evolution in more detail, we have isolated and sequenced cDNAs representing alpha 1- PI mRNAs of the mouse species Mus saxicola and Mus minutoides and have compared these with a number of other mammalian alpha 1-PI mRNAs. Relative to other mammalian mRNAs, the extent of nonsynonymous substitution is generally high throughout the alpha 1-PI mRNA molecule, indicating greater overall rates of amino acid substitution. Within and among mouse species, the 5'-half of the mRNA, but not the 3'-half, has been homogenized by concerted evolution. Finally, the reactive center is under diversifying or positive Darwinian selection in murid rodents (rats, mice) and guinea pigs yet is under purifying selection in primates and artiodactyls. The significance of these findings to alpha 1-PI function and the possible selective forces driving evolution of serpins in general are discussed.      

Mitochondrial DNA polymorphisms in subterranean mole-rats of the Spalax ehrenbergi superspecies in Israel, and its peripheral isolates

Patterns of mitochondrial DNA (mtDNA) variation were examined in 133 mole-rats constituting all four chromosomal species (2n = 52, 2n = 54, 2n = 58, and 2n = 60) of the Spalax ehrenbergi superspecies in Israel, as well as the peripheral isolates of 2n = 60. In the main range of the complex, a total of 28 mtDNA haplotypes were found in 64 mole-rats, with most haplotypes being unique to either a single chromosomal species or population. mtDNA divergence increased from low to high diploid number in a north-to-south direction in Israel. Overall levels of mtDNA diversity were unexpectedly the highest in the 2n = 60, the youngest species of the complex. The mtDNA haplotypes can be separated into two major groups, 2n = 52-54 and 2n = 58-60, and a phylogenetic analysis for each group revealed evidence of a few haplotypes not sorted by diploid number. The overall patterns of mtDNA divergence seen within and among the four chromosomal species are consistent with the parapatric mode of speciation as suggested from previous studies of allozyme and DNA hybridization. In a separate data set the patterns of mtDNA variation were examined across the main geographic range and across peripheral semi-isolates and isolates of the 2n = 60 chromosomal species. Fifteen haplotypes were found in 69 mole-rats. High levels of mtDNA diversity characterized the main range, semi-isolated, and even some desert isolated populations. The peripheral isolates contain much mtDNA diversity, including novel haplotypes.