Age-specific responses to climate identified in the growth of Quercus alba

The objective of this research was to determine whether the dendroclimatic responses of young Quercus alba (aged 29–126 years) differ from those of old Q. alba (149–312 years). We collected Q. alba increment cores across a range of size classes from Buffalo Mountain Natural Area Preserve, an oak-hickory forest in southcentral Virginia, USA. Tree cores were crossdated and raw ring widths were detrended to remove the influence of increasing circumference with age, microsite, and local stand dynamics. Standardized ring widths were averaged to develop two master chronologies from the 20 oldest and youngest trees. Ring-width indices were correlated with temperature, precipitation, and Palmer Drought Severity Index (PDSI). Annual tree-ring growth in old and young Q. alba was significantly correlated with precipitation from the previous growing season, but was not significantly correlated with temperature. Only the old trees showed a significant correlation between annual ring width and PDSI. These results may indicate that growth in old trees is more sensitive to drought than in young trees. If future climate change includes the predicted increase in mid-growing season droughts, tree-level responses are likely to be age-dependent with older trees experiencing relatively greater reductions in growth.     

Phylogenetics and taxonomy of the New World leafy spurges,Euphorbia section Tithymalus (Euphorbiaceae)

The 480 species of leafy spurges, Euphorbia subgenus Esula, represent the main temperate radiation in the large genus Euphorbia. This group is distributed primarily in temperate Eurasia, but with smaller, disjunct centres of diversity in the mountains of the Old World tropics, in temperate southern Africa and in the New World. The majority of New World diversity (32 species) occurs in a single section, section Tithymalus. We analysed sequences of the nrITS and plastid ndhF, trnH‐psbA, trnS‐trnG and trnD‐trnT regions to reconstruct the phylogeny of section Tithymalus and to examine the origins and diversification of the species native to the New World. Our results indicate that the New World species of section Tithymalus form a clade that is sister to the widespread, weedy E. peplus. The New World species fall into two primary groups: a ‘northern annual clade’ from eastern North America and a diverse clade of both annual and perennial species that is divided into three subgroups. Within the second group, there is a small ‘southern annual clade’ from Texas and northern Mexico, a perennial ‘Brachycera clade’ from the western United States and northern Mexico, and a perennial ‘Esuliformis clade’ from montane areas of Mexico, Guatemala, Honduras and the Caribbean island of Hispaniola. Ancestral state reconstructions indicate that the annual habit probably evolved in the ancestor of E. peplus and the New World clade, with a subsequent reversal to the perennial habit. In conjunction with this phylogenetic framework, the New World species of section Tithymalus are comprehensively reviewed. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 191–228.     

Cellular reactions of O6-methylguanine, a product of some alkylating carcinogens

Cultures of a purine-requiring mutant of Chinese hamster ovary cells (CHO-104b), randomly bred hamster embryo cells, or Escherichia coli B_s−1 were treated with non-toxic doses of ³H-labelled O⁶-methylguanine. DNA and RNA were isolated and subjected to enzymic digestion to nucleosides at pH8. The products of digestion were analysed by ion-exchange chromatography on columns of Dowex 50 (NH₄⁺ form) at pH8.9. No ³H-labelled O⁶-methylguanosine was detected in nucleic acid digests. ³H-labelled O⁶-methylguanine was O-demethylated yielding [³H]guanine in CHO-104b cells. Radioactivity in nucleic acid digests was associated with thymidine, guanosine, deoxyguanosine and an unidentified early-eluting product. Reports of similar unidentified products from nucleic acids labelled with various agents are discussed.     

Conformational changes of yeast plasma membrane H(+)-ATPase during activation by glucose: role of threonine-912 in the carboxy-terminal tail

Yeast Pma1 H(+)-ATPase, which belongs to the P-type family of cation-transporting ATPases, is activated up to 10-fold by growth on glucose, and indirect evidence has linked the activation to Ser/Thr phosphorylation within the C-terminal tail. We have now used limited trypsinolysis to map glucose-induced conformational changes throughout the 100 kDa ATPase. In the wild-type enzyme, trypsin cleaves first at Lys-28 and Arg-73 in the extended N-terminal segment (sites T1 and T2); subsequent cleavages occur at Arg-271 between the A domain and M3 (site T3) and at Lys-749 or Lys-754 in the M6-M7 cytoplasmic loop (site T4). Activation by glucose leads to a striking increase in trypsin sensitivity. At the C-terminal end of the protein, the Arg- and Lys-rich tail is shielded from trypsin in membranes from glucose-starved cells (GS) but becomes accessible in membranes from glucose-metabolizing cells (GM). In the presence of orthovanadate, Lys-174 at the boundary between M2 and the A domain also becomes open to cleavage in GM but not GS samples (site T5). Significantly, this global conformational change can be suppressed by mutations at Thr-912, a consensus phosphorylation site near the C-terminus. Substitution by Ala at position 912 leads to a GS-like (trypsin-resistant) state, while substitution by Asp leads to a GM-like (trypsin-sensitive) state. Thus, the present results help to dissect the intramolecular movements that result in glucose activation.     

Fucose, placental evolution and the glycocode

A family of 13 fucosyltransferase genes has evolved to catalyze the addition of fucose in various linkage positions to nascent glycoproteins. Null mutations in mice are unearthing unsuspected functions for glycoprotein fucosylation that affect embryo implantation and growth of the conceptus. Furthermore, as we show here, histological studies demonstrate that a variety of fucosylated structures are found within the glycan-rich interface between trophectoderm and uterine epithelium. We suggest that conservation or change in fucosyltransferase gene expression over evolutionary time has played a role in determining the stability of the maternal-fetal interface and therefore in shaping reproductive compatibility and, in turn, speciation.     

A note on the use of microwaves in an improved non-radioactive DNA hybridization procedure for the detection of bacteria in foodstuffs

Foodstuffs were artificially contaminated with Escherichia coli carrying plasmid pBR322, dot blotted onto nylon membranes and briefly subjected to microwaves in the presence of 1·5 mol/l NaC1/0·5 mol/l NaOH. Subsequent hybridization with a biotin-labelled probe specific for pBR322 enabled the detection of cell concentrations > 10⁴ cells/dot blot, equivalent to 2 × 10⁷ cells/g food tested. This shortened and simplified method was effective for all ten foods tested, generated low background levels and should be applicable to a wide range of bacteria.     

Binding-protein expression is subject to temporal,developmental and stress-induced regulation in terminally differentiated soybean organs

Binding protein (BiP) is a widely distributed and highly conserved endoplasmic-reticulum luminal protein that has been implicated in cotranslational folding of nascent polypeptides, and in the recognition and disposal of misfolded polypeptides. Analysis of cDNA sequences and genomic blots indicates that soybeans (Glycine max L. Merr.) possess a small gene family encoding BiP. The deduced sequence of BiP is very similar to that of other plant BiPs. We have examined the expression of BiP in several different terminally differentiated soybean organs including leaves, pods and seed cotyledons. Expression of BiP mRNA increases during leaf expansion while levels of BiP protein decrease. Leaf BiP mRNA is subject to temporal control, exhibiting a large difference in expression in a few hours between dusk and night. The expression of BiP mRNA varies in direct correlation with accumulation of seed storage proteins. The hybridization suggests that maturing-seed BiP is likely to be a different isoform from vegetative BiPs. Levels of BiP protein in maturing seeds vary with BiP mRNA. High levels of BiP mRNA are detected after 3 d of seedling growth. Little change in either BiP mRNA or protein levels was detected in maturing soybean pods, although BiP-protein levels decrease in fully mature pods. Persistent wounding of leaves by whiteflies induces massive overexpression of BiP mRNA while only slightly increasing BiP-protein levels. In contrast single-event puncture wounding only slightly induces additional BiP expression above the temporal variations. These observations indicate that BiP is not constitutively expressed in terminally differentiated plant organs. Expression of BiP is highest during the developmental stages of leaves, pods and seeds when their constituent cells are producing seed or vegetative storage proteins, and appears to be subject to complex regulation, including developmental, temporal and wounding.The mention of vendor or product does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over vendors of similar products not mentioned.Abbreviations BiP binding protein The sequences reported in this paper have been submitted to Gen-Bank and are identified with the accession numbers BiP-A (U08384), BiP-B (U08383), BiP-C (U08382) and -1,3 glucanase (U08405).     

Estimating product and energy substitution benefits in national‐scale mitigation analyses for Canada

The potential of forests and the forest sector to mitigate greenhouse gas (GHG) emissions is widely recognized, but challenging to quantify at a national scale. Mitigation benefits through the use of forest products are affected by product life cycles, which determine the duration of carbon storage in wood products and substitution benefits where emissions are avoided using wood products instead of other emissions‐intensive building products and energy fuels. Here we determined displacement factors for wood substitution in the built environment and bioenergy at the national level in Canada. For solid wood products, we compiled a basket of end‐use products and determined the reduction in emissions for two functionally equivalent products: a more wood‐intensive product vs. a less wood‐intensive one. Avoided emissions for end‐use products basket were weighted by Canadian consumption statistics to reflect national wood uses, and avoided emissions were further partitioned into displacement factors for sawnwood and panels. We also examined two bioenergy feedstock scenarios (constant supply and constrained supply) to estimate displacement factors for bioenergy using an optimized selection of bioenergy facilities which maximized avoided emissions from fossil fuels. Results demonstrated that the average displacement factors were found to be similar: product displacement factors were 0.54 tC displaced per tC of used for sawnwood and 0.45 tC tC^?1 for panels; energy displacement factors for the two feedstock scenarios were 0.47 tC tC^?1 for the constant supply and 0.89 tC tC^?1 for the constrained supply. However, there was a wide range of substitution impacts. The greatest avoided emissions occurred when wood was substituted for steel and concrete in buildings, and when bioenergy from heat facilities and/or combined heat and power facilities was substituted for energy from high‐emissions fossil fuels. We conclude that (1) national‐level substitution benefits need to be considered within a systems perspective on climate change mitigation to avoid the development of policies that deliver no net benefits to the atmosphere, (2) the use of long‐lived wood products in buildings to displace steel and concrete reduces GHG emissions, (3) the greatest bioenergy substitution benefits are achieved using a mix of facility types and capacities to displace emissions‐intensive fossil fuels.     

Activation of two forms of locomotion by a previously identified trigger interneuron for swimming in the medicinal leech

Higher-order projection interneurons that function in more than one behavior have been identified in a number of preparations. In this study, we document that stimulation of cell Tr1, a previously identified trigger interneuron for swimming in the medicinal leech, can also elicit the motor program for crawling in isolated nerve cords. We also show that motor choice is independent of the firing frequency of Tr1 and amount of spiking activity recorded extracellularly at three locations along the ventral nerve cord prior to Tr1 stimulation. On the other hand, during Tr1 stimulation there is a significant difference in the amount of activity elicited in the ventral nerve cord that correlates with the motor program activated. On average, Tr1 stimulation trials that lead to crawling elicit greater amounts of activity than in trials that lead to swimming.