The novel flightless-I gene brings together two gene families, actin- binding proteins related to gelsolin and leucine-rich-repeat proteins involved in Ras signal transduction

The Drosophila melanogaster gene flightless-I, involved in gastrulation and muscle degeneration, has Caenorhabditis elegans and human homologues. In these highly conserved genes, two previously known gene families have been brought together, families encoding the actin- binding proteins related to gelsolin and the leucine-rich-repeat (LRR) group of proteins involved in protein-protein interactions. Both these gene families exhibit characteristics of molecular changes involving replication slippage and exon shuffling. Phylogenetic analyses of 19 amino acid sequences of 6 related protein types indicate that actin- associated proteins related to gelsolin are monophyletic to a common ancestor and include flightless proteins. Conversely, comparison of 24 amino acid sequences of LRR proteins including the flightless proteins indicates that flightless proteins are members of a structurally related subgroup. Included in the flightless cluster are human and mouse rsp-1 proteins involved in suppressing v-Ras transformation of cells and the membrane-associated yeast (Saccharomyces cerevisae) adenylate cyclase whose analogous LRRs are required for interaction with Ras proteins. There is a strong possibility that ligands for this group could be related and that flightless may have a similar role in Ras signal transduction. It is hypothesized that an ancestral monomeric gelsolin precursor protein has undergone at least four independent gene reorganization events to account for the structural diversity of the extant family of gelsolin-related proteins and that gene duplication and exon shuffling events occurred prior to or at the beginning of multicellular life, resulting in the evolution of some members of the family soon after the appearance of actin-type proteins.      

Magnesium-induced inner membrane aggregation in heart mitochondria: prevention and reversal by carboxyatractyloside and bongkrekic acid

Mg(2+) at an optimal concentration of 2mM (ph 6.5) induces large increases (up to 30 percent) in the optical density of bovine heart mitochondria incubated under conditions of low ionic strength (< approx. 0.01). The increases are associated with aggregation (sticking together) of the inner membranes and are little affected by changes in the energy status of the mitochondria. Virtually all of a number of other polyvalent cations tested and Ag(+) induce increases in mitochondrial optical density similar to those induced by Mg(2+), their approximate order of concentration effectiveness in respect to Mg(2+) being: La(3+) > Pb(2+) = Cu(2+) > Cd(2+) > Zn(2+) > Ag(+) > Mn(2+) > Ca(2+) > Mg(2+). With the exception of Mg(2+), all of these cations appear to induce swelling of the mitochondria concomitant with inner membrane aggregation. The inhibitors of the adenine nucleotide transport reaction carboxyatratyloside and bongkrekic acid are capable of preventing and reversing Mg(2+)-induced aggregation at the same low concentration required for complete inhibition of phosphorylating respiration, suggesting that they inhibit the aggregation by binding to the adenine nucleotide carrier. The findings are interpreted to indicate (a) that the inner mitochondrial membrane is normally prevented from aggregating by virtue of its net negative outer surface change, (b) that the cations induce the membrane to aggregate by binding at its outer surface, decreasing the net negative charge, and (c) that carboxyatractyloside and bongkrekic acid inhibit the aggregation by binding to the outer surface of the membrane, increasing the net negative charge.     

Rates and patterns of mitochondrial DNA sequence evolution in fringilline finches (fringilla spp.) and the greenfinch (Carduelis chloris)

Rates and patterns of evolution in partial sequences of five mitochondrial genes (cytochrome b, ATPase 6, NADH dehydrogenase subunit 5, tRNA(Glu), and the control region) were compared among taxa in the passerine bird genera Fringilla and Carduelis. Rates of divergence do not vary significantly among genes, even in comparisons with the control region. Rate variation among lineages is significant only for the control region and NADH dehydrogenase subunit 5, and patterns of variation are consistent with the expectations of neutral theory. Base composition is biased in all genes but is stationary among lineages, and there is evidence for directional mutation pressure only in the control region. Despite these similarities, patterns of substitution differ among genes, consistent with alternative regimes of selective constraint. Rates of nonsynonymous substitution are higher in NADH dehydrogenase subunit 5 than in other protein-coding genes, and transitions exist in elevated proportions relative to transversions. Transitions appear to accumulate linearly with time in tRNA(Glu), and despite exhibiting the highest overall rate of divergence among species, there are no transversional changes in this gene. Finally, for resolving phylogenetic relationships among Fringilla taxa, the combined protein-coding data are broadly similar to those of the control region in terms of phylogenetic informativeness and statistical support.      

Comparing contributions of soil versus root colonization to variations in stomatal behavior and soil drying in mycorrhizal Sorghum bicolor and Cucurbita pepo

In prior studies we learned that colonization of soil can be as important as colonization of roots in determining mycorrhizal influence on the water relations of host plants. Here we use a path analysis modeling approach to test (a) whether quantity of hyphae in soil contributes to variations in stomatal behavior and soil drying, and (b) whether soil colonization or root colonization has a stronger influence on these stomatal and soil drying responses. Experiments were performed on Sorghum bicolor and Cucurbita pepo, with soils and roots colonized by a mixture of Glomus intraradices and Gigaspora margarita. Soil colonization generally made more significant contributions to stomatal conductance than did root colonization. Soil colonization did not make significant direct contributions to soil water potential measures (soil water potential at stomatal closure or soil drying rate), whereas root colonization did contribute a potentially important path to each. The findings further support a role for mycorrhization of the soil itself in contributing to the regulation of stomatal behavior of host plants.     

Orchard pollination in Capitol Reef National Park,Utah, USA. Honey bees or native bees?

Capitol Reef National Park in central Utah, USA surrounds 22 managed fruit orchards started over a century ago by Mormon pioneers. Honey bees are imported for pollination, although the area in which the Park is embedded has over 700 species of native bees, many of which are potential orchard pollinators. We studied the visitation of native bees to apple, pear, apricot, and sweet cherry over 2 years. Thirty species of bees visited the flowers but, except for pear flowers, most were uncommon compared to honey bees. Evidence that honey bees prevented native bees from foraging on orchard crop flowers was equivocal: generally, honey bee and native bee visitation rates to the flowers were not negatively correlated, nor were native bee visitation rates positively correlated with distance of orchards from honey bee hives. Conversely, competition was tentatively suggested by much larger numbers of honey bees than natives on the flowers of apples, apricots and cherry; and by the large increase of native bees on pears, where honey bee numbers were low. At least one-third of the native bee species visiting the flowers are potential pollinators, including cavity-nesting species such as Osmia lignaria propinqua, currently managed for small orchard pollination in the US, plus several fossorial species, including one rosaceous flower specialist (Andrena milwaukiensis). We suggest that gradual withdrawal of honey bees from the Park would help conserve native bee populations without decreasing orchard crop productivity, and would serve as a demonstration of the commercial value of native pollinators.     

Pollination biology of a disjunct population of the endangered sandhills endemic <Emphasis Type="Italic">Penstemon haydenii</Emphasis> S. Wats. (Scrophulariaceae) in Wyoming,USA

We studied the breeding system and flower visitors of the endangered plant, Penstemon haydenii, at several south-central Wyoming, USA occurrences. In agreement with earlier studies of the species 300 km to the east in Nebraska, we found Wyoming plants to be self-incompatible and pollinator-dependent for sexual reproduction. Flower visitors were several species of native bees in the families Apidae (particularly bumblebees), Halictidae (small sweat bees), and Megachilidae (especially in the genus Osmia); and the masarid wasp Pseudomasaris vespoides. Especially important was Osmia brevis, an abundant megachilid bee, and one of only two species (the sweat bee Lasioglossum (Dialictus) pruinosum was the other) present at all five sites. As in Nebraska, fruit set did not differ between our experimental cross-pollination treatment and an open-pollinated control. However, unlike Nebraska, open-pollinated treatments in Wyoming produced significantly fewer seeds per fruit than the experimental out-crossing treatment. We discuss several possible explanations for seed limitation: (1) a scarcity of pollinators early in the flowering season; (2) resource competition for developing ovules on open-pollinated inflorescences but not on experimental inflorescences; (3) the deposition of self pollen through intra-inflorescence and intra-genet pollinator movements; (4) few S-alleles and mating types in the Wyoming metapopulation compared to the Nebraska metapopulation, from which it likely derives.     

Disposable plastic rodent feeders for use in an automated filling system

Rodent feeder devices are often made of reusable, sanitizable materials such as glass and stainless steel. As part of an initiative at the authors' facility to automate the feeder filling process, disposable plastic feeding cups were proposed as a preferable alternative to standard feeders for use in a filling machine. The authors tested and validated designs for rodent meal-type feeder assemblies that incorporated plastic cups. Assemblies included stainless steel shields that prevented rats and mice from chewing on the plastic cups while allowing them full access to food. The use of disposable feeder cups that are filled and sealed automatically reduces labor associated with sanitation, enables staff to prepare feeders for several weeks in advance and limits personnel contact with feed that may contain harmful chemicals.     

Hydraulic conductance and water potential gradients in squash leaves showing mycorrhiza-induced increases in stomatal conductance

Stomatal conductance (g _s) and transpiration rates vary widely across plant species. Leaf hydraulic conductance (k _leaf) tends to change with g _s, to maintain hydraulic homeostasis and prevent wide and potentially harmful fluctuations in transpiration-induced water potential gradients across the leaf (ΔΨ _leaf). Because arbuscular mycorrhizal (AM) symbiosis often increases g _s in the plant host, we tested whether the symbiosis affects leaf hydraulic homeostasis. Specifically, we tested whether k _leaf changes with g _s to maintain ΔΨ _leaf or whether ΔΨ _leaf differs when g _s differs in AM and non-AM plants. Colonization of squash plants with Glomus intraradices resulted in increased g _s relative to non-AM controls, by an average of 27% under amply watered, unstressed conditions. Stomatal conductance was similar in AM and non-AM plants with exposure to NaCl stress. Across all AM and NaCl treatments, k _leaf did change in synchrony with g _s (positive correlation of g _s and k _leaf), corroborating leaf tendency toward hydraulic homeostasis under varying rates of transpirational water loss. However, k _leaf did not increase in AM plants to compensate for the higher g _s of unstressed AM plants relative to non-AM plants. Consequently, ΔΨ _leaf did tend to be higher in AM leaves. A trend toward slightly higher ΔΨ _leaf has been observed recently in more highly evolved plant taxa having higher productivity. Higher ΔΨ _leaf in leaves of mycorrhizal plants would therefore be consistent with the higher rates of gas exchange that often accompany mycorrhizal symbiosis and that are presumed to be necessary to supply the carbon needs of the fungal symbiont.     

Comparison of linkage disequilibrium and haplotype diversity on macro- and microchromosomes in chicken

Background

Toll like receptors (TLR) play the central role in the recognition of pathogen associated molecular patterns (PAMPs). Mutations in the TLR1, TLR2 and TLR4 genes may change the ability to recognize PAMPs and cause altered responsiveness to the bacterial pathogens.

Results

The study presents association between TLR gene mutations and increased susceptibility to Mycobacterium avium subsp. paratuberculosis (MAP) infection. Novel mutations in TLR genes (TLR1- Ser150Gly and Val220Met; TLR2 – Phe670Leu) were statistically correlated with the hindrance in recognition of MAP legends. This correlation was confirmed subsequently by measuring the expression levels of cytokines (IL-4, IL-8, IL-10, IL-12 and IFN-γ) in the mutant and wild type moDCs (mocyte derived dendritic cells) after challenge with MAP cell lysate or LPS. Further in silico analysis of the TLR1 and TLR4 ectodomains (ECD) revealed the polymorphic nature of the central ECD and irregularities in the central LRR (leucine rich repeat) motifs.

Conclusion

The most critical positions that may alter the pathogen recognition ability of TLR were: the 9^th amino acid position in LRR motif (TLR1–LRR10) and 4^th residue downstream to LRR domain (exta-LRR region of TLR4). The study describes novel mutations in the TLRs and presents their association with the MAP infection.