First chromosome scale genomes of ithomiine butterflies (Nymphalidae: Ithomiini): Comparative models for mimicry genetic studies

The ithomiine butterflies (Nymphalidae: Danainae) represent the largest known radiation of Müllerian mimetic butterflies. They dominate by number the mimetic butterfly communities, which include species such as the iconic neotropical Heliconius genus. Recent studies on the ecology and genetics of speciation in Ithomiini have suggested that sexual pheromones, colour pattern and perhaps hostplant could drive reproductive isolation. However, no reference genome was available for Ithomiini, which has hindered further exploration on the genetic architecture of these candidate traits, and more generally on the genomic patterns of divergence. Here, we generated high-quality, chromosome-scale genome assemblies for two Melinaea species, M. marsaeus and M. menophilus, and a draft genome of the species Ithomia salapia. We obtained genomes with a size ranging from 396 to 503 Mb across the three species and scaffold N50 of 40.5 and 23.2 Mb for the two chromosome-scale assemblies. Using collinearity analyses we identified massive rearrangements between the two closely related Melinaea species. An annotation of transposable elements and gene content was performed, as well as a specialist annotation to target chemosensory genes, which is crucial for host plant detection and mate recognition in mimetic species. A comparative genomic approach revealed independent gene expansions in ithomiines and particularly in gustatory receptor genes. These first three genomes of ithomiine mimetic butterflies constitute a valuable addition and a welcome comparison to existing biological models such as Heliconius, and will enable further understanding of the mechanisms of adaptation in butterflies.     

Effects of UV-B on flavonoids, ferulic acid, growth and photosynthesis in barley primary leaves

Barley (Hordeum vulgare L.) was grown with UV-B (280–320 nm) at levels simulating 25 nr 5% ozone depletion on the date of the summer solstice al 40°N latitude, with UV-A (320–400 nm), or with no supplemental irradiation. In plant growth chambers providing 300 μmol m^?2 s^?1 photosynthetically active radiation (PAR). UV-B-grown leaves elongated more slowly than controls but reached the same final length 1 day later. Leal specific fresh weight (mass leaf area^?1) was significantly increased by UV-B after the 7th day of growth. IV-B did not significantly affect leaf area, fresh weight, dry weight, total chlorophylls, total carotenoids or photosynthetic quantum efficiency. CO₂ assimilation was decreased by UV-B only at internal CO₂ levels above 250 μl l^?1. By the 8th day of growth, UV-B increased flavonoid (saponarin and lutonarin) accumulation in both the lower epidermis and the mesophyll: about 40% of the saponarin and 20% of the lutonarin were in the lower epidermis under all experimental conditions. Glasshouse conditions proved too variable for reproducible determination of growth and photosynthesis but were reliable for determining developmental changes in flavonoid (saponarin and lutonarin) accumulation and provided up to 800 μmol m^?2 s^?1 PAR. In the glasshouse UV-B-grown leaves had more flavonoids than controls al all stages from 5 to 30 days after planting: ca 509 more saponarin and 100% more lutonarin. Levels of soluble (vacuolar) ferulic acid esters were similar under all conditions on day 5. and on day 20 or later, but were significantly higher in UV-B-grown plants on days 10 and 15. UV-B decreased insoluble (cell-wall-bound) ferulic acid esters on a whole leaf basis but significantly increased this fraction in the lower epidermis. UV-A had no significant effects on growth, photosynthesis or ferulic acid, but it slightly increased flavonoid accumulation. The results are discussed in terms of secondary phenolics as a tissue-specific, developmentally regulated adaptive response to UV-B.     

Exchanging sequence domains between S-RNases from Nicotiana alata disrupts pollen recognition

In self-incompatible plants of the Solanaceae, the specificity of pollen rejection is controlled by a single multiallelic S-locus. Pollen tube growth is inhibited in the style when its single S-allele matches either S-allele present in the diploid pistil. Each S-allele encodes an S-RNase with a unique sequence. S-RNases are secreted into the extracellular matrix of the transmitting tract which guides pollen tubes toward the ovary. Although it is known that S-RNases are the determinants of S-allele specificity in the pistil, it is not known how allele-specific information is encoded in the sequence. Therefore, we exchanged domains between S-RNases with different recognition specificities and expressed the chimeric proteins in transgenic plants to determine their effects on pollination behavior. Nine chimeric constructs were prepared in which domains from Nicotiana alata S_A2- and S_C10-RNases were exchanged. Among these nine constructs, the entire S-RNase sequence was sampled by exchanging single variable domains as well as larger blocks of contiguous sequences. The chimeric S-RNases retained enzymatic activity and were expressed at levels comparable to control transformants expressing S_A2- and S_C10-RNase. However, none of the chimeric S-RNases caused rejection of either S_A2- or S_C10-pollen. We conclude that the recognition function of S-RNases can be disrupted by alterations in many parts of the sequence. It appears that the recognition function of S-RNase is not localized to a specific domain.     

Aggregation of β-Amyloid Peptide Is Promoted by Membrane Phospholipid Metabolites Elevated in Alzheimer's Disease Brain

Abstract: Increased amounts of β-amyloid (Aβ) peptide deposits are found in Alzheimer's disease brain. These amyloid deposits have been implicated in the pathophysiology of this common dementing illness. Aβ peptides have been shown to be toxic to neurons in cell culture, and this toxicity is critically dependent on the aggregation of the peptide into cross-β-pleated sheet fibrils. Also, in vivo and postmortem NMR studies have shown changes in certain brain membrane phospholipid metabolites in normal aging and more extensive alterations in patients with Alzheimer's disease. The finding that membrane phospholipids affect the aggregation of Aβ suggests that the abnormalities in membrane metabolism found in Alzheimer's disease could affect the deposition of Aβ in vivo. Therefore, we examined the effect of membrane phospholipid metabolites that are altered in Alzheimer's disease brain on the aggregation of Aβ(1–40) using a light scattering method. Certain metabolites (glycerophosphocholine, glycerophosphoethanolamine, and α-glycerophosphate) augment the aggregation of Aβ. Other membrane phospholipid metabolites (phosphocholine, phosphoethanolamine, and inositol-1-phosphate) have no effect. We conclude that increased membrane phospholipid metabolite concentrations may play a role in the deposition of Aβ seen in normal aging and the even greater deposition of Aβ observed in Alzheimer's disease.     

Sensitization of rhabdo-, lenti-, and spumaviruses to human serum by galactosyl(alpha1-3)galactosylation.

Vesicular stomatitis virus, human immunodeficiency virus type 2, and human foamy virus, which were produced by cell lines expressing galactosyl(alpha1-3)galactosyl (alphaGal) sugars, were found to be less stable in human serum than those from alphaGal-negative cells, indicating that galactosyl(alpha1-3)galactosylation sensitizes these viruses as well as mammalian type C oncoviruses (Rother et al., J. Exp. Med. 182:1345-1355, 1995; Takeuchi et al., Nature (London) 379:85-88, 1996) to complement killing via natural anti-alphaGal antibodies. Thus, virus killing mediated by anti-alphaGal antibodies may play a role as a barrier to animal-to-human infection of various enveloped viruses. Virus vectors for human in vivo gene therapy based on the viruses mentioned above should be produced from alphaGal-negative cells.     

RAPID AXOPLASMIC TRANSPORT IN DYSTROPHIC MICE

Abstract— Rapid axoplasmic transport was studied in dystrophic mice of the 129/ReJ-dy strain. Proteins transported in vivo through α-motoneurons of the sciatic nerve were labeled by injections of [³H] or [³⁵S] amino acids into the ventral horn of the lumbar spinal cord. Following an 18 h incubation, axoplasmic transport was quantitated by summing the radioactivity in the 10 mm length of sciatic nerve proximal to a ligation. Although the amount of transported radioactivity was small, transport appeared depressed when adult dystrophic mice were compared to controls. Transport was also studied in the sensory fibers of the sciatic nerve under in vitro conditions, resulting in high levels of transported radioactivity. In this system transport was strongly depressed. The severity of the deficiency varied with age, being small in animals with early clinical signs and becoming maximal (80–90%) in animals over 60 days of age. Proteins transported by adult dy/dy and +/+ animals were compared by gel electrophoresis using double-label techniques. Transport of nearly all proteins was depressed in dy/dy mice, although the possibility exists that small differences occur. The data suggest that the dystrophic state produces a significant deficiency in rapid axoplasmic transport in both motor and sensory fibers, and may interfere with transport processes in all neurons. Since rapid axoplasmic transport has been associated with membranes, the data are consistent with a general alteration of cellular membranes in dystrophic animals.     

A noncycling activity assay for the omega subunit of Escherichia coli RNA polymerase.

We describe a new method for quantitatively assaying the omega subunit of Escherichia coli RNA polymerase. The assay is based on the ability of RNA polymerase holoenzyme to catalyze the continuous synthesis of the dinucleotide pApU on a poly[d(A-T)] . poly[d(A-T)] template when supplied with AMP and UTP as substrates. Core enzyme, lacking omega subunit, catalyzed this reaction at a rate less than 1% that of holoenzyme. The omega subunit was not released from the enzyme/DNA complex during dinucleotide synthesis. Using this assay, a titration of a fixed concentration of core enzyme was observed with increasing concentrations of added omega subunit. Below a 1:1 omega:core ratio the measured activity increased linearly with omega concentration, whereas above a 1:1 ratio the activity remained constant. An immediate application of the assay is in determining the concentration of active omega, or equivalently of active holoenzyme, in any RNA polymerase preparation.     

Effect of Oryzalin and 1,1-Dimethylpiperidinium Chloride on Cotton and Tomato Roots Infected with the Root-knot Nematode,Meloidogyne incognita

Oryzalin (3,5-dinitro-N4,N4-dipropyl-sulfanilamide) and BAS 083 (l,l-dimethylpiperdinium chloride) reduced root-knot infection in tomato roots when respectively applied as a soil drench at 20 ppm and 10,000 ppm. Oryzalin reduced knot counts with various intervals between treatment and inoculation. BAS 083 reduced knot counts only when applied before inoculation. Oryzalin was shown not to be a contact nematicide, and BAS 083 was only a weak one. Neither compound reduced penetration by infective larvae. Postinfection reduction in knot counts by Oryzalin and BAS 083 resulted, in part, from activation of natural defense mechanisms of the host. Giant-cell development in cotton roots inoculated with nematodes was inhibited by Oryzalin. Lateral root development was inhibited by BAS 083.     

Inhibition by styrylpyridines of purified rat and bovine brain choline acetyltransferase

Nine styrylpyridine analogs were tested as inhibitors of choline acetyltransferase which had been highly purified from rat cerebrum and bovine caudate nuclei. In general, concentrations required to achieve 50% inhibiion (I₅₀ values) were in the micromolar range. For some analogs, I₅₀ values were similar to those obtained previously by other investigators who used less purified enzyme preparations. With certain analogs, however, the measured values of I₅₀ changed as the transferase became more purified, which may indicate the presence in the extract of other molecules which can interact with the enzyme. The methods used in purification of the enzyme suggest that the molecule which modifies the activity of CAT is probably a protein. The mode of inhibition by naphthylvinylpyridinium was found to be uncompetitive with respect to both choline and acetyl coenzyme A for both the rat and bovine transferases.     

C-glycosylflavone accumulation in Sandoz 6706-treated barley seedlings is a photocontrolled response

Sandoz 6706 pretreatment of white light grown barley seedlings causes a 60% increase in saponarin (6-C-glucosyl-7-O-glucosylapigenin) but a 300% increase in lutonarin (3′-hydroxysaponarin). Norflurazon has little effect on saponarin levels but is almost as effective as Sandoz 6706 in enhancing lutonarin net synthesis. Barley roots contain saponarin and lutonarin only after herbicide treatment. Mung bean seedlings respond to Sandoz 6706 by accumulating higher levels of rutin and delphinidin 3-glucoside. The results are discussed in relation to the site of action of the herbicides, the High Energy photoresponse, and control of flavonoid 3′-hydroxylation.