Reliable flow cytometric estimation of nuclear DNA content in coffee trees

La cytométrie en flux a permis d'obtenir des mesures précises de la taille des génomes de deux taxons (#C. liberica dewevrei et #C. pseudozanguebariae). L'iodure de propidium (IP) et le #Petunia hybrida ont été utilisés, respectivement, comme fluorochrome et standard interne. La linéarité de la relation entre la quantité d'ADN et le signal fluorescent digitalisé a été vérifiée. Cinq résultats majeurs ressortent : des conditions expérimentales optimales ont été définies pour l'estimation de la position des pics (moyenne et mode), le temps de coloration (au moins deux minutes), le voltage du photomultiplicateur (557 V) et de la concentration en IP (330 microg/ml) ; les effets du voltage et de la concentration en IP ont été paramétrés ; deux biais dus aux variations de voltage et de concentration en IP ont été décelés dans l'estimation de la quantité d'ADN par noyau. Ces biais ont été minimisés ; les tailles de génome de #C. liberica dewevrei et de #C. pseudozanguebariae$ ont été estimées avec précision (respectivement 2C = 1,421 plus ou moins 0,005 pg et 2C = 1,129 plus ou moins 0,005 pg) ; des variations entre génotypes ont été mises en évidence dans chaque taxon. (Résumé d'auteur

A complementary approach to estimate the internal pressure of fission gas bubbles by SEM-SIMS-EPMA in irradiated nuclear fuels

International audienceThe behaviour of gases produced by fission is of great importance for nuclear fuel in operation. Within this context, a decade ago, a general method for the characterisation of the fission gas including gas bubbles in an irradiated UO$_2$ nuclear fuel was developed and applied to determine the bubbles internal pressure. The method consists in the determination of the pressure, over a large population of bubbles, using three techniques: SEM, EPMA and SIMS. In this paper, a complementary approach using the information given by the same techniques is performed on an isolated bubble under the surface and is aiming for a better accuracy compared to the more general measurement of gas content. SEM and EPMA enable the detection of a bubble filled with xenon under the surface. SIMS enables the detection of the gas filling the bubble. The quantification is achieved using the EPMA data as reference at positions where no or nearly no bubbles are detected

White-gutted soldiers: simplification of the digestive tube for a non-particulate diet in higher Old World termites (Isoptera: Termitidae)

Previous observations have noted that in some species of higher termites the soldier caste lacks pigmented particles in its gut and, instead, is fed worker saliva that imparts a whitish coloration to the abdomen. In order to investigate the occurrence of this trait more thoroughly, we surveyed a broad diversity of termite specimens and taxonomic descriptions from the Old World subfamilies Apicotermitinae, Cubitermitinae, Foraminitermitinae, Macrotermitinae, and Termitinae. We identified 38 genera that have this “white-gutted” soldier (WGS) trait. No termite soldiers from the New World were found to possess a WGS caste. Externally, the WGS is characterized by a uniformly pale abdomen, hyaline gut, and proportionally smaller body-to-head volume ratio compared with their “dark-gutted” soldier (DGS) counterparts found in most termitid genera. The WGS is a fully formed soldier that, unlike soldiers in other higher termite taxa, has a small, narrow, and decompartmentalized digestive tube that lacks particulate food contents. The presumed saliva-nourished WGS have various forms of simplified gut morphologies that have evolved at least six times within the higher termites

Ultrastructure of the Intramandibular Gland of Workers and Queens of the Stingless Bee, Melipona quadrifasciata

The intramandibular glands of workers and queens of Melipona quadrifasciata Lepeletier (Hymenoptera: Apidae), at different ages and from different functional groups, were studied using light and transmission electron microscopy. The results demonstrated that these glands are composed of two types of secretory structures: 1.A hypertrophied epidermis on the dorsal side of the mandible that is an epithelial gland. 2. Free secretory cells filling the inner spaces of the appendices that constitute a unicellular gland. The epithelial gland is larger in the young (1-2-day-old workers), and the gland becomes involuted during the nurse worker stage. The unicellular glands of the workers posses some secretion during all of the studied phases, but secretory activity is more intensive in the foraging workers. Vesicles of secretion are absent in the unicellular glands of queens. These results demonstrate that these glands show functional adaptations in different castes corresponding to the functions of each caste

The conserved C-terminus of the PcrA/UvrD helicase interacts directly with RNA polymerase

Copyright: © 2013 Gwynn et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by a Wellcome Trust project grant to MD (Reference: 077368), an ERC starting grant to MD (Acronym: SM-DNA-REPAIR) and a BBSRC project grant to PM, NS and MD (Reference: BB/I003142/1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewedPublisher PD

DNA cruciform arms nucleate through a correlated but non-synchronous cooperative mechanism

Inverted repeat (IR) sequences in DNA can form non-canonical cruciform structures to relieve torsional stress. We use Monte Carlo simulations of a recently developed coarse-grained model of DNA to demonstrate that the nucleation of a cruciform can proceed through a cooperative mechanism. Firstly, a twist-induced denaturation bubble must diffuse so that its midpoint is near the centre of symmetry of the IR sequence. Secondly, bubble fluctuations must be large enough to allow one of the arms to form a small number of hairpin bonds. Once the first arm is partially formed, the second arm can rapidly grow to a similar size. Because bubbles can twist back on themselves, they need considerably fewer bases to resolve torsional stress than the final cruciform state does. The initially stabilised cruciform therefore continues to grow, which typically proceeds synchronously, reminiscent of the S-type mechanism of cruciform formation. By using umbrella sampling techniques we calculate, for different temperatures and superhelical densities, the free energy as a function of the number of bonds in each cruciform along the correlated but non-synchronous nucleation pathways we observed in direct simulations.Comment: 12 pages main paper + 11 pages supplementary dat

Annual transcriptome of a key zooplankton species, the copepod Calanus finmarchicus

The copepod Calanus finmarchicus (Crustacea, Copepoda) is a key zooplanktonic species with a crucial position in the North Atlantic food web and significant contributor to ocean carbon flux. Like many other high latitude animals, it has evolved a programmed arrested development called diapause to cope with long periods of limited food supply, while growth and reproduction are timed to take advantage of seasonal peaks in primary production. However, anthropogenic warming is inducing changes in the expected timing of phytoplankton blooms, suggesting phenological mismatches with negative consequences for the N. Atlantic ecosystem. While diapause mechanisms are mainly studied in terrestrial arthropods, specifically on laboratory model species, such as the fruit fly Drosophila, the molecular investigations of annual rhythms in wild marine species remain fragmentary. Here we performed a rigorous year-long monthly sampling campaign of C. finmarchicus in a Scottish Loch (UK; 56.45°N, 5.18°W) to generate an annual transcriptome. The mRNA of 36 samples (monthly triplicate of 25 individuals) have been deeply sequenced with an average depth of 137 ± 4 million reads (mean ± SE) per sample, aligned to the reference transcriptome, and filtered. We detail the quality assessment of the datasets and provide a high-quality resource for the investigation of wild annual transcriptomic rhythms (35,357 components) in a key diapausing zooplanktonic species