BPAG1a and b Associate with EB1 and EB3 and Modulate Vesicular Transport, Golgi Apparatus Structure, and Cell Migration in C2.7 Myoblasts

BPAG1a and BPAG1b (BPAG1a/b) constitute two major isoforms encoded by the dystonin (Dst) gene and show homology with MACF1a and MACF1b. These proteins are members of the plakin family, giant multi-modular proteins able to connect the intermediate filament, microtubule and microfilament cytoskeletal networks with each other and to distinct cell membrane sites. They also serve as scaffolds for signaling proteins that modulate cytoskeletal dynamics. To gain better insights into the functions of BPAG1a/b, we further characterized their C-terminal region important for their interaction with microtubules and assessed the role of these isoforms in the cytoskeletal organization of C2.7 myoblast cells. Our results show that alternative splicing does not only occur at the 5′ end of Dst and Macf1 pre-mRNAs, as previously reported, but also at their 3′ end, resulting in expression of additional four mRNA variants of BPAG1 and MACF1. These isoform-specific C-tails were able to bundle microtubules and bound to both EB1 and EB3, two microtubule plus end proteins. In the C2.7 cell line, knockdown of BPAG1a/b had no major effect on the organization of the microtubule and microfilament networks, but negatively affected endocytosis and maintenance of the Golgi apparatus structure, which became dispersed. Finally, knockdown of BPAG1a/b caused a specific decrease in the directness of cell migration, but did not impair initial cell adhesion. These data provide novel insights into the complexity of alternative splicing of Dst pre-mRNAs and into the role of BPAG1a/b in vesicular transport, Golgi apparatus structure as well as in migration in C2.7 myoblasts

Deep sequencing of subseafloor eukaryotic rRNA reveals active fungi across marine subsurface provinces

© The Author(s), 2013. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS ONE 8 (2013): e56335, doi:10.1371/journal.pone.0056335.The deep marine subsurface is a vast habitat for microbial life where cells may live on geologic timescales. Because DNA in sediments may be preserved on long timescales, ribosomal RNA (rRNA) is suggested to be a proxy for the active fraction of a microbial community in the subsurface. During an investigation of eukaryotic 18S rRNA by amplicon pyrosequencing, unique profiles of Fungi were found across a range of marine subsurface provinces including ridge flanks, continental margins, and abyssal plains. Subseafloor fungal populations exhibit statistically significant correlations with total organic carbon (TOC), nitrate, sulfide, and dissolved inorganic carbon (DIC). These correlations are supported by terminal restriction length polymorphism (TRFLP) analyses of fungal rRNA. Geochemical correlations with fungal pyrosequencing and TRFLP data from this geographically broad sample set suggests environmental selection of active Fungi in the marine subsurface. Within the same dataset, ancient rRNA signatures were recovered from plants and diatoms in marine sediments ranging from 0.03 to 2.7 million years old, suggesting that rRNA from some eukaryotic taxa may be much more stable than previously considered in the marine subsurface.This work was performed with funding from the Center for Dark Energy Biosphere Investigations (C-DEBI) to William Orsi (OCE-0939564) and The Ocean Life Institute (WHOI) to Virginia Edgcomb (OLI-27071359)

Glomales species associated with surface and deep rhizosphere of Faidherbia albida in Senegal

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Glomales species associated with surface and deep rhizosphere of Faidherbia albida in Senegal

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Biodiversité et variabilité génétique des Glomales associés à Acacia albida Del. au Sénégal

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Evaluation of commercial arbuscular mycorrhizal inoculants

In order to improve the use of commercial inoculants, 12 arbuscular mycorrhizal fungi (AMF) inoculants were evaluated in a two-step experiment under greenhouse conditions using maize. First, commercial mycorrhizal inoculants were propagated in a trap pot culture experiment under sterilized sand to evaluate their potential for maize (Zea may L.) root colonization as compared with an indigenous soil inoculum and to survey the AMF species present in the products. Three inoculants significantly increased root colonization levels compared with a soil inoculum. Instead of 12 declared AMF species, 13 fungal strains were extracted from the pot culture survey, including five undeclared species, while four declared species did not produce spores. In a second experiment, commercial products were inoculated into soil to assess their impact on maize growth and yield. Six weeks after planting, seven inoculants increased root colonization levels compared with control soil, while only three inoculants increased slightly the shoot biomass of maize plants. These experiments highlight the need to pre-evaluate commercial mycorrhizal inoculants on a selected crop and regional soil before launching large-scale field use.Afin d'optimiser l'usage des mycorhizes en agriculture, douze inoculants commerciaux de champignon mycorhizien arbusculaire (CMA) furent évalués en serres sur le maïs. Dans un premier temps, les inoculants furent propagés en pots sur sable stérilisé afin d’évaluer leur potentiel sur la colonisation racinaire du maïs par rapport à celui d'un sol agricole du Kenya et d'inventorier les espèces CMA contenues dans les inoculants. Trois inoculants augmentèrent le taux de colonisation racinaire comparé au sol agricole. Treize espèces AMF furent isolées des inoculants dont 5 non déclarées. Quatre des 12 espèces annoncées n'ont pas sporulé. Dans une seconde expérience, les inoculants furent utilisés en combinaison avec le sol agricole afin d’évaluer leur impact sur le rendement du maïs. Six semaines après le semis, 7 inoculants augmentèrent le taux de colonisation racinaire par rapport au sol témoin alors que 3 inoculants entraînèrent une légère augmentation de la biomasse aérienne. Ces évaluations démontrent la nécessité d'effectuer une pré-évaluation des inoculants commerciaux sur une culture et un sol donnés avant de les implanter à grande échelle