A high-efficiency CRISPR/Cas9 system for targeted mutagenesis in Cotton (Gossypium hirsutum L.)

The complex allotetraploid genome is one of major challenges in cotton for repressing gene expression. Developing site-specific DNA mutation is the long-term dream for cotton breeding scientists. The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system is emerging as a robust biotechnology for targeted-DNA mutation. In this study, two sgRNAs, GhMYB25-like-sgRNA1 and GhMYB25-like-sgRNA2, were designed in the identical genomic regions of GhMYB25-like A and GhMYB25-like D, which were encoded by cotton A subgenome and the D subgenome, respectively, was assembled to direct Cas9-mediated allotetraploid cotton genome editing. High proportion (14.2–21.4%) CRISPR/Cas9-induced specific truncation events, either from GhMYB25-like A DNA site or from GhMYB25-like D DNA site, were detected in 50% examined transgenic cotton through PCR amplification assay and sequencing analyses. Sequencing results also demonstrated that 100% and 98.8% mutation frequency were occurred on GhMYB25-like-sgRNA1 and GhMYB25-like-sgRNA2 target site respectively. The off-target effect was evaluated by sequencing two putative off-target sites, which have 3 and 1 mismatched nucleotides with GhMYB25-like-sgRNA1 and GhMYB25-like-sgRNA2, respectively; all the examined samples were not detected any off-targetcaused mutation events. Thus, these results demonstrated that CRISPR/Cas9 is qualified for generating DNA level mutations on allotetraploid cotton genome with high-efficiency and high-specificity.ECU Open Access Publishing Support Fun

Prognostic model to predict postoperative acute kidney injury in patients undergoing major gastrointestinal surgery based on a national prospective observational cohort study.

Background: Acute illness, existing co-morbidities and surgical stress response can all contribute to postoperative acute kidney injury (AKI) in patients undergoing major gastrointestinal surgery. The aim of this study was prospectively to develop a pragmatic prognostic model to stratify patients according to risk of developing AKI after major gastrointestinal surgery. Methods: This prospective multicentre cohort study included consecutive adults undergoing elective or emergency gastrointestinal resection, liver resection or stoma reversal in 2-week blocks over a continuous 3-month period. The primary outcome was the rate of AKI within 7 days of surgery. Bootstrap stability was used to select clinically plausible risk factors into the model. Internal model validation was carried out by bootstrap validation. Results: A total of 4544 patients were included across 173 centres in the UK and Ireland. The overall rate of AKI was 14·2 per cent (646 of 4544) and the 30-day mortality rate was 1·8 per cent (84 of 4544). Stage 1 AKI was significantly associated with 30-day mortality (unadjusted odds ratio 7·61, 95 per cent c.i. 4·49 to 12·90; P < 0·001), with increasing odds of death with each AKI stage. Six variables were selected for inclusion in the prognostic model: age, sex, ASA grade, preoperative estimated glomerular filtration rate, planned open surgery and preoperative use of either an angiotensin-converting enzyme inhibitor or an angiotensin receptor blocker. Internal validation demonstrated good model discrimination (c-statistic 0·65). Discussion: Following major gastrointestinal surgery, AKI occurred in one in seven patients. This preoperative prognostic model identified patients at high risk of postoperative AKI. Validation in an independent data set is required to ensure generalizability

Una asociación florística de helechos-bennettitales en acumulaciones travertínicas del Titónico-Berriasiense (Formación Aguilar, Burgos-Palencia, España) y sus implicaciones paleoclimáticas y vegetacionales

[EN] This report describes a macrofloral fossil assemblage discovered in travertine deposits of the Tithonian-Berriasian Aguilar Formation (provinces of Palencia and Burgos, N Spain). The assemblage includes megaremains of a single species of Filicales (Cladophlebis denticulata) and of eleven taxa tentatively identified as Bennettitales (Otozamites mundae, Otozamites sp., Ptilophyllum pecten, Ptilophyllum pectiniformis, Ptilophyllum sp. cf. pectiniformis, Ptilophyllum acutifolium, Ptilophyllum sp., Pterophyllum cycadites, Pterophyllum sp. cf. thomasii, Zamites pumilio, and cf. Pseudocycas sp.). Specimens were preserved as impressions coated with a microbial film up to 5 mm thick made up of: bacteria and cyanobacteria. Based on the available sedimentological and palaeobotanical data, a dry-savannah vegetation and an arid climate were deduced. Late Jurassic macrofloras are globally scarce and have not previously been reported in Spain. Hence, the Aguilar Formation macroflora provides valuable information on the coeval phytogeography and palaeoclimate of southwestern Europe. Furthermore, the lack of floras observed so far in palaeotravertine deposits older than Pliocene in age makes this macroflora exceptional.[ES] Se describe por vez primera vez una macroflora fósil acumulada en facies travertínicas de edad Titónico-Berriasiense, correspondientes a la Formación Aguilar (provincias de Burgos y Palencia). Esta macroflora incluye una especie de Filicales (Cladophlebis denticulata) y once taxa atribuibles al Orden Bennettitales (Otozamites mundae, Otozamites sp., Ptilophyllum pecten, Ptilophyllum pectiniformis, Ptilophyllum sp. cf. pectiniformis, Ptilophyllum acutifolium, Ptilophyllum spp., Pterophyllum cycadites, Pterophyllum sp. cf. thomasi, Zamites pumilio, and cf. Pseudocycas sp.). Los ejemplares corresponden a impresiones de restos de hojas cubiertos por un velo microbiano bien preservado, de cerca de 5 mm, formado por bacterias y cianobacterias. La composición de la flora y los datos sedimentológicos permiten inferir una vegetación de sabana que se desarrollo bajo un clima árido. Las macrofloras del Jurásico Superior son escasas a escala mundial, no habiendo sido descrita ninguna en España. Por todo ello, la macroflora de la Formación Aguilar aporta una relevante información que puede contribuir a una mejor comprensión fitogeográfica y paleoclimática del suroeste de Europa en dicha edad. Además, la ausencia, a nivel global, de floras acumuladas en travertinos anteriores al Plioceno, permite considerar esta macroflora como excepcional..V. Pujalte acknowledges the support of grant CGL2005- 02770/BTE awarded by the Ministerio de Educación y Ciencia for the fieldwork needed for this study.Peer reviewe

Multiphase Phanerozoic Subsidence and Uplift History Recorded in the Congo Basin: A Complex Successor Basin

International audienceThe Congo Basin of central Africa is a large iconic Phanerozoic sedimentary basin whose origin and tectonic evolution are poorly understood, mostly because of a lack of modern stratigraphic data, reflecting a long hiatus in field investigations during the past five decades. It is usually assumed that the Congo Basin experienced a long and continuous history of slow subsidence since the late Precambrian (e.g. 2–4 m/Ma), linked to steady-state mantle processes. Here, we used revised sedimentological and stratigraphic data of the four historic deep boreholes drilled in the center of the basin to calculate a new first-order model for its subsidence and uplift history. Because the sedimentary sequences of this basin are largely terrestrial, we apply a new backstripping method especially designed for continental domain. The results reveal two main episodes of subsidence: initially rapid subsidence during the Carboniferous-Triassic (10–20 m/Ma), and then slower subsidence during the Jurassic-Cretaceous (5–10 m/Ma), punctuated by several uplifts at 160–180 Ma (e.g. ‘Karoo’), 120–140 Ma (e.g. ‘Paraná-Etendeka’), and again in the Cenozoic, ca. 30–50 Ma (e.g. ‘Ethiopian’). This complex, multiphase subsidence and uplift history of the Congo Basin can be linked to evolving far-field geodynamic processes that first led the formation of Pangea (large-scale compression) during the late Paleozoic, and then to its break-up associated with successive outpourings of Large Igneous Provinces (or hotspot plumes) and the opening of the Indian and South Atlantic Oceans around Africa