Barcoding moléculaire

vignette|400px|Principes du barcoding de l'ADN Le barcoding moléculaire, DNA barcoding, parfois francisé en codage à barres de l'ADN, est une technique de catalogage et d'identification moléculaire permettant la caractérisation génétique d'un individu ou d'un échantillon d'individu à partir d'une courte séquence d'ADN (marqueur distinctif) choisie en fonction du groupe étudié. À la fin des années 2010, des séquenceurs d'ADN rapides, portables et bon marché apparaissent sur le marché permettant par exemple de travailler en pleine mer ou en pleine jungle où des millions d'espèces sont encore à identifier. Cette technologie contribue aussi bien à classer des individus d'espèces inconnues qu'à distinguer de nouvelles espèces ou détecter l'origine et l'identité d'un échantillon. Paul Hebert, de l'Université de Guelph au Canada, inventeur du concept, a estimé qu'on l'on pourra un jour suivre l'évolution de la vie comme on suit aujourd'hui la météo. , selon Hebert, a

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DNA barcoding of formalin‑fixed aquatic oligochaetes for biomonitoring

Régis Lionel Vivien

Background: Oligochaetes are valuable bioindicators of the quality of watercourse and lake sediments. The morphological identification of aquatic oligochaetes is difficult, prompting the development of new molecular oligochaete indices based on DNA barcoding and Next-generation sequencing of sorted specimens. In general, the samples for DNA barcoding are fixed in absolute ethanol. However, in the case of aquatic oligochaetes, this medium is not appropriate as it can induce a modification of specimen abundances and of the composition of communities. Therefore, we investigated the possibility to amplify and sequence aquatic oligochaetes fixed in formalin for a short time. We performed guanidine extraction and polymerase chain reaction (PCR) amplification/sequencing of the cytochrome c oxydase I (COI) gene on tissue fragments fixed in formalin for different periods of time (from 1 h to 1 week) and in ethanol. Results: The large majority of aquatic oligochaete specimens fixed in formalin for up to 1 week could be successfully amplified and all obtained sequences were of high quality. The amplification and sequencing success rate of formalin-fixed samples and ethanol-fixed samples was similar. These results suggest that formalin fixation of aquatic oligochaete tissues for a short time does not cause serious damages to DNA and inhibit PCR amplification. Conclusion: The possibility to fix aquatic oligochaetes with formalin before genetic analyses is very promising for diversity monitoring, for construction of a comprehensive DNA barcode library and for development of an index based on Next-generation sequencing analysis of samples composed of sorted specimens.

2016

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Testing diﬀerent (e)DNA metabarcoding approaches to assess aquatic oligochaete diversity and the biological quality of sediments

Régis Lionel Vivien

Aquatic oligochaetes are important bioindicators of sediment quality in watercourses and lakes, but their morphological identiﬁcation to the species level is challenging and sometimes impossible. The use of DNA barcoding and metabarcoding could greatly facilitate the identiﬁcations of specimens and improve ecological diagnoses based on oligochaete communities. The aim of this study was to test how well metabarcoding approaches based on high throughout sequencing (HTS) could perform in assessing oligochaete species diversity and the biological quality of sediments. We analysed oligochaete communities at several sites in Swiss rivers, comparing morphological data to metabarcoding of mixed specimen, sieved sediment and total sediment samples. We ampliﬁed the cytochrome c oxidase (COI) marker using universal primers and/or speciﬁc primers to metazoans. Our results showed that metabarcoding analysis of mixed specimens allowed a more representative assessment of the biodiversity than the metabarcoding of sieved or total sediments. Although community structures obtained with the morphological and metabarcoding analyses were diﬀerent in term of presence/ absence of species and species abundances, the ecological diagnoses based on these two approaches largely agreed.

2019

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Cytochrome c oxidase barcodes for aquatic oligochaete identification: development of a Swiss reference database

Régis Lionel Vivien

Introduction. Aquatic oligochaetes represent valuable indicators of the quality of sediments of watercourses and lakes, but their difficult identification based on morphological criteria compromises their more common use for eco-diagnostic analyses. This issue could be overcome by using DNA barcodes for species identification. A 10% threshold of cytochrome c oxidase (COI) divergence was proposed for differentiating between oligochaete species based on molecular and morphological data. A Swiss database of COI sequences of aquatic oligochaetes was initiated in 2012. The aim of this study is to complement the Swiss oligochaete database of COI sequences and to confirm the relevance of this threshold for species delimitation. Methods. We sequenced the COI sequence of 216 specimens collected in different regions of Switzerland and ITS2 region of some lineages whose delimitation with COI data was doubtful. Results. We distinguished 53 lineages, among which 34 were new for Switzerland and 17 sequenced for the first time. All the lineages were separated by more than 10% of COI variation, with the exception of some species within Nais and Uncinais. In these two genera, the threshold was lowered to 8% to be congruent with the morphological analysis. The total number of lineages reported so far for Switzerland is 75, including 59 morphospecies or unidentified species and 16 cryptic species. Discussion. Our study shows that the threshold of 10% of COI divergence is generally appropriate to distinguish aquatic oligochaete lineages, but that it must be adjusted for some species. The database reported here will be complemented in the future in parallel to the development of genetic oligochaete indices.

Peerj Inc2017

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BIO-205: Cellular and molecular biology I

The course covers the regulation of gene expression, which translates the information contained in the genome into function, by adjusting the levels and activities of mRNAs and proteins to the needs of specific cells, tissues and environments. A particular emphasis is given on experimental methods.

BIO-471: Cancer biology I

The course covers in detail molecular mechanisms of cancer development with emphasis on cell cycle control, genome stability, oncogenes and tumor suppressor genes.

BIO-310: Immunology

Ce cours décrit le fonctionnement du système immunitaire humain et les bases immunologiques de la vaccination, de la transplantation, de l'immunothérapie, et de l'allergie. Il présente aussi le rôle des cellules souches dans la formation des tissus, notamment la lignée hématopoïétique.

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Botanique

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Nematoda

Les nématodes (Nematoda), ou vers ronds et effilés, constituent un embranchement de vers non segmentés. Classés parmi les ecdysozoaires, ils sont recouverts d'une épaisse cuticule. Ils représ