TILLING - a shortcut in functional genomics

Recent advances in large-scale genome sequencing projects have opened up new possibilities for the application of conventional mutation techniques in not only forward but also reverse genetics strategies. TILLING (Targeting Induced Local Lesions IN Genomes) was developed a decade ago as an alternative to insertional mutagenesis. It takes advantage of classical mutagenesis, sequence availability and high-throughput screening for nucleotide polymorphisms in a targeted sequence. The main advantage of TILLING as a reverse genetics strategy is that it can be applied to any species, regardless of its genome size and ploidy level. The TILLING protocol provides a high frequency of point mutations distributed randomly in the genome. The great mutagenic potential of chemical agents to generate a high rate of nucleotide substitutions has been proven by the high density of mutations reported for TILLING populations in various plant species. For most of them, the analysis of several genes revealed 1 mutation/200–500 kb screened and much higher densities were observed for polyploid species, such as wheat. High-throughput TILLING permits the rapid and low-cost discovery of new alleles that are induced in plants. Several research centres have established a TILLING public service for various plant species. The recent trends in TILLING procedures rely on the diversification of bioinformatic tools, new methods of mutation detection, including mismatch-specific and sensitive endonucleases, but also various alternatives for LI-COR screening and single nucleotide polymorphism (SNP) discovery using next-generation sequencing technologies. The TILLING strategy has found numerous applications in functional genomics. Additionally, wide applications of this throughput method in basic and applied research have already been implemented through modifications of the original TILLING strategy, such as Ecotilling or Deletion TILLING

Ecological signature of the end-Triassic biotic crisis: what do bivalves have to say?

In order to understand the causes underlying the Triassic-Jurassic (T/J) mass extinction, we tested different bivalve features for extinction selectivity, i.e. shell mineralogy, age at the Rhaetian and three main autoecologic traits (feeding mechanism, tiering and motility/attachment). Also, diversity and turnover rates throughout the Triassic and the Early Jurassic were analysed in detail. The dataset employed for this analysis was a precise database at genus level including data from Induan to Sinemurian times. Results point to a true mass extinction for bivalves around the T/J boundary. This extinction was not ageselective at the boundary. Certain analyses suggested that shell mineralogy was a character significantly increasing survival odds, but this relationship seems to reflect selectivity on autoecologic traits. There was no difference in extinction proportions between both feeding types (i.e. deposit feeders and filter feeders); among the other traits, deep burrowers, epifaunal-motile and endobyssate forms seem to have been favoured, while shallow burrowers (and probably reclined forms) were more heavily affected. This pattern suggests an environmental stress at the boundary with some particular issues affecting the different life modes. Models linking magmatism in the Central Atlantic Magmatic Province with the end- Triassic mass extinction are a plausible scenario for this kind of perturbation