The European Directive in 1998 led to the increasing use of biological insecticides such as cry proteins produced by the bacterium Bacillus thuringiensis israelensis (Bti) that kill mosquito larvae after being ingested. Considering the interest in Bti as more environmentally sustainable biocide, it is important to examine environmental fate and impact of Bti especially taking into account the need of this information to fulfil the REACH criteria. Chandrashekhar (PhD 2014, North Maharashtra University, India) will employed an innovative 'Environmental Metabolic Footprinting' approach, at the University of Perpignan via Domitia, France and participate in a multidisciplinary project that spans the interface between chemistry and biology. To dynamically characterize biomarkers of Bti pollution found among metabolites issued from the sediment matrix meta-metabolome will require him to develop and optimize detection protocols using LC-MS platform. In addition, metabarcoding approach will allow to understand microbial community responses to the Bti pollution. Emphasis will be placed on better standardisation, data interpretation and evaluation that will build confidence in the value of “omics technologies – this being essential to increase their (regulatory) use. Through secondment to University Joseph Fourier, Grenoble he will deepen his knowledge base by evaluating fate of Bti in the sediment through cutting edge biological analysis tools. Taken together, these activities will advance our understanding of environmental risks associated with Bti, and pave the way for the development and adaptation to new environmental monitoring tool. EnvFate will thus increase the European research visibility to promote sustainable development, ensure the protection of environment, one of the priority areas of the H2020 program. Moreover, through the planned training and mentoring activities, Chandrashekhar will develop in to an outstanding early career researcher of great potential.

Developing an integrative research allowing to elucidate both the links between genotype, phenotype and fitness and the genomic bases of the variation form a major challenge in our understanding of speciation and the emergence of biodiversity. In DiversiFly, I propose to implement such approach by generating, analyzing and comparing phenotypic (morphometry, coloration, odor), genomic and ecological data on Ophrys orchids. This genus displays one of the highest rates of diversification and hybrid zones formed in the wild by some of its species makes Ophrys a promising model to better understand the causes of adaptive radiations: phenomena of intense and rapid diversification in response to ecological selection pressures. Through its approaches, DiversiFly aims at combining studies led at different evolutionary scales, working on endemic and threatened species. In a first part, the parallel study of two hybrid zones will allow us to determine what are the phenotypic traits (morphology, coloration, odor) predominantly involved in adaptation and reproductive isolation within the O. insectifera clade. Through cline analyses, outlier research and association studies, we will then confront phenotypic and genomic data (transcriptomes and GBS data) in order to look for the genomic bases of traits of interest. The links between phenotypes and individual fitness will be evaluated based on life history traits related to reproductive success and survival. As all individuals will be marked, phenotyping and fitness will be evaluated each year over the four years of the project to investigate their stability. In a second part, we will link micro and ‘macro-’ evolutionary scales by working on the whole genus Ophrys. We will use floral transcriptomes on species, representative of the diversity of the genus in order to identify genes involved in shaping floral phenotype, so particular of the Ophrys. Following a comparative approach, we will base our study on both sequence variation and gene levels of expression. Our results will have a significant impact in the field of ecology and evolutionary biology, contributing to the emergence of a new plant model, with a complex genome, to study speciation and evolutionary diversification. They will also contribute to fields such as genomics, conservation biology, systematics and taxonomy. We also believe that with its flowers mimicking insects, Ophrys forms an excellent model for scientific communication and towards general audience.