Regulating resistance: CncC:Maf, antioxidant response elements and the overexpression of detoxification genes in insecticide resistance

Although genetic and genomic tools have greatly furthered our understanding of resistance-associated mutations in molecular target sites of insecticides, the genomic basis of transcriptional regulation of detoxification loci in insect pests and vectors remains relatively unexplored. Recent work using RNAi, reporter assays and comparative genomics are beginning to reveal the molecular architecture of this response, identifying critical transcription factors and their binding sites. Central to this is the insect ortholog of the mammalian transcription factor Nrf2, Cap ‘n’ Collar isoform-C (CncC) which as a heterodimer with Maf-S regulates the transcription of phase I, II and III detoxification loci in a range of insects, with CncC knockdown or upregulation directly affecting phenotypic resistance. CncC:Maf binds to specific antioxidant response element sequences upstream of detoxification genes to initiate transcription. Recent work is now identifying these binding sites for resistance-associated loci and, coupled with genome sequence data and reporter assays, enabling identification of polymorphisms in the CncC:Maf binding site which regulate the insecticide resistance phenotype. © 2018 Elsevier Inc

The genome of the venomous snail Lautoconus ventricosus shed light on the origin of conotoxin diversity

Background: Venoms are deadly weapons to subdue prey or deter predators that have evolved independently in many animal lineages. The genomes of venomous animals are essential to understand the evolutionary mechanisms involved in the origin and diversification of venoms. Results: Here, we report the chromosome-level genome of the venomous Mediterranean cone snail, Lautoconus ventricosus (Caenogastropoda: Conidae). The total size of the assembly is 3.59 Gb; it has high contiguity (N50 = 93.53 Mb) and 86.6 Mb of the genome assembled into the 35 largest scaffolds or pseudochromosomes. On the basis of venom gland transcriptomes, we annotated 262 complete genes encoding conotoxin precursors, hormones, and other venom-related proteins. These genes were scattered in the different pseudochromosomes and located within repetitive regions. The genes encoding conotoxin precursors were normally structured into 3 exons, which did not necessarily coincide with the 3 structural domains of the corresponding proteins. Additionally, we found evidence in the L. ventricosus genome for a past whole-genome duplication event by means of conserved gene synteny with the Pomacea canaliculata genome, the only one available at the chromosome level within Caenogastropoda. The whole-genome duplication event was further confirmed by the presence of a duplicated hox gene cluster. Key genes for gastropod biology including those encoding proteins related to development, shell formation, and sex were located in the genome. Conclusions: The new high-quality L. ventricosus genome should become a reference for assembling and analyzing new gastropod genomes and will contribute to future evolutionary genomic studies among venomous animals.This work was funded by the Spanish Ministry of Science and Innovation (CGL2016-75255-C2-1-P [AEI/FEDER, UE] and PID2019-103947GB-C22/AEI/10.13039/501100011033 to R.Z.; BES-2017-081195 to J.R.P.-B.; BES-2014-069575 to S.A.; IJCI-2016-29566 to I.I.). I.I. acknowledges the support from the European Research Council during the latest stages of the project (Grant Agreement No. 852725; ERC-StG "TerreStriAL" to Jan de Vries, University of Goettingen)

Enzymatic hydrolysis of cellulosic fiber

Low cost cellulosic wastes like paper sludge, municipal wastes, solid wastes from food, packing etc. contain a high amount of cellulose which can be converted to bioethanol by two steps: (1) solubilization of cellulosic fibers to monosaccharides (2) conversion of monosachharides to bioethanol via fermentation. At present the implementation of this technology has been deterred by high cost for enzymes. Enzymatic hydrolysis of cellulosic fibers shows a biphasic behavior with an initial fast step followed by a slow step leading to low cellulose conversion rates. Low hydrolytic conversion rates necessitate the use of a high enzyme dosage to obtain meaningful cellulose conversion rates which make the implementation of this entire technology economically infeasible. The objective of this study is to get a better understanding of the mechanism of enzymatic hydrolysis of fibers to glucose and to investigate the effect of cationic polymers on enzymatic hydrolysis rates. To achieve the first objective, we performed experiments so as to study changes in morphological and physiochemical properties like fiber length, percentage of fines, crystallinity index, kink angle, kink index, mean curl, total organic carbon and glucose production with time. We used bleached kraft softwood, hardwood, and unbleached softwood fiber as cellulosic substrate and pergalase as cellulase enzyme. All of the experiments were carried out at experimental conditions of a temperature of 50 .C and a pH of 5.0 which maximize enzymatic activity. We studied the impact of recycling and refining on hydrolysis rates by measuring total organic carbon and glucose production. We found that refining increases enzymatic conversion rates by about as much as 20 %, however refining being energy intensive makes its implementation economically unfavorable. We found a novel way of enhancing hydrolysis rates by the use of cationic polyacrylamides. The effect of cationic polacrylamides was studied on both hardwood and softwood fibers at similar experimental conditions. Cationic polyacrylamides produced a maximum rate increase of 20 % in hydrolytic conversion rates for hardwood fibers. Even though, the increase in hydrolysis rates for softwood fibers was smaller than hardwood fibers, it was still significant. We further studied the effect of parameters like polymer concentration, cationicity and molecular weight to find a relation between properties of polymers and the increase in enzymatic hydrolysis.M.S.Committee Chair: Banerjee Sujit; Committee Member: Deng Yulin; Committee Member: Haynes Dann

Genetic variation in reproductive investment across an ephemerality gradient in Daphnia pulex

Species across the tree of life can switch between asexual and sexual reproduction. In facultatively sexual species, the ability to switch between reproductive modes is often environmentally dependent and subject to local adaptation. However, the ecological and evolutionary factors that influence the maintenance and turnover of polymorphism associated with facultative sex remain unclear. We studied the ecological and evolutionary dynamics of reproductive investment in the facultatively sexual model species, Daphnia pulex. We found that patterns of clonal diversity, but not genetic diversity varied among ponds consistent with the predicted relationship between ephemerality and clonal structure. Reconstruction of a multi-year pedigree demonstrated the coexistence of clones that differ in their investment into male production. Mapping of quantitative variation in male production using lab-generated and field-collected individuals identified multiple putative quantitative trait loci (QTL) underlying this trait, and we identified a plausible candidate gene. The evolutionary history of these QTL suggests that they are relatively young, and male limitation in this system is a rapidly evolving trait. Our work highlights the dynamic nature of the genetic structure and composition of facultative sex across space and time and suggests that quantitative genetic variation in reproductive strategy can undergo rapid evolutionary turnover