7th International Workshop on the Molecular Biology and Genetics of the Lepidoptera: August 20–26, 2006 Orthodox Academy of Crete, Kolympari, Crete, Greece

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Improvement of the assembly of heterozygous genomes of non-model organisms, a case study of the genomes of two Spodoptera frugiperda host strains

International audienceThe extraction of biological information from the draft genomes of non-model organisms may result in unattainable, incomplete, or even wrong conclusions. In particular, the combination of a high level of heterozygosity and short reads sequencing may have major impact in the annotation of genes [1,2]. This wrong gene content assessment is usually the consequence of the high fragmentation of the genome sequence but it may also come from an overestimation of the genome size. The latter because the assembly of an heterozygous region for which there is a significant divergence between the two haplotypes leads sometimes to the construction of two different contigs, instead of one consensus sequence. To date, new assemblers such as Platanus [3], have been developed in regard to heterozygous data. But, the complete re-assembly of a genome leading to new automatic and manual annotations process is very cost-effective, and may still produce erroneous scaffolds and annotations. Thus, we set up a « soft » method to detect and correct false duplications due to heterozygosity in draft assemblies. In addition, to the identification and removal of the allelic regions (i.e. unmerged haplotypes), our protocol is able to relocate and merge supernumerary gene annotations.We applied this method as a pre-requisite for the comparison of the genomes of 2 Spodoptera frugiperda (Lepidoptera: Noctuidae) strains, in the frame of the WGS project supported by the Fall armyworm International Public Consortium (FAW-IPC). This moth is a well-known pest of crops throughout the Western hemisphere. This species consists of two strains adapted to different larval host-plants: the first feeds preferentially on corn, cotton and sorghum whereas the second is more associated with rice and several pasture grasses. While, the paired-end reads of the rice-variant have been directly assembled using Platanus [3], we cleaned up and corrected the first release of the corn-variant, leading to a drastic reduction of the genome assembly, with the removal of 88Mbp (17%) and the increase of the N50 from 39,593 to 52,781bp. The suppressed fragments included 3,746 gene predictions; about 80% of them have been either relocated or merged with their complementary allele. Subsequently, in order to identify new candidate genes or genomic regions involved in the host-plant adaptation, we compared the genomes and proteomes of the 2 different strains in order to identify orthologous genes, collinear regions and genome rearrangements, taking into consideration the inflated occurrence of splitted genes due to the high fragmentation of the genome

Spodoptera frugiperda (Lepidoptera: Noctuidae) host-plant variants: two host strains or two distinct species?

International audienceThe moth Spodoptera frugiperda is a well-known pest of crops throughout the Americas, which consists of two strains adapted to different host-plants: the first feeds preferentially on corn, cotton and sorghum whereas the second is more associated with rice and several pasture grasses. Though morphologically indistinguishable, they exhibit differences in their mating behavior, pheromone compositions, and show development variability according to the host-plant. Though the latter suggest that both strains are different species, this issue is still highly controversial because hybrids naturally occur in the wild, not to mention the discrepancies among published results concerning mating success between the two strains. In order to clarify the status of the two host-plant strains of S. frugiperda, we analyze features that possibly reflect the level of post-zygotic isolation: (1) first generation (F1) hybrid lethality and sterility; (2) patterns of meiotic segregation of hybrids in reciprocal second generation (F2), as compared to the meiosis of the two parental strains. We found a significant reduction of mating success in F1 in one direction of the cross and a high level of microsatellite markers showing transmission ratio distortion in the F2 progeny. Our results support the existence of post-zygotic reproductive isolation between the two laboratory strains and are in accordance with the marked level of genetic differentiation that was recovered between individuals of the two strains collected from the field. Altogether these results provide additional evidence in favor of a sibling species status for the two strains

Establishment and analysis of a reference transcriptome for Spodoptera frugiperda

International audienceBackground Spodoptera frugiperda (Noctuidae) is a major agricultural pest throughout the American continent. The highly polyphagous larvae are frequently devastating crops of importance such as corn, sorghum, cotton and grass. In addition, the Sf9 cell line, widely used in biochemistry for in vitro protein production, is derived from S. frugiperda tissues. Many research groups are using S. frugiperda as a model organism to investigate questions such as plant adaptation, pest behavior or resistance to pesticides.ResultsIn this study, we constructed a reference transcriptome assembly (Sf_TR2012b) of RNA sequences obtained from more than 35 S. frugiperda developmental time-points and tissue samples. We assessed the quality of this reference transcriptome by annotating a ubiquitous gene family - ribosomal proteins - as well as gene families that have a more constrained spatio-temporal expression and are involved in development, immunity and olfaction. We also provide a time-course of expression that we used to characterize the transcriptional regulation of the gene families studied.ConclusionWe conclude that the Sf_TR2012b transcriptome is a valid reference transcriptome. While its reliability decreases for the detection and annotation of genes under strong transcriptional constraint we still recover a fair percentage of tissue-specific transcripts. That allowed us to explore the spatial and temporal expression of genes and to observe that some olfactory receptors are expressed in antennae and palps but also in other non related tissues such as fat bodies. Similarly, we observed an interesting interplay of gene families involved in immunity between fat bodies and antennae

Correlation of LNCR rasiRNAs Expression with Heterochromatin Formation during Development of the Holocentric Insect Spodoptera frugiperda

Repeat-associated small interfering RNAs (rasiRNAs) are derived from various genomic repetitive elements and ensure genomic stability by silencing endogenous transposable elements. Here we describe a novel subset of 46 rasiRNAs named LNCR rasiRNAs due to their homology with one long non-coding RNA (LNCR) of Spodoptera frugiperda. LNCR operates as the intermediate of an unclassified transposable element (TE-LNCR). TE-LNCR is a very invasive transposable element, present in high copy numbers in the S. frugiperda genome. LNCR rasiRNAs are single-stranded RNAs without a prominent nucleotide motif, which are organized in two distinct, strand-specific clusters. The expression of LNCR and LNCR rasiRNAs is developmentally regulated. Formation of heterochromatin in the genomic region where three copies of the TE-LNCR are embedded was followed by chromatin immunoprecipitation (ChIP) and we observed this chromatin undergo dynamic changes during development. In summary, increased LNCR expression in certain developmental stages is followed by the appearance of a variety of LNCR rasiRNAs which appears to correlate with subsequent accumulation of a heterochromatic histone mark and silencing of the genomic region with TE-LNCR. These results support the notion that a repeat-associated small interfering RNA pathway is linked to heterochromatin formation and/or maintenance during development to establish repression of the TE-LNCR transposable element. This study provides insights into the rasiRNA silencing pathway and its role in the formation of fluctuating heterochromatin during the development of one holocentric organism

Molecular determinism of ecological adaptation and speciation in two strains of the Lepidoptera Spodoptera frugiperda

Molecular determinism of ecological adaptation and speciation in two strains of the Lepidoptera [i]Spodoptera frugiperda[/i]. Congrès de génomique environnemental

Extreme gene family expansion underlies adaptation for polyphagy in Spodoptera frugiperda

Extreme gene family expansion underlies adaptation for polyphagy in [i]Spodoptera frugiperda[/i]. International Meeting on Spodoptera litura genome projec

Progress in the Fall armyworm (FAW, Spodoptera frugiperda) whole genome sequencing project by the Fall Armyworm International Public Consortium (FAW-IPC)

Progress in the Fall armyworm (FAW, [i]Spodoptera frugiperda[/i]) whole genome sequencing project by the Fall Armyworm International Public Consortium (FAW-IPC). International Meeting on Spodoptera litura genome projec