Annotation of transposable elements in the transcriptome of the Neotropical brown stink bug Euschistus heros and its chromosomal distribution.

Transposable elements (TEs) are DNA sequences capable of moving within the genome. Their distribution is very dynamic among organisms, and despite advances, there are still gaps in the understanding of the diversity and evolution of TEs in many insect species. In the case of Euschistus heros, considered the main stink bug in the soybean crop in Brazil, little is known about the participation of these elements. Therefore, the objective of the current work was to identify the different groups of transposable elements present in the E. heros transcriptome, evidencing their chromosomal distribution. Through RNA-Seq and de novo assembly, 60,009 transcripts were obtained, which were annotated locally via Blastn against specific databases. Of the 367 transcripts identified as TEs, 202 belong to Class II, with emphasis on the TIR order. Among Class I elements or retrotransposons, most were characterized as LINE. Phylogenetic analyses were performed with the protein domains, evidencing differences between Tc1-mariner sequences, which may be related to possible horizontal transfer events. The transposable elements that stood out in the transcriptome were selected for fluorescent in situ hybridization. DNA transposon probes hAT, Helitron, and Tc1-mariner showed mostly scattered signals, with the presence of some blocks. Retrotransposon probes Copia, Gypsy, Jockey, and RTE showed a more pulverized hybridization pattern, with the presence of small interstitial and/or terminal blocks. Studies like this one, integrating functional genomics and molecular cytogenetic tools, are essential to expanding knowledge about transcriptionally active mobile elements, and their behavior in the chromosomes.

Transposable elements in the transcriptome of the velvetbean caterpillar Anticarsia gemmatalis Hübner, 1818 (Lepidoptera: Erebidae).

Transposable elements (TEs) are DNA sequences that possess the ability to move from one genomic location to another. These sequences contribute to a significant fraction of the genomes of most eukaryotes and can impact their architecture and regulation. In this paper, we present the first data related to the identification and characterization of TEs present in the transcriptome of Anticarsia gemmatalis. Approximately, 835 transcripts showed significant similarity to TEs and (or) characteristic domains. Retrotransposons accounted for 71.2% (595 sequences) of the identified elements, while DNA transposons were less abundant, with 240 annotations (28.8%). TEs were classified into 30 superfamilies, with SINE3/5S and Gypsy being the most abundant. Based on the sequences of TEs found in the transcriptome, we were able to locate conserved regions in the chromosomes of this species. The analysis of differential expression of TEs in susceptible and resistant strains, challenged and not challenged with Bacillus thuringiensis (Bt) from in silico analysis, indicated that exposure to Bt can regulate the transcription of mobile genetic elements in the velvetbean caterpillar. Thus, these data contribute significantly to the knowledge of the structure and composition of these elements in the genome of this species, and suggest the role of stress on their expression.

Transcriptional Analysis of Microsatellites in Velvetbean Caterpillar Anticarsia gemmatalis Hübner, 1818.

Brazil is the largest producer of soybeans in the world. The vast extent of soybean plantations across the Brazilian territory exposes this crop to attack by several insects, including the velvetbean caterpillar, Anticarsia gemmatalis. One of the alternatives used to control this insect are the toxins produced by Bacillus thuringiensis (Bt). However, in some cases, resistance to these toxins has been reported in the laboratory. Despite the ecological and economic impact of the velvetbean caterpillar, there are few studies on the genetic structure of this species, especially with regard to microsatellites. In this paper, we carried out a comparative transcriptional analysis of microsatellites in resistant (RES) and susceptible (SUS) strains of A. gemmatalis challenged and not challenged with Bt toxins. According to the number of sequences analyzed in each group, a 7.9% simple sequence repeat (SSR) rate was identified for the SUS library, and 7.4% for SUSBt. For the RES group, this value was 8.5% and for the RESBt 7.7%. Most of the fragments found showed a shorter repeat pattern, located in mono- and trinucleotide motifs. Among the 128 types of SSR motifs, it was possible to notice a large amount of adenine and thymine in relation to guanine and cytosine, which was also seen in chromosomes after staining with base-specific fluorochromes DAPI/CMA3, highlighting DAPI-positive regions. Although the participation of microsatellites in the resistance mechanism of A. gemmatalis to Bt is not clear, the results obtained in this work contribute to a better understanding of the repetitive DNA found in transcribed regions of a non-model organism.

Testes morphology and the identification of transcripts of the hormonal pathways of the velvetbean caterpillar Anticarsia gemmatalis Hübner, 1818 (Lepidoptera: Erebidae).

Anticarsia gemmatalis is one of the main defoliating pests of soybeans in Brazil. In the current study, we characterized the histomorphology of the testes and the spermatogenesis process in A. gemmatalis. We also identified transcripts involved in the biosynthesis, metabolism, and signaling of juvenile and ecdysteroid hormones, in order to provide information about potential mechanisms of regulation of hormonal pathways in this species. Our analyses revealed that the A. gemmatalis larvae have a pair of kidney-shaped testicles. These are divided into four testicular follicles, where there are germ cell cysts at different stages of development. In the pupal stage, the testicles are fused, so adults have a single spherical testis, with a variable number of follicles. The A. gemmatalis has centripetal spermatogenesis and exhibits spermatic dimorphism. We identified 31 transcripts that encode proteins involved in juvenile hormone and ecdysteroid pathways, such as mevalonate kinase, CYP14A1, ecdysone receptor, among others. Our results on the morphology of the testes and spermatogenesis process, as well as identification of the genes involved in hormonal pathways in A. gemmatalis, provide important data for understanding the biology of this agricultural pest, which can be used as a basis for further research in other economically important lepidopterans.

Transcriptional profiling analysis of susceptible and resistant strains of Anticarsia gemmatalis and their response to Bacillus thuringiensis.

Anticarsia gemmatalis is one of the main defoliators of soybean in Brazil. Bacillus thuringiensis (Bt) transgenic crops are used for their management. In this paper we used RNA-seq to explore the response of A. gemmatalis to Bt HD73, as well as to detect transcriptional differences after Bt infection between resistant and susceptible strains. A total of 3853 and 6224 differentially expressed genes (DGEs) were identified in susceptible and resistant larvae after Bt exposure, respectively. We identified 2143 DEGs between susceptible and resistant larvae and 1991 between susceptible and resistant larvae Bt exposed. Immunity-related genes, Bt toxins receptors, proteases, genes involved in metabolic processes, transporters, cuticle proteins and mobile elements have been identified. qRT-PCR data demonstrated upregulation of five genes in susceptible strain after Bt exposure. These results provide insights to understand the molecular and cellular mechanisms of response to Bt that could be used in strategies to control agricultural pests.

Chromosome mapping in Abracris flavolineata (De Geer, 1773) (Orthoptera) from the Iguaçu National Park - Foz do Iguaçu, Paraná, Brazil.

In this paper, we present the cytomolecular analysis of a population of Abracris flavolineata collected in the largest fragment of the Brazilian Atlantic forest, the Iguaçu National Park. The diploid number in males was 23 (22+X0), with two large pairs (1-2), 7 medium (3-9), 2 small (10-11) and the X chromosome of medium size. Heterochromatic blocks were evident in the pericentromeric regions of all chromosomes. Heterogeneity in the distribution of heterochromatin was observed, with a predominance of DAPI+ blocks. However, some chromosomes showed CMA3+ blocks and other DAPI+/CMA3+ blocks. The 18S rDNA sites were distributed on the short arms of 5 pairs. In two of these pairs, such sites were in the same chromosome bearing 5S rDNA, and one of the bivalents, they were co-located. Histone H3 genes were found on one bivalent. The results added to the existing cytogenetic studies provided evidence of great karyotypic plasticity in the species. This pliancy may be the result of vicariant events related to the geographical distribution of different populations of A. flavolineata.

Analysis of the karyotype structure in Ricolla quadrispinosa (Linneus, 1767): inferences about the chromosomal evolution of the tribes of Harpactorinae (Heteroptera, Reduviidae).

The subfamily Harpactorinae is composed of six tribes. Phylogenetic studies bring together some of Harpactorinae tribes, but by and large the data on evolutionary relationships of the subfamily are scarce. Chromosome studies are of great importance for understanding the systematics of different groups of insects. For Harpactorinae, these studies are restricted to some subfamilies and involved only conventional chromosome analysis. This work analyzed cytogenetically Ricolla quadrispinosa (Linneus, 1767). The chromosome number was determined as 2n = 24 + X1X2Y in males. In metaphase II the autosomal chromosomes were organized in a ring with the pseudo-trivalent of sex chromosomes in its center. After C-banding followed by staining with DAPI, AT-rich blocks in autosomes were observed and the negatively heteropycnotic sex chromosomes. The data obtained, together with existing data for other species of the group, indicated that different chromosomal rearrangements are involved in the evolution of the species. In addition, a proposal of karyotype evolution for the subfamily, based on existing phylogenetic studies for the group is presented.

Cryptic species of the genus Pimelodella (Siluriformes: Heptapteridae) from the Miranda River, Paraguay River basin, Pantanal of Mato Grosso do Sul, Central Brazil

Specimens of Pimelodella captured in the Miranda River, Pantanal of Mato Grosso do Sul State, present morphological features that could indicate at least four species. Therefore, karyotype analysis and molecular biology provided evidence that they were only two species, one showing 2n = 46, and the other, 2n = 52 chromosomes, with only 18% genetic similarity. The morphological analysis evidenced that the dorsal filament is a male characteristic and that the upper lobe of the caudal fin was variable and might or might not be elongated in both species. With respect to morphometric characters, the formation of two groups was evident, but with a small overlap of specimens between them. Among the species with filaments on the dorsal fin observed in the Pantanal, the one with the lesser length of adipose fin base is P. griffini, which corresponds to that with 2n = 46 chromosomes, whereas the species P. taenioptera has 2n = 52 chromosomes. Thus, the accurate detection of these cryptic taxonomic units was only possible with the use of various analysis techniques. Furthermore, it is worth noting that the identification of cryptic species is important for obtaining correct estimates of fish diversity in the Pantanal.

Exemplares de Pimelodella capturados no rio Miranda, Pantanal do Mato Grosso do Sul, apresentavam características morfológicas que poderiam indicar, pelo menos, quatro espécies. Entretanto, com a análise cariotípica e da biologia molecular ficou evidente que se tratava de apenas duas espécies, uma apresentando 2n = 46 e a outra, 2n = 52 cromossomos, e com apenas 18% de similaridade genética. Pela análise morfológica foi observado que o filamento dorsal é uma característica de machos, e o lobo superior da nadadeira caudal se mostrou variável, podendo, ou não, ser alongado em ambas espécies. Com relação aos caracteres morfométricos, também houve a formação de dois grupos, mas com uma pequena sobreposição de exemplares entre eles. Das espécies com filamento na nadadeira dorsal apontadas para o Pantanal, a que possui menor comprimento da base da nadadeira adiposa é P. griffini, o que corresponde àquela com 2n = 46 cromossomos e, ao contrário, a espécie com 2n = 52 cromossomos, é P. taenioptera. Assim, apenas com o emprego de diversas técnicas de análise foi possível o reconhecimento seguro dessas unidades taxonômicas que se mostravam crípticas. Ressalta-se, ainda, que a identificação de espécies crípticas é importante para que estimativas da diversidade de peixes do Pantanal sejam feitas corretamente.