Comparative transcriptional analysis between susceptible and resistant populations of Aedes (Stegomyia) aegypti (Linnaeus, 1762) after malathion exposure.

Aedes aegypti is an important vector of arboviruses, including dengue, chikungunya and Zika. The application of synthetic insecticides is a frequently used strategy to control this insect. Malathion is an organophosphate insecticide that was widely used in Brazil in the 1980s and 1990s to control the adult form of A. aegypti. In situations where resistance to currently used insecticides is detected, the use of malathion may be resumed as a control measure. Many studies have confirmed resistance to malathion, however, comparative studies of differential gene expression of the entire transcriptome of resistant and susceptible insects are scarce. Therefore, understanding the molecular basis of resistance to this insecticide in this species is extremely important. In this paper, we present the first transcriptomic description of susceptible and resistant strains of A. aegypti challenged with malathion. Guided transcriptome assembly resulted in 39,904 transcripts, where 2133 differentially expressed transcripts were detected, and three were validated by RT-qPCR. Enrichment analysis for these identified transcripts resulted in 13 significant pathways (padj < 0.05), 8 associated with down-regulated and 5 with up-regulated transcripts in treated resistant insects. It was possible to divide the transcripts according to the mechanism of action into three main groups: (i) genes involved in detoxification metabolic pathways; (ii) genes of proteins located in the membrane/extracellular region; and (iii) genes related to DNA integration/function. These results are important in advancing knowledge of genes related to resistance mechanisms in this insect, enabling the development of effective technologies and strategies for managing insecticide resistance.

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.

Chromosomal distribution of major rDNA and genome size variation in Belostoma angustum Lauck, B. nessimiani Ribeiro & Alecrim, and B. sanctulum Montandon (Insecta, Heteroptera, Belostomatidae).

Known as "electric-light bugs", belostomatids potentially act as agents of biological control. The Belostoma genus has holokinetic chromosomes, interspecific variation in diploid number, sex chromosome system and DNA content. Thus, the chromosomal complement, the accumulation of constitutive heterochromatin and the distribution of rDNA clusters by fluorescence in situ hybridization (FISH) in Belostoma angustum (BAN), Belostoma sanctulum (BSA), and Belostoma nessimiani (BNE) were evaluated. In addition, a comparative analysis of the DNA content of these species and B. estevezae (BES) was performed. BES has the highest Belostoma DNA content, while BSA has the lowest. BAN showed 2n = 29 + X1X2Y, while BSA and BNE had 2n = 14 + XY. BSA showed 18S rDNA markings on sex chromosomes, while BNE and BAN did on autosomes. The difference between BSA and BNE occurs because of the possible movement of the rDNA cluster in BNE. We suggest the occurrence of fusion in the autosomes of BSA and BNE, and fragmentation in the sex chromosomes in BAN. Also, the genome size of 1-2 pg represents a haploid DNA content of a common ancestor, from which the genomes of BES and BAN had evolved by gene duplication and heterochromatinization events.

Influence of temperature variation on gene expression and cocoon production in Bombyx mori Linnaeus, 1758 (Lepidoptera: Bombycidae).

Silkworms (Bombyx mori) are lepidopterans of economic importance for global silk production. However, factors that directly affect the yield and quality of silkworm cocoon production, such as diseases and temperature fluctuations, cause great economic losses. Knowing how they respond to rearing temperature during the most critical stage of their life cycle (i.e., fifth instar) could provide information on their adaptation and improve silk production. In the current work, we analyzed transcriptional data from two groups of B. mori that were reared at 26 °C and 34 °C throughout the fifth instar. The silkworms and cocoons were weighed. In total, 3115 transcripts were differentially expressed (DE; including 1696 down-regulated and 1419 up-regulated) among the 29,157 sequences found by transcriptome assembly. We emphasize the genes associated with immunological response, transcription factors, silk biosynthesis, and heat shock proteins, among the DE transcripts in response to the temperature conditions. Silkworms reared at 34 °C presented a reduced mean body weight (-0.944 g in comparison to the 26 °C group), which had a direct impact on the weight of cocoons formed and the silk conversion rate. These changes were statistically significant when compared to silkworms reared at 26 °C. Mortality rates (6 and 9 %, at 26 °C and 34 °C, respectively) were similar to those obtained in breeding fields. The findings provide information on the biological processes involved in the temperature response mechanism of silkworms, as well as information that may be used in future climatization processes at rearing facilities and in breeding for improved thermotolerance.

New cytogenetic data for three species of Pentatomidae (Heteroptera): Dichelops melacanthus (Dallas, 1851), Loxa viridis (Palisot de Beauvois, 1805), and Edessa collaris (Dallas, 1851).

In this paper, we present new cytogenetic data for three species of the family Pentatomidae: Dichelops melacanthus (Dallas, 1851), Loxa viridis (Palisot de Beauvois, 1805), and Edessa collaris (Dallas, 1851). All studied species presented holocentric chromosomes and inverted meiosis for the sex chromosomes. D. melacanthus has 2n = 12 (10A + XY); L. viridis showed 2n = 14 (12A + XY); and E. collaris showed 2n = 14 (12A + XY). C-banding was performed for the first time in these species and revealed terminal and interstitial heterochromatic regions on the autosomes; DAPI/CMA3 staining showed different fluorescent patterns. In all species, fluorescence in situ hybridization (FISH) with 18S rDNA probe identified signals on one autosomal bivalent, this being the first report of FISH application in the species D. melacanthus and L. viridis. The results obtained add to those already existing in the literature, enabling a better understanding of the meiotic behavior of these insects.

Unrevealing the Karyotypic Evolution and Cytotaxonomy of Armored Catfishes (Loricariinae) with Emphasis in Sturisoma, Loricariichthys, Loricaria, Proloricaria, Pyxiloricaria, and Rineloricaria.

This study provides new insight into the chromosomal diversification in Loricariinae. We analyzed nine species from different Brazilian hydrographic basins, using conventional and molecular cytogenetic methods, aiming to understand the karyotypic diversification, and contribute with cytotaxonomic markers in this group considered one of the most diverse of Loricariidae. Our results evidenced a high karyotypic variability in diploid number (2n) ranging from 2n = 54 (Loricariichthys platymetopon and Loricariichthys anus), 2n = 60 (Rineloricaria reisi and Rineloricaria parva), 2n = 62 (Proloricaria prolixa), 2n = 64 (Loricaria cataphracta complex species), 2n = 66 (Sturisoma barbatum), and 2n = 68 (Pyxiloricaria menezesi). Different patterns of 18S and 5S ribosomal DNA (rDNA) were also identified, while slight divergences in heterochromatin distribution were observed. This high variability is probably related with independent events of Robertsonian translocations, pericentric inversions, and different mechanisms of rDNA sites dispersion (nonreciprocal translocation and transposable element [TEs] co-localization). In addition, our study provides a set of efficient chromosomal markers for the characterization of all analyzed species, and certainly, in future analyzes, will contribute as a useful cytotaxonomic tool in groups where the traditional taxonomy based on morphological data are not sufficient to clarify their relationship.

Cytogenetic markers applied to cytotaxonomy in two soybean pests: Anticarsia gemmatalis (Hübner, 1818) and Chrysodeixis includens (Walker, 1858).

Anticarsia gemmatalis (Hübner, 1818) and Chrysodeixis includens (Walker, 1858) are species of Lepidoptera that cause great damages in the soybean plantations of Brazil. Despite the importance they have in this regard, there are no studies on the chromosomal organization of these species and recently, A. gemmatalis, which belonged to the Noctuidae family, was allocated to the Erebidae family. Therefore, the objective of this paper was to analyze, through conventional and molecular cytogenetic markers, both species of Lepidoptera. A 2n = 62 was observed, with ZZ/ZW sex chromosome system and holokinetic chromosomes for both species. There was homogeneity in the number of 18S rDNA sites for both species. However, variations in heterochromatin distribution were observed between both species. The cytogenetic analyses enabled separation of the species, corroborating the transference of A. gemmatalis, from the family Noctuidae to the family Erebidae, suggesting new cytotaxonomic characteristics.

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.