Comparative analysis of Spodoptera frugiperda (J. E. Smith) (Lepidoptera, Noctuidae) corn and rice strains microbiota revealed minor changes across life cycle and strain endosymbiont association.

Background: Spodoptera frugiperda (FAW) is a pest that poses a significant threat to corn production worldwide, causing millions of dollars in losses. The species has evolved into two strains (corn and rice) that differ in their genetics, reproductive isolation, and resistance to insecticides and Bacillus thuringiensis endotoxins. The microbiota plays an important role in insects' physiology, nutrient acquisition, and response to chemical and biological controls. Several studies have been carried out on FAW microbiota from larvae guts using laboratory or field samples and a couple of studies have analyzed the corn strain microbiota across its life cycle. This investigation reveals the first comparison between corn strain (CS) and rice strain (RS) of FAW during different developmental insect stages and, more importantly, endosymbiont detection in both strains, highlighting the importance of studying both FAW populations and samples from different stages. Methods: The composition of microbiota during the life cycle of the FAW corn and rice strains was analyzed through high-throughput sequencing of the bacterial 16S rRNA gene using the MiSeq system. Additionally, culture-dependent techniques were used to isolate gut bacteria and the Transcribed Internal Spacer-ITS, 16S rRNA, and gyrB genes were examined to enhance bacterial identification. Results: Richness, diversity, and bacterial composition changed significantly across the life cycle of FAW. Most diversity was observed in eggs and males. Differences in gut microbiota diversity between CS and RS were minor. However, Leuconostoc, A2, Klebsiella, Lachnoclostridium, Spiroplasma, and Mucispirilum were mainly associated with RS and Colidextribacter, Pelomonas, Weissella, and Arsenophonus to CS, suggesting that FAW strains differ in several genera according to the host plant. Firmicutes and Proteobacteria were the dominant phyla during FAW metamorphosis. Illeobacterium, Ralstonia, and Burkholderia exhibited similar abundancies in both strains. Enterococcus was identified as a conserved taxon across the entire FAW life cycle. Microbiota core communities mainly consisted of Enterococcus and Illeobacterium. A positive correlation was found between Spiroplasma with RS (sampled from eggs, larvae, pupae, and adults) and Arsenophonus (sampled from eggs, larvae, and adults) with CS. Enterococcus mundtii was predominant in all developmental stages. Previous studies have suggested its importance in FAW response to B. thuringensis. Our results are relevant for the characterization of FAW corn and rice strains microbiota to develop new strategies for their control. Detection of Arsenophonus in CS and Spiroplasma in RS are promising for the improvement of this pest management, as these bacteria induce male killing and larvae fitness reduction in other Lepidoptera species.

Two host-plant strains in the fall armyworm.

The fall armyworm (Spodoptera frugiperda) is one of the major pest insects damaging diverse crops including cotton, corn, rice, and sorghum. Fall armyworms have been identified as two morphologically indistinguishable strains, the corn strain, and the rice strain, named after their preferred host-plants. Although initially recognized as host-plant strains, there has been an ongoing debate regarding whether the corn and rice strains should be considered as such. In this article, we present arguments based on recent population genomics studies supporting that these two strains should be considered to be host-plant strains. Furthermore, host-plant adaptation appears to be a driving evolutionary force responsible for incipient speciation in the fall armyworm.

Microbiome analysis of Spodoptera frugiperda (Lepidoptera, Noctuidae) larvae exposed to Bacillus thuringiensis (Bt) endotoxins.

Background: Spodoptera frugiperda (or fall armyworm, FAW) is a polyphagous pest native to Western Hemisphere and recently discovered in the Eastern Hemisphere. In Colombia, S. frugiperda is recognized as a pest of economic importance in corn. The species has genetically differentiated into two host populations named "corn" and "rice" strains. In 2012, a study made in central Colombia demonstrated that the corn strain is less susceptible to Bacillus thuringiensis (Bt) endotoxins (Cry1Ac and Cry 1Ab) than the rice strain. In this country, Bt transgenic corn has been extensively produced over the last 15 years. Since gut microbiota plays a role in the physiology and immunity of insects, and has been implicated in promoting the insecticidal activity of Bt, in this study an analysis of the interaction between Bt endotoxins and FAW gut microbiota was made. Also, the detection of endosymbionts was performed here, as they might have important implications in the biological control of a pest. Methods: The composition and diversity of microbiomes associated with larval specimens of S. frugiperda(corn strain) was investigated in a bioassay based on six treatments in the presence/absence of Bt toxins and antibiotics (Ab) through bacterial isolate analyses and by high throughput sequencing of the bacterial 16S rRNA gene. Additionally, species specific primers were used, to detect endosymbionts from gonads in S. frugiperda corn strain. Results: Firmicutes, Proteobacteria and Bacteroidota were the most dominant bacterial phyla found in S. frugiperda corn strain. No significant differences in bacteria species diversity and richness among the six treatments were found. Two species of Enterococcus spp., E. mundtii and E. casseliflavus were detected in treatments with Bt and antibiotics, suggesting that they are less susceptible to both of them. Additionally, the endosymbiont Arsenophonus was also identified on treatments in presence of Bt and antibiotics. The results obtained here are important since little knowledge exists about the gut microbiota on this pest and its interaction with Bt endotoxins. Previous studies made in Lepidoptera suggest that alteration of gut microbiota can be used to improve the management of pest populations, demonstrating the relevance of the results obtained in this work.

Two New Strains of Wolbachia Affecting Natural Avocado Thrips.

Wolbachia is an obligate intracellular bacterium with a high frequency of infection and a continental distribution in arthropods and nematodes. This endosymbiont can induce various reproductive phenotypes in their hosts and has been previously found naturally in several pests including thrips (Thripidae). These insects cause physical fruit damage and economic losses in avocado. The presence of Wolbachia was evaluated for the first time in avocado thrips populations of Frankliniella sp. and Scirtothrips hansoni sp.n. from eastern Antioquia. DNA from adult thrips individuals was used to assess the detection of Wolbachia by amplifying a fragment (600 bp) of the Wolbachia major surface protein (wsp) gene. Results confirmed the presence of two new Wolbachia strains in these two thrips species, with a higher percentage of natural infection in S. hansoni sp.n. The first Wolbachia species was found in Frankliniella sp. and belongs to supergroup A and the second was detected in S. hansoni sp.n. and is part of supergroup B. Wolbachia was more frequently found in females (32.73%), and only found in one male. Analysis of phylogenetic relationships, suggests that the two new Wolbachia sequences (wFran: Frankliniella and wShan: Scirtothrips hansoni) detected here represent two new groups for this endosymbiont. The haplotype network shows the presence of two possible haplotypes for each strain. Future studies to evaluate the possible use of Wolbachia as a control agent in avocado thrips are necessary. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12088-021-00951-5.

Mitochondrial genome characterization of Tecia solanivora (Lepidoptera: Gelechiidae) and its phylogenetic relationship with other lepidopteran insects.

The complete mitogenome of the potato tuber moth Tecia solanivora (Lepidoptera: Gelechiidae) was sequenced, annotated, characterized and compared with 140 species of the order Lepidoptera. The circular genome is 15,251 bp, containing 37 genes (13 protein-coding genes (PCGs), two rRNA genes, 22 tRNA genes and an A+T-rich region). The gene arrangement was identical to other lepidopteran mitogenomes but different from the ancestral arrangement found in most insects for the tRNA-Met gene (A+T-region, tRNA-I, tRNA-Q, tRNA-M). The mitogenome of T. solanivora is highly A+T-biased (78.2%) and exhibits negative AT- and GC-skews. All PCGs are initiated by canonical ATN start codons, except for Cytochrome Oxidase subunit 1 (COI), which is initiated by CGA. Most PCGs have a complete typical stop codon (TAA). Only NAD1 has a TAG stop codon and the COII and NAD5 genes have an incomplete stop codon consisting of just a T. The A+T-rich region is 332 bp long and contains common features found in lepidopteran mitogenomes, including the 'ATAGA' motif, a 17 bp poly (T) stretch and a (AT)8 element preceded by the 'ATTTA' motif. Other tandem repeats like (TAA)4 and (TAT)7 were found, as well as (T)6 and (A)10 mononucleotide repeat elements. Finally, this mitogenome has 20 intergenic spacer regions. The phylogenetic relationship of T. solanivora with 28 other lepidopteran families (12 superfamilies) showed that taxonomic classification by morphological features coincides with the inferred phylogeny. Thus, the Gelechiidae family represents a monophyletic group, suggesting that T. solanivora and Pectinophora gossypiella have a recent common ancestor.

Habitat association and seasonality in a mosaic and bimodal hybrid zone between Chorthippus brunneus and C. jacobsi (Orthoptera: Acrididae).

Understanding why some hybrid zones are bimodal and others unimodal can aid in identifying barriers to gene exchange following secondary contact. The hybrid zone between the grasshoppers Chorthippus brunneus and C. jacobsi contains a mix of allopatric parental populations and inter-mingled bimodal and unimodal sympatric populations, and provides an ideal system to examine the roles of local selection and gene flow between populations in maintaining bimodality. However, it is first necessary to confirm, over a larger spatial scale, previously identified associations between population composition and season and habitat. Here we use cline-fitting of one morphological and one song trait along two valley transects, and intervening mountains, to confirm previously identified habitat associations (mountain versus valley) and seasonal changes in population composition. As expected from previous findings of studies on a smaller spatial scale, C. jacobsi dominated mountain habitats and mixed populations dominated valleys, and C. brunneus became more prevalent in August. Controlling for habitat and incorporating into the analysis seasonal changes in cline parameters and the standard errors of parental trait values revealed wider clines than previous studies (best estimates of 6.4 to 24.5 km in our study versus 2.8 to 4.7 km in previous studies) and increased percentage of trait variance explained (52.7% and 61.5% for transects 1 and 2 respectively, versus 17.6%). Revealing such strong and consistent patterns within a complex hybrid zone will allow more focused examination of the causes of variation in bimodality in mixed populations, in particular the roles of local selection versus habitat heterogeneity and gene flow between differentiated populations.

[Genetic variability of Brycon henni (Characiformes: Characidae) in the middle basin of Nare and Guatapé Rivers, Magdalena River system, Colombia]. / Variabilidad genética de Brycon henni (Characiformes: Characidae) en la cuenca media de los ríos Nare y Guatapé, sistema Río Magdalena, Colombia.

Brycon henni is a native species in Magdalena's River basin, and because of its cultural and economic importance, is strongly overexploited. This study aimed to describe the genetic variability and population structure of this species from Nare and Guatapé rivers basins. A total of 195 individuals were collected and DNA extractions were obtained from muscle and blood tissue. Fourteen primers were evaluated with the RAPD technique, being four of them polymorphic, and produced 66 different fragments (63% polymorphism). Besides, using the molecular variance (AMOVA) analysis, the population structure was described for all sites (phi(ST) = 0.297, phi(ST) = 0.163; Nare and Guatapé river basins, respectively), and suggested the importance of the migratory behaviour of the species in the genetic differentiation. Genetic distances among sampled sites showed that most of the genetic differentiation occurred between sites Guatapé and El Cardal with respect to the others. A Mantel Test demonstrated a correlation between genetic and geographic distances (r = 0.431 both of the basins evaluated; r = 0.377, Nare river middle basin), and suggested isolation by distance. The outcomes obtained in this study have valuable implications in species conservation and the genetic variability of natural populations of B. henni, and should be complemented with morphological analyses.

Variabilidad genética de Brycon henni (Characiformes: Characidae) en la cuenca media de los ríos Nare y Guatapé, sistema Río Magdalena, Colombia

Genetic variability of Brycon henni (Characiformes: Characidae) in the middle basin of Nare and Guatapé rivers, Magdalena River system, Colombia. Brycon henni is a native species in Magdalena´s River basin, and because of its cultural and economic importance, is strongly overexploited. This study aimed to describe the genetic variability and population structure of this species from Nare and Guatapé rivers basins. A total of 195 individuals were collected and DNA extractions were obtained from muscle and blood tissue. Fourteen primers were evaluated with the RAPD technique, being four of them polymorphic, and produced 66 different fragments (63% polymorphism). Besides, using the molecular variance (AMOVA) analysis, the population structure was described for all sites ( ΦST=0.297, ΦST=0.163; Nare and Guatapé river basins, respectively), and suggested the importance of the migratory behaviour of the species in the genetic differentiation. Genetic distances among sampled sites showed that most of the genetic differentiation occurred between sites Guatapé and El Cardal with respect to the others. A Mantel Test demonstrated a correlation between genetic and geographic distances (r=0.431 both of the basins evaluated; r=0.377, Nare river middle basin), and suggested isolation by distance. The outcomes obtained in this study have valuable implications in species conservation and the genetic variability of natural populations of B. henni, and should be complemented with morphological analyses. Rev. Biol. Trop. 59 (1): 269-282. Epub 2011 March 01.

Brycon henni es una especie nativa de la cuenca del río Magdalena, Colombia, donde es fuertemente explotada, por lo cual en este trabajo se estudió la variabilidad genética y estructura poblacional en individuos provenientes de la cuenca media de los ríos Nare y Guatapé, Antioquia. Se recolectaron 195 ejemplares y se les extrajo ADN genómico total del tejido muscular y/o sanguíneo. Se evaluaron 14 cebadores para la técnica RAPD (Random Amplifyed Polimorphic DNA), siendo cuatro de ellos polimórficos, los cuales generaron 66 fragmentos diferentes (63% polimorfismo). Por medio de un análisis de varianza molecular (AMOVA), se determinó la estructuración poblacional para todos los sitios evaluados y la cuenca del río Nare ( ΦST =0.297 y ΦST =0.163, respectivamente). Las distancias genéticas de Nei mostraron que todas las poblaciones diferían entre sí y que la mayor diferenciación genética se dio para las poblaciones del río Guatapé y la quebrada El Cardal. Un test de Mantel mostró correlación entre las distancias genéticas y geográficas (ambas cuencas, r=0.431; cuenca del río Nare, r=0.377). Esto sugiere aislamiento por distancia y separación de grupos genéticos producidos por los embalses sobre los ríos Nare y Guatapé, lo cual puede tener implicaciones relevantes en la conservación de la variabilidad genética de las poblaciones naturales de esta especie en el oriente antioqueño.