Complete dosage compensation and sex-biased gene expression in the moth Manduca sexta.

Sex chromosome dosage compensation balances homogametic sex chromosome expression with autosomal expression in the heterogametic sex, leading to sex chromosome expression parity between the sexes. If compensation is incomplete, this can lead to expression imbalance and sex-biased gene expression. Recent work has uncovered an intriguing and variable pattern of dosage compensation across species that includes a lack of complete dosage compensation in ZW species compared with XY species. This has led to the hypothesis that ZW species do not require complete compensation or that complete compensation would negatively affect their fitness. To date, only one study, a study of the moth Bombyx mori, has discovered evidence for complete dosage compensation in a ZW species. We examined another moth species, Manduca sexta, using high-throughput sequencing to survey gene expression in the head tissue of males and females. We found dosage compensation to be complete in M. sexta with average expression between the Z chromosome in males and females being equal. When genes expressed at very low levels are removed by filtering, we found that average autosome expression was highly similar to average Z expression, suggesting that the majority of genes in M. sexta are completely dosage compensated. Further, this compensation was accompanied by sex-specific gene expression associated with important sexually dimorphic traits. We suggest that complete dosage compensation in ZW species might be more common than previously appreciated and linked to additional selective processes, such as sexual selection. More ZW and lepidopteran species should now be examined in a phylogenetic framework, to understand the evolution of dosage compensation.

Hawkmoth fauna (Sphingidae, Lepidoptera) in a semi-deciduous rainforest remnant: composition, temporal fluctuations, and new records for Northeastern Brazil.

We carried out a qualitative and quantitative inventory of the hawkmoth fauna (Sphingidae) of an area of semi-deciduous seasonal rainforest in the state of Pernambuco (Tapacurá Ecological Station), northeastern Brazil. Hawkmoths were sampled monthly from October 2004 to February 2007 (27 months). We recorded 31 species from 16 genera, three tribes, and three families. Macroglossinae was the most abundant subfamily and represented ca. 71% of all species. Out of the 277 individuals collected, 88.4% were males. Five new records were made for northeastern Brazil: Enyo gorgon (Cramer, 1777), Perigonia stulta (Herrich-Schäffer, [1854]), Eupyrrhoglossum sagra (Poey, 1832), Nyceryx coffaeae (Walker, 1856) and Xylophanes chiron (Drury, 1773). Eight further species were recorded for the first time for the Pernambuco Endemism Center, showing the important role played by Tapacurá Station in preserving the biodiversity of this insect group. Species richness and abundance were directly related to rainfall: about 70% of all individuals were captured during the rainy season. Changes in Sphingidae populations may, however, be caused by other factors that directly affect either larvae and adults of those insects, such as matrix effect and forest fragment size, which influence migration processes and the presence of predators.

Evolution of Manduca sexta hornworms and relatives: biogeographical analysis reveals an ancestral diversification in Central America.

The hawkmoth genus Manduca is a diverse group of very large, conspicuous moths that has served as an important model across many biological disciplines. Two species in particular, the tobacco hornworm (Manduca sexta) and the tomato hornworm (Manduca quinquemaculatus) have been researched extensively. Studies across biological fields have referred to these two species as being closely related or even sister species, but the extent to which these two model organisms are related remains largely unknown. We conducted a comprehensive multi-gene phylogenetic analysis of Manduca, based on both an ML and Bayesian framework, which resulted in a monophyletic Manduca but only when two other genera, Dolba and Euryglottis are included. We tentatively conclude that the sister group to Manduca sexta comprises the Caribbean M. afflicta and M. johanni, and the sister lineage to this clade includes M. quinquemaculatus and the Hawaiian M. blackburni. Thus, M. sexta and M. quinquemaculatus are closely related, but are not sister species. Biogeographical analyses reveal an ancestral center of diversification in Central America, and Manduca appears to have subsequently colonized North and South America. Our phylogeny provides an important foundation for comparative studies of two model organisms and their relatives.

Identification of chemosensory receptor genes in Manduca sexta and knockdown by RNA interference.

BACKGROUND: Insects detect environmental chemicals via a large and rapidly evolving family of chemosensory receptor proteins. Although our understanding of the molecular genetic basis for Drosophila chemoreception has increased enormously in the last decade, similar understanding in other insects remains limited. The tobacco hornworm, Manduca sexta, has long been an important model for insect chemosensation, particularly from ecological, behavioral, and physiological standpoints. It is also a major agricultural pest on solanaceous crops. However, little sequence information and lack of genetic tools has prevented molecular genetic analysis in this species. The ability to connect molecular genetic mechanisms, including potential lineage-specific changes in chemosensory genes, to ecologically relevant behaviors and specializations in M. sexta would be greatly beneficial. RESULTS: Here, we sequenced transcriptomes from adult and larval chemosensory tissues and identified chemosensory genes based on sequence homology. We also used dsRNA feeding as a method to induce RNA interference in larval chemosensory tissues. CONCLUSIONS: We report identification of new chemosensory receptor genes including 17 novel odorant receptors and one novel gustatory receptor. Further, we demonstrate that systemic RNA interference can be used in larval olfactory neurons to reduce expression of chemosensory receptor transcripts. Together, our results further the development of M. sexta as a model for functional analysis of insect chemosensation.

Molecular cloning and functional expression of a proline transporter from Manduca sexta.

We report the molecular cloning of a L-proline transporter, MasPROT cDNA and its splice variants MasPROT.16 and MasPROT.2 from the central nervous system of Manduca sexta. Sequence analysis revealed that MasPROT belongs to a family of high affinity Na+/Cl- dependent neurotransmitter transporters. The deduced amino acid (aa) sequence of 556 aa having an estimated molecular mass of 58.9 kDa is predicted to have 12 putative transmembrane domains (TMD) and a characteristic large extracellular loop between TMD3 and TMD4. Sequence comparison to other members of the family indicates that it falls into the glycine-proline transporter subfamily. Transiently expressed MasPROT cDNA in Xenopus oocytes exclusively transported proline. Northern analysis shows that it is expressed predominantly in central nervous system, however, low levels are present in midgut, hindgut and Malpighian tubules. Two mRNA transcripts of sizes 3.6 and 8 Kb were found in all tissues except hindgut, where only a smaller transcript exists. RT-PCR and Southern blot analysis revealed the presence of MasPROT transcripts in flight muscles but not in leg muscles. Our preliminary data suggests that this transporter is an insect homologue of mammalian proline transporters. MasPROT.16 is a short splice variant encoding for 174 amino acids and shares 138 amino acids from the N terminus of MasPROT. MasPROT.2 is a long splice variant that contains six introns that coincide precisely with the previously mapped exon/intron boundaries of the members of this superfamily.