Identification and Analysis of MicroRNAs Associated with Wing Polyphenism in the Brown Planthopper, Nilaparvata lugens

Many insects are capable of developing two types of wings (i.e., wing polyphenism) to adapt to various environments. Though the roles of microRNAs (miRNAs) in regulating animal growth and development have been well studied, their potential roles in modulating wing polyphenism remain largely elusive. To identify wing polyphenism-related miRNAs, we isolated small RNAs from 1st to 5th instar nymphs of long-wing (LW) and short-wing (SW) strains of the brown planthopper (BPH), Nilaparvata lugens. Small RNA libraries were then constructed and sequenced, yielding 158 conserved and 96 novel miRNAs. Among these, 122 miRNAs were differentially expressed between the two BPH strains. Specifically, 47, 2, 27 and 41 miRNAs were more highly expressed in the 1st, 3rd, 4th and 5th instars, respectively, of the LW strain compared with the SW strain. In contrast, 47, 3, 29 and 25 miRNAs were more highly expressed in the 1st, 3rd, 4th and 5th instars, respectively, of the SW strain compared with the LW strain. Next, we predicted the targets of these miRNAs and carried out Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis. We found that a number of pathways might be involved in wing form determination, such as the insulin, MAPK, mTOR, FoxO and thyroid hormone signaling pathways and the thyroid hormone synthesis pathway. Thirty and 45 differentially expressed miRNAs targeted genes in the insulin signaling and insect hormone biosynthesis pathways, respectively, which are related to wing dimorphism. Among these miRNAs, Nlu-miR-14-3p, Nlu-miR-9a-5p and Nlu-miR-315-5p, were confirmed to interact with insulin receptors (NlInRs) in dual luciferase reporter assays. These discoveries are helpful for understanding the miRNA-mediated regulatory mechanism of wing polyphenism in BPHs and shed new light on how insects respond to environmental cues through developmental plasticity.

Ultrabithorax is a key regulator for the dimorphism of wings, a main cause for the outbreak of planthoppers in rice

Rice planthoppers, the most devastating rice pests, occur in two wing forms: the short-wing form for rapid population growth and long-wing form for long-distance migration, which together create the mechanism for outbreak. Here we show that Ultrabithorax (Ubx) is a key regulator for switching between the long- and short-wing forms of rice planthoppers. Ubx is expressed in both forewing and hindwing pads, which is different from the canonical model of Ubx expression. In brown planthoppers, expression of Ubx (NlUbx) is regulated by nutritional status of the rice host. High-quality young plants induce NlUbx expression leading to the short-wing form; low-quality ripe plants reduce NlUbx expression resulting in long-wing form. We also showed that NlUbx is regulated by the insulin receptors NlInR1 and NlInR2. The default expression of NlInR1 inhibits NlUbx resulting in long-wings, while high-quality hosts induce NlInR2 expression, which represses NlInR1 thus promoting NlUbx expression to produce short-wings.

Evolution at two time frames: polymorphisms from an ancient singular divergence event fuel contemporary parallel evolution

When species occur in repeated ecologically distinct habitats across their range, adaptation may proceed surprisingly fast and result in parallel evolution. There is increasing evidence that such cases of rapid parallel evolution are fueled by standing genetic variation, but the origin of this genetic variation remains poorly understood. In Pogonus chalceus beetles, short- and long-winged ecotypes have diverged in response to contrasting hydrological regimes and can be repeatedly found along the Atlantic European coast. By analyzing genomic variation across the beetles’ distribution, we reveal that genomically widespread short-wing selected alleles evolved during a singular divergence event, estimated at ~0.19 Mya. The ancient and differentially selected alleles are currently polymorphic in all populations across the range, allowing for the fast evolution of one ecotype from a small number of random individuals, as low as 5 to 15, of the populations of the other ecotype. Our results suggest that cases of fast parallel ecological divergence might be the result of evolution at two different time frames: divergence in the past, followed by repeated selection on the divergently evolved alleles after admixture. We suggest that this mechanism may be common and potentially further driven by periods of geographic isolation imposed by large-scale environmental changes such as glacial cycles.