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Eyespots on the wings of nymphalid butterflies represent colorful examples of pattern formation, yet the developmental origins and mechanisms underlying eyespot center differentiation are still poorly understood. Using CRISPR-Cas9 we re-examine the function of Distal-less (Dll) as an activator or repressor of eyespots, a topic that remains controversial. We show that the phenotypic outcome of CRISPR mutations depends upon which specific exon is targeted. In Bicyclus anynana, exon 2 mutations are associated with both missing and ectopic eyespots, and also exon skipping. Exon 3 mutations, which do not lead to exon skipping, produce only null phenotypes, including missing eyespots, lighter wing coloration and loss of scales. Reaction-diffusion modeling of Dll function, using Wnt and Dpp as candidate morphogens, accurately replicates these complex crispant phenotypes. These results provide new insight into the function of Dll as a potential activator of eyespot development, scale growth and melanization, and suggest that the tuning of Dll expression levels can generate a diversity of eyespot phenotypes, including their appearance on the wing.

Supplementary Video: CRISPR-Cas9 Mediated Genome Editing in Bicyclus anynana Butterflies

The role of secondary circulations has recently been studied in the context of well-defined surface heterogeneity in a semiarid ecosystem where it was found that energy balance closure over a desert–forest system and the structure of the boundary layer was impacted by advection and flux divergence. As a part of the CliFF (“Climate feedbacks and benefits of semi-arid forests”, a collaboration between KIT, Germany, and the Weizmann Institute, Israel) campaign, we studied the boundary layer dynamics and turbulent transport of energy corresponding to this effect in Yatir Forest situated in the Negev Desert in Israel. The forest surrounded by small shrubs presents a distinct feature of surface heterogeneity, allowing us to study the differences between their interactions with the atmosphere above by conducting measurements with two eddy covariance (EC) stations and two Doppler lidars. As expected, the turbulence intensity and vertical fluxes of momentum and sensible heat are found to be higher above the forest compared to the shrubland. Turbulent statistics indicative of nonlocal motions are also found to differ over the forest and shrubland and also display a strong diurnal cycle. The production of turbulent kinetic energy (TKE) over the forest is strongly mechanical, while buoyancy effects generate most of the TKE over the shrubland. Overall TKE production is much higher above the forest compared to the shrubland. The forest is also found to be more efficient in dissipating TKE. The TKE budget appears to be balanced on average both for the forest and shrubland, although the imbalance of the TKE budget, which includes the role of TKE transport, is found to be quite different in terms of diurnal cycles for the forest and shrubland. The difference in turbulent quantities and the relationships between the components of TKE budget are used to infer the characteristics of the turbulent transport of energy between the desert and the forest.