In the eastern United States, 74 species of Anillini in two genera have been described, with most belonging to Anillinus Casey. Until now, no systematic framework has existed for this large genus, hampering integrative studies. Using DNA sequences from 101 Nearctic species, we present a well-resolved molecular phylogeny supporting a sound systematic framework. Sixteen species groups of Appalachian Anillinus are diagnosed, in part using newly recognized variation in the number of modified male protarsi and the state of the spermathecal duct. We present the first descriptions of Nearctic anilline larvae, which possess none of the synapomorphies of previously described anilline larvae. Within Anillinus, two major clades are mostly consistent with setation of the right paramere: a \"hairy clade\" with more than four setae, and a \"quadrisetose clade.\" Throughout the phylogeny, microhabitat use varies within each clade, and several endogean lineages are phylogenetically isolated. Our work increases the South Carolina fauna by nearly five-fold. Nine new species are described, Serranillusmonadnock sp. nov., Anillinuscastaneus sp. nov., Anillinuschoestoea sp. nov., Anillinusdentatus sp. nov., Anillinusjancae sp. nov., Anillinusmica sp. nov., Anillinusmicamicus sp. nov., Anillinusseneca sp. nov., and Anillinussimplex sp. nov. Several species are newly reported from South Carolina, bringing the total to 20 described species representing seven species groups. Two endemic groups inhabit deep clay soils in the Piedmont and possess unique male sexual characters. The Anillini are a unique component of Nearctic biodiversity, with great potential as a model system for studies of biogeography, secondary male sexual modification, and endogean adaptations.

UNASSIGNED: Troglocoelotes Zhao & S. Li, 2019 is the only known genus of Coelotinae of which all species have deep morphological adaptations to the subterranean environment, such as depigmentation of body, degenerated or absent eyes and, frequently, with attenuated bodies and/or appendages. Four species of Troglocoelotes have been reported from Guizhou Province, China.
UNASSIGNED: A new funnel-web spider of the genus Troglocoelotes is described and illustrated on the basis of a single female specimen from Tongren City, Guizhou: Troglocoelotessinanensis sp. nov. Additionally, photos of the collection site and a distribution map are also provided.

Introduction: CRISPR/Cas9 is a gene-editing technology which could specifically cleave dsDNA and induce target gene mutation. CRISPR/Cas9 has been widely used in gene functional studies in many fields, such as medicine, biology, and agriculture due to its simple design, low cost, and high efficiency. Although it has been well developed in model fish and freshwater fish for gene function analysis, it is still novel in the studies dealing with economic crustacean species. Methods: In this study, we established a CRISPR/Cas9 system based on microinjection for M. nipponense, an important economic crustacean aquaculture species. The vitellogenin (Vg) gene and the eyeless (Ey) gene were selected as the targeted genes for mutation. Two sgRNAs were designed for Mn-Vg and Mn-Ey gene editing, respectively. Results and Discussion: For sg-Vg-1, the gastrula survival ratio was 8.69%, and the final hatching ratio was 4.83%. The blastula mutant ratio was 10%, and the hatching individual mutant ratio was 30%. For sg-Vg-2, the gastrula survival ratio was 5.85%, and the final hatching ratio was 3.89%. The blastula mutant ratio was 16.67%, and no mutant sequences were detected in hatching individuals. For sg-Ey-1, the gastrula survival ratio was 6.25%, and the final hatching ratio was 2.34%. The blastula mutant ratio was 10.00%, and the hatching individual mutant ratio was 66.67%. For sg-Ey-2, the gastrula survival ratio was 6.00%, and the final hatching ratio was 2.67%. No mutant sequence was detected in both blastula stage and hatching individuals. There were no significant morphological changes observed in the Mn-Vg group. Two deformed types were detected in sg-Ey-1-injected embryos. An evident developmental delay of the compound eye was detected in Ey-sg1-H1 in the zoea stage. The compound eyes of the Ey-sg1-H2 embryo could not form well-defined spheres, and the whole compound eye appeared to diffuse at the end of the late zoea stage. The establishment of a gene-editing platform based on CRISPR/Cas9 will not only provide an efficient and convenient method for gene function analysis but also provide a powerful tool for molecular-assisted breeding of Macrobrachium nipponense.

In Drosophila, expression of eyeless (ey) gene is restricted to the developing eyes and central nervous system. However, the flanking genes, myoglianin (myo), and bent (bt) have different temporal and spatial expression patterns as compared to the ey. How distinct regulation of ey is maintained is mostly unknown. Earlier, we have identified a boundary element intervening myo and ey genes (ME boundary) that prevents the crosstalk between the cis-regulatory elements of myo and ey genes. In the present study, we further searched for the cis-elements that define the domain of ey and maintain its expression pattern. We identify another boundary element between ey and bt, the EB boundary. The EB boundary separates the regulatory landscapes of ey and bt genes. The two boundaries, ME and EB, show a long-range interaction as well as interact with the nuclear architecture. This suggests functional autonomy of the ey locus and its insulation from differentially regulated flanking regions. We also identify a new Polycomb Response Element, the ey-PRE, within the ey domain. The expression state of the ey gene, once established during early development is likely to be maintained with the help of ey-PRE. Our study proposes a general regulatory mechanism by which a gene can be maintained in a functionally independent chromatin domain in gene-rich euchromatin.

The transcription factor Pax6 is considered the master control gene for eye formation because (1) it is present within the genomes and retina/lens of all animals with a visual system; (2) severe retinal defects accompany its loss; (3) Pax6 genes have the ability to substitute for one another across the animal kingdom; and (4) Pax6 genes are capable of inducing ectopic eye/lens in flies and mammals. Many roles of Pax6 were first elucidated in Drosophila through studies of the gene eyeless (ey), which controls both growth of the entire eye-antennal imaginal disc and fate specification of the eye. We show that Ey also plays a surprising role within cells of the peripodial epithelium to control pattern formation. It regulates the expression of decapentaplegic (dpp), which is required for initiation of the morphogenetic furrow in the eye itself. Loss of Ey within the peripodial epithelium leads to the loss of dpp expression within the eye, failure of the furrow to initiate, and abrogation of retinal development. These findings reveal an unexpected mechanism for how Pax6 controls eye development in Drosophila.

A new linyphiid genus Callosagen. n., with two new species Callosa ciliatasp. n. (♂♀, type species) and Callosa baiseensissp. n. (♂♀), from southwest China are described. Detailed description of genitalic characters and somatic features is provided, as well as light microscopy and SEM micrographs of each species. Callosagen. n. was found in caves in Yunnan and Guangxi, and its copulatory organs are similar to those of Bathyphantes and Porrhomma, but differ greatly in details. The monophyly and placement of Callosagen. n. are supported by the results of molecular analysis.

Cell proliferation is coupled with nutrient availability. If nutrients become limited, proliferation ceases, because growth factor and/or PI3-kinase activity levels become attenuated. Here, we report an exception to this generality within a subpopulation of Drosophila neural stem cells (neuroblasts). We find that most neuroblasts enter and exit cell cycle in a nutrient-dependent manner that is reversible and regulated by PI3-kinase. However, a small subset, the mushroom body neuroblasts, which generate neurons important for memory and learning, divide independent of dietary nutrient conditions and PI3-kinase activity. This nutrient-independent proliferation is regulated by Eyeless, a Pax-6 orthologue, expressed in mushroom body neuroblasts. When Eyeless is knocked down, mushroom body neuroblasts exit cell cycle when nutrients are withdrawn. Conversely, when Eyeless is ectopically expressed, some non-mushroom body neuroblasts divide independent of dietary nutrient conditions. Therefore, Eyeless uncouples MB neuroblast proliferation from nutrient availability, allowing preferential neurogenesis in brain subregions during nutrient poor conditions.

Paired box 6 (Pax6) is considered to be the master control gene for eye development in all seeing animals studied so far. In vertebrates, it is required not only for lens/retina formation but also for the development of the CNS, olfactory system, and pancreas. Although Pax6 plays important roles in cell differentiation, proliferation, and patterning during the development of these systems, the underlying mechanism remains poorly understood. In the fruit fly, Drosophila melanogaster, Pax6 also functions in a range of tissues, including the eye and brain. In this report, we describe the function of Pax6 in Drosophila eye-antennal disc development. Previous studies have suggested that the two fly Pax6 genes, eyeless (ey) and twin of eyeless (toy), initiate eye specification, whereas eyegone (eyg) and the Notch (N) pathway independently regulate cell proliferation. Here, we show that Pax6 controls eye progenitor cell survival and proliferation through the activation of teashirt (tsh) and eyg, thereby indicating that Pax6 initiates both eye specification and proliferation. Although simultaneous loss of ey and toy during early eye-antennal disc development disrupts the development of all head structures derived from the eye-antennal disc, overexpression of N or tsh in the absence of Pax6 rescues only antennal and head epidermis development. Furthermore, overexpression of tsh induces a homeotic transformation of the fly head into thoracic structures. Taking these data together, we demonstrate that Pax6 promotes development of the entire eye-antennal disc and that the retinal determination network works to repress alternative tissue fates, which ensures proper development of adult head structures.

A major problem in developmental genetics is how HOX transcription factors, like Proboscipedia (PB) and Ultrabithorax (UBX), regulate distinct programs of gene expression to result in a proboscis versus a haltere, respectively, when the DNA-binding homeodomain (HD) of HOX transcription factors recognizes similar DNA-binding sequences. Indeed, the lack of DNA-binding specificity is a problem for all transcription factors (TFs), as the DNA-binding domains generally recognize small targets of five to six bases in length. Although not the initial intent of the study, I found extensive non-specificity of TF function. Multiple TFs including HOX and HD-containing and non-HD-containing TFs induced both wingless and eyeless phenotypes. The TFs Labial (LAB), Deformed (DFD), Fushi tarazu (FTZ), and Squeeze (SQZ) induced ectopic larval thoracic (T) 1 beard formation in T2 and T3. The TF Doublesex male (DSXM) rescued the reduced maxillary palp pb phenotype. These examples of non-specificity of TF function across classes of TFs, combined with previous observations, compromise the implicit, initial assumption often made that an intrinsic mechanism of TF specificity is important for function. Interestingly, the functional complementation of the pb phenotype may suggest a larger role for regulation of expression of TFs in restriction of function as opposed to an intrinsic specificity of TF function. These observations have major ramifications for analysis of functional conservation in evolution and development.

Two new blind species of Sinella are described from Nanjing, China. Sinella quinseta sp. n. from Purple Mountain possesses unique 5+5 central macrochaetae on Abd. II, and can be distinguished from other species of the genus by the postlabial chaetae and the dorsal chaetotaxy. Sinella qixiaensis sp. n. from Qixia Mountain is characterized by the paddle-like S-chaetae of Ant. III organ and the smooth straight chaetae on the manubrium and base of dens; it differs from two closely related species by the smooth manubrial chaetae, the labial chaetae, the Ant. III organ, and the macrochaetae on Abd. II. Sinella fuyanensis Chen & Christiansen and Sinella quinocula Chen & Christiansen were also newly recorded from Nanjing.