HINTW,a W‐chromosome HINT gene in chick,is expressed ubiquitously and is a robust female cell marker applicable in intraspecific chimera studies

Grafting and transplantation experiments in embryology require proper distinction between host and donor tissues. For the avian model this has traditionally been achieved by using two closely related species (e.g., chick and quail) followed by species‐specific antibody staining. Here, we show that an in situ hybridization probe against the HINTW gene is a robust and reliable marker for female‐derived chicken cells. At all pre‐circulation stages tested, all cells in female embryos, independently confirmed by PCR analysis, were strongly positive for HINTW, whereas all male embryos were negative. This probe is broadly applicable in intra‐specific chick/chick chimera studies, and as a proof of principle, we utilized this probe to detect female cells in three experimental settings: (1) to mark female donor cells in a node transplantation assay; (2) to distinguish female cells in male/female twins generated by the Cornish pasty culture; and (3) to detect female half of the embryo in artificially generated bilateral gynandromorphs. A rapid, PCR based pre‐screening step increases the efficiency of obtaining desired donor/host sex combination from 25% to 100%. For most avian chimera studies, this female‐specific in situ probe is a low cost alternative to the commonly used QCPN antibody and to ubiquitous‐GFP chicken strains which are not widely available to the research community. genesis 52:424–430, 2014. © 2014 Wiley Periodicals, Inc.     

Spatial and temporal influences of conifer planted forests on the orchard pest Plautia stali (Hemiptera: Pentatomidae)

Land use changes have generally been considered a local matter, but they are becoming an issue of much wider concern as changes in the use of non-agricultural lands occasionally generate nationwide agricultural pest problems. Plautia stali Scott is a serious pest in orchards, and the widespread distribution of this pest is likely associated with the expansion of planted coniferous forests in Japan. An experiment was conducted on 29 study sites in diverse areas of planted coniferous forest in central Japan, and the abundance of P. stali was monitored over four seasons for 3 years. Results revealed that the optimum radius required to cause an effect on the area surrounding the conifer planted forest was up to 4 km—specifically, 2,000, 400, 1,500, and 200 m in April, June, August, and October, respectively. Positive correlations were observed between conifer planted forests and the collected numbers of P. stali in April, June, and August. These temporal scales provide a direct evaluation of the landscape effects of a conifer planted forest on population dynamics, and this information will contribute to the development and improvement of area-wide P. stali management.     

Flowering induced by 5-azacytidine, a DNA demethylating reagent in a short-day plant, Perilla frutescens var. crispa

Treatment with 5-azacytidine, a DNA demethylating reagent, induced flowering in Perilla frutescens (L.) Britton var. crispa (Thunb. ex Murray) Decne. ex L. H. Bailey, an absolute short-day plant under long days. The 5-azacytidine treatment induced slight suppression of vegetative growth but had no obvious effect on any other phenotypes. The Southern hybridization analysis of the genomic DNA isolated from the leaves of 5-azacytidine-treated plants and digested with restriction enzyme, methylation-insensitive Msp I or methylation-sensitive Hpa II with P. frutescens 25S-18S rDNA intergenic spacer probe indicated that the 5-azacytidine treatment caused demethylation of the genomic DNA. The 5-azacytidine-induced flowering was delayed as compared with the short day-induced flowering. Flowers were formed even at the lower nodes which had not been directly treated with 5-azacytidine. The results suggest that DNA demethylation induced flowering by inducing the production of a transmissible flowering stimulus in P. frutescens .     

An alien Sennertia mite (Acari: Chaetodactylidae) associated with an introduced Oriental bamboo‐nesting large carpenter bee (Hymenoptera: Apidae: Xylocopa) invading the central Honshu Island,Japan

Since 2006, an introduced Oriental bamboo‐nesting large carpenter bee, Xylocopa tranquebarorum, has been recorded from the central Honshu Island, Japan, which is inhabited only by the endemic subspecies, Xylocopa appendiculata circumvolans. Carpenter bees (tribes Xylocopini and Ceratinini) have ecological associations with specific Sennertia spp. in all geographic regions of their distribution, thus it is worried that the introduced carpenter bee has brought non‐indigenous mites into Japan. In their native ranges, X. a. circumvolans and X. tranquebarorum each has specific Sennertia mite faunas: the four Japanese Alloxylocopa bees including X. a. circumvolans have associations with S. alfkeni, while X. tranquebarorum has association with S. potanini in China (except Taiwan) and with S. horrida in South to East Asia including Taiwan. In the present study, we examined phoretic mite fauna on the introduced X. tranquebarorum, and determined whether the mites are indigenous or not based on morphological character and two gene sequences (mitochondrial cytochrome oxidase subunit I gene and nuclear ribosomal internal transcribed spacer). It was found from the result of this study that the non‐indigenous Sennertia mite has invaded Japan with the introduced X. tranquebarorum. We discuss geographic origin of the introduced X. tranquebarorum based on associated mite fauna and potential ecological risk caused by the introduced Xylocopa‐Sennertia association.     

Application of the LAMP assay for the detection of the Argentine ant, <Emphasis Type="Italic">Linepithema humile</Emphasis> (Hymenoptera: Formicidae), from captures of pan traps

We applied the loop-mediated isothermal amplification (LAMP) assay to monitor invasions of Linepithema humile (Mayr), the Argentine ant, a notorious invasive insect worldwide. Species-specific LAMP primers were designed on the basis of the partial sequence of the cytochrome c oxidase subunit I region of L. humile. The species specificity and sensitivity of these primers were determined in the laboratory and considered adequate for practical use. We also confirmed that the assay successfully detected L. humile from captures of pan traps, which contained L. humile and several non-target ant species. The assay detected the target species even when the captures contained only a leg or an antenna. Since the LAMP assay is simple and rapid, this assay will contribute to the early detection and accurate identification of L. humile.     

Species status of Incisitermes spp. (Isoptera: Kalotermitidae) in Japan

The species status of Japanese populations of Incisitermes immigrans from Iwo and Minami Daito Islands was examined using mitochondrial 16S molecular barcode sequences. The molecular sequences of these two populations were compared to those of other Incisitermes spp. deposited in the GenBank database using a maximum likelihood phylogenetic analysis. This analysis suggested that the Minami Daito population is indeed I. schwarzi, as suspected previously, while the sequence of the Iwo Island population was identical to that of authentic I. immigrans. In addition to I. minor, which is recorded from middle and southern Japan, the presence of three Incisitermes species in Japan was confirmed.     

Function of the cypX and moxY Genes in Aflatoxin Biosynthesis in Aspergillus parasiticus

The pathway oxoaverantin (OAVN) → averufin (AVR) → hydroxyversicolorone (HVN) → versiconal hemiacetal acetate (VHA) is involved in aflatoxin biosynthesis, and the cypX and moxY genes, which are present in the aflatoxin gene cluster, have been previously suggested to be involved in this pathway. To clarify the function of these two genes in more detail, we disrupted the genes in aflatoxigenic Aspergillus parasiticus NRRL 2999. The cypX-deleted mutant lost aflatoxin productivity and accumulated AVR in the mycelia. Although this mutant converted HVN, versicolorone (VONE), VHA, and versiconol acetate (VOAc) to aflatoxins in feeding experiments, it could not produce aflatoxins from either OAVN or AVR. The moxY-deleted mutant also lost aflatoxin productivity, whereas it newly accumulated HVN and VONE. In feeding experiments, this mutant converted either VHA or VOAc to aflatoxins but did not convert OAVN, AVR, HVN, or VONE to aflatoxins. These results demonstrated that cypX encodes AVR monooxygenase, catalyzing the reaction from AVR to HVN, and moxY encodes HVN monooxygenase, catalyzing a Baeyer-Villiger reaction from HVN to VHA as well as from VONE to VOAc. In this work, we devised a simple and rapid method to extract DNA from many fungi for PCR analyses in which cell disruption with a shaker and phenol extraction were combined.