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Furins are a family of proteases involved in the processing of many proproteins including growth regulators, immune proteins, and toxins. However, very few studies have been conducted on furin-like PCs in insects. Herein, we isolated and characterized full-length furin-like cDNAs (Psfurin-1 and Psfurin-2) from larvae of Protaetia brevitarsis seulensis (Kolbe). To search for furin-like PC cDNAs, we used high-throughput RNA sequencing (RNA-seq) expression profiling of data. The Psfurin-1 and Psfurin-2 transcripts, 2,370 and 3,702 nucleotides in length, respectively, encode peptides of 790 and 1,234 amino acids with calculated molecular masses of 88.3 and 135.6 kDa. We identified a consensus site for proprotein convertase that includes Asp, His, Asn, and Ser catalytic residues, and revealed the presence of a subtilisin-like serine protease/putative cell envelope proteinase domain related to the immune system. The predicted structures of the two proteins are very similar, but the cell envelope protease/immune-related domain differs somewhat. To examine the possible immune-related expression of Psfurin-1 and 2 transcripts, late larvae were injected with the Gram-negative bacterium Escherichia coli or the yeast Saccharomyces cerevisiae. Expression of Psfurin-1 mRNA in E. coli K12-challenged larvae was ~15-fold higher at 8 h and ~20-fold higher at 12 h than in LB medium-challenged control larvae. In the case of yeast-challenged larvae, expression of Psfurin-1 mRNA was not as elevated as in E. coli K12-challenged larvae, but expression was 4–5-fold higher than in LB medium-challenged larvae. Expression of Psfurin-2 mRNA in E. coli K12-challenged and yeast-challenged larvae was also significantly higher than in LB medium-challenged controls from 4 to 48 h, but expression levels were lower than for Psfurin-1 mRNA in both E. coli K12- and yeast-challenged larvae.

Dwarfism in crops is a useful breeding trait. In this study, we aimed to identify a candidate gene controlling semi-dwarfism in watermelon (Citrullus lanatus) using a combination of genetic mapping and quantitative trait loci (QTL)-seq. We evaluated phenotypes using an F2 and F2:3 population derived from a cross between a “Bush Sugar Baby” (BSB, semi-dwarf type) and a PCL-J1 (normal type) cultivar. Results indicated that a single recessive gene confers semi-dwarfism in BSB. We constructed a genetic map using 180 F2 plants and 336 single-nucleotide polymorphisms (SNPs), detected using genotyping-by-sequencing, and mapped the semi-dwarfism locus, sdw-1, between the SNP markers linked by 9.6 cM (0.99 Mb) on Chr. 9. QTL analysis pointed to the same genomic location for sdw-1 using single-marker analysis. Further, based on the QTL-seq, we identified a significant genomic region for sdw-1 that matched with the sdw-1-flanking region in the genetic linkage map. This 1-Mb region was narrowed down by mapping three SNP markers developed from the QTL-seq data. The sdw-1 locus was mapped to the 0.44-Mb genomic region, which harbored 13 genes. One of the 13 genes, ClCG09G018320 (ATP-binding cassette (ABC) transporter B family member 19), showed the SNPs in its coding sequence, and cleaved amplified APS markers developed from those SNPs co-segregated with sdw-1 in fine mapping using 620 F2 plants. Our results implied that a frameshift mutation in the ABC transporter gene and the resulting alteration in auxin transportation are the most likely to be responsible for semi-dwarfism in BSB.

In this study, more than 1,000 cricket (Gryllus bimaculatus) hemocytes were classified based on their size and morphology. These hemocytes were classified into six types: granulocytes, plasmatocytes, prohemocytes, spherulocytes, coagulocytes, and oenocytoids. Hemocyte cultures was observed in real time to determine which hemocytes were associated with cellular immune responses against potential pathogens. Granulocytes were identified as the professional immune cell that mediates nodulation, encapsulation, and phagocytosis of pathogens. Granulocytes have been shown to actively produce various sticky nets (amoeba-like hairs and extracellular traps) from their plasma membranes that they use to gather other hemocytes and to implement cellular immune responses. The activation of lysosomes in granulocytes started at 4 h, peaked at 12 h, and returned to baseline by 24 h post-infection. At 48 h post-infection, cells could be found within the cytoplasm of granulocytes and reactivated lysosomes surrounding these cells were visible. This result seems to reflect a phenomenon in which necrotic granulocytes are removed by other healthy granulocytes. This unique mechanism of cellular immunity is therefore a way to efficiently and effectively remove pathogens and simultaneously maintain healthy hemocytes.

Fourteen watermelon cultivars with different fruit flesh colors (red, salmon yellow, orange, and canary yellow) were analyzed for carotenoid contents (prolycopene, lycopene, β-carotene, ζ-carotene, and neoxanthin). Genes encoding the carotenoid biosynthesis enzymes carotenoid isomerase (encoded by CRTISO), which catalyzes the isomerization of prolycopene to lycopene, and β-carotene hydroxylase (CHYB), which catalyzes the conversion of β-carotene to xanthophyll, were also analyzed. High-performance liquid chromatography showed that the salmon yellow and orange flesh accumulated either prolycopene (orange-P flesh) or β-carotene (orange-β flesh), whereas lycopene and neoxanthin were the main carotenoids in the red and canary yellow flesh, respectively. Quantitative real-time polymerase chain reaction indicated that CRTISO and CHYB were mainly expressed during fruit maturation, regardless of the flesh color, and there was no significant association between differential gene expression and flesh color. Importantly, transcript sequencing revealed a non-synonymous single-nucleotide mismatch (T > C1976) in exon 13 of CRTISO between orange-P-fleshed and other cultivars, suggesting CRTISO as a candidate gene for high prolycopene accumulation. However, in β-carotene-accumulating cultivars, there were no mutations in CHYB transcripts. A cleaved amplified polymorphic sequence marker was developed for T > C1976, and its applicability for marker-assisted selection of orange-P flesh was validated in 105 watermelon accessions.

The pulmonary and intestinal systems have several characteristics in common. It is believed that these similarities somehow function to cause pulmonary–intestinal crosstalk during inflammation. Many studies have shown that pulmonary disease occurs in association with inflammatory bowel disease more often than is commonly recognized. Bambusae Caulis in Taeniam, a medicinal herb originated from the inner bark of Phyllostachys nigra var. henosis (Milford) Rendle (Poaceae), has been used to cure fever, diarrhea, and chest inflammation in Korea as well as in China. Cigarette smoke is a well-known risk factor for several inflammatory disorders. In this study, we induced pulmonary and bowel inflammation in mice using cigarette smoke and investigated whether Bambusae Caulis in Taeniam extract modulates the inflammatory response in both the lung and the bowel. C57BL/6 mice were exposed to cigarette smoke for 90 min per day for three weeks, and Bambusae Caulis in Taeniam extract was administered via oral injection 2 h before cigarette smoke exposure. The bronchoalveolar lavage cells were counted and hematoxylin and eosin staining were performed. Levels of inflammatory mediators in lung and large intestine were determined by enzyme-linked immunosorbent assay, real-time polymerase chain reaction, and Western blotting. Our results showed that Bambusae Caulis in Taeniam attenuated cigarette smoke-induced inflammatory response in both the lung and the bowel of mice by inhibiting the production of pro-inflammatory cytokines, chemokines, and protease as well as NF-κB signaling factor. Therefore, we suggest that Bambusae Caulis in Taeniam extract might be a candidate therapeutic agent for inhibiting pulmonary and intestinal inflammation.