SummaryA hot pepper (Capsicum annuum) cDNA clone encoding pathogenesis-related protein 10 (CaPR-10) was isolated by differential screening of a cDNA library prepared from pepper leaves inoculated with tobacco mosaic virus pathotype (TMV-P 0 ). CaPR-10 transcripts were induced in the incompatible interaction with TMV-P 0 or Xanthomonas campestris pv. vesicatoria (Xcv) but not induced in the compatible interaction. Characterization of enzymatic properties of CaPR-10 indicated that the recombinant protein exhibits a ribonucleolytic activity against TMV RNA, as well as against pepper total RNA, and shows its putative antiviral activity in several conditions. The CaPR-10 protein existed at very low level in leaf tissue but was dramatically induced as soon as plants were inoculated with TMV-P 0 , and this was correlated with the increase of its ribonucleolytic activity. Immunoblot analysis and pull-down assays using proteins extracted from pepper leaves showed that TMV-P 0 inoculation led to the phosphorylation of CaPR-10, a modi®cation that should affect its capacity for RNase function. We present data that the induction and subsequent phosphorylation of CaPR-10 increased its ribonucleolytic activity to cleave invading viral RNAs, and this activity should be important to its antiviral pathway during viral attack in vivo.

Plant WRKY transcription factors were previously implicated in the alteration of gene expression in response to various pathogens. The WRKY proteins constitute a large family of plant transcription factors, whose precise functions have yet to be elucidated. Using a domain-specific differential display procedure, we isolated a WRKY gene, which is rapidly induced during an incompatible interaction between hot pepper and Tobacco mosaic virus (TMV) or Xanthomonas campestris pv . vesicatoria (Xcv). The full-length cDNA of CaWRKY-a (Capsicum annuum WRKY-a) encodes a putative polypeptide of 546 amino acids, containing two WRKY domains with a zinc finger motif. The expression of CaWRKY-a could be rapidly induced by not only chemical elicitor such as salicylic acid (SA) or ethephon but also wounding treatments. The nuclear localization of CaWRKY-a was determined in transient expression system using tobacco BY-2 cells by polyethylene glycol (PEG)-mediated transformation experiment. With oligonucleotide molecules containing the putative W-box sequences as a probe, we confirmed that CaWRKY-a protein had W-box-binding activity. These results suggest that CaWRKY-a might be involved as a transcription factor in plant defense-related signal transduction pathways.

To understand the molecular defense mechanism controlling the hypersensitive response (HR) better, we examined the hot pepper plant (Capsicum annuum L. cv. Bugang), which exhibits an HR in response to infection by Tobacco mosaic virus pathotype P0 (TMV-P0). A full-length cDNA clone was isolated by differential screening of a cDNA library that was constructed with mRNA extracted from hot pepper leaves during the resistance response to TMV-P0. The predicted amino acid sequence of the cDNA clone exhibited a high sequence similarity to germin-like protein (GLP). The CaGLP1 (Capsicum annuum GLP1) cDNA contains a single open reading frame of 660 bp encoding 220 amino acid residues. Upon inoculation with TMV or Xanthomonas, CaGLP1 transcripts were specifically accumulated in the incompatible interaction but not in the compatible interaction. In plants treated with salicylic acid (SA) or ethephon, which are signal molecules in the defense-related signal transduction pathway, CaGLP1 transcripts were accumulated rapidly. As far as we know, this is the first report that plant GLPs can be specifically induced during a defense response against viral infection. These data suggest that in the hot pepper plant CaGLP1 may be involved in the defense response to viral pathogens, and thus be classified as a new family of PR proteins, 'PR-16'.

Although several antiviral agents have become available for coronavirus disease 2019 (COVID-19) treatment, oral drugs are still limited. Camostat mesylate, an orally bioavailable serine protease inhibitor, has been used to treat chronic pancreatitis in South Korea, and it has an in vitro inhibitory potential against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

: This study was indicated to enhance the anti-inflammation activities by the fermentation of the fruits of Aronia melanocarpa (Michx.) Elliott. The extracts by 70% ethanol (EE) showed better biological activities than those by hot water (WE) from campared result of the effect of extraction solvents. Then, the extract from 70% ethanol extraction was further fermented by lactic acid, denoted as FEE. For antioxidant activities, the FEE had showed the highest value as 0.832 of reducing powder, in comparison with those of EE and WE. Cytotoxicity of the water extraction (WE) was measured for 12.06% in addition of 1.0 ㎎/㎖ of FEE. For anti-inflammation activities, NO production from the macrophage, RAW 264.7 was observed as 7.24 μM and 8.52 μM from FEE and EE, respectively. Prostaglandin E 2 (PGE 2 ) production from human fibroblast cell, CCD-986sk, was also estimated for 152 pg/㎖ in addition of 1.0 ㎎/㎖ of the FEE. The lowest production of both IL-6 and TNF-α were 3.5 pg/㎖ and 865.5 pg/㎖, respectively in addition of 1.0 ㎎/㎖ of the FEE, whereas 74.5 pg/㎖ and 982.4 pg/㎖ in treated with same concenrations of the EE. It was also found that the FEE was higher amounts than other extracts through HPLC analysis of the anthocyanins. These results strongly indicate that fermentation process of the lactic acid could enhance anti-inflammation activities of extracts by increasing the amounts of the anthocyanins, especially cyanidin-galactoside. Our results suggest that the application of the fermentation process for other medicinal herbs can be improved their biological activities.