First report of alstroemeria mosaic virus infecting Alstroemeria in Korea.

Alstroemeria, a member of the Alstroemriaceae family, is a popular cut flower plant with a long-base life and a wide variety of flower colors. It is widely cultivated in many countries, especially in Central and South America. However, numerous viruses such as alstroemeria carlavirus (AlCV), alstroemeria mosaic virus (AlMV), cucumber mosaic virus (CMV), tomato spotted wilt virus (TSWV), alstroemeria streak virus (AlSV), and impatiens necrotic virus (INSV) can infect Alstroemeria and significantly decrease its yield (Kim, 2020). Among these viruses, AlMV is well known to cause an endemic viral disease in the Netherlands (Corine M. et al. 1992). AlMV is a member of the genus potyvirus in the family Potyviridae, one of the most widely distributed families of plant viruses. In 2021, symptomatic alstroemeria plants showing interveinal leaf streaking with elongated light green and chlorosis of leaves were identified from farms in a greenhouse in Gwangju, South Korea. Potyvirus-like particles (approximately 750-800 nm in length) were observed from sap of the symptomatic plants by electron microscope (Supplementary Fig. 1). To confirm virus infection, total RNA was extracted from an alstroemeria leaf using a Beniprep® Super Plant RNA extraction kit (IVT7005, Invirustech Co., Korea). A cDNA library was synthesized and analyzed by high throughput sequencing (HTS) using an Illumina NovaSeq6000 S4 sequencer. A total of 48,072,240 raw reads were obtained after quality filtering with FastQC. Remaining sequences were de novo assembled into contigs with a Trinity assembler. Nucleotide blast analysis of contigs against NCBI viral reference database revealed that 24 assembled contigs (> 1,000 bp) were sequences of AlMV. To confirm AlMV detection, raw reads were mapped to known AlMV complete genome (9,774 bp) using Bowtie2 program. Results showed that a total of 4,698,112 reads were mapped. A consensus sequence (9,778 bp, accession no. LC709275) was then obtained. To verify the presence of AlMV, RT-PCR assay was conducted with AlMV's CP gene-specific primers: AlMV-F (5'-CACGAGGCTGTGAAACAAGC -3') and AlMV-R (5'- CCAGGCGACACGGCTAAATA-3'). PCR products of the expected size (538 bp) were cloned, sequenced, and subjected to GenBank BLASTn search. A 538 bp partial CP sequence was used for BLAST analysis which revealed that it shared 100% identities with the consensus sequence (LC709275) and 96.99~98.76% nucleotide identities with four AlMV isolates (MK440140, NC043135, MT892648, DQ295032). Phylogenetic analysis based on partial CP sequences of representative members of potyviruses (family Potyviridae) using 1,000 bootstrap replicates based on either neighbor-joining or Kimura 2 parameter methods in MEGA-X revealed that AlMV isolate JNU-2 was grouped together with the four known AlMV isolates (Supplementary Fig. 2). To determine the incidence of AlMV in a greenhouse, 30 alstroemeria samples were collected and tested by RT-PCR. Results showed that 23 samples were positive for AlMV by PCR-gel electrophoresis and Sanger sequencing, suggesting a high incidence of AlMV infection. To the best of our knowledge, this is the first report of natural infection with AlMV in Alstroemeria in Korea. Further surveys of AlMV infection in greenhouses will help us prevent the spread of this viral disease in Alstroemeria.

Metabolite and Elastase Activity Changes in Beach Rose (Rosa rugosa) Fruit and Seeds at Various Stages of Ripeness.

Rose hips are the fruits of the beach rose (Rosa rugosa). To determine the optimal harvest time and to obtain the maximum functional compounds, rose hips at various stages of ripeness (immature, early, mid, and late) were harvested, and the flesh tissue and seeds were separated. The rose hip flesh showed the highest total phenolic content at the mid-ripeness stage (8.45 ± 0.62 mg/g gallic acid equivalent concentration (dry weight)). The early-, mid-, and late-ripeness stages of rose hip flesh did not show significantly different 2,2-diphenyl-1-picrylhydrazyl antioxidant capacities. The elastase inhibitory activity of the 95% ethanol extract from the rose hip seeds was highest at the mid-ripeness stage; however, the elastase inhibitory activity of the rose hip tissue was not significantly different from that of the seeds. Pathway analysis using MetaboAnalyst showed that sucrose, fructose, and glucose gradually increased as the fruit ripened. Ursolic acid was detected in the seeds but not in the flesh. Of the fatty acids, linoleic acid concentrations were highest in rose hip seeds, followed by linolenic acid, oleic acid, and palmitic acid. Fatty acids and ursolic acid might be the active compounds responsible for elastase inhibitory activity and can be utilized as a functional cosmetic material.

Ultrasound-assisted extraction and antioxidant activity of phenolic and flavonoid compounds and ascorbic acid from rugosa rose (Rosa rugosa Thunb.) fruit.

The present study aimed to extract total phenolic compounds (TPC), total flavonoid compounds (TFC), and ascorbic acid (AA) from the fruit of rugosa rose (Rosa rugosa Thunb.) by ultrasound-assisted extraction (UAE), and to evaluate their antioxidant activities. UAE significantly increased the extract yield compared with that obtained using the conventional method. TPC, TFC, and AA were extracted, depending on the extraction conditions (temperature, time, and ethanol concentration), in the range of 50.73-96.69, 15.93-31.88, and 3.06-6.08 mg/g, respectively. TPC and TFC were effectively extracted at a relatively high temperature (50 °C) than AA was (30 °C). The solvent condition used to extract TPC, TFC, and AA was 50% ethanol. The UAE condition for the highest antioxidant activity was obtained 30 °C, 30 min, and 50% ethanol, which were the same condition for the highest AA extraction. Among the extracts, AA showed a strong correlation with antioxidant activity at p-value of 0.001.

Construction of a high-resolution linkage map of Rfd1, a restorer-of-fertility locus for cytoplasmic male sterility conferred by DCGMS cytoplasm in radish (Raphanus sativus L.) using synteny between radish and Arabidopsis genomes.

Cytoplasmic male sterility caused by Dongbu cytoplasmic and genic male-sterility (DCGMS) cytoplasm and its nuclear restorer-of-fertility locus (Rfd1) with a linked molecular marker (A137) have been reported in radish (Raphanus sativus L.). To construct a linkage map of the Rfd1 locus, linked amplified fragment length polymorphism (AFLP) markers were screened using bulked segregant analysis. A 220-bp linked AFLP fragment sequence from radish showed homology with an Arabidopsis coding sequence. Using this Arabidopsis gene sequence, a simple PCR marker (A220) was developed. The A137 and A220 markers flanked the Rfd1 locus. Two homologous Arabidopsis genes with both marker sequences were positioned on Arabidopsis chromosome-3 with an interval of 2.4 Mb. To integrate the Rfd1 locus into a previously reported expressed sequence tag (EST)-simple sequence repeat (SSR) linkage map, the radish EST sequences located in three syntenic blocks within the 2.4-Mb interval were used to develop single nucleotide polymorphism (SNP) markers for tagging each block. The SNP marker in linkage group-2 co-segregated with male fertility in an F(2) population. Using radish ESTs positioned in linkage group-2, five intron length polymorphism (ILP) markers and one cleaved amplified polymorphic sequence (CAPS) marker were developed and used to construct a linkage map of the Rfd1 locus. Two closely linked markers delimited the Rfd1 locus within a 985-kb interval of Arabidopsis chromosome-3. Synteny between the radish and Arabidopsis genomes in the 985-kb interval were used to develop three ILP and three CAPS markers. Two ILP markers further delimited the Rfd1 locus to a 220-kb interval of Arabidopsis chromosome-3.

Comparison of the content of bioactive substances and the inhibitory effects against rat plasma oxidation of conventional and organic hot peppers (Capsicum annuum L.).

The aim of this study was to evaluate the chemical compositions and antioxidative activities of hot pepper fruits cultivated with strict management by organic and conventional agricultural practices. The ascorbic acid content in the organically grown hot pepper (OGP) was significantly higher than that of conventionally grown hot pepper (CGP) in both green and red fruits. The content of other bioactive compounds such as flavonoids (apigenin, luteolin, quercetin) and total phenolics in OGP was typically higher than in CGP regardless of fruit color. In addition, the ABTS(+) radical-scavenging activity of OGP red fruits was significantly higher than that of CGP red fruits. Moreover, regardless of the color of the fruits, a higher antioxidative activity was observed in blood plasma from rats administered the OGP fruit extracts than in blood plasma from rats administered the CGP fruit extracts. It was hypothesized that the higher antioxidant activity of the OGP fruits may have resulted from the higher antioxidant content in the OGP fruits. These results suggest that consumption of pepper fruits may increase antioxidant activity in the blood, and OGP fruits may be more effective in increasing this antioxidant activity than CGP fruits.

Anti-inflammatory activity of a water extract of Acorus calamus L. leaves on keratinocyte HaCaT cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Acorus calamus L., sweet flag, is a well-known medicinal plant that grows worldwide wildly along swamps, rivers, and lakes. AIM OF THE STUDY: The aim of this study was to evaluate the anti-inflammatory activity of Acorus calamus leaf (ACL) extract and to explore its mechanism of action on human keratinocyte HaCaT cells. MATERIALS AND METHODS: HaCaT cells treated with polyinosinic:polycytidylic acid (polyI:C) and peptidoglycan (PGN) induced the inflammatory reactions. The anti-inflammatory activities of ACL were investigated using RT-PCR, ELISA assay, immunoblotting, and immunofluorescence staining. RESULTS: HaCaT cells induced the pro-inflammatory cytokines, interleukin-8 (IL-8) and/or interleukin-6 (IL-6) expressions after treatment with polyI:C or PGN. ACL inhibited the expression of IL-8 and IL-6 RNA and protein levels, and attenuated the activation of NF-kappaB and IRF3 after polyI:C treatment. ACL also inhibited expression of IL-8 and activation of NF-kappaB following PGN induction. CONCLUSIONS: These results suggest that ACL inhibits the production of pro-inflammatory cytokines through multiple mechanisms and may be a novel and effective anti-inflammatory agent for the treatment of skin diseases.

The reference genetic linkage map for the multinational Brassica rapa genome sequencing project.

We describe the construction of a reference genetic linkage map for the Brassica A genome, which will form the backbone for anchoring sequence contigs for the Multinational Brassica rapa Genome Sequencing Project. Seventy-eight doubled haploid lines derived from anther culture of the F(1) of a cross between two diverse Chinese cabbage (B. rapa ssp. pekinensis) inbred lines, 'Chiifu-401-42' (C) and 'Kenshin-402-43' (K) were used to construct the map. The map comprises a total of 556 markers, including 278 AFLP, 235 SSR, 25 RAPD and 18 ESTP, STS and CAPS markers. Ten linkage groups were identified and designated as R1-R10 through alignment and orientation using SSR markers in common with existing B. napus reference linkage maps. The total length of the linkage map was 1,182 cM with an average interval of 2.83 cM between adjacent loci. The length of linkage groups ranged from 81 to 161 cM for R04 and R06, respectively. The use of 235 SSR markers allowed us to align the A-genome chromosomes of B. napus with those of B. rapa ssp. pekinensis. The development of this map is vital to the integration of genome sequence and genetic information and will enable the international research community to share resources and data for the improvement of B. rapa and other cultivated Brassica species.

Production of indole-3-acetic acid in the plant-beneficial strain Pseudomonas chlororaphis O6 is negatively regulated by the global sensor kinase GacS.

Certain plant growth-promoting bacteria, such as Pseudomonas fluorescens 89B61 and Bacillus pumilus SE34, secreted high levels of indole-3-acetic acid (IAA) in tryptophan-amended medium in stationary phase as determined by chromogenic analysis and high-performance liquid chromatography. Two other growth-promoting strains, P. chlororaphis O6 and Serratia marcescens 90-166, did not produce these high levels of IAA. However, when the gacS mutant of P. chlororaphis O6 was grown in tryptophan-supplemented medium, IAA was detected in culture filtrates. IAA production by the gacS mutant in P. chlororaphis O6 was repressed in the tryptophan medium by complementation with the wild-type gacS gene. Thus, the global regulatory Gac system in P. chlororaphis O6 acts as a negative regulator of IAA production from trypophan.