Characterization of a novel Toll/interleukin-1 receptor (TIR)-TIR gene differentially expressed in common bean (Phaseolus vulgaris cv. Othello) undergoing a defence response to the geminivirus Bean dwarf mosaic virus

Common bean ( Phaseolus vulgaris L.) cultivar (cv.) Othello develops a hypersensitive response-associated vascular resistance to infection by Bean dwarf mosaic virus (BDMV), a single-stranded DNA virus (genus Begomovirus , family Geminiviridae ). A PCR-based cDNA subtraction approach was used to identify genes involved in this resistance response. Eighteen clones, potentially involved with BDMV resistance, were identified based upon being up-regulated in BDMV-infected tissues and/or having sequence similarity with known resistance-associated genes. Analysis of these clones revealed potential genes involved in pathogen defence, including pathogenesis-related protein genes and resistance gene analogues (RGAs). Further characterization of one RGA, F1-10 , revealed that it encodes a predicted protein with a double Toll/interleukin-1 receptor (TIR) motif. Full-length ( F1-10 ) and spliced ( F1-10sp ) forms of the RGA were strongly up-regulated in BDMV-infected cv. Othello hypocotyl tissues by 4 days post-inoculation, but not in equivalent mock-inoculated tissues. In agroinfiltration experiments, F1-10 , but not F1-10sp , mediated resistance to BDMV in the susceptible common bean cv. Topcrop. By contrast, transgenic Nicotiana benthamiana lines expressing F1-10 or F1-10sp were not resistant to BDMV. Interestingly, when these transgenic lines were inoculated with the potyvirus Bean yellow mosaic virus, some F1-10 lines showed a more severe symptom phenotype compared with non-transgenic control plants. Based on these findings, F1-10 was named: Phaseolus vulgaris VIRUS response TIR-TIR GENE 1 ( PvVTT1 ).     

Change of Haemolymph Ferritin of Sweet Potato Hornworm, Agrius convolvuli by Heavy Metals

Agrius convolvuli haemolymph ferritin was purified by KBr density gradient ultracentrifugation and anion exchange column chromatography. The 670 kDa ferritin was composed of two subunits of 26 kDa and 31 kDa. It was also shown that the protein had an isoelectric point (pI) of pH 7.4. The N‐terminal amino acid sequences of the two subunits were NH2‐DNXYQDVSLDXSQAXNXL (26 kDa subunit) and NH2‐TQXHVNPVNIQRDXVTMHXS (31 kDa subunit). The sequential analysis showed that they had high similarity to lepidopteran ferritin subunits, S‐ and G‐type, respectively. Using electron microscope, it was observed that the protein had a core whose size was about 7 nm. In the amino acid composition of the protein, Glu (13.22%), Asp (10.43%), Pro (9.69%), Leu (9.63%), Ala (9.55%) and Gly (8.49%) were in relatively high contents while Tyr (1.21%), His (2.58%) and Arg (3.10 %) were in low. It was shown that the amount of ferritin in A. convolvuli haemolymph was increased by injection of eight different heavy metal ions, FeCl₃, HgCl₂, CuSO₄, ZnSO₄, MnCl₂, MgCl₂, CrCl₃ and CdCl₂. Among the ions, Fe³⁺, Hg²⁺, Zn⁴⁺, Mn²⁺ and Cd.²⁺ significantly induced the amount of the protein.     

Phenotypic and Genetic Diversity of Erwinia carotovora ssp. carotovora Strains from Asia

A collection of 87 strains of the soft rot pathogen Erwinia carotovora ssp. carotovora (Ecc) isolated from various host plants in Japan, Korea and Thailand was characterized by bacteriological, pathological and genetic properties. On the basis of pathogenicity on the potato, tomato, onion and cucumber, strains were divided into four groups. They were also characterized by PCR‐restriction fragment length polymorphisms (RFLP) of 16S ribosomal DNA (rDNA), 16S‐23S rDNA intergenic spacer regions (ISRs) and a pel gene encoding pectate lyase. By analysis of 16S rDNA RFLP generated by Hinf I, Ecc strains were differentiated into two groups where it was discovered that most strains from Korea and Japan belonged to the same group. In the analysis of ISRs RFLP with MboI, two patterns were found. All Thai strains showed the same pattern. In the analysis of the pel gene RFLP with Sau3AI, all strains were separated into two independent patterns except for one strain. The strain (MAFF 301937) isolated from the mulberry showed a unique RFLP pattern of the pel gene. In cluster analysis based on 26 phenotypic characters, Ecc strains were composed of two groups, A and B. Group A contained typical Ecc strains which provided negative reactions in testing the production of reducing substances from sucrose and acids from α‐methyl glucoside. All Thai strains and most of the Korean strains belonged to group A, whereas group B contained atypical Ecc strains, which were isolated in Japan and Korea; the properties of this group were similar to those of E. carotovora ssp. atroseptica. The research reported here was undertaken to provide information on the strains of E. carotovora ssp. carotovora in Asia.     

Rapid and Efficient Purification of Haemolymph Ferritin from Cricket, Gryllus bimaculatus

The Gryllus bimaculatus ferritin was purified from the haemolymph by a consecutive four‐step procedures consisting of 50% ammonium sulfate fractionation, anion exchange column chromatography using HiTrap^TM Q column (1.6 x 4 cm, Amersham Bioscience), 70°C heat treatment for 10 min, acid treatment of 0.1 M sodium acetate buffer (pH 6.0), and gel filtration column chromatography using G4000SW column (0.75 x 60 cm, Tosoh, Japan) connected on FPLC system. The purified ferritin was found to have two major subunits of 32 and 30 kDa and three minor subunits of 28, 27, and 25 kDa by 2D electrophoresis analysis. Amino acid composition analysis showed that there are high contents of Asp, Glu, Met, Leu, and Lys residues in ferritin while low contents of Cys, Tyr, and Trp residues in the protein. G. bimaculatus haemolymph ferritin could be classified as a methionine‐rich protein.     

Characteristics of Vaginal Microbiome in Women with Pelvic Inflammatory Disease in Korea

Human vaginal microorganisms play an important role in maintaining good health throughout the human life cycle. An imbalance in the vaginal microbiota is associated with an increased risk of pelvic inflammatory disease (PID). This study aimed to characterize and compare vaginal microbial profiles of premenopausal Korean women with and without PID. 74 Korean premenopausal female vaginal samples were obtained; 33 were from healthy women (a control group) and 41 from PID patients. Vaginal fluid samples were collected from the vaginal wall and posterior cervix and then analyzed by 16S ribosomal ribonucleic acid (rRNA) gene-based amplicon sequencing. Results showed a significant difference between the vaginal microbial communities of the two groups (Jensen-Shannon, p = 0.014; Bray-Curtis, p = 0.009; Generalized UniFrac, p = 0.007; UniFrac, p = 0.008). Lactobacillus accounted for the highest percentage (61.0%) of the control group but was significantly decreased (34.9%) in PID patients; this was the most significant difference among all bacterial communities (p = 0.028, LDA effect size = 5.129). In addition, in the PID patient group, species diversity significantly increased (Simpson, p = 0.07) as the proportion of various pathogens increased evenly, resulting in a polymicrobial infection. Similarly, lactate, which constituted the highest percentage of the organic acids in the control group, was significantly decreased in the PID patient group (p = 0.04). The present study’s findings will help understand PID from the microbiome perspective and are expected to contribute to the development of more efficient PID diagnosis and treatment modalities.     

麦类作物遗传转化

麦类作物包括小麦(Triticum aestivum L.)、硬粒小麦(Triticum turgidum con v.durum Dest.e.m)、大麦(Hordeum vulgare L.)、黑麦(Secale cereal L.)、燕麦(Avena sativa L.)及小大麦(×Tritordeum Ascherson et Graebuer.).自从基因枪被发明以来,科学家们已经利用来自麦类作物的幼胚、 盾片、成熟种子胚、花粉粒、花药、幼穗、叶基组织、发芽种子幼苗的顶端分生组织及其愈伤组织或培养物作为外植体,通过基因枪、农杆菌介导、 PEG法、电激法、微注射法、硅化纤维素介导、幼穗注射法等技术先后将一些选择标记基因、报告基因和有用的目的基因如抗真菌、抗虫、 籽粒品质、抗干旱基因等转化到麦类作物中.转基因植物表现为抗性增强或籽粒的加工品质提高和营养成份增加.被转化的基因通常以单位点多拷贝的形式随机整合到受体细胞的基因组中,并以孟德尔规律遗传.整合位点一般分布在染色体的近端粒区域,整合的拷贝数大多为5～10个拷贝,最高可达到50个拷贝.在转化过程中,被转化的质粒上的片段包括选择标记基因、目标基因、甚至质粒的抗生素基因和其他无关序列,随机地连接并形成多个分子量大小不等,组成成分不同的分子簇,或首先由其中一个分子簇整合到植物基因组中,这会导致在整合位点附近产生"热点",易于其他分子簇在此处整合,从而完成两期整合;或被转化的质粒上的选择标记基因、目标基因、质粒的抗生素基因和其他无关序列、植物基因组DNA等片段共同形成各种不同类型的分子簇,当植物细胞染色体复制时,在复制叉处整合到植物基因组中.转基因可以在各种水平上表达,也会时常发生基因沉默,这会导致转基因植物DNA水平上表达但在蛋白质水平上不表达,后代偏向分离,沉默的转基因重新表达.转基因的位置效应、甲基化和启动子都会诱发转基因沉默.在麦类作物中,35S启动子易于导致转基因沉默,应尽量减少使用.转基因还导致被转化麦类作物在农艺性状和细胞学上的变异.目前,麦类作物遗传转化已经成为一种常规的技术,转基因麦类作物正开始进入商业应用阶段.相信多种转化新技术的应用和发展将会培育出高产、稳产、优质、低投入的各类品种和种质.     

Description of three liocranid spider species from Korea (Araneae: Liocranidae)

Three liocranid spider species are described with illustrations: a new species, Agroeca bonghwaensis n. sp. and two species new to Korean spider fauna, Agroeca montana Hayashi, 1986 and Scotina palliardii (Koch, 1881). The occurrence of the genus Scotina Menge, 1873 is also recorded from Korea for the first time.     

A Clinical Trial to Evaluate the Efficacy of α-Viniferin in Staphylococcus aureus – Specific Decolonization without Depleting the Normal Microbiota of Nares

Staphylococcus aureus is currently a significant multidrug-resistant bacterium, causing severe healthcare-associated and community-acquired infections worldwide. The current antibiotic regimen against this pathogen is becoming ineffective due to resistance, in addition, they disrupt the normal microbiota. It highlights the urgent need for a pathogen-specific drug with high antibacterial efficacy against S. aureus. α-Viniferin, a bioactive phytochemical compound, has been reported to have excellent anti-Staphylococcus efficacy as a topical agent. However, so far, there were no clinical trials that have been conducted to elucidate its efficacy. The present study aimed to investigate the antibacterial efficacy of α-viniferin against S. aureus in a ten-day clinical trial. Based on the results, α-viniferin showed 50% minimum inhibitory concentrations (MIC₅₀ values) of 7.8 μg/ml in culture broth medium. α-Viniferin was administered in the nares three times a day for ten days using a sterile cotton swab stick. Nasal swab specimens were collected before (0 days) and after finishing the trial (10^th day), and then analyzed. In the culture and RT-PCR-based analysis, S. ureus was reduced significantly: 0.01. In addition, 16S ribosomal RNA-based amplicon sequencing analysis showed that S. aureus reduced from 51.03% to 23.99% at the genus level. RNA-seq analysis was also done to gain insights into molecular mechanisms of α-viniferin against S. aureus, which revealed that some gene groups were reduced in 5-fold FC cutoff at two times MIC conditions. The study results demonstrate α-viniferin as a potential S. aureus-specific drug candidate.