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Hydrogen Peroxide Prompted Lignification Affects Pathogenicity of Hemi-biotrophic Pathogen Bipolaris sorokiniana to Wheat

  • Poudel, Ajit;Navathe, Sudhir;Chand, Ramesh;Mishra, Vinod K.;Singh, Pawan K.;Joshi, Arun K.
    • The Plant Pathology Journal
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    • v.35 no.4
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    • pp.287-300
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    • 2019
  • Spot blotch caused by Bipolaris sorokiniana has spread to more than 9 million ha of wheat in the warm, humid areas of the Eastern Gangetic Plains (EGP) of South Asia and is a disease of major concern in other similar wheat growing regions worldwide. Differential lignin content in resistant and susceptible genotypes and its association with free radicals such as hydrogen peroxide ($H_2O_2$), superoxide ($O_2{^-}$) and hydroxyl radical ($OH^-$) were studied after inoculation under field conditions for two consecutive years. $H_2O_2$ significantly influenced lignin content in flag leaves, whereas there was a negative correlation among lignin and $H_2O_2$ to the Area Under Disease Progress Curve (AUDPC). The production of $H_2O_2$ was higher in the resistant genotypes than susceptible ones. The $O_2{^-}$ and $OH^-$ positively correlated with AUDPC but negatively with lignin content. This study illustrates that $H_2O_2$ has a vital role in prompting lignification and thereby resistance to spot blotch in wheat. We used cluster analysis to separate the resistant and susceptible genotypes by phenotypic and biochemical traits. $H_2O_2$ associated lignin production significantly reduced the number of appressoria and penetration pegs. We visualized the effect of lignin in disease resistance using differential histochemical staining of tissue from resistant and susceptible genotypes, which shows the variable accumulation of hydrogen peroxide and lignin around penetration sites.

CRISPR/Cas9-mediated generation of a Plac8 knockout mouse model

  • Lee, HyunJeong;Kim, Joo-Il;Park, Jin-Sung;Roh, Jae-il;Lee, Jaehoon;Kang, Byeong-Cheol;Lee, Han-Woong
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.279-287
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    • 2018
  • Placenta specific 8 (PLAC8, also known as ONZIN) is a multi-functional protein that is highly expressed in the intestine, lung, spleen, and innate immune cells, and is involved in various diseases, including cancers, obesity, and innate immune deficiency. Here, we generated a Plac8 knockout mouse using the CRISPR/Cas9 system. The Cas9 mRNA and two single guide RNAs targeting a region near the translation start codon at Plac8 exon 2 were microinjected into mouse zygotes. This successfully eliminated the conventional translation start site, as confirmed by Sanger sequencing and PCR genotyping analysis. Unlike the previous Plac8 deficient models displaying increased adipose tissue and body weights, our male Plac8 knockout mice showed rather lower body weight than sex-matched littermate controls, though the only difference between these two mouse models is genetic context. Differently from the previously constructed embryonic stem cell-derived Plac8 knockout mouse that contains a neomycin resistance cassette, this knockout mouse model is free from a negative selection marker or other external insertions, which will be useful in future studies aimed at elucidating the multi-functional and physiological roles of PLAC8 in various diseases, without interference from exogenous foreign DNA.

Promoted Growth and Development of Carnation Plantlets In Vitro by Ventilation and Combined Red and Blue Light

  • Nguyen, Quan Hoang;Thi, Luc The;Park, Yoo Gyeong;Jeong, Byoung Ryong
    • FLOWER RESEARCH JOURNAL
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    • v.26 no.4
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    • pp.166-178
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    • 2018
  • In this study, the principal objective was to investigate the effect of light quality and vessel ventilation on the growth and development, physiology, activities of antioxidant enzymes, and contents of mineral nutrients of carnation (Dianthus caryophyllus L.) 'Marble Beauty'. Single node cuttings stuck into the plant growth regulator (PGR)-free MS medium in containers covered with caps with or without a ventilation filter were cultured first four weeks under white and then additional four weeks under either white (control), blue, red, or red + blue light emitting diodes (LEDs) for 56 days. Interestingly, a ventilated culture condition not only reduced the percentage of the hyperhydricity, but also increased the total chlorophyll content (Chl a + Chl b) of the plantlets as compared to the non-ventilated condition. In addition, blue LEDs produced plantlets with the greatest number of shoots and red LEDs produced plantlets with the greatest shoot length. The quality of plantlets was improved under a ventilation condition. Besides, under a ventilated condition, red + blue LEDs raised APX activity, and blue LEDs not only raised the activity of the CAT, but also increased tissue contents of such elements as K, Ca, Mg, Zn, Mn and Fe. The red LEDs increased contents of B and Si under a ventilated condition, and Na accumulation under a non-ventilated condition. Thus, including blue or red LEDs as the light source in a ventilated culture condition will produce plantlets of carnation 'Marble Beauty' in vitro with improved quality.

Nanoengineered, cell-derived extracellular matrix influences ECM-related gene expression of mesenchymal stem cells

  • Ozguldez, Hatice O.;Cha, Junghwa;Hong, Yoonmi;Koh, Ilkyoo;Kim, Pilnam
    • Biomaterials Research
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    • v.22 no.4
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    • pp.337-345
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    • 2018
  • Background: Human mesenchymal stem cells (hMSCs) are, due to their pluripotency, useful sources of cells for stem cell therapy and tissue regeneration. The phenotypes of hMSCs are strongly influenced by their microenvironment, in particular the extracellular matrix (ECM), the composition and structure of which are important in regulating stem cell fate. In reciprocal manner, the properties of ECM are remodeled by the hMSCs, but the mechanism involved in ECM remodeling by hMSCs under topographical stimulus is unclear. In this study, we therefore examined the effect of nanotopography on the expression of ECM proteins by hMSCs by analyzing the quantity and structure of the ECM on a nanogrooved surface. Methods: To develop the nanoengineered, hMSC-derived ECM, we fabricated the nanogrooves on a coverglass using a UV-curable polyurethane acrylate (PUA). Then, hMSCs were cultivated on the nanogrooves, and the cells at the full confluency were decellularized. To analyze the effect of nanotopography on the hMSCs, the hMSCs were re-seeded on the nanoengineered, hMSC-derived ECM. Results: hMSCs cultured within the nano-engineered hMSC-derived ECM sheet showed a different pattern of expression of ECM proteins from those cultured on ECM-free, nanogrooved surface. Moreover, hMSCs on the nano-engineered ECM sheet had a shorter vinculin length and were less well-aligned than those on the other surface. In addition, the expression pattern of ECM-related genes by hMSCs on the nanoengineered ECM sheet was altered. Interestingly, the expression of genes for osteogenesis-related ECM proteins was downregulated, while that of genes for chondrogenesis-related ECM proteins was upregulated, on the nanoengineered ECM sheet. Conclusions: The nanoengineered ECM influenced the phenotypic features of hMSCs, and that hMSCs can remodel their ECM microenvironment in the presence of a nanostructured ECM to guide differentiation into a specific lineage.

Evaluation of Clinical Usefulness of Herbal Mixture HO-Series for Improving Hangover (복합생약 HO-Series의 숙취개선 임상적 유용성 평가)

  • Chang, Bo Yoon;Bae, Jin Hye;Kim, Da Eun;Kim, Dae Sung;Cho, Hyoung Kwon;Kim, Sung Yeon
    • Korean Journal of Pharmacognosy
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    • v.51 no.4
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    • pp.278-290
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    • 2020
  • The purpose of this study is to investigate the hangover relieving effect of HO-series. HO-S1 is an herbal mixture, which consists of extracts from Flower of Pueraria lobata Ohwi, Glycyrrhiza glabra Linné, Fruit of Lycium chinense Miller, Poria cocos Wolf, Acanthopanax sessiliflorum Seeman, Scutellaria baicalensis Georgi, Atractylodes lancea De Candlle and Zingiber officinale Roscoe. HO-S2 is a candidate that has been performed to ultra filtration based on HO-S1. HO-S3 is a mixture of amino acids and vitamins based on HO-S2. HO-01 is the final beverage base produced based on HO-S3. The antioxidant activity of HO-series was similar to that of vitamin C or trolox. The production of t-BHP induced reactive oxygen species(ROS) was significantly blocked in the presence of HO-series. In vivo study, AUC of alcohol and acetaldehyde concentrations in HO-S2 and HO-S3 treated groups significantly decreased. Hepatic alcohol dehydrogenase(ADH) and acetaldehyde dehydrogenase(ALDH) activity were significantly higher in HO-S2 and HO-S3 treated groups. And 2E1 activity and glutathione were significantly elevated, while the malondialdehyde level was not significantly in liver tissue. After alcohol exposure, the sensitivity scores of blood alcohol and acetaldehyde concentration and hangover symptoms were significantly decreased in the HO-01 intake group compared with the non-intake group. ALDH activity was significantly increased in the HO-01 intake group. HO-series have antioxidant activity and a protective effect from ROS. HO-S2, HO-S3 and HO-01 are potentially highly beneficial in relieving hangover, as it scavenges reactive free radicals and boosts the endogenous antioxidant system.

Development and Field-evaluation of Automatic Spreader for Bedding Materials in Duck Houses (오리사 바닥 깔짚자동살포장치 개발 및 실증)

  • Kwon, Kyeong-seok;Woo, Jae-seok;Noh, Je-hee;Oh, Sang-ik;Kim, Jong-bok;Kim, Jung-kon;Yang, Kayoung;Jang, Donghwa;Choi, Sungmin
    • Journal of The Korean Society of Agricultural Engineers
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    • v.63 no.1
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    • pp.37-48
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    • 2021
  • The automatic-spreader of bedding materials was developed to reduce labor cost, and to achieve successful biosecurity in duck houses. Algorithm of the device was designed to realize a concept of the automatic unmanned operation including entire processes such as loading and spreading of the bedding materials. From the field measurement, the relationship between the expected water content reduction and weight of bedding materials per unit area according to the operation condition of the devices was induced. In the case of the measurement of particulate matters during the process of spreading works, the results of using both conventional manual-spreader and automatic-spreader were exceeded the allowable limit of inhalable and respirable dust, respectively. But, workers using automatic-spreader could be free from harmful aero-condition because they did not stay inside the facility during the spreading works. In addition, from the Duck hepatitis virus PCR test, influence on pulmonary tissue of the duck was not found. It could be expected that the development of the automatic-spreader of bedding materials for duck house can contribute to the advancement of duck breeding facilities.

The Danger-Associated Peptide PEP1 Directs Cellular Reprogramming in the Arabidopsis Root Vascular System

  • Dhar, Souvik;Kim, Hyoujin;Segonzac, Cecile;Lee, Ji-Young
    • Molecules and Cells
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    • v.44 no.11
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    • pp.830-842
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    • 2021
  • When perceiving microbe-associated molecular patterns (MAMPs) or plant-derived damage-associated molecular patterns (DAMPs), plants alter their root growth and development by displaying a reduction in the root length and the formation of root hairs and lateral roots. The exogenous application of a MAMP peptide, flg22, was shown to affect root growth by suppressing meristem activity. In addition to MAMPs, the DAMP peptide PEP1 suppresses root growth while also promoting root hair formation. However, the question of whether and how these elicitor peptides affect the development of the vascular system in the root has not been explored. The cellular receptors of PEP1, PEPR1 and PEPR2 are highly expressed in the root vascular system, while the receptors of flg22 (FLS2) and elf18 (EFR) are not. Consistent with the expression patterns of PEP1 receptors, we found that exogenously applied PEP1 has a strong impact on the division of stele cells, leading to a reduction of these cells. We also observed the alteration in the number and organization of cells that differentiate into xylem vessels. These PEP1-mediated developmental changes appear to be linked to the blockage of symplastic connections triggered by PEP1. PEP1 dramatically disrupts the symplastic movement of free green fluorescence protein (GFP) from phloem sieve elements to neighboring cells in the root meristem, leading to the deposition of a high level of callose between cells. Taken together, our first survey of PEP1-mediated vascular tissue development provides new insights into the PEP1 function as a regulator of cellular reprogramming in the Arabidopsis root vascular system.

Efficacy of bivalent vaccines of porcine circovirus type 2 and Mycoplasma hyopneumoniae in specific pathogen-free pigs challenged with porcine circovirus type 2d

  • Lim, Jeonggyo;Jin, Myongha;Yoon, Injoong;Yoo, Han Sang
    • Journal of Veterinary Science
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    • v.23 no.3
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    • pp.49.1-49.13
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    • 2022
  • Background: Porcine circovirus type 2 (PCV2) and Mycoplasma hyopneumoniae (MHP) are economically significant pathogens in the pig industry. The use of combined vaccines against PCV2 and MHP is one of the most effective ways of protecting pigs from both diseases, and it has become a part of general management. Objectives: This study evaluated the efficacy of two new bivalent vaccines of PCV2 and MHP (Myco-X and Myco-XD) in SPF pigs. Myco-X and Myco-XD are a combined vaccine of MHP with PCV2b and PCV2d, respectively. Methods: Sixteen pigs were divided into four groups: Myco-X-vaccinated challenged, Myco-XD-vaccinated challenged, unvaccinated challenged, and unvaccinated unchallenged. Two milliliters of Myco-X were administered intramuscularly, and 0.5 mL of Myco-XD was injected intradermally at 3 wk of age. The pigs were challenged with virulent PCV2d via the intramuscular and intranasal route 4 wk post-vaccination. Results: All vaccinated pigs showed effective reduction of the clinical signs, the PCV2d load in the blood and nasal swab samples, as well as lung and lymphoid tissue lesions in the challenge test. Compared to unvaccinated challenged animals, the vaccinated challenged animals showed significantly higher (p < 0.05) levels of anti-PCV2 IgG, PCV2d-specific interferon-γ (IFN-γ), and anti-MHP IgG. Conclusions: Based on clinical, microbiological, serological, and pathological assessments, this study confirmed that both combined vaccines could protect pigs against PCV2 infection caused by PCV2d. On the other hand, further research on the efficacy evaluation of these new vaccines against the MHP challenge and PCV2d/MHP co-challenge is needed.

Successful Management of Subcutaneous Abscess in a Captive Leopard Gecko (Eublepharis macularius)

  • Win, Phyo Wai;Rhim, Haerin;Kim, Myeongsu;Gim, Seulgi;Han, Jae-Ik
    • Journal of Veterinary Clinics
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    • v.39 no.5
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    • pp.272-276
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    • 2022
  • An 8 month old leopard gecko (Eublepharis macularius) with a large nodule was referred to our hospital. During the physical examination, the nodule had an unclear boundary from the top of the left eye to the front of the left ear and prevented the opening of the left eye. A hard, cheese-like, yellow, pus-filled nodule was observed. A cytological examination of a pus swab sample revealed pyogranulomatous inflammation with rod-shaped bacteria. Ofloxacin was chosen as the empirical topical antimicrobial drug for treatment. The swab samples were inoculated in trypticase soy agar with 5% sheep blood and incubated at 37℃ for 24 h. Gram-negative bacteria were identified via Gram staining, and the Kirby-Bauer antimicrobial susceptible disk diffusion test against 24 antibiotics according to protocol M100-Ed32 of CLSI showed that the fluoroquinolone group (ciprofloxacin and enrofloxacin) was susceptible to the isolated bacteria. Molecular identification based on 16S ribosomal RNA gene sequencing confirmed that the isolated bacteria had a 99.85% nucleotide similarity with Serratia surfactantfaciens (GenBank accession no. CP014948). After 1 week, the boundaries of the nodule became clear; thus, the abscess was physically removed by expanding the hole formed above the eye for drainage, and flushing was repeated. After another 1 week, new tissue restoration without scarring was observed. This is a rare case report of the successful management of a subcutaneous abscess and scar-free healing in a lizard.

The effect of Panax notoginseng saponins on oxidative stress induced by PCV2 infection in immune cells: in vitro and in vivo studies

  • Wang, Qiu-Hua;Kuang, Na;Hu, Wen-yue;Yin, Dan;Wei, Ying-Yi;Hu, Ting-Jun
    • Journal of Veterinary Science
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    • v.21 no.4
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    • pp.61.1-61.16
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    • 2020
  • Background: Panax notoginseng saponins (PNS) are bioactive substances extracted from P. notoginseng that are widely used to treat cardiovascular and cerebrovascular diseases and interstitial diseases. PNS have the functions of scavenging free radicals, anti-inflammation, improving blood supply for tissue and so on. Objectives: The aim of this study was to investigate the effects of PNS on the oxidative stress of immune cells induced by porcine circovirus 2 (PCV2) infection in vitro and in vivo. Methods: Using an oxidative stress model of PCV2 infection in a porcine lung cell line (3D4/2 cells) and mice, the levels of nitric oxide (NO), reactive oxygen species (ROS), total glutathione (T-GSH), reduced glutathione (GSH), and oxidized glutathione (GSSG) and the activities of xanthine oxidase (XOD), myeloperoxidase (MPO) and inducible nitric oxide synthetase (iNOS) were determined to evaluate the regulatory effects of PNS on oxidative stress. Results: PNS treatment significantly reduced the levels of NO and ROS, the content of GSSG and the activities of XOD, MPO, and iNOS (p < 0.05), while significantly increasing GSH and the ratio of GSH/GSSG in infected 3D4/2 cells (p < 0.05).Similarly, in the in vivo study, PNS treatment significantly decreased the level of ROS in spleen lymphocytes of infected mice (p < 0.05), increased the levels of GSH and T-GSH (p < 0.05), significantly decreased the GSSG level (p < 0.05), and decreased the activities of XOD, MPO, and iNOS. Conclusions: PNS could regulate the oxidative stress of immune cells induced by PCV2 infection in vitro and in vivo.