scholarly journals Novel Diagnosis for Citrus Stubborn Disease by Detection of a Spiroplasma citri-Secreted Protein

2014 ◽  
Vol 104 (2) ◽  
pp. 188-195 ◽  
Author(s):  
Jinxia Shi ◽  
Deborah Pagliaccia ◽  
Robyn Morgan ◽  
Yongli Qiao ◽  
Songqin Pan ◽  
...  
Keyword(s):  
The United States ◽  
Fruit Production ◽  
Plant Diseases ◽  
Microbial Pathogens ◽  
Mass Spectrometry Analysis ◽  
Bacterial Disease ◽  
Secreted Protein ◽  
Spiroplasma Citri ◽  
Spectrometry Analysis ◽  
Citrus Stubborn Disease

Citrus stubborn disease (CSD), first identified in California, is a widespread bacterial disease found in most arid citrus-producing regions in the United States and the Mediterranean Region. The disease is caused by Spiroplasma citri, an insect-transmitted and phloem-colonizing bacterium. CSD causes significant tree damage resulting in loss of fruit production and quality. Detection of CSD is challenging due to low and fluctuating titer and sporadic distribution of the pathogen in infected trees. In this study, we report the development of a novel diagnostic method for CSD using an S. citri-secreted protein as the detection marker. Microbial pathogens secrete a variety of proteins during infection that can potentially disperse systemically in infected plants with the vascular flow. Therefore, their distribution may not be restricted to the pathogen infection sites and could be used as a biological marker for infection. Using mass spectrometry analysis, we identified a unique secreted protein from S. citri that is highly expressed in the presence of citrus phloem extract. ScCCPP1, an antibody generated against this protein, was able to distinguish S. citri-infected citrus and periwinkle from healthy plants. In addition, the antiserum could be used to detect CSD using a simple direct tissue print assay without the need for sample processing or specialized lab equipment and may be suitable for field surveys. This study provides proof of a novel concept of using pathogen-secreted protein as a marker for diagnosis of a citrus bacterial disease and can probably be applied to other plant diseases.

scholarly journals Identification of Novel Surface Proteins of Anaplasma phagocytophilum by Affinity Purification and Proteomics

2007 ◽  
Vol 189 (21) ◽  
pp. 7819-7828 ◽  
Author(s):  
Yan Ge ◽  
Yasuko Rikihisa
Keyword(s):  
Anaplasma Phagocytophilum ◽  
Affinity Purification ◽  
The United States ◽  
Capillary Liquid Chromatography ◽  
Etiologic Agent ◽  
Surface Proteins ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis ◽  
Major Surface

ABSTRACT Anaplasma phagocytophilum is the etiologic agent of human granulocytic anaplasmosis (HGA), one of the major tick-borne zoonoses in the United States. The surface of A. phagocytophilum plays a crucial role in subverting the hostile host cell environment. However, except for the P44/Msp2 outer membrane protein family, the surface components of A. phagocytophilum are largely unknown. To identify the major surface proteins of A. phagocytophilum, a membrane-impermeable, cleavable biotin reagent, sulfosuccinimidyl-2-[biotinamido]ethyl-1,3-dithiopropionate (Sulfo-NHS-SS-Biotin), was used to label intact bacteria. The biotinylated bacterial surface proteins were isolated by streptavidin agarose affinity purification and then separated by electrophoresis, followed by capillary liquid chromatography-nanospray tandem mass spectrometry analysis. Among the major proteins captured by affinity purification were five A. phagocytophilum proteins, Omp85, hypothetical proteins APH_0404 (designated Asp62) and APH_0405 (designated Asp55), P44 family proteins, and Omp-1A. The surface exposure of Asp62 and Asp55 was verified by immunofluorescence microscopy. Recombinant Asp62 and Asp55 proteins were recognized by an HGA patient serum. Anti-Asp62 and anti-Asp55 peptide sera partially neutralized A. phagocytophilum infection of HL-60 cells in vitro. We found that the Asp62 and Asp55 genes were cotranscribed and conserved among members of the family Anaplasmataceae. With the exception of P44-18, all of the proteins were newly revealed major surface-exposed proteins whose study should facilitate understanding the interaction between A. phagocytophilum and the host. These proteins may serve as targets for development of chemotherapy, diagnostics, and vaccines.

scholarly journals Formation of Strecker Aldehydes in Casein Digestion

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 500-500
Author(s):  
Yuyin Zhou ◽  
Chi Chen
Keyword(s):  
Degradation Products ◽  
In Vitro Digestion ◽  
The United States ◽  
Mass Spectrometry Analysis ◽  
United States Department ◽  
Gastric Digestion ◽  
Spectrometry Analysis ◽  
Strecker Aldehydes ◽  
Intestinal Digestion

Abstract Objectives This study investigated the occurrence of Strecker degradation during in vitro digestion. Methods Casein was first incubated with artificial gastric fluid containing porcine pepsin and HCl (pH = 2) for 60 min. After adjusting pH to 7 with sodium hydroxide, casein gastric digesta was then incubated with artificial intestinal fluid containing porcine pancreatin (pH = 7) for 120 min. Digesta samples were collected at 0, 10, 20, 30, and 60 min of gastric digestion, and then 10, 20, 30, 60, and 120 min of intestinal digestion. Free amino acids and aldehydes in digesta samples were analyzed by the liquid chromatography-mass spectrometry analysis. Results Multiple aldehydes were detected in gastric digestion samples, and their concentrations were further increased by intestinal digestion. Among them, isovaleraldehyde, isobutyraldehyde, phenylacetaldehyde, and acetaldehyde, are the Strecker degradation products of leucine, valine, phenylalanine, and alanine, respectively. Without digestive enzymes, casein incubation did not produce Strecker aldehydes. Conclusions In vitro digestion of proteins can produce Strecker aldehydes. Funding Sources This research was partially supported by the Agricultural Experiment Station project MIN-18-125 (C. C.) from the United States Department of Agriculture (USDA).

scholarly journals Molecular identification of actinomycetes with antimicrobial, antioxidant and anticancer properties

2019 ◽  
Vol 10 (2) ◽  
pp. 218-231
Author(s):  
Mohamed S Abdel-Aziz ◽  
Amal S Hathout ◽  
Aziza A El-Neleety ◽  
Ahmed A Hamed ◽  
Bassem A Sabry ◽  
...  
Keyword(s):  
Molecular Identification ◽  
Wide Spectrum ◽  
Salmonella Typhi ◽  
Antimicrobial Activities ◽  
Culture Broth ◽  
Microbial Pathogens ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Spectrometry Analysis

The objectives of this study were to isolate and identify the actinomycetes strains from the soil and marine sediments and to evaluate the antimicrobial, antioxidant and cytotoxic activity of their bioactive secondary metabolites. Eight actinomycetes strains were isolated from soil and marine sediment samples collected from different areas in Egypt. Only three actinomycetes exhibited a wide spectrum of antimicrobial activities. They were active in vitro against microbial pathogen viz: Staphylococcus aureus, Escherichia coli, Salmonella typhi, Aspergillus parasiticus, Fusarium solani and Fusarium oxysporum. These promising isolates were selected and identified using molecular identification technique and identified as Streptomyces spp. The crude extracts from the three Streptomyces exhibited potent antimicrobial activities against a set of microbial pathogens as well as antioxidant and anticancer activity in human hepatocellular carcinoma cell line (HepG2). The crude extract of Streptomyces isolate D showed antitumor activity with lC50 0.85 µg/ mL. Forty compounds were identified from the two most promising ethyl acetate extracts of culture broth of Streptomyces sp. (D-EGY) by Gas Chromatography-Mass Spectrometry analysis. It could be concluded that the streptomycetes isolated from the Egyptian environment are promising candidates as antimicrobial, antioxidant and anticancer.

scholarly journals New Perspectives on the Epidemiology of Citrus Stubborn Disease in California Orchards

2010 ◽  
Vol 11 (1) ◽  
pp. 37
Author(s):  
Alexandre F. S. Mello ◽  
Raymond K. Yokomi ◽  
Ulrich Melcher ◽  
Jianchi Chen ◽  
Edwin Civerolo ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Fruit Quality ◽  
Pathogen Detection ◽  
Disease Incidence ◽  
Sampling Technique ◽  
The United States ◽  
Citrus Industry ◽  
Spiroplasma Citri ◽  
Specific Pcr ◽  
Citrus Stubborn Disease

Although citrus stubborn disease (CSD), caused by the phloem resident mollicute Spiroplasma citri, is a significant threat to California citrus industry, our knowledge of its epidemiology is mostly anecdotal. We optimized multiple pathogen-detection protocols, measured disease incidence in two plots of commercial California groves, assessed pathogen impact on fruit quality and yield, and evaluated genetic diversity among S. citri isolates. Fruit columellas and receptacles were more suitable than leaves or bark for bacterial cultivation. Using cultivation and S. citri-specific PCR for detection, the incidence of CSD in two orchards, respectively, ranged from 46 to 85% and 1 to 4%, depending on the sampling technique. Yield and quality of fruits produced by trees that were mildly or severely CSD-symptomatic were compared to those of S. citri-free trees in one California orchard in 2006 and 2007. These infected trees had reduced fruit quality and up to 32% lower yield relative to S. citri-free trees. Using RAPD markers to compare 35 S. citri isolates collected 20 years ago from the United States and Mediterranean region with 34 isolates recently collected from California, significant genetic diversity was identified but was not correlated with the time or location of collection. Our findings suggest that CSD incidence in the commercial groves evaluated could be as high as 85% and its impact on yield and fruit quality are significant. Accepted for publication 1 March 2010. Published 26 May 2010.

Hydrophobic Particle Films Improve Tree Fruit Productivity

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 548a-548 ◽  
Author(s):  
D.M. Glenn ◽  
G. Puterka ◽  
T. Baugher ◽  
T. Unruh ◽  
S. Drake
Keyword(s):  
West Virginia ◽  
Production Systems ◽  
Fruit Size ◽  
The United States ◽  
Fruit Production ◽  
Red Color ◽  
Tree Fruit ◽  
Fruit Productivity ◽  
Leaf Level ◽  
Hydrophobic Particle

Hydrophobic particle film technology (HPF) is a developing pest control system for tree fruit production systems. Studies were established in Chile, and Washington, Pennsylvania, and West Virginia in the United States, to evaluate the effect of HPF technology on tree fruit yield and quality. Studies in Chile, Washington, and West Virginia demonstrated increased photosynthetic rate at the leaf level. Yield was increased in peaches (Chile) and apples (West Virginia), and fruit size was increased in apples (Washington and Pennsylvania). Increased red color in apple was demonstrated at all sites with reduced russetting and `Stayman' cracking in Pennsylvania. HPF technology appears to be an effective tool in reducing water and heat stress in tree fruit resulting in increased fruit quality.

Spectrophotometry Measurement of Polynuclear Aromatic Hydrocarbons in Hamilton Harbour Sediments

1991 ◽  
Vol 26 (1) ◽  
pp. 1-16 ◽  
Author(s):  
T.P. Murphy ◽  
H. Brouwer ◽  
M.E. Fox ◽  
E. Nagy
Keyword(s):  
Aromatic Hydrocarbons ◽  
Total Concentration ◽  
Coal Tar ◽  
Sediment Cores ◽  
Polynuclear Aromatic Hydrocarbons ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Near Surface ◽  
Spectrometry Analysis ◽  
Hamilton Harbour

Abstract Eighty-one sediment cores were collected to determine the extent of coal tar contamination in a toxic area of Hamilton Harbour. Over 800 samples were analyzed by a UV spectrophotometric technique that was standardized with gas chromatography/mass spectrometry analysis. The coal tar distribution was variable. The highest concentrations were near the Stelco outfalls and the Hamilton-Wentworth combined sewer outfalls. The total concentration of the 16 polynuclear aromatic hydrocarbons (PAHs) in 48,300 m3 of near-surface sediments exceeded 200 µg/g.

Sign in / Sign up

Export Citation Format

Share Document