scholarly journals Efficiency of detecting resting spores of Synchytrium endobioticum (potato wart disease) in infested sites of Ukraine and Georgia

2021 ◽  
Vol 8 (2) ◽  
pp. 36-48
Author(s):  
A. Zelya ◽  
L. Janse ◽  
J. Janse ◽  
S. Ghoghoberidze ◽  
G. Zelya ◽  
...  
Keyword(s):  
Organic Matter ◽  
Extraction Method ◽  
Sodium Iodide ◽  
Detection Efficiency ◽  
Extraction Methods ◽  
Inoculum Level ◽  
Potato Wart ◽  
Synchytrium Endobioticum ◽  
Resting Spores ◽  
High Level

Aim. To determine the occurrence and persistence of Synchytrium endobioticum, resting spore contamination in a small survey of (known infested) potato plots in Ukraine and Georgia; to compare the detection efficiency for resting spores (winter sporangia) of S. endobioticum using an extraction method, routinely applied in Ukraine, based on the use of sodium iodide (NaI) and an extraction method largely based on EPPO Standard PM 7/28(2) (2017), using kaolin and calcium chloride (CaCl2) for extraction. Methods. The examination of fields, aimed at detecting of S. endobioticum in 22 infested plots in Georgia, was conducted following the standard European Plant Protection Organisation (EPPO) phytosanitary procedure PM 3/59(3) (2017), and in Ukraine in 11 plots according to the Ukrainian Standard ‘Methodological recommendations on sampling during quarantine inspection and evaluation’ (Omeluta V P et al, 1996). Resting spores were extracted using kaolin and CaCl2 (following largely EPPO Standard PM 7/28(2) 2017), and floatation in a solution of NaI (Zelya et al, 2005), respectively. The content of soil organic matter (SOM), collected in potato wart infested plots in Ukraine and Georgia, was determined according to the ‘Method of laboratory determination of the content of organic matter’, largely following the method of Tyurin to determine total soil organic carbon (Jankauskas B et al, 2006). The persistence or decline of potato wart in infested plots was evaluated, based on our own observations and the data of the national phytosanitary services (https://dpss.gov.ua; http://agr.georgia.gov). The results of the study were subjected to statistical analysis, using Statistica 5 software. Results. It was found that under a low up to high level of organic matter in soil samples (2.0–2.7 and 3.1–3.9 %, respectively) and a moderate to high level of inoculum in soil (3–15 up to 41–65 resting spores/g soil, respectively) there were no significant differences in the efficiency of two extraction methods under consideration: the floatation in a solution of NaI and the application of kaolin and CaCl2. However, at a low number of resting spores present in soil (1–2 resting spores/g soil), the efficiency of the method using NaI decreased statistically significant by 20–30 % as opposed to the method using kaolin/CaCl2. A relatively high level of soil contamination with resting spores was found in 11 investigated plots of 4 Ukrainian regions: 41–46 resting spores/g soil in the Ivano-Frankivsk Region, 49 in the Lviv Region, 40–65 in the Zakarpattia and 52–65 Chernivtsi Regions. The majority of the 22 investigated Georgian plots showed a low inoculum level (1–7 resting spores/g soil) and only in one village their level amounted to 15 resting spores/g soil (Uchguli village, Mestia municipality). Conclusions. The method applying sodium iodide was found to be comparable to the method applying kaolin and CaCl2 under conditions of moderate to high inoculum levels (15–65 resting spores/g soil) but the latter method was more efficient under conditions of high content of organic matter and very low inoculum level (

scholarly journals An Alternative Bioassay for Synchytrium endobioticum Demonstrates the Expression of Potato Wart Resistance in Aboveground Plant Parts

2019 ◽  
Vol 109 (6) ◽  
pp. 1043-1052 ◽  
Author(s):  
B. T. L. H. van de Vossenberg ◽  
M. P. E. van Gent-Pelzer ◽  
M. Boerma ◽  
L. P. van der Gouw ◽  
T. A. J. van der Lee ◽  
...  
Keyword(s):  
Molecular Techniques ◽  
Breeding Programs ◽  
Potato Wart ◽  
Potato Varieties ◽  
Synchytrium Endobioticum ◽  
Plant Parts ◽  
Resting Spores ◽  
Plant Pathogen Interaction ◽  
Speed Up ◽  
Polymerase Chain

The obligate biotrophic chytrid species Synchytrium endobioticum is the causal agent of potato wart disease. Currently, 39 pathotypes have been described based on their interaction with a differential set of potato varieties. Wart resistance and pathotyping is performed using bioassays in which etiolated tuber sprouts are inoculated. Here, we describe an alternative method in which aboveground plant parts are inoculated. Susceptible plants produced typical wart symptoms in developing but not in fully expanded aboveground organs. Colonization of the host by S. endobioticum was verified by screening for resting spores by microscopy and by molecular techniques using TaqMan polymerase chain reaction and RNAseq analysis. When applied to resistant plants, none of these symptoms were detectable. Recognition of S. endobioticum pathotypes by differentially resistant potato varieties was identical in axillary buds and the tuber-based bioassays. This suggests that S. endobioticum resistance genes are expressed in both etiolated “belowground” sprouts and green aboveground organs. RNAseq analysis demonstrated that the symptomatic aboveground materials contain less contaminants compared with resting spores extracted from tuber-based assays. This reduced microbial contamination in the aboveground bioassay could be an important advantage to study this obligate biotrophic plant–pathogen interaction. Because wart resistance is active in both below- and aboveground organs, the aboveground bioassay can potentially speed up screening for S. endobioticum resistance in potato breeding programs because it omits the requirement for tuber formation. In addition, possibilities arise to express S. endobioticum effectors in potato leaves through agroinfiltration, thereby providing additional phenotyping tools for research and breeding. [Formula: see text] Copyright © 2019 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

scholarly journals First Report of Synchytrium endobioticum (Potato Wart Disease) Pathotype 18(T1) in Poland

Plant Disease ◽  
2014 ◽  
Vol 98 (5) ◽  
pp. 688-688 ◽  
Author(s):  
J. Przetakiewicz
Keyword(s):  
Western Europe ◽  
Plant Pathogens ◽  
Ornamental Plants ◽  
Wart Disease ◽  
Potato Wart ◽  
Synchytrium Endobioticum ◽  
Potato Wart Disease ◽  
The United Kingdom ◽  
Resting Spores ◽  
The 19Th Century

Synchytrium endobioticum (Schilb.) Perc. is a soil-borne biotrophic fungus causing potato wart disease (PWD) of cultivated potato, one of the most important crops in Poland. S. endobioticum infects epidermal cells of young potato organs, such as eyes, sprouts, young tubers, stolons, stems, leaves, and even flowers, but never roots. S. endobioticum survives in the soil as winter (resting) spores, which germinate, infect the plant, and produce secondary sporangia (summer spores); infection results in galls on the stolons and tubers, in which the pathogen multiplies. Its long persistence in soil and the severe losses it inflicts to potato crops have prompted its inclusion into the A2 quarantine list. The fungus originates from the Andean zones of South America, from where it spread to North America and Europe at the end of the 19th century. S. endobioticum was first reported in the United Kingdom in 1876. This pathogen is quite variable. The first discovery of a new pathotype in Europe occurred in former East Germany in 1941, and there have been 38 S. endobioticum pathotypes identified so far. Pathotypes 1(D1), 2(G1), 6(O1), 8(F1), and 18(T1) are of greatest relevance in Europe. In 2008, the Polish Inspectorate of Plant Health and Seed Inspection (PIORiN) collected soil samples from Mazowieckie Region in Central Poland. Microscopic examinations revealed the presence of viable resting spores of S. endobioticum in a soil sample collected from a crop plantation of ornamentals intended for export. One kilogram of soil contained an average of 300 viable spores. A bioassay (pot tests), recommended by the EPPO standard PM 3/59 (1), showed no wart symptoms because of the very low sporangium density of S. endobioticum. However, concentrating S. endobioticum inoculum by centrifugation and using for a bioassay modified Potocek's tube test (1) allowed us to obtain fresh galls with summer sporangia. The first symptoms of PWD were visible on sprouts of extremely susceptible potato genotypes 7 weeks after inoculation. To identify the pathotype of S. endobioticum, 10 differential potato cultivars (Deodara, Tomensa, Eersteling, Producent, Combi, Saphir, Delcora, Miriam, Karolin, and Ulme) were inoculated with fresh galls of S. endobioticum using the Glynne-Lemmerzahl method (2), according to EPPO standard PM 7/28. Galls were formed on all cultivars except Saphir, Karolin, and Ulme (resistant cultivars). This virulence profile was identical to that of European pathotype 18(T1) of S. endobioticum. This is the first detection in Poland of pathotype 18(T1), which is one of the most virulent pathotypes of this fungus. It should be noted that in 2004, prior to planting, the field was investigated by PIORiN and found to be free of S. endobioticum. The winter sporangia were found on the field with ornamental plants originating from Western Europe, where pathotype 18(T1) is still occurring. S. endobioticum is a classic example of the distribution of plant pathogens by man. Although an infection source has not been determined, the field was probably infested by soil connected with roots of the plantlets. This is an example of alternative ways for S. endobioticum spreading without potato as a main host. References: (1) EPPO. Bull. OEPP/EPPO Bull. 29:225, 1999. (2) EPPO. Bull. OEPP/EPPO Bull. 34:213, 2004.

scholarly journals A new method for extraction of exopolymers from activated sludges

1998 ◽  
Vol 37 (4-5) ◽  
pp. 367-370 ◽  
Author(s):  
Joana Azeredo ◽  
Rosário Oliveira ◽  
Valentina Lazarova
Keyword(s):  
Organic Matter ◽  
Organic Carbon ◽  
Total Organic Carbon ◽  
Total Protein ◽  
Extraction Method ◽  
Combined Treatment ◽  
Extraction Methods ◽  
New Method ◽  
Preliminary Extraction ◽  
Activated Sludge Systems

The exopolymeric matrix that surrounds the biomass greatly contributes to the efficiency of activated sludge systems. To study the composition of this matrix a preliminary extraction method is required. In this work three extraction methods (vapor, sonication and combined treatment by sonication and Dowex resin) were used and compared with a new method which uses glutaraldehyde. The extraction effectiveness was estimated by measuring the total protein, the total organic carbon in the extracted solutions and by determining the monosaccharide constituents of the extracted polysaccharides. The proposed new method proved to be the most suitable because it extracted a great quantity of organic matter without disrupting the cells.

scholarly journals The phytosanitary term of old sources potato wart Synchytrium endobioticum (Schilb.) Perc. in Ukraine

2020 ◽  
pp. 9-15
Author(s):  
A. Zelia ◽  
V. Hunchak ◽  
A. Melnyk ◽  
T. Andriichuk ◽  
H. Popesku ◽  
...  
Keyword(s):  
Standard Method ◽  
Sodium Iodide ◽  
Potato Cultivar ◽  
Potato Wart ◽  
Synchytrium Endobioticum ◽  
Carpathian Region ◽  
Anti Cancer ◽  
The Common ◽  
Selection Of

Goal. To do analyze phitosanitary state of wart potato sources in Carpathian region of Ukraine with determining infectious load of soil. Identify existing anti-cancer patatoes in Ukraine. Methods. The selection of soil’s samples for zoosporangia of wart pathogen was provided by standard method as per DSTU 3355-96. The sample’s selection points fixed by GPS-navigator Garmin eTrex Legend. The potato wart zoosporangia extraction was conducted by floatation in solution of sodium iodide. The solution Coomassi blue G-250 used for zoosporangia viability determining. Results. Of potato wart pathogens infectious load in Carpathian region of Ukraine 2016—2018 showed the different level of zoosporangia soil’s infection in researched areas. In the hearths of the Chernivtsi region the infectious load was reached 56—68 zoosporangia/1g in sources of Carpathian region, 42—48 zoosporangioa was in the sources of Zakarpattia region, 40—45/1g zoosporangia was determined in Ivano-Frankivsk region, 49 zoosporangia /1g was determined in town Turka Lviv region. The common (D1) pathotype and four aggressive pathotypes of wart potato: 11 (M1) - Mizhgirrya, 13 (R2) - Rachiv, 18 (Ya) - Yasynya and 22 (B1) - Bystrytsya were determined by pathotypes identification in old sources of Zakarpattia, Ivano-Frankivsk, Lviv and Chernivtsi region Conclusions. Analysis of phitosanitary state of wart potato sources in Carpathian region of Ukraine showed the different level of soil’s infectious load in researched areas. There is a common pathotype (D1) and 4 aggressive pathogen disease in Ukraine for present time. The identification of Ukrainian wart pothotypes is planned to identify by European potato cultivar test-assortment usage in perspective.

Sequence of events in the germination of the resting spore of Synchytrium endobioticum, European pathotype 2, the causal agent of potato wart disease

1986 ◽  
Vol 64 (9) ◽  
pp. 2144-2150 ◽  
Author(s):  
Michael C. Hampson
Keyword(s):  
Life History ◽  
Causal Agent ◽  
Wart Disease ◽  
Potato Wart ◽  
Synchytrium Endobioticum ◽  
Potato Wart Disease ◽  
Resting Spore ◽  
Sequence Of Events ◽  
Resting Spores ◽  
History Of

Populations of freshly harvested to 4-year-old resting spores of Synchytrium endobioticum (Schilb.) Perc. from sand compost and aqueous immersion cultures were observed, using normal and television microscopy. Germination vesicles were found in aging cultures: at 49 days in sand compost or peaking at the 11th day in aqueous immersion. Freshly harvested spores gave rise to the largest quantity of vesicles in aqueous immersion. The ability to produce vesicles fell off in rapid exponential fashion with time after spore harvest. Release of the sporangia from the enveloping vesicles and discharge of zoospores from sporangia were studied in detail through the use of videotape recordings. This is the first reported description of these phenomena in the life history of S. endobioticum.

Characterization of dissolved organic matter (DOM) extracted from soils by hot water percolation (HWP)

2010 ◽  
Vol 59 (1) ◽  
pp. 99-108 ◽  
Author(s):  
M. Takács ◽  
Gy. Füleky
Keyword(s):  
Organic Matter ◽  
Humic Acid ◽  
Natural Organic Matter ◽  
Extraction Methods ◽  
Soil Samples ◽  
Hot Water ◽  
Soil Extracts ◽  
Water Percolation ◽  
Soil Humic Acid

The Hot Water Percolation (HWP) technique for preparing soil extracts has several advantages: it is easily carried out, fast, and several parameters can be measured from the same solution. The object of this study was to examine the possible use of HWP extracts for the characterization of soil organic matter. The HPLC-SEC chromatograms, UV-VIS and fluorescence properties of the HWP extracts were studied and the results were compared with those of the International Humic Substances Society (IHSS) Soil Humic Acid (HA), IHSS Soil Fulvic Acid (FA) and IHSS Suwannee Natural Organic Matter (NOM) standards as well as their HA counterparts isolated by traditional extraction methods from the original soil samples. The DOM of the HWP solution is probably a mixture of organic materials, which have some characteristics similar to the Soil FA fractions and NOM. The HWP extracted organic material can be studied and characterized using simple techniques, like UV-VIS and fluorescence spectroscopy.

scholarly journals Evaluation of DNA Extraction Methods Developed for Forensic and Ancient DNA Applications Using Bone Samples of Different Age

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 146
Author(s):  
Catarina Xavier ◽  
Mayra Eduardoff ◽  
Barbara Bertoglio ◽  
Christina Amory ◽  
Cordula Berger ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Ancient Dna ◽  
Extraction Method ◽  
Fragment Size ◽  
Molecular Genetic ◽  
Extraction Methods ◽  
Degraded Dna ◽  
Size Analysis ◽  
Dna Fragments ◽  
Dna Amount

The efficient extraction of DNA from challenging samples, such as bones, is critical for the success of downstream genotyping analysis in molecular genetic disciplines. Even though the ancient DNA community has developed several protocols targeting small DNA fragments that are typically present in decomposed or old specimens, only recently forensic geneticists have started to adopt those protocols. Here, we compare an ancient DNA extraction protocol (Dabney) with a bone extraction method (Loreille) typically used in forensics. Real-time quantitative PCR and forensically representative typing methods including fragment size analysis and sequencing were used to assess protocol performance. We used four bone samples of different age in replicates to study the effects of both extraction methods. Our results confirm Loreille’s overall increased gain of DNA when enough tissue is available and Dabney’s improved efficiency for retrieving shorter DNA fragments that is beneficial when highly degraded DNA is present. The results suggest that the choice of extraction method needs to be based on available sample, degradation state, and targeted genotyping method. We modified the Dabney protocol by pooling parallel lysates prior to purification to study gain and performance in single tube typing assays and found that up to six parallel lysates lead to an almost linear gain of extracted DNA. These data are promising for further forensic investigations as the adapted Dabney protocol combines increased sensitivity for degraded DNA with necessary total DNA amount for forensic applications.

scholarly journals A Research on Landslides Automatic Extraction Model Based on the Improved Mask R-CNN

2021 ◽  
Vol 10 (3) ◽  
pp. 168
Author(s):  
Peng Liu ◽  
Yongming Wei ◽  
Qinjun Wang ◽  
Jingjing Xie ◽  
Yu Chen ◽  
...  
Keyword(s):  
Remote Sensing Data ◽  
Extraction Methods ◽  
New Method ◽  
Detection Accuracy ◽  
Sufficient Information ◽  
Automatic Extraction ◽  
Emergency Rescue ◽  
Feature Pyramid ◽  
High Level ◽  
Extraction Model

Landslides are the most common and destructive secondary geological hazards caused by earthquakes. It is difficult to extract landslides automatically based on remote sensing data, which is import for the scenario of disaster emergency rescue. The literature review showed that the current landslides extraction methods mostly depend on expert interpretation which was low automation and thus was unable to provide sufficient information for earthquake rescue in time. To solve the above problem, an end-to-end improved Mask R-CNN model was proposed. The main innovations of this paper were (1) replacing the feature extraction layer with an effective ResNeXt module to extract the landslides. (2) Increasing the bottom-up channel in the feature pyramid network to make full use of low-level positioning and high-level semantic information. (3) Adding edge losses to the loss function to improve the accuracy of the landslide boundary detection accuracy. At the end of this paper, Jiuzhaigou County, Sichuan Province, was used as the study area to evaluate the new model. Results showed that the new method had a precision of 95.8%, a recall of 93.1%, and an overall accuracy (OA) of 94.7%. Compared with the traditional Mask R-CNN model, they have been significantly improved by 13.9%, 13.4%, and 9.9%, respectively. It was proved that the new method was effective in the landslides automatic extraction.

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