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Zróżnicowanie morfologiczne i molekularne polskich izolatów nicienia Bursaphelenchus mucronatus

100%

Filipiak A.

Progress in Plant Protection

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2011

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tom 51

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nr 3

Bursaphelenchus mucronatus is morphologically and genetically closely related to B. xylophilus – the causal agent of pine wilt disease and a quarantine organism in many countries. B. mucronatus is widely distributed in Asia and Europe with two distinct European and East Asian genotypes recorded among isolated populations. The main objective of this study was to examine the morphological and molecular variation among populations of B. mucronatus isolated from geographically distant regions. Although typical morphological characters for both, European and East Asian phenotypes were observed, no clear distinction among examined populations could be made based on morphological characters only. Molecular analysis confirmed the earlier identification of examined isolates as B. mucronatus but it also did not allow clear distinction between European and East Asian genotypes among examined populations.

2

The use of real-time polymerase chain reaction with high resolution melting (real-time PCR-HRM) analysis for the detection and identification of pathogenetic and nonpathogenetic populations of the quarantine nematode Bursaphelenchus xylophilus

100%

Filipiak A.

Progress in Plant Protection

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2017

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tom 57

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nr 3

3

Grozne szkodniki porzeczki czarnej i czerwonej

100%

Filipiak A.

Warzywa

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2008

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nr 09

51-54

4

Rola i zadania Stowarzyszenia Mlynarzy RP

100%

Filipiak A.

Biuletyn Informacyjny. Centralne Laboratorium Technologii Przetwórstwa i Przechowalnictwa Zbóż

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1998

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nr 1-2

36

5

Pryszczarki w sadzie

100%

Filipiak A.

Ochrona Roślin

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2007

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tom 52

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nr 12

20-23

6

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Pathogenicity of selected isolates of the quarantine pinewood nematode Bursaphelenchus xylophilus to Scots pine (Pinus sylvestris L.)

100%

Filipiak A.

Journal of Plant Protection Research

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2015

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tom 55

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nr 4

The pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease (PWD). This nematode is considered to be an indigenous to North America and was introduced to Japan in the late 19th century. Subsequently, it has spread throughout Japan and in many other countries, China, Taiwan, and South Korea. In 1999, B. xylophilus was discovered in Portugal, and in 2008 in Spain. So far the studies have revealed that the pathogenicity of B. xylophilus varies between different isolates. The conducted study compared the pathogenicity of five isolates of B. xylophilus, originating from different parts of Japan, to 3-year-old Pinus sylvestris, and their ability to reproduce in the seedlings. The results revealed diverse virulence of B. xylophilus resulting in plant mortality. Three isolates S10, Ka4, and T4 caused 100% mortality of plants within three months while at the same time, the other two isolates, C14-5 and OKD-1 did not cause any disease symptoms on plants. After seven months, some dieback occurred on two seedlings, but similar symptoms were also found on the control plant. Moreover, a significant positive correlation was found between nematode virulence and the number of nematodes reproducing on pine seedlings.

63%

Chorazewicz J., Filipiak A.

Przegląd Geodezyjny

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2018

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tom 90

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nr 10

14

The use of the real-time PCR technique for detection of the quarantine nematode Bursaphelenchus xylophilus

63%

Filipiak A., Tomalak M.

Progress in Plant Protection

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2013

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tom 53

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nr 1

The real-time PCR (polymerase chain reaction) based on TaqMan chemistry was used to detect and distinguish Bursaphelenchus xylophilus from other wood-inhabiting nematode species of the xylophilus group. In the study reported here DNA (deoxyribonucleic acid) samples of B. xylophilus and two other most common in Europe and both morphologically and genetically similar nematode species, i.e. B. fraudulentus and B. mucronatus, were examined in detail. The study confirmed a high efficiency of the reaction primers and TaqMan probe. These components allowed detection of B. xylophilus in such small target DNA samples as 3 pg. In contrast, in none of the examined concentrations of DNA samples derived from B. mucronatus and B. fraudulentus any signal was produced. These results showed high sensitivity and specificity of the real-time PCR assay and its potential for rapid and accurate molecular identification of B. xylophilus, even in samples contaminated with DNA of other, most closely related nematode species. Therefore, we conclude that this technique could be of a great value to plant quarantine inspection.

15

Mozliwosci rozwoju turystyki i rekreacji na terenie Lesnego Kompleksu Promocyjnego Lasy Puszczy Kozienickiej

63%

Filipiak A., Luczka-Bakula W.

Ekonomia i Środowisko

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2005

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nr 1

153-167

16

Wykorzystanie modelu 3D do przeprowadzenia analizy zacienienia

63%

Chorazewicz J., Filipiak A.

Przegląd Geodezyjny

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2018

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tom 90

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nr 06

17

Efektywnosc pracy filtrow odzelaziajacych ze styropianowym zlozem plywajacym

63%

Siwiec T., Filipiak A.

Acta Scientiarum Polonorum. Formatio Circumiectus

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2003

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tom 02

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nr 2

103-116

18

Wewnątrz - i międzygatunkowa zgodność reprodukcji polskich izolatów Bursaphelenchus mucronatus oraz kwarantannowego gatunku B. xylophilus

63%

Filipiak A., Tomalak M.

Progress in Plant Protection

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2009

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tom 49

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nr 1

19

Uproszczona metoda molekularnej identyfikacji nicieni z rodzaju Bursaphelenchus, należących do grupy xylophilus, z wykorzystaniem reakcji PCR ze starterami specyficznymi

63%

Filipiak A., Tomalak M.

Progress in Plant Protection

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2010

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tom 50

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nr 1

Both native European species, Bursaphelenchus mucronatus and B. fraudulentus, are relatively common and harmless to trees. They belong to the xylophilus group, which includes also the quarantine pest B. xylophilus – the causal agent of the pine wilt disease. They can be difficult to distinguish morphologically from each other and from B. xylophilus. Therefore, reliable methods of taxonomic identification of these species are therefore of particular interest to plant quarantine services. PCR amplification with specific primers enables rapid and precise species identification, even from a single nematode. In 2004, Matsunaga and Togashi designed specific primers for B. xylophilus and B. mucronatus. The aim of this study was to design specific primers for B. fraudulentus. The specificity of the newly designed primers was tested on eight isolates of B. fraudulentus which originated from different parts in Europe (Austria, Germany, Poland, Russia and Ungarn). For all isolates, PCR amplification resulted in products of identical length of 617 bp, but no PCR products were generated for B. xylophilus and B. mucronatus. The PCR amplification with primer sets specific for B. xylophilus (XF and XR), B. mucronatus (MF and MR) and those for B. fraudulentus (FF and FR) designed in this study resulted in amplicons of different lengths (557, 210 and 617 bp, respectively), which can be easily distinguished in agarose gels.

20

The use of PCR-HRM technique for detection of the quarantine nematode Bursaphelenchus xylophilus

63%

Filipiak A., Tomalak M.

Progress in Plant Protection

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2014

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tom 54

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nr 4

High Resolution Melting Analysis (PCR-HRM – Polymerase Chain Reaction-High Resolution Melting) is a recently developed technique for fast, high-throughput post-PCR analysis of genetic mutations or variance in sequences of nucleic acid. It enables researchers to rapidly detect and categorize genetic mutations, identify new genetic variants without sequencing, or determine the genetic variation in a population prior to sequencing. PCR-HRM technique was also used to detect and distinguish Bursaphelenchus xylophilus from morphologically and genetically similar nematode species B. mucronatus. In the research reported here PCR-HRM was conducted on the rDNA of B. xylophilus (China) and Polish isolates of B. mucronatus (Mdz-01, Rad-03 and Wro-01), with the use of primers specifically designed from the ITS-1 region of B. mucronatus. The conducted study revealed that PCR-HRM is an effective essay which allows one to distinguish of B. xylophilus from B. mucronatus. This technique is very sensitive and enables detection of differences even in single nucleotides. PCR-HRM technique is also much cheaper when compared to commonly used PCR-RFLP.

Ograniczanie wyników

Zróżnicowanie morfologiczne i molekularne polskich izolatów nicienia Bursaphelenchus mucronatus

The use of real-time polymerase chain reaction with high resolution melting (real-time PCR-HRM) analysis for the detection and identification of pathogenetic and nonpathogenetic populations of the quarantine nematode Bursaphelenchus xylophilus

Grozne szkodniki porzeczki czarnej i czerwonej

Rola i zadania Stowarzyszenia Mlynarzy RP

Pryszczarki w sadzie

Pathogenicity of selected isolates of the quarantine pinewood nematode Bursaphelenchus xylophilus to Scots pine (Pinus sylvestris L.)

Grozne szkodniki maliny

Choroba wiedniecia sosny

Powszechne szkodniki maliny

Reproductive competitiveness of the pine wood nematode Bursaphelenchus mucronatus and Polish populations of B. xylophilus develop in a common environment (in vitro)

Patogenicznosc wybranych izolatow Bursaphelenchus mucronatus w stosunku do sosny

To juz 10 lat

Dane przestrzenne pozyskiwane przy pomocy BSL i ich zastosowanie w leśnictwie

The use of the real-time PCR technique for detection of the quarantine nematode Bursaphelenchus xylophilus

Mozliwosci rozwoju turystyki i rekreacji na terenie Lesnego Kompleksu Promocyjnego Lasy Puszczy Kozienickiej

Wykorzystanie modelu 3D do przeprowadzenia analizy zacienienia

Efektywnosc pracy filtrow odzelaziajacych ze styropianowym zlozem plywajacym

Wewnątrz - i międzygatunkowa zgodność reprodukcji polskich izolatów Bursaphelenchus mucronatus oraz kwarantannowego gatunku B. xylophilus

Uproszczona metoda molekularnej identyfikacji nicieni z rodzaju Bursaphelenchus, należących do grupy xylophilus, z wykorzystaniem reakcji PCR ze starterami specyficznymi

The use of PCR-HRM technique for detection of the quarantine nematode Bursaphelenchus xylophilus