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1

XLI Olimpiada WIedzy o Mleku i Mleczarstwie

100%
Budziszewska M.
Przegląd Mleczarski
|
2015
|
nr 06
2

XLII Olimpiada Wiedzy o Mleku i Mleczarstwie - Łomża 2016

86%
Budziszewska M.
Przegląd Mleczarski
|
2016
|
nr 06
3
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Analysis of the interaction between tomato torrado virus proteins using the yeast two-hybrid system

72%
Budziszewska M., Obrepalska-Steplowska A.
Journal of Plant Protection Research
|
2013
|
tom 53
|
nr 4
Ten years ago for the first time the new picorna-like virus species - Tomato torrado virus (ToTV) - was found and described on tomato plants. The isolates of this pathogen were reported in Europe, America, and Oceania including Australia. Because of its unique biological and molecular features, ToTV was classified to the new genus Torradovirus, in the Secoviridae family. In Poland, three isolates: Wal’03, Kra, and Ros ToTV were identified on greenhouse tomato cultivars. At present, the biology and the genome structure of this virus are characterised. But there is no data extending beyond the bioinformatics analyses about the function of viral proteins, polyproteins, and non-coding sequences, as well as possible interactions between viral, host and vector factors that may be important for the infection process, encapsidation, transport in plants, and transmission. In this study, we have undertaken a search for the possible protein-protein interaction of ToTV encoded proteins using the yeast two-hybrid (Y2H) system. The viral genome fragments covering full sequences for nine known proteins of ToTV were amplified using specific primers with characteristic recombination sites. This process enabled the construction of basic entry clones for each protein that further facilitated manipulations with prepared constructs using Gateway technology. Two-hybrid assays were performed in the yeast strain and tested interactions of ToTV proteins were analysed in several combinations using auxotrophy markers. Our analyses did not reveal the presence of interactions between ToTV domains. Surprisingly, no interactions were found in the case of various CP subunits as well as between CP subunits and 3A protein, that in some virus families are known to play a role in viral life cycle. This role includes virion assembly or cell-to-cell transport. The lack of interactions may be a result of the limitation of this experimental system, or suggest that these proteins may interact indirectly, or require the presence of genomic RNAs or some host factors.
4

Wstępne badania nad zdolnością czynników kodowanych przez wirusa nekrozy pomidora (ToTV) do supresji PTGS

58%
Wieczorek P., Budziszewska M., Obrepalska-Steplowska A.
Progress in Plant Protection
|
2011
|
tom 51
|
nr 3
Postranscriptional gene silencing, PTGS, is one of the important plant defense mechanisms induced during viral infection. The process is sequence-specific and results from the occurrence of dsRNA intermediate molecules generated during replication of viruses in the cytoplasm. The presence of dsRNA molecules activates the cascade of reactions in the host’s cells, leading to elimination of virus’ genetic material. In spite of its high efficiency, the PTGS can be inhibited by virus-encoded factors, usually proteins, that enable virus replication progress and spread of the pathogen in the host. In the study we used an experimental model in which stably expressed green fluorescent protein in Nicotiana benthamiana 16c plants, was locally silenced with hairpin RNA construct, whereas virus-encoded PTGS putative suppressors were delivered by infecting the plants with ToTV. We have shown that Tomato Torrado Virus-encoded factors can efficiently suppress PTGS induced in used tobacco plants infected with the ToTV. In contrast, in the leaves of N. benthamiana with induced PTGS we identified lower expression rate of the GFP when not infected with the virus. This observation indicates that ToTV encodes strong PTGS suppressor (or suppressors). Nature and mechanism of which will need to be determined in further research.
5

Complete nucleotide sequence of a Polish strain of peanut stunt virus [PSV-P] that is related to but not a typical member of subgroup I

58%
Obrepalska-Steplowska A., Budziszewska M., Pospieszny H.
Acta Biochimica Polonica
|
2008
|
tom 55
|
nr 4
731-739
Peanut stunt virus (PSV) is a common legume pathogen present worldwide. It is also infectious for many other plants including peanut and some vegetables. Viruses of this species are classified at present into three subgroups based on their serology and nucleotide homology. Some of them may also carry an additional subviral element — satellite RNA. Analysis of the full genome sequence of a Polish strain — PSV-P — associated with satRNA was performed and showed that it may be classified as a derivative of the subgroup I sharing 83.9–87.9% nucleotide homology with other members of this subgroup. A comparative study of sequenced PSV strains indicates that PSV-P shows the highest identity level with PSV-ER or PSV-J depending on the region used for analysis. Phylogenetic analyses, on the other hand, have revealed that PSV-P is related to representatives of the subgroup I to the same degree, with the exception of the coat protein coding sequence where PSV-P is clustered together with PSV-ER.
6

Analysis of expression of GAPDH gene in Solanum lycopersicum L. infected with Tomato torrado virus (ToTV)

58%
Wieczorek P., Budziszewska M., Obrepalska-Steplowska A.
Progress in Plant Protection
|
2013
|
tom 53
|
nr 2
Real-time PCR (polymerase chain reaction) is a method commonly used for analysis of genes expression. However, accurate interpretation of real-time PCR results needs additional normalization step, where the expression level of analyzed gene is corrected to the actual amount of total RNA taken for the analysis. As a normalizer any gene can be used, provided its expression is stable during the experiment. GAPDH (glyceraldehyde 3-phosphate dehydrogenase) is one of candidates that might be used as a reference gene for normalization of real-time PCR. Here we indicated a high identity of GAPDH mRNA sequence isolated from ten varieties of tomato (Solanum lycopersicum). Moreover, we reported here, that expression of the gene was rather unstable in tomato during Tomato torrado virus infection (ToTV).
7

Use of digital droplet PCR analysis in early diagnostics of winter wheat infection caused by Tilletia spp.

58%
Pieczul K., Kubiak K., Budziszewska M., Swierczynska I.
Progress in Plant Protection
|
2020
|
tom 60
|
nr 3
8

Badanie różnic w akumulacji izolatów Kra i Wal'03 wirusa nekrozy pomidora w roślinie gospodarza

58%
Budziszewska M., Wieczorek P., Obrępalska-Stęplowska A.
Progress in Plant Protection
|
2011
|
tom 51
|
nr 4
Tomato torrado virus, ToTV, is a serious pathogen of tomato causing strong necrosis on leaves and fruits, and of whole plants, leading to permanent wilting, being a cause of crops reduction. In Poland, three isolates of the ToTV were identified so far: Wal’03, Kra and Ros differing in their pathogenicity, showing that Kra and Wal’03 are the most extreme. The aim of our work was to determine whether a degree of the aggressiveness of the pathogen correlates with virus accumulation in host’s cells. Using real-time PCR approach we analyzed the relative and absolute expression of ToTV in Nicotiana benthamiana and Solanum lycopersicum plants. Analysis based on the standard curve method showed that the copy number of Kra is almost 10 000-fold higher than Wal’03 isolate in tomato tissue, whereas in tobacco, the results were normalized to reference gene: ß-actin, indicating 4-fold higher concentration of Kra in relation to Wal’03. The obtained results showed that Kra isolate was present in plants at a higher concentration that suggests its elevated replication rate responsible for higher aggressiveness.
9

Wykorzystanie techniki HRM do identyfikacji pojedynczych zmian w 3'UTR RNA polskich izolatów wirusa ToTV

58%
Wieczorek P., Budziszewska M., Obrebalska-Steplowska A.
Progress in Plant Protection
|
2010
|
tom 50
|
nr 3
Tomato torrado virus is a member of Secoviridae family, genus Torradovirus. It is a dangerous pathogen of tomato plants causing intensive necrosis of leaves and fruits, leading to plant death, and significantly decreasing production of this vegetable. In Poland three isolates of ToTV were identified: Wal’03, Kra and Ros. On the basis of the previously performed molecular and genomic analyses distinctive genetic variability was revealed within 3`UTR region of RNA1 Kra and Wal’03 isolates. On the basis of this heterogenous region a rapid protocol of PCR-HRM reaction was developed allowing the identification and differentiation of two isolates of Tomato torrado virus as well as constituting rapid test to monitor the nucleotide point mutation within this regulatory region. Since the 3`UTR region is known to play a role in the replication process hence the featured heterogeneity might have an impact on the control of viral particles accumulation.
10

Rola komorek dendrytycznych w odpowiedzi transplantacyjnej

58%
Budziszewska M., Korecka-Polak A., Korczak-Kowalska G.
Postępy Biologii Komórki
|
2009
|
tom 36
|
nr 4
745-754
11

Wirus nekrozy pomidora (Tomato torrado virus) nowy wirus przenoszony przez mączlika szklarniowego (Trialeurodes vaporariorum)

51%
Pospieszny H., Budziszewska M., Hasiow B., Obrepalska-Steplowska A., Borodynko N.
Zeszyty Problemowe Postępów Nauk Rolniczych
|
2008
|
tom 529
Dwukrotnie, w roku 2003 i 2007, z pomidora szklarniowego z objawami nekrozy liści wyizolowano wirusa sferycznego. Występowniu wirusa każdorazowo towarzyszyła obecność mączlika szklarniowego (Trialeurodes vaporariorum). Eksperymentalnie po raz pierwszy wykazano, że mączlik szklarniowy jest wektorem wirusa sferycznego. Mączlik szklarniowy przenosił wirusa bardzo efektywnie (100%). Wirus porażał zakres roślin głównie z rodziny psiankowatych. Badania w mikroskopie elektronowym wykazały, że wirus ma średnicę ok. 28 nm, występuje w soku w niskiej koncentracji w postaci pojedyńczych cząstek, a w oraganach komórkowych w postaci skupisk podobnych do kryształów. Oczyszczone preparaty wirusa sedymentowały w gradiencie gęstości sacharozy w postaci 2 stref, wynikających z 2 typów cząstek różniących się współczynnikiem sedymentacji. Genom wirusa składa się z dwóch fragmentów: RNA1 o wielkości 7800 pz i RNA2 o wielkości 5400 pz. Białko otoczki wirusowej składa się z 3 podjednostek o wielkości 35, 26 i 23 kD. Opisany w roku 2007 nowy wirus Tomato torrado virus (ToTV) wykazywał duże podobieństwo do polskiego izolatu przenoszonego przez mączlika. Na podstawie sekwencji ToTV zaprojektowano własne startery, które w RT-PCR z polskim izolatem dały produkty o wielkości ok 892 pz dla RNA1 i 573 pz dla RNA2. Produkty te poddano sekwencjonowaniu i porównanie z sekwencjami ToTV wykazało pokrewieństwo 99 i 98% odpowiednio dla RNA1 I RNA2. Podobieństwo objawów chorobowych, morfologii cząstek wirusa, organizacji genomu i wysoki stopień pokrewieństwa genetycznego pozwala uznać polski izolat wirusa sferycznego, przenoszonego przez mączlika szklarniowego, za izolat wirusa nekrozy pomidora (ToTV).
12

Wirus nekrozy pomidora [Tomato torrado virus] - nowy patogen pomidora

51%
Pospieszny H., Borodynko N., Hasiow B., Obrepalska-Steplowska A., Budziszewska M.
Progress in Plant Protection
|
2008
|
tom 48
|
nr 3
1085-1096
13

Charakterystyka morfometryczna i genetyczna polskich populacji Meloidogyne hapla oraz Meloidogyne arenaria

51%
Nowaczyk K., Dobosz R., Budziszewska M., Kornobis S., Obrepalska-Steplowska A.
Progress in Plant Protection
|
2008
|
tom 48
|
nr 1
126-130
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