A. C. Z. Machado, O. F. Dorigo, R. M. D. G Carneiro and J. V. De AraÚJo Filho
according to NaOH method (Stan-ton et al., 1998). Amplification of the 18S-ITS1-28S region of ri-bosomal DNA was performed using the Kit Taq PCR Master Mix (Promega) and the nematode universal primers rDNA2 (5'-TT-GATTACGTCCCTGCCCTTT-3') and rDNA1.58S (5'-ACGAGC-CGAGTGATCCACCG-3'). In a microcentrifuge tube were added 25μl of the Kit Taq PCR Master Mix, 1.5μl (0.3 microM) from each primer, 18μl water mili-Q and 4μl total DNA. The DNA was subjected to a PCR with the following specifications: 94 °C (2 min); followed by 40 cycles at 94 °C (1 min), 57 °C (1 min) and 72 °C (2
M. Elshishka, V. Peneva, S. Lazarova and S. Kumari
individuals of the Kaliště population (out of 10 with positive species-specific PCR products) were used for sequencing D2–D3 expansion segments of the 28S and ITS1 regions of ribosomal DNA. D2–D3 expansion segment of the 28S gene was amplified and sequenced using the following D2A primers: 5'-ACA AGT ACC GTG AGG GAA AGT TG-3' and D3B: 5'-TCG GAA GGA ACC AGC TAC TA-3' ( Nunn, 1992 ). Internal transcribed spacer 1 (ITS1) was amplified and sequenced using the primers BL18: 5'-CCC GTG GMT ACT ACC GAT T-3' + 5818: 5'-ACG ARC CGA GTG ATC CAC-3' ( Boutsika et al ., 2004a ). PCR
Anna Żółciak, Justyna Anna Nowakowska, Artur Pacia, Nenad Keča and Tomasz Oszako
ribosomal DNA (ITS1, ITS2, and 5.8 S rRNA Gene): its biological meaning and application in medical mycology. Communicating Current Research and Educational Topics and Trends in Applied Microbiology , 105, 783–787.
Kowalski, T. 2006. Chalara fraxinea sp. nov. associated with dieback of ash ( Fraxinus excelsior ) in Poland. Forest Pathology , 36, 264–270. DOI: 10.1111/j.1439-0329.2006.00453.x
Kowalski, T., Bartnik, C. 2010. Morphological variation in colonies of Chalara fraxinea isolated from ash ( Fraxinus excelsior L.) stems with symptoms of dieback and
During a survey on the occurrence of the plant parasitic nematodes of the family Longidoridae in Poland, 925 soil samples were taken. Longidorus distinctus was present in 10 (1.08 %) of these samples. In this Research Note we provide: 1) distribution map of these populations, 2) morphometric data, 3) sequence data for D2-D3 28S rDNA and (partial)18S-ITS1 -5.8S(partial) markers and 4) LdistFOR primer (5′-GGCTGTAAAGATATATGCGT-3’) effective in obtaining ITS1 sequence for the species. Morphometric similarities and dissimilarities with data on other published populations are discussed.
In the course of a taxonomic study of the genus Cephaloziella (Spruce) Schiffn. (Cephaloziellaceae, Marchantiophyta) in Asia, the new species Cephaloziella konstantinovae Mamontov & Vilnet, sp. nov., from the eastern regions of Russia and from the Republic of Mongolia was discovered. The new species is formally described and illustrated here. Morphologically it is similar to C. divaricata var. asperifolia (Taylor) Damsh., but differs in its leaf shape and thin-walled, inflated stem and leaf cells. The new species can be distinguished from other Cephaloziella taxa by the following characters: (i) female bracts entirely free from each other and from bracteole, (ii) perianth campanulate, (iii) cells of perianth mouth subquadrate, (iv) capsule spherical, (v) seta with 8–10 + 4–6-seriate morphology, and (vi) elaters with 1–2 spiral bands. Molecular phylogenetic analyses of nrITS1-5.8S-ITS2 and chloroplast trnL-F sequences from 63 samples (34 species, 23 genera) confirm the taxonomical status of the new species. Five specimens of C. konstantinovae form a clade placed sister to a clade of C. elachista (J. B. Jack) Schiffn. and C. rubella (Nees) Warnst.
M. E. Raphahlelo, I. Přikrylová, M. M. Matla, J. Theron and W. J. Luus-Powell
small subunit of ribosomal DNA (SSU rDNA) and the entire first internal transcribed spacer (ITS1) region were amplified in one round using S1 and IR8 primers ( Šimková et al ., 2003 ) according to protocol of Mendlová et al . (2012) and the large subunit of ribosomal DNA (LSU rDNA) region was amplified using primers C1 and D2 ( Hassouna et al ., 1984 ) using the protocol described in Mendlová et al . (2012) . Subsequently, 5 μΙ of PCR product was visualized on Gold View stained agarose gel (1%) and the remaining 20 μΙ was purified using the High Pure PCR
necessary for reaction: 12.5 μL Go Taq®, 10 μL NFW (Nuclease Free Water), and 1 μL each primer [10 mM] and 0,5 μL of cDNA from the single female specimen, totaling 25 μL per reaction was submitted to PCR at 94 °C for seven minutes; followed by 35 cycles at 94 °C for 60 seconds, 55 °C for 60 seconds, 72 °C for 60 seconds; and 72 °C for 10 minutes (Mrácek et al., 2006). The internal transcribed spacer (ITS-1) region of the ribosomal DNA sequence (rDNA) was amplified using the forward primer BL58 (5’-CCCGTCGMTACTACCGATT-3’) and reverse primer 5818 (5’-ACGARCCGAGTGATCCAC-3
K. Choudhary, A. Kumar Verma, S. Swaroop and N. Agrawal
.V., CHEN, C.H., LEIN, Y.T., LIN, H.E., DAI, C.F., WALLACE, C.C. (2004): Secondary structure and phylogenetic utility of the ribosomal internal transcribed spacer 2 (ITS2) in scleractinian corals. Zool. Stud., 43: 759 - 771
CHEN, D., WANG, G., YAO, W., NIE, P. (2007): Utility of ITS1-5.8S- ITS2 sequences for species discrimination and phylogenetic inference of two closely related bucephalid digeneans (Digenea: Bucephalidae): Dollfustrema vaneyi and Dollfustrema hefeiensis. Parasitol. Res., 101: 791 - 800. DOI: 10.1007/s00436-007-0549-0
. Mycological Research 109 (8): 853-859.
CECH, T. (1997): Phytophthora- Krankheit der Erle in Österreich. Forstschutz Aktuell 19 (20): 14-16.
COOKE, D. E. L. - DUNCAN, J. M. (1997): Phylogenetic analysis of Phytophthora species based on the ITS1 and ITS2 sequences of ribosomal DNA. Mycological Research 101: 667-677.
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ÉRSEK, T. - RIBEIRO, O. K. (2010): Mini Review Article: An Annotated List of New
on drug discrimination and learning in rhesus monkeys. Pharmacol Biochem Behav 64: 367-371.
Gupta PK, Yadav SK, Bhutia YD, Singh P, Rao P, Gujar NL, Ganesan K, Bhattacharya R. (2013). Synthesis and comparative bioeffi cacy of N-(1- phenethyl-4-piperidinyl)propionanilide (fentanyl) and its1-substituted analogs in Swiss albino mice. Med Chem Res 22: 3888-3896.
Hallenbeck JL. (2003). Palliative care perspective. Oxford University Press, Inc., New York Higashikawa Y, Suzuki S. (2008). Studies on 1-(2-phenethyl)-4-(N