scholarly journals Molecular Characterization of Meloidogyne hispanica (Nematoda, Meloidogynidae) by Phylogenetic Analysis of Genes Within the rDNA in Meloidogyne spp.

Plant Disease ◽  
2008 ◽  
Vol 92 (7) ◽  
pp. 1104-1110 ◽  
Author(s):  
Blanca B. Landa ◽  
Juan E. Palomares Rius ◽  
Nicola Vovlas ◽  
Regina M. D. G. Carneiro ◽  
Carla M. N. Maleita ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Nematode Species ◽  
Maximum Parsimony Analysis ◽  
Biological Traits ◽  
Root Knot Nematode ◽  
Specific Pcr ◽  
Meloidogyne Spp ◽  
Species Specific ◽  
Prunus Spp

In the past, the distribution of Meloidogyne hispanica, the Seville root-knot nematode, appeared to be restricted to the southern part of Spain and Prunus spp.; however, its distribution has been confirmed to be worldwide because it occurs in all continents (Europe, Africa, Asia, Australia, and North, Central, and South America). Differentiation of M. hispanica from other Meloidogyne spp., mainly M. arenaria, can be very difficult using morphological and biological traits data. These species are quite similar and can be regularly confused in inaccurate taxonomic comparisons. In this study, species-specific polymerase chain reaction (PCR) and phylogenetic analysis of sequences from three ribosomal (r)DNA regions (18S, internal transcribed spacer [ITS]1-5.8S-ITS2, and D2-D3 of 28S) were used to characterize three M. hispanica isolates from different geographical origins (Brazil, Portugal, and Spain). Molecular analyses showed identical sequences for all three isolates for the three rDNA regions. Maximum parsimony analysis of the three rDNA regions and the species-specific PCR demonstrated and supported the differentiation of M. hispanica from M. incognita, M. javanica, and M. arenaria and from all described root-knot nematode species.

scholarly journals First Report of the Root Knot Nematode, Meloidogyne inornata, on Common Bean in Paraná State, Brazil

Plant Disease ◽  
2013 ◽  
Vol 97 (3) ◽  
pp. 431-431 ◽  
Author(s):  
A. C. Z. Machado ◽  
O. F. Dorigo ◽  
D. Mattei
Keyword(s):  
Common Bean ◽  
Nematode Species ◽  
State University ◽  
Root Knot Nematode ◽  
First Report ◽  
North Carolina State University ◽  
Adult Females ◽  
Bean Plants ◽  
Meloidogyne Spp ◽  
Species Specific

Common bean (Phaseolus vulgaris F.) is one of the most important crops in Paraná State, which is responsible for almost 10% of the Brazilian production (4). Root knot nematodes, Meloidogyne spp., are common parasites of this crop worldwide, but damage caused by Meloidogyne inornata has not been reported. During a survey of nematode species present on common bean fields in Paraná State, Brazil, galled root samples of cultivars Tuiuiú and Eldorado were submitted, in June 2012, in the Nematology Laboratory from IAPAR, collected in the municipalities of Araucária (25°35′34″S, 49°24′36″W) and Santana do Itararé (23°45′18″S, 49°37′44″W). Plants did not exhibit any above-ground symptoms. The specimens were identified through perineal patterns and esterase phenotypes of 20 adult females extracted from dissected roots (2,3). The population densities observed in the samples were 140 and 700 J2 and eggs per gram of roots, respectively, for both samples. Characteristics were consistent with those described for M. inornata. For example, perineal patterns of M. inornata showed a high dorsal arch, with smooth to wavy striae, similar to those of M. incognita; but no punctate markings between anus and tail terminus were observed. However, from the esterase electrophoresis we obtained the I3 (Rm = 0.83, 1.15, and 1.32) phenotype, typical of M. inornata, a species-specific phenotype used to differentiate this species from M. incognita (1). Moreover, the excretory pore of adult females was located 32.1 (± 5.4) μm from the anterior end, consistent with the M. inornata description (25 to 53 μm) (1). To the best of our knowledge, this is the first report of M. inornata parasitizing common bean roots. This finding has great importance for Brazilian agriculture, since this nematode may damage common bean plants and become an additional problem for this crop. Additional work is necessary in order to elucidate the losses caused by M. inornata on common bean. References: (1) R. M. D. G. Carneiro et al. Nematology 10:123, 2008. (2) P. R. Esbenshade and A. C. Triantaphyllou J. Nematol. 22:10, 1990. (3) K. M. Hartman and J. N. Sasser. Page 115 in: An Advanced Treatise on Meloidogyne, Volume II Methodology. K. R. Barker et al., eds. Raleigh: North Carolina State University Graphics, 1985. (4) MAPA. Feijão, Ministério da Agricultura, Brasil. Retrieved from http://www.agricultura.gov.br/vegetal/culturas/feijao September 05, 2012.

scholarly journals Morphological, biochemical, and molecular characterization of Meloidogyne spp. populations from Brazilian soybean production regions

2017 ◽  
Vol 47 (5) ◽  
Author(s):  
Camilla Martins de Oliveira ◽  
Ismail Teodoro Souza Junior ◽  
Jerônimo Vieira de Araújo Filho ◽  
Marcos Augusto de Freitas ◽  
Mara Rúbia da Rocha ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Nematode Species ◽  
Economic Importance ◽  
Root Knot Nematode ◽  
Mato Grosso ◽  
Great Economic Importance ◽  
Meloidogyne Spp ◽  
Perineal Region ◽  
Mixed Populations

ABSTRACT: Soybean is a commodity of great economic importance worldwide, particularly in Brazil, world’s second largest producer. Nematodes, especially those of the Meloidogyne genus, severely limit productivity. Identification of nematode species is important for effective soybean management. Here, 26 populations of root-knot nematode (Meloidogyne spp.) from 15 municipalities in the states of Bahia, Mato Grosso, Goias, and Minas Gerais were characterized based on the morphology of the female perineal region, esterase profile, and identification based on amplification of specific regions of the population genome. Among the Meloidogyne spp. populations obtained, M. incognita and M. javanica, were identified. No mixed populations were present in the samples. Diagnosis based on molecular analysis was shown to be reliable and the fastest for characterization of nematode populations compared to other methods analyzed.

scholarly journals Development and Validation of a Nested-PCR-Denaturing Gradient Gel Electrophoresis Method for Taxonomic Characterization of Bifidobacterial Communities

2003 ◽  
Vol 69 (11) ◽  
pp. 6380-6385 ◽  
Author(s):  
R. Temmerman ◽  
L. Masco ◽  
T. Vanhoutte ◽  
G. Huys ◽  
J. Swings
Keyword(s):  
Nested Pcr ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Temporal Changes ◽  
Rrna Gene ◽  
Specific Pcr ◽  
Gradient Gel Electrophoresis ◽  
Species Specific ◽  
Taxonomic Characterization

ABSTRACT The taxonomic characterization of a bacterial community is difficult to combine with the monitoring of its temporal changes. None of the currently available identification techniques are able to visualize a “complete” community, whereas techniques designed for analyzing bacterial ecosystems generally display limited or labor-intensive identification potential. This paper describes the optimization and validation of a nested-PCR-denaturing gradient gel electrophoresis (DGGE) approach for the species-specific analysis of bifidobacterial communities from any ecosystem. The method comprises a Bifidobacterium-specific PCR step, followed by purification of the amplicons that serve as template DNA in a second PCR step that amplifies the V3 and V6-V8 regions of the 16S rRNA gene. A mix of both amplicons is analyzed on a DGGE gel, after which the band positions are compared with a previously constructed database of reference strains. The method was validated through the analysis of four artificial mixtures, mimicking the possible bifidobacterial microbiota of the human and chicken intestine, a rumen, and the environment, and of two fecal samples. Except for the species Bifidobacterium coryneforme and B. indicum, all currently known bifidobacteria originating from various ecosystems can be identified in a highly reproducible manner. Because no further cloning and sequencing of the DGGE bands is necessary, this nested-PCR-DGGE technique can be completed within a 24-h span, allowing the species-specific monitoring of temporal changes in the bifidobacterial community.

scholarly journals Isolation and Characterization of Brachyspira spp. from Dogs in the Czech Republic

2010 ◽  
Vol 79 (3) ◽  
pp. 437-442
Author(s):  
Daniel Šperling ◽  
František Čada ◽  
Alois Čížek
Keyword(s):  
Czech Republic ◽  
Biochemical Activity ◽  
Animal Shelters ◽  
The Czech Republic ◽  
Specific Pcr ◽  
Isolation And Characterization ◽  
High Prevalence ◽  
Species Specific

The objectives of this study were to establish the prevalence of intestinal Spirochetes of the genusBrachyspirain Czech dogs and to determine the susceptibility of obtainedB. pilosicoliisolates to selected antibacterial substances. Spirochetes were diagnosed microscopically in 23 out of 1139 samples of dogs’ excrements, primarily intended for a parasitological testing. The cultivation of positive samples provided 10 brachyspira isolates, which were, on the basis of their biochemical activity and the results of the species-specific PCR, identified asB. pilosicoli(9 isolates) andB. hyodysenteriae(1 isolate). These dogs came from households. All the 7 tested isolatesB. pilosicoliwere sensitive to metronidazole and doxycycline, uniformly resistant to erythromycin, partly sensitive to cefazoline, lincomicine and ampicilline except for one isolate ofB. pilosicoli, which was resistant to ampicilline. The second part of study was focused on dogs with diarrhoea that came from animal shelters, where a high prevalence of 58% (10/17) ofB. pilosicoliwas found.

A simple method for discriminating Bursaphelenchus xylophilus and B. mucronatus by species-specific polymerase chain reaction primer pairs

Nematology ◽  
2004 ◽  
Vol 6 (2) ◽  
pp. 273-277 ◽  
Author(s):  
Koji Matsunaga ◽  
Katsumi Togashi
Keyword(s):  
Pcr Amplification ◽  
Nematode Species ◽  
Bursaphelenchus Xylophilus ◽  
Polymerase Chain Reaction Primer ◽  
Dna Fragments ◽  
Simple Method ◽  
Specific Pcr ◽  
Pcr Products ◽  
Specific Pcr Primer ◽  
Species Specific

Abstract Two species-specific PCR primer pairs were developed for identifying the two nematode species, Bursaphelenchus xylophilus and B. mucronatus. The primer pairs were developed from the sequence of ribosomal DNA (rDNA) repeats to produce DNA fragments of different lengths by PCR amplification. The DNA fragments for B. mucronatus and B. xylophilus were 210 bp and 557 bp, respectively. When mixed, neither primer pair inhibited the PCR amplification of the other. Five isolates of B. xylophilus and four isolates of B. mucronatus showed different band profiles of PCR products between the two species, but identical profiles among isolates of the same species.

scholarly journals Species-Specific PCR Assays for Differentiating Heterodera filipjevi and H. avenae

Plant Disease ◽  
2013 ◽  
Vol 97 (12) ◽  
pp. 1611-1619 ◽  
Author(s):  
Guiping Yan ◽  
Richard W. Smiley ◽  
Patricia A. Okubara ◽  
Andrea M. Skantar
Keyword(s):  
Pacific Northwest ◽  
Management Practices ◽  
Pcr Amplification ◽  
Nematode Species ◽  
Heterodera Avenae ◽  
Cyst Nematodes ◽  
Specific Pcr ◽  
The Pacific ◽  
Pcr Assays ◽  
Species Specific

Heterodera avenae and H. filipjevi are economically important cyst nematodes that restrict production of cereal crops in the Pacific Northwest United States and elsewhere in the world. Identification of these two species is critical for recommending and implementing effective management practices. Primers were designed from the internal transcribed spacer (ITS) regions of H. avenae and H. filipjevi ribosomal DNA. The primers were highly specific when examined on target isolates but did not amplify DNA from nontarget Heterodera, Globodera, Meloidogyne, Pratylenchus, and other nematode species tested. Polymerase chain reaction (PCR) and amplification conditions were established, and H. avenae and H. filipjevi were clearly distinguished by PCR fragments of 242 and 170 bp, respectively. Robust PCR amplification was achieved with DNA extracted from a single egg or second-stage juvenile (J2) using a laboratory-made worm lysis buffer, and DNA from 0.5 egg or J2 using a commercial kit. The PCR assays were successfully employed for differentiation of H. filipjevi and H. avenae populations collected from eight locations in three Pacific Northwest states. This is the first report of a species-specific ITS PCR assay to detect and identify H. filipjevi. The assays for both species will enhance diagnosis of cereal cyst nematode species in infested fields.

scholarly journals CHARACTERIZATION OF RESISTANCE TO ROOT-KNOT NEMATODES IN SWEETPOTATO

HortScience ◽  
2005 ◽  
Vol 40 (3) ◽  
pp. 868e-869
Author(s):  
J.A. Thies
Keyword(s):  
Meloidogyne Incognita ◽  
Ipomoea Batatas ◽  
Nematode Species ◽  
Plant Introduction ◽  
Root Knot Nematode ◽  
Race 1 ◽  
Race 2 ◽  
The U.S ◽  
Greenhouse Tests

Thirteen sweetpotato (Ipomoea batatas) genotypes were characterized for resistance to Meloidogyne incognita, M. javanica, M. hapla, and M. arenaria races 1 and 2 in greenhouse tests. The following sweetpotato genotypes representing a range of reactions to M. incognita were evaluated: U.S. Plant Introduction (PI) 399163 (highly resistant = HR), Sumor (HR), Nemagold (HR), Excel (HR), Tinian (HR), Hernandez (resistant = R), Jewel (R), Regal (R), Porto Rico (intermediate = I), Centennial (susceptible = S), Georgia Jet (S), Sulfur (S), and Beauregard (S). Meloidogyne incognita was most pathogenic to sweetpotato of the four Meloidogyne spp. evaluated in these studies. The U.S. Plant Introduction (PI) 399163 and Sumor were resistant to M. incognita in all tests. Only two genotypes, Beauregard and Porto Rico, were susceptible to M. javanica. All genotypes evaluated were resistant to M. hapla, M. arenaria race 1, and M. arenaria race 2. Sumor, U.S. PI 399163, and Nemagold appear to provide the highest levels of resistance against the four Meloidogyne spp. used in these studies. Since M. incognita is the most commonly occurring root-knot nematode species in sweetpotato growing areas of the southern U.S. and is pathogenic to most of the commonly grown sweetpotato cultivars, efforts to develop resistant cultivars that have desirable horticultural characteristics for the U.S. market should be directed toward this root-knot nematode species.

scholarly journals Molecular Characterization of the Toxic Cyanobacterium Cylindrospermopsis raciborskii and Design of a Species-Specific PCR

2000 ◽  
Vol 66 (1) ◽  
pp. 332-338 ◽  
Author(s):  
Kim M. Wilson ◽  
Mark A. Schembri ◽  
Peter D. Baker ◽  
Christopher P. Saint
Keyword(s):  
Cylindrospermopsis Raciborskii ◽  
Morphological Identification ◽  
Common Component ◽  
Climatic Regions ◽  
Specific Pcr ◽  
Species Specific ◽  
Genetic Profiles ◽  
Sequence Profiles ◽  
Toxic Bloom

ABSTRACT Cylindrospermopsis raciborskii is a toxic-bloom-forming cyanobacterium that is commonly found in tropical to subtropical climatic regions worldwide, but it is also recognized as a common component of cyanobacterial communities in temperate climates. Genetic profiles of C. raciborskii were examined in 19 cultured isolates originating from geographically diverse regions of Australia and represented by two distinct morphotypes. A 609-bp region of rpoC1, a DNA-dependent RNA polymerase gene, was amplified by PCR from these isolates with cyanobacterium-specific primers. Sequence analysis revealed that all isolates belonged to the same species, including morphotypes with straight or coiled trichomes. Additional rpoC1 gene sequences obtained for a range of cyanobacteria highlighted clustering of C. raciborskii with other heterocyst-producing cyanobacteria (orders Nostocales andStigonematales). In contrast, randomly amplified polymorphic DNA and short tandemly repeated repetitive sequence profiles revealed a greater level of genetic heterogeneity amongC. raciborskii isolates than did rpoC1 gene analysis, and unique band profiles were also found among each of the cyanobacterial genera examined. A PCR test targeting a region of therpoC1 gene unique to C. raciborskii was developed for the specific identification of C. raciborskiifrom both purified genomic DNA and environmental samples. The PCR was evaluated with a number of cyanobacterial isolates, but a PCR-positive result was only achieved with C. raciborskii. This method provides an accurate alternative to traditional morphological identification of C. raciborskii.

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