scholarly journals Characterization of Verticillium dahliae and V. tricorpus Isolates from Lettuce and Artichoke

Plant Disease ◽  
2008 ◽  
Vol 92 (1) ◽  
pp. 69-77 ◽  
Author(s):  
Qing-Ming Qin ◽  
Gary E. Vallad ◽  
Krishna V. Subbarao
Keyword(s):  
Sequence Information ◽  
Root Weight ◽  
Internal Transcribed Spacer Region ◽  
Early Introduction ◽  
Reduced Height ◽  
Verticillium Tricorpus ◽  
Pathogenicity Tests ◽  
Plausible Mechanism ◽  
Polymerase Chain

Verticillium isolates collected from lettuce and artichoke were characterized for morphology, growth and pathogenicity. Several isolates were identified as Verticillium tricorpus based on morphological and cultural characteristics, including the production of dark resting mycelia, chlamydospores, microsclerotia, and yellow to orange pigmentation in culture. Compared with isolates of V. dahliae, these isolates also produced microsclerotia and conidia that were significantly larger and exhibited a distinct growth pattern at varying temperatures. Using database sequence information, primers were developed from the internal transcribed spacer region to produce a diagnostic 337-bp product specific to V. tricorpus and used to confirm the identification of isolates. Pathogenicity tests indicated that isolates of V. tricorpus were weak pathogens, causing a median disease severity (DS) of <1 (0-to-5 scale) on lettuce and artichoke. In contrast, isolates of V. dahliae consistently caused severe wilt with a median DS of >3.5 on lettuce and 5.0 on artichoke. Although lettuce and artichoke inoculated with isolates of V. tricorpus exhibited reduced height and fresh foliar and root weight, the reductions were not statistically significant, unlike in plants inoculated with isolates of V. dahliae. Lettuce co-inoculated with isolates of V. tricorpus and V. dahliae exhibited reduced symptoms of Verticillium wilt and improved growth relative to those inoculated with V. dahliae alone. The early introduction of V. tricorpus in soil-drench inoculations appeared to provide better relief from subsequent V. dahliae inoculation than when the two species were co-inoculated simultaneously using the root-dip method, suggesting competitive exclusion as a plausible mechanism. A spore-polymerase chain reaction assay developed using cultured spores directly as template and primers specific to V. tricorpus confirmed the presence of V. tricorpus on inoculated roots. This work demonstrates the potential use of V. tricorpus to directly reduce the effect of V. dahliae on lettuce and artichoke and, to our knowledge, is the first reported characterization of V. tricorpus isolates collected from lettuce and artichoke.

scholarly journals MOLECULAR CHARACTERIZATION OF A POLYGALACTURONASE INHIBITOR OF PEAR FRUIT

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 580c-580
Author(s):  
Henrik Stotz ◽  
Ann Powell ◽  
Susan Damon ◽  
Audrey Hentzen ◽  
Carl Greve ◽  
...  
Keyword(s):  
Blot Analysis ◽  
Cross Reactivity ◽  
Sequence Information ◽  
Pear Fruit ◽  
Higher Plant ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Glycosylation Sites ◽  
Polymerase Chain

Higher plant inhibitors of fungal polygalacturonases are potential contributors to plant defense. To test this hypothesis we have raised antibodies against the `Bartlett' pear fruit polygalacturonase inhibitor (PGIP) and cloned a pear fruit PGIP cDNA. The pear PGIP cDNA was isolated by polymerase chain reactions based on our amino acid and nucleotide sequence information. Sequence analysis predicts a gene product of 34.5 kD with an isoelectric point of 6.02 in agreement with our biochemical data. Seven potential glycosylation sites are consistent with the glycoprotein character of these PGIPs. Southern blot analysis suggests the presence of 1 or 2 genes in the pear genome. Northern blot analysis indicates the presence of a transcript of 1.5 kb. Western blot analysis shows cross-reactivity of the anti-pear PGIP antibody to various dicot species as well as corn.

scholarly journals Molecular Characterization of a Re-Emergent Brugia Malayi Parasite in Sri Lanka, Suggestive of a Novel Strain With Close Nucleotide Homology to Brugia Pahangi

2020 ◽  
Author(s):  
Chandana Harendra Mallawarachchi ◽  
Nilmini T. G. A. N Chandrasena ◽  
Ranjan Premarathna ◽  
S.M.N.S.M. Maleesha Mallawarachchi ◽  
Nilmini Y. I. S. Gunawardane ◽  
...  
Keyword(s):  
Sri Lanka ◽  
Its2 Region ◽  
Species Identity ◽  
Internal Transcribed Spacer Region ◽  
Region Gene ◽  
Specific Pcr ◽  
Baited Traps ◽  
Human Blood Samples ◽  
Polymerase Chain

Abstract Background Brugian filariasis has re-emerged in Sri Lanka after four decades of quiescence. As microscopy alone was insufficient for ascertaining the species identity of the re-emerged sub-periodic Brugia spp. parasite, molecular speciation was performed. The transmission dynamics of the parasite was studied by entomological procedures.Methods Human blood samples positive for Brugia spp. microfilariae (MF) (n=8) were collected and DNA extracted using ReliaPrep™ Blood DNA Miniprep System (modified). Polymerase chain reaction (PCR) was performed with pan-filarial primers specific for the internal transcribed spacer region 2 (ITS2) of the ribosomal DNA (rDNA) of MF. Results Of those tested, seven (87.5%) yielded a band at 615bp establishing the species identity of the re-emerged filarial parasite as B. malayi. Comparison of the ITS2 region gene sequences of B. malayi MF isolated from humans (n=2), dogs (n=3) and cats (n=6) with GenBank sequences revealed a higher sequence homology with B. pahangi than B. malayi, but phylogeny was closer to B. malayi. A total of 82 mosquitoes of genus Mansonia comprising of M. annulifera (65), M. uniformis (14) and M. indiana (3) were collected by cattle-baited traps. Mosquito dissections identified 17 infected mosquitoes: one M. uniformis (7.14%) and 16 M. annulifera (24.6%). The DNA extracts of all infected Mansonia mosquitoes elicited the 615bp band on pan-filarial primer specific PCR. Conclusions The re-emergent B. malayi is a genetic variant or a novel species closely related to B. malayi and B. pahangi. Mansonia spp. mosquitoes were vectors of this zoonotic variant B. malayi circulating among cats, dogs and humans in Sri Lanka.

scholarly journals Characterization of genomic PIG-A gene: a gene for glycosylphosphatidylinositol-anchor biosynthesis and paroxysmal nocturnal hemoglobinuria

Blood ◽  
1994 ◽  
Vol 83 (11) ◽  
pp. 3126-3131
Author(s):  
Y Iida ◽  
J Takeda ◽  
T Miyata ◽  
N Inoue ◽  
J Nishimura ◽  
...  
Keyword(s):  
Blood Cells ◽  
Paroxysmal Nocturnal Hemoglobinuria ◽  
Somatic Mutations ◽  
Surface Expression ◽  
Sequence Information ◽  
Chain Reaction ◽  
Nucleotide Sequence Information ◽  
Flanking Region ◽  
Polymerase Chain

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hemolytic anemia characterized by the presence of abnormal subpopulations of blood cells that are deficient in surface expression of glycosylphosphatidylinositol (GPI)-anchored proteins. Recent studies showed that the gene termed PIG-A, which participates in the first step of GPI-anchor biosynthesis, is mutated in the abnormal blood cells from patients with PNH. In this study the genomic PIG-A gene was cloned and characterized to obtain nucleotide sequence information for analyzing somatic mutations of PIG-A in patients with PNH. The PIG-A gene is at least 17 kb long and has six exons. The exon-intron boundaries and 583 bp of the 5′ flanking region were sequenced. The 5′ flanking region has no TATA-like sequence, but includes four CAAT boxes, two AP-2 sequences, and a CRE sequence, some of which are present in regions necessary for the promoter activity. We report pairs of oligonucleotide primers for polymerase chain reaction that should be useful to amplify and analyze various regions of the PIG-A gene in patients with PNH.

scholarly journals Molecular characterization of a re-emergent Brugia malayi parasite in Sri Lanka, suggestive of a novel strain with close nucleotide homology to Brugia pahangi

2020 ◽  
Author(s):  
Chandana Harendra Mallawarachchi ◽  
Nilmini T. G. A. N Chandrasena ◽  
Ranjan Premarathna ◽  
S.M.N.S.M. Maleesha Mallawarachchi ◽  
Nilmini Y. I. S. Gunawardane ◽  
...  
Keyword(s):  
Sri Lanka ◽  
Its2 Region ◽  
Species Identity ◽  
Internal Transcribed Spacer Region ◽  
Region Gene ◽  
Specific Pcr ◽  
Baited Traps ◽  
Human Blood Samples ◽  
Polymerase Chain

Abstract Background Brugian filariasis has re-emerged in Sri Lanka after four decades of quiescence. As microscopy alone was insufficient for ascertaining the species identity of the re-emerged sub-periodic Brugia spp. parasite, molecular speciation was performed. The transmission dynamics of the parasite was studied by entomological procedures.Methods Human blood samples positive for Brugia spp. microfilariae (MF) (n=8) were collected and DNA extracted using ReliaPrep™ Blood DNA Miniprep System (modified). Polymerase chain reaction (PCR) was performed with pan-filarial primers specific for the internal transcribed spacer region 2 (ITS2) of the ribosomal DNA (rDNA) of MF. Results Of those tested, seven (87.5%) yielded a band at 615bp establishing the species identity of the re-emerged filarial parasite as B. malayi. Comparison of the ITS2 region gene sequences of B. malayi MF isolated from humans (n=2), dogs (n=3) and cats (n=6) with GenBank sequences revealed a higher sequence homology with B. pahangi than B. malayi, but phylogeny was closer to B. malayi. A total of 82 mosquitoes of genus Mansonia comprising of M. annulifera (65), M. uniformis (14) and M. indiana (3) were collected by cattle-baited traps. Mosquito dissections identified 17 infected mosquitoes: one M. uniformis (7.14%) and 16 M. annulifera (24.6%). The DNA extracts of all infected Mansonia mosquitoes elicited the 615bp band on pan-filarial primer specific PCR. Conclusions The re-emergent B. malayi is a genetic variant or a novel species closely related to B. malayi and B. pahangi. Mansonia spp. mosquitoes were vectors of this zoonotic variant B. malayi circulating among cats, dogs and humans in Sri Lanka.

scholarly journals Biological and Molecular Characterization of Four Botryosphaeria Species Isolated from Pear Plants Showing Stem Wart and Stem Canker in China

Plant Disease ◽  
2014 ◽  
Vol 98 (6) ◽  
pp. 716-726 ◽  
Author(s):  
Lifeng Zhai ◽  
Meixin Zhang ◽  
Gang Lv ◽  
Xiaoren Chen ◽  
Nana Jia ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Phylogenetic Analyses ◽  
Stem Canker ◽  
Morphological Characterization ◽  
The Other ◽  
Internal Transcribed Spacer Region ◽  
Pathogenicity Tests ◽  
The Common ◽  
Stem Cankers

Pear stem wart and pear stem canker, which have been considered as two different fungal diseases caused by pathogens belonging to Botryosphaeria spp., commonly occur and cause serious damage in the main pear-producing areas in China. To identify the species of this genus infecting pear in China, 131 Botryosphaeria isolates were recovered from pear samples exhibiting symptoms collected from 20 different provinces and areas. Morphological characterization and phylogenetic analyses of the ribosomal DNA internal transcribed spacer region and the β-tubulin and EF1-α genes revealed that Botryosphaeria dothidea, B. rhodina, B. obtusa, and B. parva were associated with different pear stem wart and stem canker symptoms. Remarkably, all isolates of B. dothidea were obtained from the samples showing either stem wart or stem canker lesions; however, the isolates of the other three species were obtained only from the samples showing stem canker. Pathogenicity tests on the pear shoots showed that B. dothidea isolates could induce stem wart or stem canker lesions but all the isolates of the other three species could only induce stem cankers. However, the isolates of B. parva, B. rhodina, and B. obtusa exhibited higher virulence than that of the B. dothidea isolates on the pear fruit. Our results suggest that B. dothidea is the common causal agent for these two diseases (a pear stem wart and a pear-related stem canker), whereas B. parva, B. rhodina, and B. obtusa only cause pear stem canker diseases. To our knowledge, this study represents the first report for biological and molecular characterization of four Botryosphaeria spp. isolated from pear plants showing stem wart and stem canker in China.

scholarly journals Characterization of Xanthomonas hortorum pv. pelargonii Isolated from Geranium in Serbia

Plant Disease ◽  
2016 ◽  
Vol 100 (1) ◽  
pp. 164-170
Author(s):  
Jelica Balaž ◽  
Žarko Ivanović ◽  
Andrej Davidović ◽  
Renata Iličić ◽  
Jaap Janse ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Bacterial Blight ◽  
Gene Sequence ◽  
Chain Reaction ◽  
Pelargonium Zonale ◽  
Molecular Assays ◽  
Pathogenicity Tests ◽  
Polymerase Chain ◽  
Phenotypic Tests

Geranium leaves and stems with symptoms of bacterial blight were collected from commercial greenhouses during the last decade in Serbia. In total, 17 isolates with colony morphology typical for the genus Xanthomonas were characterized with pathogenicity, biochemical, serological, and molecular assays. All 17 isolates reacted positive in a polymerase chain reaction (PCR) using XcpM1 and XcpM2 primers specific for Xanthomonas hortorum pv. pelargonii. In pathogenicity tests on Pelargonium zonale (leaf and stem inoculation), all isolates caused typical symptoms on leaves starting 2 days after inoculation as sunken, water-soaked, irregular lesions, and 6 to 8 days after inoculation on stems as necrotic lesions also showing yellow exudate. Symptoms resulted in general wilting of inoculated plants 20 days after inoculation. Selected phenotypic tests indicated that all isolates showed the same results as described for the bacterium X. hortorum pv. pelargonii. Repetitive sequence-based PCR typing using BOX and ERIC revealed that all isolates showed two fingerprinting profiles but (GTG)5 and REP did not reveal differences. Multilocus sequence typing of partial sequences of rpoD, dnaK, fyuA, and gyrB genes of tested isolates and sequences obtained from GenBank of Xanthomonas pathovar pathotype strains did not reveal genetic variability among the isolates, showing the same gene sequence pattern.

scholarly journals Molecular Characterization of a Reemergent Brugia malayi Parasite in Sri Lanka, Suggestive of a Novel Strain

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
C. H. Mallawarachchi ◽  
T. G. A. N. Chandrasena ◽  
G. P. Withanage ◽  
R. Premarathna ◽  
S. M. N. S. M. Mallawarachchi ◽  
...  
Keyword(s):  
Sri Lanka ◽  
Brugia Malayi ◽  
Public Health Issue ◽  
Hybrid Strain ◽  
Species Identity ◽  
Internal Transcribed Spacer Region ◽  
Pairwise Divergence ◽  
The Mean ◽  
Polymerase Chain

Sri Lanka achieved elimination status for lymphatic filariasis in 2016; still, the disease remains a potential public health issue. The present study is aimed at identifying a subperiodic Brugia sp. parasite which has reemerged in Sri Lanka after four decades via molecular-based analysis. Polymerase chain reaction performed with pan-filarial primers specific for the internal transcribed spacer region-2 (ITS-2) of the rDNA of Brugia filarial parasites isolated from human, canine, and feline blood samples yielded a 615 bp band establishing the species identity as Brugia malayi. Comparison of the ITS2 sequences of the reemerged B. malayi isolates with GenBank sequences revealed a higher sequence homology with B. pahangi than B. malayi with similar phylogenetic evidence. However, the mean interspecies Kimura-2-parameter pairwise divergence between the generated Brugia sequences with B. malayi and B. pahangi was less than 3%. During the analysis of parsimony sites of the new ITS2 sequences, substitutions at A36T, A296G, T373A, and G482A made the sequences different from both B. pahangi and B. malayi suggesting the possibility of a new genetic variant or a hybrid strain of B. malayi and B. pahangi. Mosquito dissections and xenomonitoring identified M. uniformis and M. annulifera as vectors of this novel strain of B. malayi circulating among cats, dogs, and humans in Sri Lanka.

scholarly journals Characterization of genomic PIG-A gene: a gene for glycosylphosphatidylinositol-anchor biosynthesis and paroxysmal nocturnal hemoglobinuria

Blood ◽  
1994 ◽  
Vol 83 (11) ◽  
pp. 3126-3131 ◽  
Author(s):  
Y Iida ◽  
J Takeda ◽  
T Miyata ◽  
N Inoue ◽  
J Nishimura ◽  
...  
Keyword(s):  
Blood Cells ◽  
Paroxysmal Nocturnal Hemoglobinuria ◽  
Somatic Mutations ◽  
Surface Expression ◽  
Sequence Information ◽  
Chain Reaction ◽  
Nucleotide Sequence Information ◽  
Flanking Region ◽  
Polymerase Chain

Abstract Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hemolytic anemia characterized by the presence of abnormal subpopulations of blood cells that are deficient in surface expression of glycosylphosphatidylinositol (GPI)-anchored proteins. Recent studies showed that the gene termed PIG-A, which participates in the first step of GPI-anchor biosynthesis, is mutated in the abnormal blood cells from patients with PNH. In this study the genomic PIG-A gene was cloned and characterized to obtain nucleotide sequence information for analyzing somatic mutations of PIG-A in patients with PNH. The PIG-A gene is at least 17 kb long and has six exons. The exon-intron boundaries and 583 bp of the 5′ flanking region were sequenced. The 5′ flanking region has no TATA-like sequence, but includes four CAAT boxes, two AP-2 sequences, and a CRE sequence, some of which are present in regions necessary for the promoter activity. We report pairs of oligonucleotide primers for polymerase chain reaction that should be useful to amplify and analyze various regions of the PIG-A gene in patients with PNH.

scholarly journals Characterization of the Colletotrichum Species Causing Anthracnose in Andean Blackberry in Colombia

Plant Disease ◽  
2014 ◽  
Vol 98 (11) ◽  
pp. 1503-1513 ◽  
Author(s):  
Lucía Afanador-Kafuri ◽  
Alonso González ◽  
Lederson Gañán ◽  
Juan Fernando Mejía ◽  
Nadya Cardona ◽  
...  
Keyword(s):  
Colletotrichum Gloeosporioides ◽  
Morphological Characteristics ◽  
High Sensitivity ◽  
Chain Reaction ◽  
Internal Transcribed Spacer Region ◽  
Colletotrichum Spp ◽  
Polymerase Chain ◽  
Polymerase Chain Reaction Primers ◽  
Multilocus Phylogeny

Since 1992, anthracnose of Andean blackberry (Rubus glaucus) has generated losses as high as 40% for farmers in Colombia. In this study, our goal was to characterize 240 Colletotrichum isolates from Andean blackberry in eight areas of Colombia. These isolates were evaluated according to morphological characteristics, sensitivity to benomyl, pathogenicity, and genetic variability. Identification of the genus Colletotrichum was achieved by using species complex-specific polymerase chain reaction primers. A multilocus phylogeny approach was used to identify isolates to the species level with sequences from the ribosomal internal transcribed spacer region and partial sequences of the actin, β-tubulin 2, calmodulin, chitin synthase 1, glutamine synthetase, and glyceraldehyde-3-phosphate dehydrogenase genes. Most of the isolates were identified as Colletotrichum gloeosporioides sensu lato, were associated with the Castilla ecotype, showed high sensitivity to benomyl, and were highly aggressive. Isolates identified as C. acutatum sensu lato were found mainly on the Thornless ecotype, were highly resistant to benomyl, and showed intermediate aggressiveness. Only three isolates were identified as C. boninense sensu lato. The species identified included C. fructicola, C. kahawae subsp. ciggaro, C. godetiae, C. karstii, C. brassicicola, and undetermined Colletotrichum spp. This study is the first report of these species associated with anthracnose in Andean blackberry.

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