scholarly journals First Report on Infection of Eucalyptus pellita Seeds by Ralstonia solanacearum

2020 ◽  
Vol 3 (1) ◽  
pp. 94
Author(s):  
Bayo A. Siregar ◽  
Abdul Gafur ◽  
Pranita Nuri ◽  
H. Halimah ◽  
Budi Tjahjono ◽  
...  
Keyword(s):  
Bacterial Wilt ◽  
Molecular Detection ◽  
Direct Detection ◽  
Selective Media ◽  
First Report ◽  
Eucalyptus Pellita ◽  
Bacterial Inoculum ◽  
Pcr Method ◽  
Species Specific ◽  
Physical And Chemical

Bacterial wilt is one of major threats to eucalyptus plantations which may cause significant losses. Until now, study about bacterial wilt on Eucalyptus pellita in Indonesia has been very limited, especially about the presence of the pathogen on or in the seeds. This study aims to provide evidence of the existence of the R. solanacearum bacterium on or in E. pellita seeds. Detection of seed-borne bacteria is determined by several approaches such as (i) direct detection using universal and selective media in the laboratory, (ii) the nursery test, and (iii) species-specific molecular detection. The results of our study indicate that R. solanacearum can be detected from eucalyptus seeds using universal and selective media in the laboratory, nursery test, and molecular-based detection using the Enrichment PCR method. The bacterial inoculum is also proven to exist both on the surface of and inside the eucalyptus seeds. This is the first report that R. solanacearum is a seed-borne pathogen in E. pellita seeds. Previous studies in different agricultural systems show that the effective method used to control the pathogen is through seed treatments using biological, physical, and chemical approaches.

Occurrence of the Lance Nematode Hoplolaimus stephanus Infecting Bentgrass Agrostis stolonifera in Georgia, USA

2021 ◽  
Author(s):  
Ganpati B Jagdale ◽  
Gema Takbir Takbir Nugraha ◽  
Katherine Martin ◽  
Alfredo D D Martinez-Espinoza ◽  
Abolfazl Hajihassani
Keyword(s):  
South Carolina ◽  
Dna Sequence ◽  
Creeping Bentgrass ◽  
Agrostis Stolonifera ◽  
High Population ◽  
Specific Primer ◽  
Sequence Analyses ◽  
First Report ◽  
Pcr Method ◽  
Species Specific

A high population of lance nematodes Hoplolaimus spp. were found associated with creeping bentgrass (Agrostis stolonifera L.) in May 2019 in Georgia, USA. The nematode was pathogenic to bentgrass as its population increased by over 3-fold 180 days after inoculation under greenhouse conditions. Morphological measurements of body and stylet lengths of both mature females and males were similar to a grass population of H. stephanus from South Carolina. DNA sequence analyses of the D1-D3 expansion segments of the 28s gene identified the nematode as H. stephanus. The DNA sequence of the nematode was 99.7% identical to a H. stephanus isolate from South Carolina. Also, the PCR method using a species-specific primer set confirmed the identity of H. stephanus. To the best of our knowledge, this is the first report of H. stephanus Sher, 1963, infecting creeping bentgrass in Georgia.

scholarly journals First Report of Cucurbit chlorotic yellows virus infecting Cucumber Plants in Spain

Plant Disease ◽  
2021 ◽  
Author(s):  
Robert Chynoweth ◽  
Daniel Jimenez ◽  
Daniele Liberti ◽  
Daniel Bellon-Dona ◽  
Alejandro Carralero ◽  
...  
Keyword(s):  
Molecular Detection ◽  
Geographical Area ◽  
Rdrp Gene ◽  
Rt Pcr ◽  
Specific Primers ◽  
First Report ◽  
Lettuce Infectious Yellows Virus ◽  
Pcr Method ◽  
Beet Pseudo Yellows Virus ◽  
Cucurbit Chlorotic Yellows Virus

During the winter 2018, symptoms of leaf chlorotic spots (Figure 1) followed by symptoms of leaf interveinal chlorosis (Figure 2) and severe chlorosis in basal leaves were observed in cucumber cv Laredo (Cucumis sativus) plants in three separated greenhouses, sited in distinct locations in southern Spain. In all cases, Bemisia tabaci populations were observed on infected plants. The symptomology observed was similar to that caused by whitefly transmitted Cucurbit yellow stunting disorder virus (CYSDV, genus Crinivirus, family Closteroviridae), which is usually found infecting cucumber plants in this geographical area (1). Samples from four different cucumber plants of distinct greenhouses were collected and tested for the presence of CYSDV. Total RNA was extracted from the samples using the NucleoSpin RNA Plant kit (Macherey-Nagel, Germany). Molecular detection of CYSDV was performed using the multiplex and degenerate primer RT-PCR method (2), specific to the region of the highly conserved RNA-dependent RNA polymerase (RdRp) gene of criniviruses, which also detects other criniviruses such as Lettuce infectious yellows virus (LIYV) and Beet pseudo-yellows virus (BPYV). Results indicated that the viral species CYSDV, LIYV and BPYV were not detected in the four cucurbit plant samples. In 2004, an emergent crinivirus (Cucurbit chlorotic yellows virus, CCYV), inducing symptoms similar to those caused by CYSDV, was described infecting cucurbits in Japan (3). Recently, CCYV was detected in 2011 in Greece (4) and in 2014 in Egypt (5) and Saudi Arabia (6). Therefore, the four RNA samples were tested for the presence of the CCYV by a RT-PCR method previously described (7). Specific primers were designed to amplify 336 nt of the capsid protein (CP) gene and 680 nt of the RdRp gene, located on CCYV genomic RNA 1 and RNA 2, respectively. In all cases, clear cDNA bands of both expected sizes were detected for each cucumber sample that were then purified and sequenced via Sanger technology. BLAST analysis of those sequences showed 99% identity with the nucleotide sequence of the CP and RpRd genes from the CCYV isolates from Greece (LT992911, LT992910), China (KY400633.1, KX118632) and Taiwan (JF502222). To our knowledge, this is the first report of CCYV infecting cucurbits in Spain. Probably CCYV has been spread throughout the Mediterranean basin, remaining undetected due to the yellowing symptom similarities between CYSDV and CCYV. Detection of the emergent virus CCYV in Spain represents a new threat for the horticultural area of southern Europe.

scholarly journals Diversity of Ralstonia pseudosolanacearum, the causal agent of bacterial wilt on Eucalyptus pellita in Indonesia

2021 ◽  
Vol 22 (6) ◽  
Author(s):  
Bayo Alhusaeri Siregar ◽  
Giyanto Giyanto ◽  
Sri Hendrastuti Hidayat ◽  
Iskandar Zulkarnain Siregar ◽  
Budi Tjahjono
Keyword(s):  
Bacterial Wilt ◽  
Species Complex ◽  
Phenotypic Diversity ◽  
Morphological Characteristics ◽  
Genotypic Diversity ◽  
Causal Agent ◽  
Gene Sequences ◽  
Eucalyptus Pellita ◽  
Ralstonia Pseudosolanacearum ◽  
Species Specific

Abstract. Siregar BA, Giyanto, Hidayat SH, Siregar IZ, Tjahjono B. 2021. Diversity of Ralstonia pseudosolanacearum, the causal agent of bacterial wilt on Eucalyptus pellita in Indonesia. Biodiversitas 22: 2538-2545. The Ralstonia species complex was initially classified into five races and five biovars but the classification could not accommodate the isolates' phylogenetic history or geographic origins. A phylotype and sequevar system is based on the geographic distribution and characteristics of endoglucanase (egl) and hypersensitive response and pathogenicity (hrp) gene sequences. This study aims to describe pathogen diversity of the causal agent of bacterial wilt on Eucalyptus pellita F. Muell. Pathogens were isolated from wilting seedlings and trees in several Eucalyptus plantations. The phenotypic diversity analysis included biovar, exopolysaccharide quantification and virulence tests, while genotypic diversity included phylotypes and sequevar determination based on egl gene sequences. A total of 35 strains were isolated from the field and nurseries of Eucalyptus in various locations. All isolates were confirmed as Ralstonia species complex based on morphological characteristics and molecular studies using species-specific primers. These isolates were dominantly classified as biovar 3 and 4 and had a high variation on virulence and EPS production. Based on the egl sequence's alignment, 29 strains of Phylotype I are grouped into four sequevar references (sequevars 14, 17, 18, 30) and new a sequevar 58. This study shows that strains of R. pseudosolanacearum causing bacterial wilt on E. pellita in Indonesia have high phenotypic and genotypic diversities.

scholarly journals First Report of Chlamydia abortus in Farmed Fur Animals

2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Zhaocai Li ◽  
Ping Liu ◽  
Xiaoan Cao ◽  
Zhongzi Lou ◽  
Kinga Zaręba-Marchewka ◽  
...  
Keyword(s):  
Economic Loss ◽  
23S Rrna ◽  
Specific Gene ◽  
First Report ◽  
Veterinary Importance ◽  
Potential Risks ◽  
Species Specific ◽  
Fur Animals ◽  
Globally Distributed ◽  
Raccoon Dogs

Chlamydia (C.) abortus, a globally distributed obligate intracellular bacterium, has attracted increasing interest according to its veterinary importance and zoonotic nature. C. abortus can infect a variety of animals and cause foetal loss in livestock resulting in economic loss. In this study, the samples collected from two farms of foxes (n=20), raccoon dogs (n=15) and minks (n=20), were investigated by Chlamydiaceae- and Chlamydia species-specific real-time PCR. The results showed that all the tested foxes (20/20) and raccoon dogs (15/15) harbored Chlamydia spp., while 5% of minks (1/20) were positive for Chlamydia spp. C. abortus was identified in all positive samples as the dominant Chlamydia species, with C. pecorum DNA coexistence in some of the rectal samples (7/20) taken from foxes. Phylogenetic analysis based on specific gene fragments of 16S rRNA, IGS-23S rRNA, and ompA revealed that all sequences obtained in this study were assigned to the Chlamydiaceae family with high similarity to C. abortus S26/3 and B577 previously identified in ruminants. This is the first report confirming that farmed foxes, raccoon dogs, and minks carry C. abortus. Further studies are needed to fully elucidate the epidemiology and pathogenicity of this pathogen in farmed fur animals as well as the potential risks to public health.

scholarly journals PseudomonasDiversity in Crude-Oil-Contaminated Intertidal Sand Samples Obtained after thePrestigeOil Spill

2010 ◽  
Vol 77 (3) ◽  
pp. 1076-1085 ◽  
Author(s):  
Magdalena Mulet ◽  
Zoyla David ◽  
Balbina Nogales ◽  
Rafael Bosch ◽  
Jorge Lalucat ◽  
...  
Keyword(s):  
Carbon Sources ◽  
Species Level ◽  
Gene Library ◽  
Direct Plating ◽  
Selective Media ◽  
Dna Library ◽  
Culture Independent ◽  
Northwestern Spain ◽  
Pcr Method ◽  
Culture Dependent

ABSTRACTThe Galicia seashore, in northwestern Spain, was one of the shorelines affected by thePrestigeoil spill in November 2002. The diversity of autochthonousPseudomonaspopulations present at two beaches (Carnota municipality) was analyzed using culture-independent and culture-dependent methods. The first analysis involved the screening of anrpoDgene library. The second involved the isolation of 94Pseudomonasstrains that were able to grow on selective media by direct plating or after serial enrichments on several carbon sources: biphenyl, gentisate, hexadecane, methylnaphthalene, naphthalene, phenanthrene, salicylate, xylene, and succinate. Eight denitrifyingPseudomonasstrains were also isolated by their ability to grow anaerobically with nitrate. The calculated coverage index forPseudomonasspecies was 89% when clones and isolates were considered together, and there were 29 phylospecies detected. The most abundant were members of the speciesP. stutzeri,P.putida,P. anguilliseptica, andP. oleovorans. Thirty-one isolates could not be identified at the species level and were considered representatives of 16 putative novelPseudomonasspecies. One isolate was considered representative of a novelP. stutzerigenomovar. Concordant results were obtained when the diversities of the cloned DNA library and the cultured strains were compared. The clone library obtained by therpoDPCR method was a useful tool for evaluatingPseudomonascommunities and also for microdiversity studies ofPseudomonaspopulations.

scholarly journals MOLECULAR DETECTION AND CHARACTERIZATION OF STAPHYLOCOCCUS AUREUS ISOLATED FROM RAW MILK SOLD IN DIFFERENT MARKETS OF BANGLADESH

2017 ◽  
Vol 14 (2) ◽  
pp. 277-282 ◽  
Author(s):  
M. A. Islam ◽  
S. M. L. Kabir ◽  
M. T. Rahman
Keyword(s):  
Molecular Detection ◽  
Culture Media ◽  
Raw Milk ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Selective Media ◽  
Milk Samples ◽  
Antimicrobial Sensitivity ◽  
Antimicrobial Sensitivity Test

The study was intended for molecular detection of S. aureus isolated from raw cow’s milk. A total of 20 milk samples were collected from different upazila markets of Jamalpur, Tangail, Kishoreganj and Netrokona districts of Bangladesh. Milk samples were cultured onto various culture media for the isolation of bacteria. The isolated bacteria were identified by studying cultural properties on different selective media, biochemical tests, and finally by PCR. Out of 20 samples, 15 (75%) milk samples were found to be positive for S. aureus. S. aureus specific 16S rRNA gene was amplified from all isolates and identified as S. aureus. Antimicrobial sensitivity test was carried out to ascertain the susceptibility of the organism to various antibiotics. Its results showed that the S. aureus isolates were resistant to amoxicillin (100%), erythromycin (73.33%) and tetracycline (73.33%) but sensitive to azithromycin (93.33%), ciprofloxacin (93.33%), gentamicin (100%), norfloxacin (86.67%) and streptomycin (86.67%).

scholarly journals First Report of Phlebotomine Sand Flies (Diptera: Psychodidae) in Kansas and Missouri, and a PCR Method to Distinguish Lutzomyia shannoni From Lutzomyia vexator

2012 ◽  
Vol 49 (6) ◽  
pp. 1460-1465 ◽  
Author(s):  
Ju-Lin Weng ◽  
Samantha L. Young ◽  
David M. Gordon ◽  
David Claborn ◽  
Christine Petersen ◽  
...  
Keyword(s):  
Sand Flies ◽  
Phlebotomine Sand Flies ◽  
First Report ◽  
Pcr Method

scholarly journals First Report of Bacterial Wilt of Amaranth (Amaranthus cruentus) Caused by Ralstonia solanacearum in Benin

Plant Disease ◽  
2019 ◽  
Vol 103 (3) ◽  
pp. 578-578 ◽  
Author(s):  
R. Sikirou ◽  
M.-E. E. A. Dossoumou ◽  
B. Zocli ◽  
V. Afari-Sefa ◽  
J. Honfoga ◽  
...  
Keyword(s):  
Bacterial Wilt ◽  
Ralstonia Solanacearum ◽  
Amaranthus Cruentus ◽  
First Report

scholarly journals First Report of Orange Rust of Sugarcane Caused by Puccinia kuehnii in Colombia

Plant Disease ◽  
2012 ◽  
Vol 96 (1) ◽  
pp. 143-143 ◽  
Author(s):  
M. Cadavid ◽  
J. C. Ángel ◽  
J. I. Victoria
Keyword(s):  
Internal Transcribed Spacer ◽  
Pcr Primers ◽  
The United States ◽  
Optical Microscope ◽  
First Report ◽  
5.8S Rdna ◽  
Specific Pcr ◽  
Plant Pathol ◽  
Species Specific ◽  
New Cultivars

Symptoms of sugarcane orange rust were first observed in July 2010 on sugarcane (interspecific hybrid of Saccharum L. species) cv. CC 01-1884 planted in the La Cabaña Sugar Mill, Puerto Tejada, Colombia. Morphological features of uredinial lesions and urediniospores inspected with an optical microscope and scanning electron microscopy were distinct from common rust of sugarcane caused by Puccinia melanocephala Syd. & P. Syd., revealing spores identical morphologically to those described for the fungus P. kuehnii (Kruger) E. Butler, causal agent of sugarcane orange rust (1,3). Uredinial lesions were orange and distinctly lighter in color than pustules of P. melanocephala. Urediniospores were orange to light cinnamon brown, mostly ovoid to pyriform, variable in size (27.3 to 39.2 × 16.7 to 21.2 μm), with pronounced apical wall and moderately echinulate with spines evenly distributed. Paraphyses, telia, and teliospores were not observed. Species-specific PCR primers designed from the internal transcribed spacer (ITS)1, ITS2, and 5.8S rDNA regions of P. melanocephala and P. kuehnii were used to differentiate the two species (2). The primers Pm1-F and Pm1-R amplified a 480-bp product from P. melanocepahala DNA in leaf samples with symptoms of common rust. By contrast, the primers Pk1-F and Pk1-R generated a 527-bp product from presumed P. kuehnii DNA in leaf samples with signs of orange rust, confirming the identity as P. kuehnii. The Centro de Investigación de la Caña de Azúcar de Colombia (Cenicaña) started a survey of different cultivars in nurseries and experimental and commercial fields in the Cauca River Valley and collected leaf samples for additional analyses. Experimental cvs. CC 01-1884, CC 01-1866, and CC 01-1305 were found to be highly susceptible to orange rust and were eliminated from regional trials, whereas commercial cvs. CC 85-92 and CC 84-75, the most widely grown cultivars, were resistant. With the discovery of orange rust of sugarcane in Colombia, Cenicaña has incorporated orange rust resistance in the selection and development of new cultivars. To our knowledge, this is the first report of P. kuehnii on sugarcane in Colombia. Orange rust has also been reported from the United States, Cuba, Mexico, Guatemala, Nicaragua, El Salvador, Costa Rica, Panama, Ecuador, and Brazil. References: (1) J. C. Comstock et al. Plant Dis. 92:175, 2008. (2) N. C. Glynn et al. Plant Pathol. 59:703, 2010. (3) E. V. Virtudazo et al. Mycoscience 42:167, 2001.

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