Polymerase chain reaction-based detection ofFusarium circinatum, the causal agent of pitch canker disease

2008 ◽  
Vol 8 (6) ◽  
pp. 1270-1273 ◽  
Author(s):  
T. D. RAMSFIELD ◽  
K. DOBBIE ◽  
M. A. DICK ◽  
R. D. BALL
Keyword(s):  
Polymerase Chain Reaction ◽  
Causal Agent ◽  
Pitch Canker ◽  
Chain Reaction ◽  
Canker Disease ◽  
Polymerase Chain

scholarly journals Molecular Identification of Bacterial Pathogen Infecting Coconut Leaf Beetle

Buletin Palma ◽  
2016 ◽  
Vol 16 (2) ◽  
pp. 147
Author(s):  
JELFINA C. ALOUW ◽  
DIANA NOVIANTI ◽  
MELDY L.A. HOSANG
Keyword(s):  
Polymerase Chain Reaction ◽  
16S Rrna ◽  
Serratia Marcescens ◽  
Molecular Identification ◽  
Ribosomal Rna ◽  
Causal Agent ◽  
16S Ribosomal Rna ◽  
Font Size ◽  
Chain Reaction ◽  
Polymerase Chain

<p><span style="font-size: medium;">ABSTRACT </span></p><p>Many species of microorganisms can cause diseases and mortality of insect pests. Accurate detection and identification of the entomophatogens are essential for development of biological control agent to the pest. Brontispa longissima, a serious and invasive pest of coconut, was infected by bacterium causing mortality of the larvae and pupae in coconut field. Objective of the research was to identify bacterium as a causal agent of the field-infected B. longissima using molecular  technique.  Research  was  conducted  between  April  and  August 2011.  Molecular  identification  using polymerase chain reaction (PCR) amplification of 16s ribosomal RNA of the infected larvae and sequencing of the gene showed that Serratia marcescens is the causal agent of the disease.</p><p>Keywords: Brontispa longissima, coconut, 16s rRNA, Serratia marcescens.</p><p> </p><p><span style="font-size: medium;">Identifikasi Molekular Bakteri Pathogen yang Menginfeksi Hama Daun Kelapa <br />Brontispa longissima(Coleoptera:Chrysomelidae)</span></p><p><span style="font-size: medium;">ABSTRAK </span></p><p>Banyak mikroorganisme dapat menimbulkan penyakit pada serangga hama.  Deteksi dan identifikasi yang akurat dari  pathogen  penyebab  penyakit  pada  serangga (entomopathogen)  hama  merupakan  tahap  yang  penting  dalam  pengembangan pengendalian biologi untuk hama tersebut.  Brontispa longissima sebagai hama penting dan bersifat  invasif pada tanaman kelapa diinfeksi oleh sejenis bakteri yang menyebabkan kematian larva dan pupa dari serangga  tersebut di lapangan. Penelitian ini bertujuan untuk mengidentifikasi organisme penyebab penyakit pada hama B. longissima dengan menggunakan teknik molekuler. Penelitian dilaksanakan pada bulan April sampai dengan Agustus  2011. Identifikasi bakteri dilakukan dengan mengamplifikasi 16s ribosomal RNA dari larva yang terinfeksi dengan menggunakan PCR (polymerase chain reaction).  Hasil analisis sekuens nukleotida 16s ribosomal RNA dari larva yang terinfeksi menunjukkan bahwa Serratia marcescens adalah bakteri penyebab dari penyakit tersebut.</p><p>Kata kunci: Brontispa longissima, kelapa, 16s rRNA, Serratia marcescens.</p>

scholarly journals Low incidence of peach latent mosaic viroid in peach mother blocks in Serbia

2014 ◽  
Vol 29 (2) ◽  
pp. 109-114
Author(s):  
Darko Jevremovic ◽  
Svetlana Paunovic
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Mosaic Disease ◽  
Causal Agent ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Low Incidence

Peach latent mosaic viroid (PLMVd) is the causal agent of peach latent mosaic disease that is common on peaches and nectarines worldwide. Most of the isolates do not cause any symptoms on the foliage and the disease may be latent for years. A survey to investigate the presence of PLMVd in selected peach mother blocks in 9 Serbian districts was carried out in 2011 through 2013. A total of 315 trees/samples originating from 43 mother blocks, representing 35 peach and nectarine varieties and 2 rootstocks, were tested by Reverse Transcription - Polymerase Chain Reaction (RT-PCR). PLMVd was detected in 13 samples (4.13%) belonging to 7 varieties and one vineyard peach rootstock. Infected samples were found in 7 mother blocks from 3 districts. Our results indicated a low incidence of PLMVd in the analyzed peach mother blocks.

scholarly journals Light Microscopy and PCR Based Detection of the Causal Agent of Leaf Mosaic of Jute

2013 ◽  
Vol 22 (1-2) ◽  
pp. 19-25
Author(s):  
KMG Dastogeer ◽  
MA Ali ◽  
M Ashrafuzzaman
Keyword(s):  
Polymerase Chain Reaction ◽  
Light Microscope ◽  
Inclusion Bodies ◽  
Causal Agent ◽  
Healthy Plant ◽  
Infected Leaf ◽  
Chain Reaction ◽  
Degenerate Primers ◽  
Leaf Tissues ◽  
Polymerase Chain

Leaf mosaic transmitted by whitefly is a devastating disease of jute. It is thought to be caused by a virus belonging to begomovirus genus under geminivirus family. To confirm the identity of the causal agent, infected and healthy leaves were studied using light microscope and by using polymerase chain reaction (PCR) technique of DNA. The inclusion bodies were observed under light microscope as large, blue-violet, prominent inclusion bodies in the nucleus of the infected leaf tissues. In molecular detection technique DNA from infected and healthy plants was extracted and analyzed by polymerase chain reaction (PCR) using degenerate primers PALIv1978/PARIc496. PCR fragment of the expected size 1.2kb for the common region (CR) in the geminivirus were obtained from infected plants. DNA collected from healthy plant did not show any band during electrophoresis. Therefore, it can be concluded that leaf mosaic of jute is cause by a virus.DOI: http://dx.doi.org/10.3329/pa.v22i1-2.16463 Progress. Agric. 22(1 & 2): 19 - 25, 2011

scholarly journals A Polymerase Chain Reaction Protocol for the Detection of Xanthomonas albilineans, the Causal Agent of Sugarcane Leaf Scald Disease

Plant Disease ◽  
1997 ◽  
Vol 81 (2) ◽  
pp. 189-194 ◽  
Author(s):  
Y.-B. Pan ◽  
M. P. Grisham ◽  
D. M. Burner
Keyword(s):  
Polymerase Chain Reaction ◽  
Blot Hybridization ◽  
Causal Agent ◽  
Rrna Genes ◽  
23S Rrna ◽  
Dot Blot ◽  
Chain Reaction ◽  
Leaf Scald ◽  
Polymerase Chain ◽  
Xanthomonas Albilineans

A polymerase chain reaction (PCR) protocol was developed that amplified a 360-bp DNA product unique to Xanthomonas albilineans (Xa), the causal agent of sugarcane leaf scald disease. The assay utilizes previously described PCR primers that target the intergenic transcribed spacer (ITS) region between the 16S and 23S rRNA genes. Primer pair Ala4/L1 allowed amplification of a 360-bp DNA fragment from 71 Xa strains including representatives of serovars I, II, and III. Fragments of different sizes were also amplified from three unidentified saprophytic bacteria from sugarcane. Xa could be detected at a lower bacterial concentration with the PCR protocol than with a serological dot blot assay. With PCR, as little as 1.25 pg of Xa genomic DNA (125 fg if followed by Southern blot hybridization), or as few as 0 to 5 CFU of Xa per reaction were detected from infected sugarcane sap and leaf diffusate. Five CFU of Xa per reaction were detected from suspension culture. The PCR protocol provides a rapid, reliable, and economical tool for routine detection and identification of Xa.

Multiplex real-time polymerase chain reaction (PCR) for detection ofPhytophthora ramorum, the causal agent of sudden oak death

2009 ◽  
Vol 31 (2) ◽  
pp. 195-210 ◽  
Author(s):  
Guillaume Bilodeau ◽  
Gervais Pelletier ◽  
Françoise Pelletier ◽  
Richard C. Hamelin ◽  
C. André Lévesque
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Causal Agent ◽  
Sudden Oak Death ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Identification and Detection of Phoma tracheiphila, Causal Agent of Citrus Mal Secco Disease, by Real-Time Polymerase Chain Reaction

Plant Disease ◽  
2006 ◽  
Vol 90 (12) ◽  
pp. 1523-1530 ◽  
Author(s):  
G. Licciardello ◽  
F. M. Grasso ◽  
P. Bella ◽  
G. Cirvilleri ◽  
V. Grimaldi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Real Time Pcr ◽  
Causal Agent ◽  
Conventional Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Polymerase Chain ◽  
Phoma Tracheiphila ◽  
Mal Secco

Phoma tracheiphila is the causal agent of a tracheomycotic disease of citrus called mal secco causing the dieback of twigs and branches. This pathogen is of quarantine concern; therefore, fast and reliable protocols are required to detect it promptly. A specific primer pair and a dual-labeled fluorogenic probe were used in a real-time polymerase chain reaction (PCR) with the Cepheid Smart Cycler II System (Transportable Device TD configuration) to detect this fungus in citrus samples. Real-time PCR assay was compared to modified conventional PCR assay. The sensitivity of the former was evaluated by testing P. tracheiphila DNA dilutions, and the minimum amount detectable was about 500 fg, whereas the linear quantification range was within 100 ng to 1 pg. Conventional PCR sensitivity was 10 pg. Conventional and real-time PCR successfully detected the fungus in woody samples of naturally infected lemon and artificially inoculated sour orange seedlings. Nevertheless, real-time PCR was about 10- to 20-fold more sensitive than conventional PCR, and preliminary results indicate that the former technique achieves quantitative monitoring of the fungus in tissues. Simple and rapid procedures to obtain suitable DNA samples from fungal cultures and citrus woody samples for PCR assays enable diagnosis to be completed in a short time.

scholarly journals Rapid Diagnosis of Pseudomonas syringae pv. papulans, the Causal Agent of Blister Spot of Apple, by Polymerase Chain Reaction Using Specifically Designed hrpL Gene Primers

2002 ◽  
Vol 92 (10) ◽  
pp. 1077-1083 ◽  
Author(s):  
Mohamed Kerkoud ◽  
Charles Manceau ◽  
Jean Pierre Paulin
Keyword(s):  
Polymerase Chain Reaction ◽  
Pseudomonas Syringae ◽  
Polymerase Chain Reaction Amplification ◽  
Restriction Enzymes ◽  
Causal Agent ◽  
Chain Reaction ◽  
Diseased Fruit ◽  
Polymerase Chain ◽  
Primer Sets ◽  
Consensus Primers

The identification and detection of Pseudomonas syringae pv. papulans, the causal agent of blister spot of apple, on apple leaves and fruit was achieved by polymerase chain reaction amplification of a specific DNA fragment of the hrpL sequence. The consensus primers hrpL1 and hrpL2 were designed based on the alignment of pseudomonad hrpL gene sequences available in nucleic acid data banks. This primer set produced a 631-bp amplicon from 37 of the 57 pseudomonads strains tested. These strains belonged to genomospecies 1 and 2, as described by Gardan et al. (8). The amplicon obtained from 30 of these strains was digested with eight restriction enzymes. Three different restriction patterns were produced from strains belonging to genomospecies 1, resulting in A1 and A2 patterns, while strains belonging to genomospecies 2 were characterized by a B pattern. Patterns A1 and A2 differed at only two sites, a Bsp 143I site located at nucleotide 360 and a MseI site located at nucleotides 22–24. Group A2 consisted solely of P. syringae pv. papulans strains. The hrpL gene in P. syringae pv. papulans strain CFBP3323 was sequenced. Two primer sets, Pap1/Pap2 and Pap1/Pap3, were designed and tested for specificity to P. syringae pv. papulans. These primers amplified expected fragments of 242 and 303 bp, respectively. Pap1/Pap2 amplified a fragment only with P. syringae pv. papulans DNA, while Pap1/Pap3 amplified all tested strains belonging to genomospecies 1. A diagnostic procedure using the Pap1/Pap2 primer set was successful for the detection of P. syringae pv. papulans in diseased fruit and artificially inoculated leaves.

Sign in / Sign up

Export Citation Format

Share Document