In planta detection of Macrophomina phaseolina from jute (Corchorus olitorius) by a sodium acetate-based direct PCR method

2014 ◽  
Vol 42 (5) ◽  
pp. 673-676 ◽  
Author(s):  
Chinmay Biswas ◽  
Piyali Dey ◽  
Kunal Mandal ◽  
Subrata Satpathy ◽  
P. G. Karmakar
Keyword(s):  
Sodium Acetate ◽  
Macrophomina Phaseolina ◽  
Corchorus Olitorius ◽  
In Planta ◽  
Direct Pcr ◽  
Pcr Method

Direct PCR-based genetic mapping of rice telomeric repeat associated sequences

Genome ◽  
1998 ◽  
Vol 41 (2) ◽  
pp. 193-198 ◽  
Author(s):  
Lishuang Shen ◽  
Lihuang Zhu
Keyword(s):  
Genetic Mapping ◽  
Oryza Sativa L ◽  
Telomeric Repeat ◽  
Doubled Haploid Population ◽  
Direct Pcr ◽  
Japonica Variety ◽  
Rapd Primer ◽  
Associated Sequences ◽  
Haploid Population ◽  
Pcr Method

Direct PCR-based genetic mapping of telomeric repeat associated sequences (TASs) was achieved using a RAPD primer mediated asymmetric PCR method. Twenty-two TAS loci were mapped in a rice doubled haploid population derived from a cross between an indica variety (Zhaiyeqing8) and a japonica variety (Jingxi17). Of these, 11 loci were mapped to the most distal position of seven chromosome arms and lengthened the linkage groups by 7.4-22.6 cM, five were mapped to the approximate positions of the centromeric regions, and six were mapped to other interstitial chromosomal regions.Key words: rice, Oryza sativa L., genetic mapping, telomeric repeat, telomeric repeat associated sequences, RAPD primer mediated PCR.

Immunolocalization of a β-1,4-endoglucanase from Macrophomina phaseolina expressed in planta

1997 ◽  
Vol 43 (5) ◽  
pp. 491-495 ◽  
Author(s):  
Richard W. Jones ◽  
Haiyin Wang
Keyword(s):  
Specific Antibody ◽  
Macrophomina Phaseolina ◽  
Plant Production ◽  
Symptom Expression ◽  
Stem Tissue ◽  
In Planta ◽  
Charcoal Rot

The in planta endoglucanase production by Macrophomina phaseolina was analyzed by probing tissue blots with a peptide-specific antibody. Endoglucanase (EGL 1) was readily detected at 60 h after microsclerotial inoculation to corn and tobacco stems. Production continued through the study, along with growth of the fungus in stem tissue. Endoglucanase was rapidly transported through the xylem, resulting in distribution to distal portions of the plant. Production at the site of infection was correlated with symptom expression, suggesting a role for endoglucanases in disease progression.Key words: cellulase, charcoal rot.

scholarly journals Whole blood direct PCR method combined with HLA PCR−SSP typing

1998 ◽  
Vol 5 (2) ◽  
pp. 91-95
Author(s):  
Keiko Osawa ◽  
Naoyuki Nishimura ◽  
Taeko K. Naruse ◽  
Hidetoshi Inoko
Keyword(s):  
Whole Blood ◽  
Direct Pcr ◽  
Pcr Method

Immuno-capture PCR method for detecting Acidovorax avenae subsp. citrulli from watermelon

2007 ◽  
Vol 4 (2) ◽  
pp. 173-179 ◽  
Author(s):  
Wang Xiao ◽  
Zhang Le ◽  
Xu Fu-Shou ◽  
Zhao Li-Han ◽  
Xie Guan-Lin
Keyword(s):  
Polymerase Chain Reaction ◽  
Low Cost ◽  
Chain Reaction ◽  
Direct Pcr ◽  
Causal Organism ◽  
Bacterial Fruit Blotch ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Honeydew Melon ◽  
Melon Seeds

AbstractAn immuno-capture polymerase chain reaction (IC-PCR) method for detection of Acidovorax avenae subsp. citrulli (AAC), the causal organism of bacterial fruit blotch (BFB) of watermelon, was developed by combining the immunosorbent enrichment (ISE) method with classical PCR and comparing with the direct PCR and growth check methods. The results showed that all A. avenae subsp. citrulli strains tested have produced 360 bp specific fragments using IC-PCR and direct PCR methods, while other strains from 10 different genera showed negative PCR results. The minimum detection concentration was about 50–100 cfu/ml and 104 cfu/ml, respectively. The IC-PCR sensitivity was 100 times higher than that of direct PCR. The examination of seven batches of different melon seeds from the markets by IC-PCR showed that one cantaloupe, two honeydew melon and two watermelon seed varieties carried the pathogen, indicating that the IC-PCR is an accurate, sensitive, rapid and low-cost technique.

scholarly journals Detection, Survival, and Sources of Inoculum for Bacterial Diseases of Leafy Crucifers in Oklahoma

Plant Disease ◽  
2002 ◽  
Vol 86 (8) ◽  
pp. 883-888 ◽  
Author(s):  
Youfu Zhao ◽  
John P. Damicone ◽  
Carol L. Bender
Keyword(s):  
Pseudomonas Syringae ◽  
Xanthomonas Campestris ◽  
Bacterial Pathogens ◽  
Pcr Amplification ◽  
Soil Surface ◽  
In Planta ◽  
Bacterial Diseases ◽  
Pcr Product ◽  
Pathogenicity Tests ◽  
Pcr Method

Xanthomonas campestris pv. campestris, X. campestris pv. armoraciae, and Pseudomonas syringae pv. maculicola are bacterial pathogens that cause leaf spot diseases on leafy crucifers in Oklahoma. Polymerase chain reaction (PCR) amplification of the cfl gene from the gene cluster encoding the phytotoxin coronatine was used to identify coronatine-producing strains of P. syringae, and the expected 0.65-kb PCR product was detected in 19 strains of P. syringae pv. maculicola originating from diseased crucifers in Oklahoma. A simple, rapid PCR method based on primers from the cfl gene was developed to detect coronatine-producing strains of P. syringae in planta. Pathogenicity tests confirmed the cfl-positive strains to be P. syringae pv. maculicola. To monitor the survival of X. campestris pv. armoraciae and P. syringae pv. maculicola in the field, turnip and collards were inoculated with rifampicin-resistant strains and were buried beneath the soil or left on the soil surface. Both pathogens were recovered from turnip and collard debris up to 2 months following burial, but neither pathogen was recovered from soil after the debris had decomposed. However, both pathogens were recovered from plant debris left on the soil surface for up to 5 months. Four production fields were surveyed for sources of inoculum of the bacterial pathogens from October 1999 to May 2000. X. campestris pv. campestris was isolated from the weed shepherd's purse (Capsella bursa-pastoris) in all fields, and from volunteer turnip and kale in three fields. X. campestris pv. campestris and P. syringae pv. maculicola were isolated from surface debris and regrowth from crop stubble left in one field after harvest in the fall. X. campestris pv. campestris was detected in 6 of 51 lots of crucifer seed assayed. X. campestris pv. armoraciae and P. syringae pv. maculicola were not recovered from weeds, volunteer plants, or seed lots.

scholarly journals A quantitative real-time PCR method for in planta monitoring of Phytophthora infestans growth

2010 ◽  
Vol 51 (6) ◽  
pp. 603-610 ◽  
Author(s):  
B. Llorente ◽  
F. Bravo-Almonacid ◽  
C. Cvitanich ◽  
E. Orlowska ◽  
H.N. Torres ◽  
...  
Keyword(s):  
Phytophthora Infestans ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
In Planta ◽  
Pcr Method

scholarly journals Detection of Salmonella typhimurium ATCC 14028 in Powder Prepared Traditional Medicines Using Real-Time PCR

2021 ◽  
Vol 4 (3) ◽  
pp. 178-183
Author(s):  
Alfi Sophian ◽  
Ratna Purwaningsih ◽  
Muindar Muindar ◽  
Eka Putri Juniarti Igirisa ◽  
Muhammad Luthfi Amirullah
Keyword(s):  
Salmonella Typhimurium ◽  
Real Time ◽  
Real Time Pcr ◽  
Positive Control ◽  
Negative Control ◽  
Pcr Analysis ◽  
Direct Pcr ◽  
Traditional Medicines ◽  
Pcr Method ◽  
Alternative Testing

The detection of Salmonella typhimurium ATCC 14028 using real-time PCR on powdered traditional medicinal products was carried out in the microbiology and molecular biology testing laboratory of the Food and Drug Administration in Gorontalo. This research aims to provide a reference for alternative testing methods in testing the products of traditional powder preparations on the market. The sample consisted of 10 traditional powder preparations spiked with positive control of S. typhimurium ATCC 14028 phase 2. The method used in the study was real-time PCR analysis using the SYBR® Green method, while DNA isolation using the direct PCR method. Data analysis was performed by analyzing the sample's melting temperature (Tm) curve and comparing it with positive control. The results showed that S. typhimurium ATCC 14028 was detected in samples at an average Tm value of 84.18°C, with ranges of 84.0-84.5°C. For positive control, the Tm value was at 85.2°C, while for the negative control, the Tm value was not detected. Based on these data, it can be concluded that S. typhimurium ATCC 14028 in traditional medicine products powder preparations can be detected using real-time PCR.

scholarly journals Development of rapid direct PCR assays to identify downy and powdery mildew and grey mould in Vitis vinifera tissues

OENO One ◽  
2014 ◽  
Vol 48 (4) ◽  
pp. 261 ◽  
Author(s):  
Katia Gindro ◽  
Nicole Lecoultre ◽  
Luca Molino ◽  
Jean-Pierre De Joffrey ◽  
Sylvain Schnee ◽  
...  
Keyword(s):  
Vitis Vinifera ◽  
Downy Mildew ◽  
Fungal Pathogens ◽  
Pcr Amplification ◽  
Microscopic Analysis ◽  
Grey Mould ◽  
Dna Purification ◽  
Direct Pcr ◽  
Fungal Propagules ◽  
Pcr Method

<p style="text-align: justify;"><strong>Aims</strong>: The development of a rapid and reliable direct PCR method to detect fungal propagules in grapevine tissues without prior DNA purification steps, and illustration of its potential use with different examples.</p><p style="text-align: justify;"><strong>Methods and results</strong>: Different grapevine samples crushed in the presence of polyvinylpolypyrrolidone (PVPP) were used as templates for direct PCR amplification with primers specifie for <em>Erysiphe necator</em>, <em>Plasmopara viticola</em>, <em>Botrytis cinerea</em> and <em>Vitis vinifera</em>. Sequencing of the PCR products confirmed the specificity of the amplifications. The sensitivity tested using conidia/sporangia dilution series was high, ranging from five sporangia for <em>P. viticola</em> to one conidium for <em>E. necator</em>. The potential of this technique is illustrated through the study of four epidemiological questions. Fungal propagules were observed in dormant buds using microscopy, but the responsible species could not be identified. Direct PCR revealed the presence of <em>E. necator</em> and<em> B. cinerea</em> in 29 % and 65 % of the buds, respectively. Downy mildew could be detected in asymptomatic leaves sampled in fields after potentially infectious events. In bunch, microscopic analysis of rachis sections showed the presence of hyphae growing in the green tissue. Direct PCR identified the presence of <em>P. viticola</em>.</p><p style="text-align: justify;"><strong>Conclusion</strong>: A direct PCR method without DNA purification was demonstrated to be a simple and reliable method for the detection and identification of fungal pathogens in grapevine tissues. This method, together with microscopy, is a very interesting tool that can be used to study various epidemiological problems in the grapevine, including important unanswered questions such as the route of infection that leads to brown rot caused by downy mildew.</p><p style="text-align: justify;"><strong>Significance and impact of the study</strong>: Direct PCR was shown to be a simple and versatile technique for the study of epidemiological questions in the grapevine. This technique could be extended to other pathosystems with minor adaptations.</p>

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