IDENTIFICATION OF PFIESTERIA PISCICIDA (DINOPHYCEAE) AND PFIESTERIA-LIKE ORGANISMS USING INTERNAL TRANSCRIBED SPACER-SPECIFIC PCR ASSAYS1

2003 ◽  
Vol 39 (4) ◽  
pp. 754-761 ◽  
Author(s):  
R. Wayne Litaker ◽  
Mark W. Vandersea ◽  
Steven R. Kibler ◽  
Kimberly S. Reece ◽  
Nancy A. Stokes ◽  
...  
Plant Disease ◽  
2012 ◽  
Vol 96 (1) ◽  
pp. 143-143 ◽  
Author(s):  
M. Cadavid ◽  
J. C. Ángel ◽  
J. I. Victoria

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.


2003 ◽  
Vol 39 (s1) ◽  
pp. 36-36 ◽  
Author(s):  
R. W. Litaker ◽  
M. W. Vandersea ◽  
S. R. Kibler ◽  
P. A. Tester ◽  
K. S. Reece ◽  
...  

2000 ◽  
Vol 38 (11) ◽  
pp. 4080-4085 ◽  
Author(s):  
Heekyung Park ◽  
Hyunjung Jang ◽  
Cheolmin Kim ◽  
Byungseon Chung ◽  
Chulhun L. Chang ◽  
...  

We evaluated the usefulness of PCR assays that target the internal transcribed spacer (ITS) region for identifying mycobacteria at the species level. The conservative and species-specific ITS sequences of 33 species of mycobacteria were analyzed in a multialignment analysis. One pair of panmycobacterial primers and seven pairs of mycobacterial species-specific primers were designed. All PCRs were performed under the same conditions. The specificities of the primers were tested with type strains of 20 mycobacterial species from the American Type Culture Collection; 205 clinical isolates of mycobacteria, including 118Mycobacterium tuberculosis isolates and 87 isolates of nontuberculous mycobacteria from 10 species; and 76 clinical isolates of 28 nonmycobacterial pathogenic bacterial species. PCR with the panmycobacterial primers amplified fragments of approximately 270 to 400 bp in all mycobacteria. PCR with the M. tuberculosiscomplex-specific primers amplified an approximately 120-bp fragment only for the M. tuberculosis complex. Multiplex PCR with the panmycobacterial primers and the M. tuberculosiscomplex-specific primers amplified two fragments that were specific for all mycobacteria and the M. tuberculosis complex, respectively. PCR with M. avium complex-, M. fortuitum-, M. chelonae-, M. gordonae-, M. scrofulaceum-, andM. szulgai-specific primers amplified specific fragments only for the respective target organisms. These novel primers can be used to detect and identify mycobacteria simultaneously under the same PCR conditions. Furthermore, this protocol facilitates early and accurate diagnosis of mycobacteriosis.


2020 ◽  
Author(s):  
Tiantian Geng ◽  
Zhenyu Lei ◽  
Cheng Ye ◽  
Bang Shen ◽  
Rui Fang ◽  
...  

Abstract Background:Coccidiosis is an intestinal parasitic disease that causes huge economic losses to the poultry industry globally. At present, the primary control strategy is administration of anticoccidial drugs with feed. However, overuse of anticoccidials, such as sulfachloropyrazine sodium (SC), has resulted in an increase in the emergence of drug resistance.Methods: We aimed to evaluate coccidiosis prevalence and SC resistance in field isolates to provide reasonable guidance on the use of anticoccidial drugs in the Hubei and Henan provinces. We collected 318 fresh fecal samples from 137 chicken farms. We used internal transcribed spacer 1 (ITS1) sequence of ribosomal DNA to identify the species from 94 samples that were collected from different farms and to assess drug resistance.Results: As shown by genus-specific PCR results, the positivity rate of Eimeria was 97.17 % (309/318), and the most common species were E. mitis (66.67%), E. tenella (46.86%), and E. necatrix (41.51%). Animal experiment demonstrated that 25 strains were completely resistant to SC, among which 16 were from Henan and nine were from Hubei. Twenty-four strains were partially resistant, among which 8 and 16 strains were identified from Hubei and Henan, respectively.Conclusions: In summary, these data indicated that chicken coccidia is ubiquitous and SC resistance is widespread, in the Hubei and Henan provinces. The results provide important insights into the control of chicken coccidiosis in this region.


2016 ◽  
Vol 8 (2) ◽  
pp. 864-867
Author(s):  
R. Yonzone ◽  
B. Kumar ◽  
P.P. Singh ◽  
M.S. Hunjan ◽  
B. Das

This study was aim to identify the pathogen associated with the red stripe/top rot disease. The isolates were collected from major sugarcane growing districts of Punjab. Molecular identification of the pathogen was carried out to confirm the associated organism with this disease. Of the different sets of specific PCR based molecular markers were used, primer sets SeQ1 (Aaa) and SeQ2 (Aaa) amplified the expected 550bp of ITS (Internal Transcribed Spacer) region of the rDNA which revealed the pathogen as Acidovorax avenae pv. avenae, whereas the primer set Aaaf3 and Aaar2 are specific to strains that infect rice crop did not amplify any fragment. Our studies confirmed Acidovorax avenae pv. avenae as the causal bacterium associated with the red stripe/top rot of sugarcane in Punjab.


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