scholarly journals Identification of Colletotrichum Species Associated with Chili Anthracnose in Indonesia by Morphological Characteristics and Species-Specific Primers

2017 ◽  
Vol 12 (1) ◽  
pp. 7-15 ◽  
Author(s):  
Widodo . ◽  
Sri Hendrastuti Hidayat
Keyword(s):  
Morphological Characteristics ◽  
Specific Primers ◽  
Colletotrichum Species ◽  
Species Specific

scholarly journals Diversity of Colletotrichum Species causing Anthracnose Disease from Mango cv. Nam Dork Mai See Tong based on ISSR-PCR

2021 ◽  
Author(s):  
S. Bincader ◽  
R. Pongpisutta ◽  
C. Rattanakreetakul
Keyword(s):  
Species Complex ◽  
Pcr Amplification ◽  
Molecular Techniques ◽  
Specific Primers ◽  
Cophenetic Correlation ◽  
Anthracnose Disease ◽  
Colletotrichum Species ◽  
Morphological Studies ◽  
Pathogenicity Tests ◽  
Species Specific

Background: Anthracnose disease caused by the genus Colletotrichum is one of the crucial problems occurring in the field, along with postharvest diseases and affects mango quality in Thailand. In particular, the Nam Dork Mai See Tong cultivar, which is highly susceptible to the disease, is an important product for exportation. Methods: In this research, thirty-seven Colletotrichum species isolate were obtained from anthracnose disease in mango cv. Nam Dork Mai See Tong in three provinces in Thailand. Morphological studies and molecular techniques using species-specific primers were investigated; moreover, the diversity of pathogens was analyzed using PCR amplification of inter simple sequence repeats (ISSRs) with 6 primers, including pathogenicity tests. Result: Morphological studies and molecular detection with species-specific primers revealed that 32 isolates belonged to the C. gloeosporioides species complex and 5 isolates to the C. acutatum species complex. The genetic diversity of pathogens was analyzed. PCR amplification using 6 ISSR primers produced 35 polymorphic bands. These bands were used to construct UPGMA, in which cluster analysis divided the 37 isolates into 3 main groups and 8 subgroups at 61-73% Jaccard similarity coefficient with cophenetic correlation (r) = 0.6781. The ISSR technique showed the greatest genetic variation among isolates collected from different locations. Hence, a study based on ISSR markers was profitable to investigate the phylogenetic relationship of the genus Colletotrichum. Pathogenicity tests revealed that PC006 (Ca) and CS005 (Cg) showed the highest aggressiveness, with disease incidences of 84.74 and 80.90%, respectively. This study indicates that the diversity of pathogenic Colletotrichum species related to mango plantations in Thailand is increasing.

Development of a species-specific PCR to detect the cereal cyst nematode, Heterodera latipons

Nematology ◽  
2013 ◽  
Vol 15 (6) ◽  
pp. 709-717 ◽  
Author(s):  
Fateh Toumi ◽  
Fateh Toumi ◽  
Lieven Waeyenberge ◽  
Fateh Toumi ◽  
Lieven Waeyenberge ◽  
...  
Keyword(s):  
Morphological Characteristics ◽  
Molecular Techniques ◽  
Actin Gene ◽  
Specific Primers ◽  
Second Stage ◽  
Specific Pcr ◽  
Pcr Products ◽  
Heterodera Latipons ◽  
Species Specific ◽  
Very High

Several Heterodera species can reduce the yield of wheat and barley, among which H. avenae, H. filipjevi and H. latipons are economically the most important. Their identification, based on morphological characteristics, is not straightforward but can be made easier using molecular techniques. In this study, we developed species-specific primers for the detection of H. latipons. The actin gene of eight Heterodera species was partially sequenced and, after purifying and sequencing the PCR products, all sequences were aligned to find unique sites. The alignment showed moderate to very high similarities between the species. However, a small fragment of the actin gene was suitable for the construction of a potentially useful species-specific primer for H. latipons. The optimised PCR was subsequently tested with several populations of 14 Heterodera species and a single population of Punctodera punctata. Heterodera latipons was represented by 16 populations originating from six different countries. The primer set (Hlat-act), designed using AlleleID 7.73, was shown to be very specific. To test its sensitivity further, the PCR was conducted on DNA extracted from five second-stage juveniles (J2) of H. latipons mixed with five or 100 J2 belonging to H. avenae. The PCR was able to detect up to 1:10 dilution of the DNA obtained from five J2. The results showed that a specific and sensitive H. latipons species-specific PCR was constructed.

scholarly journals A multiplex polymerase chain reaction based method for rapid identification of two species of the genus Scolytus Geoffroy (Col: Curculionidae: Scolytinae) in Iran

2016 ◽  
Vol 48 (1) ◽  
pp. 11 ◽  
Author(s):  
S. Amini ◽  
R. Hosseini
Keyword(s):  
Polymerase Chain Reaction ◽  
Bark Beetle ◽  
Multiplex Polymerase Chain Reaction ◽  
Fragment Size ◽  
Morphological Characteristics ◽  
Beetle Species ◽  
Chain Reaction ◽  
Specific Primers ◽  
Polymerase Chain ◽  
Species Specific

Molecular identification is going to be more widespread in taxonomic studies of insects when traditional tools are problematic and time consuming. Identification of bark beetles, as one of the most important pests of forests, based on morphological characteristics is difficult because of their small size and morphological similarities. In the current study, species-specific primers were desi gned to identify two most abundant and morphologically similar bark beetle species <em>Scolytus ensifer</em> Eichhoff 1881 and <em>S. ecksteini</em> Butovitsch 1929, both found on <em>Ulmus minor</em> Miller in north of Iran. These species-specific primers successfully produced a fragment size with 318 bp and 465 bp of mitochondrial cytochrome oxidase 1 (<em>CO1</em>) gene in <em>S. ensifer </em>and<em> S. ecksteini</em> respectively. The results revealed tha t the multiplex polymerase chain reaction using the species-specific primers could amplify a unique band to distinguish these two species so confirmed this method as a convenient and quick tool to identify those two bark beetle species.

Species-specific duplex PCR for the detection of Pratylenchus penetrans

Nematology ◽  
2009 ◽  
Vol 11 (6) ◽  
pp. 847-857 ◽  
Author(s):  
Lieven Waeyenberge ◽  
Nicole Viaene ◽  
Maurice Moens
Keyword(s):  
Internal Control ◽  
Dna Sequences ◽  
Rrna Gene ◽  
Duplex Pcr ◽  
Specific Primers ◽  
5.8S Rrna Gene ◽  
5.8S Rrna ◽  
Wide Range ◽  
Pcr Products ◽  
Species Specific

Abstract ITS1, the 5.8S rRNA gene and ITS2 of the rDNA region were sequenced from 20 different Pratylenchus species. Additionally, the same region was sequenced from seven populations of P. penetrans. After purifying, cloning and sequencing the PCR products, all sequences were aligned in order to find unique sites suitable for the design of species-specific primers for P. penetrans. Since ITS regions showed variability between and even within populations of P. penetrans, only three small DNA sequences were suitable for the construction of three potentially useful species-specific primers. New species-specific primers were paired with existing universal ITS primers and tested in all possible primer combinations. The best performing primer set, supplemented with a universal 28S rDNA primer set that served as an internal control, was tested in duplex PCR. The ideal annealing temperature, Mg2+ concentration and primer ratios were then determined for the most promising primer set. The optimised duplex PCR was subsequently tested on a wide range of different Pratylenchus spp. and 25 P. penetrans populations originating from all over the world. To test the sensitivity, the duplex PCR was conducted on DNA extracted from a single P. penetrans nematode mixed with varying amounts of nematodes belonging to another Pratylenchus species. Results showed that a reliable and sensitive P. penetrans species-specific duplex PCR was constructed.

scholarly journals Characterization of the Bull's Eye Rot of Apple in Chile

Plant Disease ◽  
2013 ◽  
Vol 97 (4) ◽  
pp. 485-490 ◽  
Author(s):  
Sylvana Soto-Alvear ◽  
Mauricio Lolas ◽  
Inés M. Rosales ◽  
Eduardo R. Chávez ◽  
Bernardo A. Latorre
Keyword(s):  
Morphological Characteristics ◽  
The United States ◽  
Apple Fruit ◽  
Maritime Transportation ◽  
Tubulin Genes ◽  
Internal Transcribed Spacer Sequences ◽  
Apple Production ◽  
Species Specific ◽  
Bull's Eye

Apple fruit in Chile are primarily produced for export to Asia, Europe, and the United States, which typically requires 15 to 40 days of maritime transportation. Therefore, Chilean apple production must fulfill the sanitization requirements imposed by the receiving countries. Under these circumstances, it was important to clarify the etiology of bull's eye rot that can severely affect ‘Cripps Pink’ apple and other late-harvest cultivars in Chile. Based on morphological characteristics and the partial sequence analysis of the internal transcribed spacer sequences and β-tubulin genes, Neofabraea alba was identified as the causal agent of the bull's eye rot of Chilean apple. These results were further corroborated using species-specific primers. The incidence of bull's eye rot varied considerably; for instance, in 2009, 0.0 to 58.7% in 38 Cripps Pink orchards surveyed in the relatively arid and humid apple-growing areas of Chile, respectively. There was no evidence for the presence of N. malicorticis or N. perennans, which are commonly identified as causal agents of bull's eye rot in other apple-producing countries. Altogether, these data suggest that N. alba might represent the predominant and possibly the only cause of bull's-eye rot of Chilean apple.

Species‐specific primers of chitin synthase 1 gene for the differentiation of the Trichophyton mentagrophytes complex

Mycoses ◽  
1999 ◽  
Vol 42 (1‐2) ◽  
pp. 71-74 ◽  
Author(s):  
R. Kano ◽  
Y. Nakamura ◽  
T. Watari ◽  
S. Watanabe ◽  
H. Takahashi ◽  
...  
Keyword(s):  
Trichophyton Mentagrophytes ◽  
Chitin Synthase ◽  
Specific Primers ◽  
Trichophyton Mentagrophytes Complex ◽  
Species Specific

scholarly journals Multiplex PCR with 16S rRNA Gene-Targeted Primers of Bifidobacterium spp. To Identify Sources of Fecal Pollution

2004 ◽  
Vol 70 (5) ◽  
pp. 3171-3175 ◽  
Author(s):  
X. Bonjoch ◽  
E. Ballesté ◽  
A. R. Blanch
Keyword(s):  
16S Rrna ◽  
Multiplex Pcr ◽  
Fecal Pollution ◽  
Sensitivity Threshold ◽  
Rrna Gene ◽  
Specific Primers ◽  
Animal Wastewater ◽  
Wastewater Samples ◽  
Species Specific ◽  
Different Origins

ABSTRACT Bifidobacteria are one of the most common bacterial types found in the intestines of humans and other animals and may be used as indicators of human fecal pollution. The presence of nine human-related Bifidobacterium species was analyzed in human and animal wastewater samples of different origins by using species-specific primers based on 16S rRNA sequences. Only B. adolescentis and B. dentium were found exclusively in human sewage. A multiplex PCR approach with strain-specific primers was developed. The method showed a sensitivity threshold of 10 cells/ml. This new molecular method could provide useful information for the characterization of fecal pollution sources.

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