RFLP Identification of Colletotrichum Species Isolated From Chilli in Thailand

Pongpisutta, R.; Winyarat, W.; Rattanakreetakul, Chainarong

doi:10.17660/actahortic.2013.973.24

Cited by 3 publications

(2 citation statements)

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“…Furthermore, RFLP or virtual RFLP techniques are considered to be essential tools to solve the problems of species differentiation in microorganisms, such as the identification of 3 species of Colletotrichum causing chili anthracnose in Thailand, pineapple spoilage fungi, and three major Meloidogyne species in Korea, as well as the classification of phytoplasma using cpn60 universal target sequences (Han et al 2004;Koffi et al 2019;Pérez-López et al 2016;Pongpisutta et al 2013) Apart from this, In silico testing has reported that this method can be solve the error of PCR-RFLP method, caused by primers that may not amplify PCR products and low PCR product concentration that affect availability of template for cleavage by restriction enzymes and affect visibility of digested product on agarose gels, including PCR master mixed such as MgCl2, Taq buffer contained with KCl or (NH4)2SO4 and primer dimers that can inhibit activity of restriction enzyme (Ramdeen and Rampersad 2014).…”

Section: Discussionmentioning

confidence: 99%

“…Genomic DNA of each isolate was extracted using the method of Pongpisutta et al (2013) and the modified protocol of Zimand et al (1994). PCR amplification was performed for the partial internal transcribed spacer (ITS region), actin gene (ACT), chitin synthase 1 gene (CHS-1) and β-tubulin (TUB) regions, as well as with specific primers for C. acutatum and C. gloeosporioides (Table 1).…”

Section: Dna Extraction and Fungal Confirmationmentioning

confidence: 99%

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Colletotrichum Species Related To “Nam Dork Mai See Tong” Mango In Central Thailand And Risk Detection of Carbendazim-Resistant Strains

Bincader

Pongpisutta

Rattanakreetakul

2021

Preprint

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Nam Dork Mai See Tong mango is a deluxe commercial fruit in Thailand, however anthracnose disease becoming a major problem affecting market-driven. Fungicide treatment is the main component in the mango anthracnose management, notwithstanding the appearance of fungal-resistance has been become a factor of fungicide limition and can affect to increasingly higher costs. Thirty-two isolates of the Colletotrichum species complex were obtained from anthracnose-diseased mango cv. Nam Dork Mai See Tong in 3 provinces of Thailand. Fungal identification based on morphological and molecular markers was investigated. All isolates were divided into 3 species: C. asianum , C. gloeosporioides and C. siamense . Pathogenicity tests revealed that all 3 species caused symptoms on artificially unwounded mango fruits and leaves. Only C. gloeosporioides have previously been reported on mango, while C. asianum and C. siamense were first reports associated with mango in Thailand. The responsiveness of Colletotrichum isolates to carbendazim was evaluated using an MIC assay. Nine isolates of C. asianum were highly resistant, while 5 and 8 isolates of C. gloeosporioides were resistant (R) and highly resistant (HR), respectively. Moreover, all isolates of C. siamense were HR. Mutations were detected by PCR of a partial sequence of the β-tubulin gene. The sequence of β-tubulin gene in HR strains showed a single nucleotide transversion of adenine to cytosine, resulting in a substitution at codon 198. However, R strains were found to have only an amino acid change at codon 200. Additionally, PCR-RFLP was applied as a rapid technique to detect carbendazim resistance. The results demonstrated that only the Bsh 1236I restriction enzyme generated two bands (200 and 300 bp) in the highly resistant strain, but these bands were absent in the sensitive (S) and R strains. Hence, PCR-RFLP technique showed the potential to specifically detect benzimidazole fungicide resistance in 3 species of Colletotrichum. Moreover, this technique is accessible and feasible for laboratory assessment

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Section: Discussionmentioning

confidence: 99%

Section: Dna Extraction and Fungal Confirmationmentioning

confidence: 99%

Colletotrichum Species Related To “Nam Dork Mai See Tong” Mango In Central Thailand And Risk Detection of Carbendazim-Resistant Strains

Bincader

Pongpisutta

Rattanakreetakul

2021

Preprint

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Decontamination of seed borne disease in pepper (Capsicum annuum L.) seed and the enhancement of seed quality by the emulated plasma technology

Ahmad

Sripong

Uthairatanakij

et al. 2022

Scientia Horticulturae

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Two Newly Identified Colletotrichum Species Associated with Mango Anthracnose in Central Thailand

Rattanakreetakul

Keawmanee

Bincader

et al. 2023

Plants

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Anthracnose caused by Colletotrichum spp. is one of the major problems in mango production worldwide, including Thailand. All mango cultivars are susceptible, but Nam Dok Mai See Thong (NDMST) is the most vulnerable. Through a single spore isolation method, a total of 37 isolates of Colletotrichum spp. were obtained from NDMST showing anthracnose symptoms. Identification was performed using a combination of morphology characteristics, Koch’s postulates, and phylogenetic analysis. The pathogenicity assay and Koch’s postulates on leaves and fruit confirmed that all Colletotrichum spp. tested were causal agents of mango anthracnose. Multilocus analysis using DNA sequences of internal transcribed spacer (ITS) regions, β-tubulin (TUB2), actin (ACT), and chitin synthase (CHS-1) was performed for molecular identification. Two concatenated phylogenetic trees were constructed using either two-loci of ITS and TUB2, or four-loci of ITS, TUB2, ACT, and CHS-1. Both phylogenetic trees were indistinguishable and showed that these 37 isolates belong to C. acutatum, C. asianum, C. gloeosporioides, and C. siamense. Our results indicated that using at least two loci of ITS and TUB2, were sufficient to infer Colletotrichum species complexes. Of 37 isolates, C. gloeosporioides was the most dominant species (19 isolates), followed by C. asianum (10 isolates), C. acutatum (5 isolates), and C. siamense (3 isolates). In Thailand, C. gloeosporioides and C. acutatum have been reported to cause anthracnose in mango, however, this is the first report of C. asianum and C. siamense associated with mango anthracnose in central Thailand.

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RFLP Identification of Colletotrichum Species Isolated From Chilli in Thailand

Cited by 3 publications

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Colletotrichum Species Related To “Nam Dork Mai See Tong” Mango In Central Thailand And Risk Detection of Carbendazim-Resistant Strains

Colletotrichum Species Related To “Nam Dork Mai See Tong” Mango In Central Thailand And Risk Detection of Carbendazim-Resistant Strains

Decontamination of seed borne disease in pepper (Capsicum annuum L.) seed and the enhancement of seed quality by the emulated plasma technology

Two Newly Identified Colletotrichum Species Associated with Mango Anthracnose in Central Thailand

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