Tetra primer ARMS-PCR for identification of SNP in β-tubulin of Botrytis cinerea, responsible of resistance to benzimidazole

Muñoz, Claudio; Talquenca, Sebastián Gómez; Volpe, Melisa Lanza

doi:10.1016/j.mimet.2009.06.007

Cited by 22 publications

(14 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…SNP signatures represent single base pair positions with different sequence signatures (alleles) in individuals of a population (Brookes 1999). SNPs have been detected in genomes of many taxa (Kim and Misra 2007) including fungi (e.g., Bain et al 2007;Kristensen et al 2007;Xu et al 2007;Lambreghts et al 2009;Munoz et al 2009). They have been widely applied in biomedical fields (Kim and Misra 2007) and have a great potential for use in ecology, evolution, and conservation biology (Morin et al 2004).…”

Section: Sequencingmentioning

confidence: 99%

Molecular ecology of fungal entomopathogens: molecular genetic tools and their applications in population and fate studies

Enkerli

Widmer

2009

BioControl

View full text Add to dashboard Cite

The power of molecular genetic techniques to address ecological research questions has opened a distinct interdisciplinary research area collectively referred to as molecular ecology. Molecular ecology combines aspects of diverse research fields like population and evolutionary genetics, as well as biodiversity, conservation biology, behavioural ecology, or species-habitat interactions. Molecular techniques detect specific DNA sequence characteristics that are used as genetic markers to discriminate individuals or taxonomic groups, for instance in analyses of population and community structures, for elucidation of phylogenetic relationships, or for the characterization and monitoring of specific strains in the environment. Here, we summarize the PCR-based molecular techniques used in molecular ecological research on fungal entomopathogens and discuss novel techniques that may have relevance to the studies of entomopathogenic fungi in the future. We discuss the flow chart of the molecular ecology approaches and we highlight some of the critical steps involved. There are still many unresolved questions in the understanding of the ecology of fungal entomopathogens. These include population characteristics and relations of genotypes and habitats as well as hostpathogen interactions. Molecular tools can provide substantial support for ecological research and offer insight into this far inaccessible systems. Application of molecular ecology approaches will stimulate and accelerate new research in the field of entomophathogen ecology.

show abstract

Section: Sequencingmentioning

confidence: 99%

Molecular ecology of fungal entomopathogens: molecular genetic tools and their applications in population and fate studies

Enkerli

Widmer

2009

BioControl

View full text Add to dashboard Cite

show abstract

“…At present, many molecular methods have been used to detect fungicide resistance in B. cinerea , including sequencing ( Leroux et al, 2010 ; Fillinger et al, 2012 ), polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP; Ishii et al, 2009 ; Van der Heyden et al, 2014 ), allele-specific PCR (AS PCR; De Miccolis et al, 2012 , 2014 ; Yin et al, 2012 ; Van der Heyden et al, 2014 ), tetra-primer amplification refractory mutation systems (ARMSs) PCR ( Munoz et al, 2009 ), real-time PCR ( Banno et al, 2008 ; Billard et al, 2012 ), and high-resolution melting (HRM) analysis ( Chatzidimopoulos et al, 2014 ). However each of these molecular assays can only detect a single allele or simultaneously detect different alleles in a single gene.…”

Section: Introductionmentioning

confidence: 99%

Suspension Array for Multiplex Detection of Eight Fungicide-Resistance Related Alleles in Botrytis cinerea

Xie

Zhao

et al. 2016

Front. Microbiol.

View full text Add to dashboard Cite

A simple and high-throughput assay to detect fungicide resistance is required for large-scale monitoring of the emergence of resistant strains of Botrytis cinerea. Using suspension array technology performed on a Bio-Plex 200 System, we developed a single-tube allele-specific primer extension assay that can simultaneously detect eight alleles in one reaction. These eight alleles include E198 and 198A of the β-Tubulin gene (BenA), H272 and 272Y of the Succinate dehydrogenase iron–sulfur subunit gene (SdhB), I365 and 365S of the putative osmosensor histidine kinase gene (BcOS1), and F412 and 412S of the 3-ketoreductase gene (erg27). This assay was first established and optimized with eight plasmid templates containing the DNA sequence variants BenA-E198, BenA-198A, SdhB-H272, SdhB-272Y, BcOS1-I365, BcOS1-365S, erg27-F412, and erg27-412S. Results indicated that none of the probes showed cross-reactivity with one another. The minimum limit of detection for these genotypes was one copy per test. Four mutant plasmids were mixed with 10 ng/μL wild-type genomic DNA in different ratios. Detection sensitivity of mutant loci was 0.45% for BenA-E198A, BcOS1-I365S, and erg27-F412S, and was 4.5% for SdhB-H272Y. A minimum quantity of 0.1 ng of genomic DNA was necessary to obtain reliable results. This is the first reported assay that can simultaneously detect mutations in BenA, SdhB, BcOS1, and erg27.

show abstract

“…Fr. [teleomorph Botryotinia fuckeliana ( d e B a r y ) W h e t z e l ] i s a phytopathogenic fungi responsible for grey mould, which is an important disease in more than 200 species of fruits, vegetables, ornamental crops and affecting a great number of organs [1] . Grey mould, cause important economic losses before and after the harvest [2] , worldwide losses from this fungus account for 20% of the harvest of the affected crops, and the cost is estimated at billion euros per year.…”

Section: Introductionmentioning

confidence: 99%

“…Grey mould, cause important economic losses before and after the harvest [2] , worldwide losses from this fungus account for 20% of the harvest of the affected crops, and the cost is estimated at billion euros per year. The market size for anti-Botrytis products has been [15][16][17][18][19][20][21][22][23][24][25] million US dollars in recent 3 years [3] . An important biological feature is that the process of infection and growth by B. cinerea is often associated with prior colonization of dead or dying plant debris as a nutrient-providing saprophytic base [4] .…”

Section: Introductionmentioning

confidence: 99%

“…cinerea is a haploid and heteroallelic ascomycete [12] . Analysis genetic such as RFLP-PCR [13,14] , ARMS-PCR [15] , allelespecific PCR [16] , RAPD-PCR [4] , nested PCR, and detection transposable elements (TEs) [13] , amongst other molecular typing techniques, have characterized to B. cinerea. They have molecular analysis identify two groups based on the presence or absence of two TEs in the genome of B. cinerea, vacuma (isolates without either TEs) and transposa (isolates with the TEs boty y flipper) [13] .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Effect of temperature on the morphological characteristics of Botrytis cinerea and its correlated with the genetic variability

2014

JCLM

View full text Add to dashboard Cite

Tetra primer ARMS-PCR for identification of SNP in β-tubulin of Botrytis cinerea, responsible of resistance to benzimidazole

Cited by 22 publications

References 10 publications

Molecular ecology of fungal entomopathogens: molecular genetic tools and their applications in population and fate studies

Molecular ecology of fungal entomopathogens: molecular genetic tools and their applications in population and fate studies

Suspension Array for Multiplex Detection of Eight Fungicide-Resistance Related Alleles in Botrytis cinerea

Effect of temperature on the morphological characteristics of Botrytis cinerea and its correlated with the genetic variability

Contact Info

Product

Resources

About