Multiplex Detection of Fusarium Species

Yli-Mattila, Tapani; Nayaka, S. Chandra; Mudili, Venkataramana; Yörük, Emre

doi:10.1007/978-1-4939-6707-0_18

Cited by 8 publications

(5 citation statements)

References 36 publications

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“…For detection of Fusarium species, Yli-Mattila et al have demonstrated a PCR-based method which can simultaneously detect different mycotoxins from different species. This multiplex genotype PCR assay is fast, cost-effective, and advantageous over other techniques …”

Section: Multiplex Detection Of Plant Pathogensmentioning

confidence: 99%

“…This multiplex genotype PCR assay is fast, cost-effective, and advantageous over other techniques. 133 Xylella fastidiosa, a plant pathogen having various subspecies, has been seen to infect up to 560 different plant species. Dupas et al have described novel tetraplex and triplex qPCR (quantitative PCR) assays for its detection.…”

Section: Multiplex Detection Of Plant Pathogensmentioning

confidence: 99%

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Emerging Extraction and Diagnostic Tools for Detection of Plant Pathogens: Recent Trends, Challenges, and Future Scope

Verma

Shukla

Kulkarni

et al. 2022

ACS Agric. Sci. Technol.

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Plant pathogens are a serious threat to agriculture for long-term viability and cost loss of billions of dollars yearly. Many pathogens have been documented in the literature that infect a variety of crops. Nevertheless, new pathogens emerge and often result in disease outbreaks, leading to million-dollar losses. Currently, several diagnostic approaches with enhanced sensitivity and specificity for the identification of widespread and/or unknown plant pathogens are constantly being developed. Whereas the extensively used approaches for plant pathogen diagnostics are mostly serological and nucleic acid-based assays, many different nucleic acid-based approaches for amplifying target DNA/RNA have also emerged over time. However, these approaches lack precision, specificity, and rapidity, making them unsuitable for on-field analysis. As a result, there is a lot of interest arising in fielddeployable point of care (POC) devices and artificial intelligence (AI)-assisted pathogens' detection accurately at an early stage within a minute. Similarly, development of a cell-lysis and purification-free DNA/RNA extraction process is also crucial for quick sample preparation for molecular diagnosis of plant pathogens at field level. In this review, we have discussed advanced tools that are trending not only to extract nucleic acids but also detect plant pathogens. We have also discussed critical challenges and future perspectives of disease diagnostic tools for plant pathogens' detection. In summary, advanced plant disease diagnostic tools can be helpful for routine monitoring of plant pathogens toward improving crop productivity and yield that can be used for improving the financial status of farmers.

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Section: Multiplex Detection Of Plant Pathogensmentioning

confidence: 99%

Section: Multiplex Detection Of Plant Pathogensmentioning

confidence: 99%

Emerging Extraction and Diagnostic Tools for Detection of Plant Pathogens: Recent Trends, Challenges, and Future Scope

Verma

Shukla

Kulkarni

et al. 2022

ACS Agric. Sci. Technol.

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“…The detection and characterisation of mycotoxigenic fungus using multiplex PCR (mPCR), which targets the genes responsible for the biosynthesis of mycotoxins, is rapid, inexpensive, and reliable (Priyanka et al 2015 ). Compared to monoplex PCR, mPCR allows for the simultaneous amplification of more than two genomic DNA regions in a single PCR reaction (Yli-Mattila et al 2017 ). The mPCR assay with competitive interior amplification control to eliminate false, negative findings was developed by utilising specific primers for each of the fungi species and optimising and validating them using standard isolates (Priyanka et al 2013 ; Neera and Murali 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Development of a real-time PCR and multiplex PCR assay for the detection and identification of mycotoxigenic fungi in stored maize grains

2023

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This study aimed to identify important mycotoxigenic fungi and accurate detection of mycotoxin in stored maize grains using molecular methods. The current study also optimised the real-time PCR (RT-PCR) assay. The melting curve was established to identify isolated fungal species of Aspergillus (4), Fusarium (3), Penicillium (3), and Alternaria (one). A multiplex polymerase chain reaction (mPCR) technique was developed for the detection and characterisation of mycotoxin producing fungi, mycotoxin metabolic pathway genes, and the determination of eleven mycotoxins in stored maize grains using high-performance liquid chromatography (HPLC). The mPCR results indicated positive signals for potentially mycotoxigenic fungal species tested of Aspergillus, Fusarium, Penicillium , and Alternaria . A protocol for multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was tested to distinguish between free and contaminated, stored maize with aflatoxin B1 (AFB1). The expression pattern of four aflatoxin biosynthetic pathway genes, AFB1 ( aflQ, aflP, aflO , and aflD ), was a good marker for contaminated, stored maize grains. HPLC analysis showed that maize grain samples were contaminated with mycotoxins, and the concentration was above the detection level. The results indicate that the polyphasic approach might provide a sensitive, rapid, and accurate method for detecting and identifying mycotoxigenic fungal species and mycotoxins in stored maize grains.

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“…The best way to identify new species and confirm the status of morphological species is to use phylogenetic species recognition, with genealogical concordance, in which several separate DNA sequences are used together (GCPSR) [8]. The sequence data can also be used for designing probes for species-specific genotyping of the isolates and multiplex detection of fungal species [9].…”

Section: Introductionmentioning

confidence: 99%

A Polyphasic Approach to Compare the Genomic Profiles of Aflatoxigenic and Non-Aflatoxigenic Isolates of Aspergillus Section Flavi

Abbas

Hussien

Yli-Mattila

2020

Toxins

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Aflatoxins (AF) are highly toxic compounds produced by Aspergillus section Flavi. They spoil food crops and present a serious global health hazard to humans and livestock. The aim of this study was to examine the phylogenetic relationships among aflatoxigenic and non-aflatoxigenic Aspergillus isolates. A polyphasic approach combining phylogenetic, sequence, and toxin analyses was applied to 40 Aspergillus section Flavi isolates collected from eight countries around the world (USA, Philippines, Egypt, India, Australia, Indonesia, China, and Uganda). This allows one to pinpoint the key genomic features that distinguish AF producing and non-producing isolates. Based on molecular identification, 32 (80%) were identified as A. flavus, three (7.5%) as A. parasiticus, three (7.5%) as A. nomius and one (2.5%) as A. tamarii. Toxin analysis showed that 22 (55%) Aspergillus isolates were aflatoxigenic. The majority of the toxic isolates (62.5%) originated from Egypt. The highest aflatoxin production potential was observed in an A. nomius isolate which is originally isolated from the Philippines. DNA-based molecular markers such as random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR) were used to evaluate the genetic diversity and phylogenetic relationships among these 40 Aspergillus isolates, which were originally selected from 80 isolates. The percentage of polymorphic bands in three RAPD and three ISSR primers was 81.9% and 79.37%, respectively. Analysis of molecular variance showed significant diversity within the populations, 92% for RAPD and 85% for ISSR primers. The average of Polymorphism Information Content (PIC), Marker Index (MI), Nei’s gene diversity (H) and Shannon’s diversity index (I) in ISSR markers are higher than those in RAPD markers. Based on banding patterns and gene diversities values, we observed that the ISSR-PCR provides clearer data and is more successful in genetic diversity analyses than RAPD-PCR. Dendrograms generated from UPGMA (Unweighted Pair Group Method with Arithmetic Mean) cluster analyses for RAPD and ISSR markers were related to the geographic origin.

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Multiplex Detection of Fusarium Species

Cited by 8 publications

References 36 publications

Emerging Extraction and Diagnostic Tools for Detection of Plant Pathogens: Recent Trends, Challenges, and Future Scope

Emerging Extraction and Diagnostic Tools for Detection of Plant Pathogens: Recent Trends, Challenges, and Future Scope

Development of a real-time PCR and multiplex PCR assay for the detection and identification of mycotoxigenic fungi in stored maize grains

A Polyphasic Approach to Compare the Genomic Profiles of Aflatoxigenic and Non-Aflatoxigenic Isolates of Aspergillus Section Flavi

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