Development of amplified fragment length polymorphism (AFLP)-derived specific primer for the detection of <i>Fusarium solani</i>
 aetiological agent of peanut brown root rot

Casasnovas, Francisco; Fantini, E.N.; Palazzini, Juan Manuel; Giaj-Merlera, G.; Chulze, Sofía Noemí; Reynoso, María Marta; Torres, Adriana Mabel

doi:10.1111/jam.12183

Cited by 13 publications

(6 citation statements)

References 40 publications

(70 reference statements)

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“…The following are available online at , Figure S1: Species specific multiplex PCR assays using DNA isolated from stored wheat grain, Table S1: Primer sets used in this study (including references [ 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 ]), Table S2: Fungal isolates used in this study.…”

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confidence: 99%

Rapid Detection and Identification of Mycotoxigenic Fungi and Mycotoxins in Stored Wheat Grain

Sudharsan

Britzi²,

Zakin

et al. 2017

Toxins

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This study aimed to assess the occurrence of toxigenic fungi and mycotoxin contamination in stored wheat grains by using advanced molecular and analytical techniques. A multiplex polymerase chain reaction (PCR) strategy was established for rapid identification of mycotoxigenic fungi, and an improved analytical method was developed for simultaneous multi-mycotoxin determination in wheat grains by liquid chromatography-tandem mass spectrometry (LC/MS/MS) without the need for any clean-up. The optimized multiplex PCR method was highly specific in detecting fungal species containing species-specific and mycotoxin metabolic pathway genes. The method was applied for evaluation of 34 wheat grain samples collected from storage warehouses for the presence of mycotoxin-producing fungi, and a few samples were found positive for Fusarium and Aspergillus species. Further chemical analysis revealed that 17 samples contained mycotoxins above the level of detection, but only six samples were found to be contaminated over the EU regulatory limits with at least one mycotoxin. Aflatoxin B1, fumonisins, and deoxynivalenol were the most common toxins found in these samples. The results showed a strong correlation between the presence of mycotoxin biosynthesis genes as analyzed by multiplex PCR and mycotoxin detection by LC/MS/MS. The present findings indicate that a combined approach might provide rapid, accurate, and sensitive detection of mycotoxigenic species and mycotoxins in wheat grains.

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confidence: 99%

Rapid Detection and Identification of Mycotoxigenic Fungi and Mycotoxins in Stored Wheat Grain

Sudharsan

Britzi²,

Zakin

et al. 2017

Toxins

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show abstract

“…AFLP fingerprinting proved to be powerful in grouping citrus cultivars according to their level of relatedness to each other. AFLP is a widely used technique in the population genetic and phylogenetic studies of plants and microorganisms (Keivani et al 2010;Al-Sadi et al 2012;Kumar et al 2012 (Nei 1978) of 12 citrus species and 6 accessions based on AFLP fingerprinting analysis using 910 polymorphic loci Casasnovas et al 2013;Al-Maamari et al 2014;Al-Sadi et al 2015). AFLP is also commonly used for studying relationship and genetic diversity in citrus species (Nartvaranant & Nartvaranant 2011;Khiavi et al 2015;Zhou et al 2017).…”

Section: Discussionmentioning

confidence: 99%

AFLP Fingerprinting Analysis of Citrus Cultivars and Wild Accessions from Oman Suggests the Presence of Six Distinct Cultivars

Al-Nadabi

Khan

Al-Yahyai

et al. 2018

Agriculture (Pol'nohospodárstvo)

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A study was conducted to evaluate genetic relatedness of 27 citrus cultivars and 6 wild citrus accessions using AFLP fingerprinting. The 27 citrus cultivars belonged to Citrus sinensis, C. aurantifolia, C. aurantium, C. paradise, C. reticulata, C. limon, C. latifolia, C. maxima, C. limettoides, C. limetta, C. medica and C. Jambhiri. The wild cultivars were obtained from Oman while the other cultivars originated from Oman and other countries. AFLP analysis using 4 primer pair combinations resolved 910 polymorphic alleles. All citrus cultivars and accessions had low genetic diversity (H = 0.0281 to 0.1300), with the percent polymorphic loci ranging from 8 to 35%. Populations of the six wild citrus accessions showed a very low level of genetic diversity (< 0.0700). Cluster analysis of the 33 cultivars and accessions showed that they share a high level of genetic similarity (81‒99%; mean = 92%). The six wild accessions clustered into two main clusters, with the analysis indicating that the six wild accessions may make up six distinct cultivars. The study provides information on the phylogeny of citrus cultivars and citrus diversity in Oman, a country through which citrus moved in the past from Asia to different African and European countries. In addition, it shows that some distinct citrus cultivars are present in this part of the world.

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“…So far, methods to detect F. solani are based on pathogen isolation, microscope observation, polymerase chain reaction (PCR), real-time qPCR (qPCR), TaqMan real-time PCR, amplified fragment length polymorphisms (AFLPs), and others [ 7 , 8 , 9 , 10 ]. However, the limitation of nucleic acid-based detection techniques is that they cannot distinguish between viable and nonviable cells, since the DNA of dead cells can exist in air or soil for a long time [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

An Improved Method for Quantification of Viable Fusarium Cells in Infected Soil Products by Propidium Monoazide Coupled with Real-Time PCR

Chen

Xie

et al. 2022

Microorganisms

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Fusarium is a soil-borne pathogen that causes root rot disease in cucumber. To date, quantitative real-time PCR (qPCR) is a common tool to detect the content of Fusarium in soil. However, qPCR cannot distinguish between viable and nonviable cells. The aim of this study was to develop a detection technique to pretreat tissue fluid with propidium monoazide (PMA) followed by extract DNA, and then to quantify viable Fusarium cells in contaminated soil. In this work, the specific primer pair F8-1/F8-2 was designed based on the translation elongation factor (EF) gene and a PMA-qPCR assay was established to amplify and quantify soils of viable Fusarium cells. The PMA pretreatment test was optimized, which indicated that the optimal PMA concentration and light exposure time were 50 mmol L−1 and 15 min, respectively. The lowest limit of viable cells in suspension detected and soil by PMA-qPCR were 82 spore mL−1 and 91.24 spore g−1, respectively. For naturally contaminated soil, viable Fusarium cells were detected in eight of the 18 samples, and the Fusarium amount ranged from 104 to 106 spore g−1. In conclusion, the PMA-qPCR method has the characteristics of high sensitivity, efficiency, and time saving, which could support nursery plants to avoid Fusarium infection and agro-industry losses.

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Development of amplified fragment length polymorphism (AFLP)-derived specific primer for the detection of Fusarium solani aetiological agent of peanut brown root rot

Cited by 13 publications

References 40 publications

Rapid Detection and Identification of Mycotoxigenic Fungi and Mycotoxins in Stored Wheat Grain

Rapid Detection and Identification of Mycotoxigenic Fungi and Mycotoxins in Stored Wheat Grain

AFLP Fingerprinting Analysis of Citrus Cultivars and Wild Accessions from Oman Suggests the Presence of Six Distinct Cultivars

An Improved Method for Quantification of Viable Fusarium Cells in Infected Soil Products by Propidium Monoazide Coupled with Real-Time PCR

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