Molecular diagnostics of the Mediterranean olive cyst nematode, Heterodera mediterranea Vovlas, Inserra &amp; Stone, 1981 using conventional and real-time PCR

Subbotin, Sergei A.; Burbridge, Julie; Palomares‐Rius, Juan E.; Castillo, Pablo

doi:10.1007/s10658-022-02598-4

Cited by 3 publications

References 10 publications

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Pioneering Nit Gene Exploitation to Develop Molecular Diagnostic Assay for Rapid Detection of Cotton Root Rot Incitant, Macrophomina phaseolina (Tassi) Goid, in Field Soil

Saini,

Kumar,

Singh

et al. 2024

Journal of Basic Microbiology

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Cotton root rot caused by Macrophomina phaseolina pose a significant threat to cotton production, leading to substantial yield and quality losses. Early and accurate diagnosis of this pathogen in soil is crucial for effective disease management. This study presents a pioneering investigation into the utilization of the nit gene encoding nitrilase for the development of a molecular diagnostic assay aimed at the rapid detection of M. phaseolina in field soils. The methodology involved the design and validation of primers targeting the Nit gene sequence, followed by the optimization of PCR conditions for efficient amplification. Leveraging state‐of‐the‐art molecular techniques, the assay offers a novel protocol to accurately identify the presence of M. phaseolina in soil with high sensitivity and specificity. The specificity of the designed primers was confirmed through PCR amplification using DNA from M. phaseolina and other related fungi. Sensitivity tests demonstrated that the PCR assay reliably detected M. phaseolina DNA at concentrations as low as 1 ng. Furthermore, the performance of the diagnostic assay was rigorously evaluated using field soil samples with a known status of M. phaseolina infection, demonstrating its reliability and efficacy in real‐world scenarios. This study introduces a novel molecular marker for the detection of M. phaseolina and offers a rapid and efficient means for screening M. phaseolina in large soil samples with minimal time and manpower.

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Pioneering Nit Gene Exploitation to Develop Molecular Diagnostic Assay for Rapid Detection of Cotton Root Rot Incitant, Macrophomina phaseolina (Tassi) Goid, in Field Soil

Saini,

Kumar,

Singh

et al. 2024

Journal of Basic Microbiology

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A diagnostic gap to fill: Development of molecular tools to distinguish the cyst-forming nematode species Heterodera carotae and Heterodera cruciferae

Fouville,

Biget,

Montarry

et al. 2024

Physiological and Molecular Plant Pathology

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A diagnostic gap to fill: Development of molecular tools to distinguish the plant-parasitic nematode speciesHeterodera carotaeandHeterodera cruciferae

Fouville,

Biget,

Montarry

et al. 2024

Preprint

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The carrot-parasitic nematode, Heterodera carotae, is an important pest causing qualitative and quantitative production losses across several countries worldwide and must be carefully identified and monitored. Indeed, in culture crops, Heterodera carotae and Heterodera cruciferae, two genetically closely related species from the Goettingiana group, can be found in mixture and are virtually morphologically non-recognizable. However, only H. carotae is able to develop on carrot. To monitor carrot production and set up suitable H. carotae control plans, simple and reliable diagnostic methods to detect and identify these two species are needed. In this study, we developed and successfully tested two real-time PCR protocols (SYBR Green singleplex and TaqMan multiplex real-time PCRs) against both sister species: H. carotae and H. cruciferae. Using two specific sets of primers targeting the two species, which were designed from sequences containing microsatellite loci, we managed, whatever the methods, to distinguish the two species. Moreover, we showed a higher specificity using the TaqMan assays (test on cysts) but a better sensitivity (tests on J2 juvenile larvae) with the SYBR Green assays. In this study, we thus proposed two useful and reliable diagnostic tools to enable farmers and scientists to rapidly detect two plant-parasitic nematodes and supervise agricultural management.

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Molecular diagnostics of the Mediterranean olive cyst nematode, Heterodera mediterranea Vovlas, Inserra & Stone, 1981 using conventional and real-time PCR

Cited by 3 publications

References 10 publications

Pioneering Nit Gene Exploitation to Develop Molecular Diagnostic Assay for Rapid Detection of Cotton Root Rot Incitant, Macrophomina phaseolina (Tassi) Goid, in Field Soil

Pioneering Nit Gene Exploitation to Develop Molecular Diagnostic Assay for Rapid Detection of Cotton Root Rot Incitant, Macrophomina phaseolina (Tassi) Goid, in Field Soil

A diagnostic gap to fill: Development of molecular tools to distinguish the cyst-forming nematode species Heterodera carotae and Heterodera cruciferae

A diagnostic gap to fill: Development of molecular tools to distinguish the plant-parasitic nematode speciesHeterodera carotaeandHeterodera cruciferae

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