A review of methods for nematode identification

Seesao, Yuwalee; Merlin, Sophie; Viscogliosi, Éric; Aliouat-Denis, Cécile-Marie; Audebert, Christophe

doi:10.1016/j.mimet.2016.05.030

Cited by 65 publications

(45 citation statements)

References 116 publications

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“…Additionally, we present evidence that LAMP is faster than traditional morphology-based microscopic methods. The entire morphological diagnosis process took about 2 days which is very consistent with other reported observational methods of RKN diagnosis that mostly rely on the distinct morphological and anatomical characteristics of second-stage juveniles, adult males, as well as perineal patterns of adult females [10, 28–30]. Our results based on the morphological characteristics of J2s in this study were in agreement with those reported for M. partityla [26, 27] and confirms the reliability of the LAMP technique for rapid identification of M. partityla .…”

Section: Discussionsupporting

confidence: 82%

Rapid detection of Pecan Root-Knot Nematode,Meloidogyne partityla, in laboratory and field conditions using loop-mediated isothermal amplification

Waliullah

Bell

Stackhouse

et al. 2020

Preprint

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2Meloidogyne partityla is the dominant root-knot nematode (RKN) species parasitizing pecan in 3 Georgia. This species is known to cause a reduction in root growth and a decline in yields from 4 mature pecan trees. Rapid and accurate diagnosis of this RKN is required to control this nematode 5 disease and reduce losses in pecan production. In this study, a loop-mediated isothermal 6 amplification (LAMP) method was developed for simple, rapid and on-site detection of M. 7 partityla in infested plant roots and validated to detect the nematode in laboratory and field 8 conditions. Specific primers were designed based on the sequence distinction of internal 9 transcribed spacer (ITS)-18S/5.8S ribosomal RNA gene between M. partityla and other 10 Meloidogyne spp. The LAMP detection technique could detect the presence of M. partityla 11 genomic DNA at a concentration as low as 1 pg, and no cross reactivity was found with DNA from 12 other major RKN species such as M. javanica, M. incognita and M. arenaria, and M. hapla. We 13 also conducted a traditional morphology-based diagnostic assay and conventional polymerase 14 chain reaction (PCR) assay to determine which of these techniques was less time consuming, more 15 sensitive, and convenient to use in the field. The LAMP assay provided more rapid results, 16 amplifying the target nematode species in less than 60 min at 65°C, with results 100 times more 17 sensitive than conventional PCR (~2-3 hrs). Morphology-based, traditional diagnosis was highly 18 time-consuming (2 days) and more laborious than conventional PCR and LAMP assays. These 19 features greatly simplified the operating procedure and made the assay a powerful tool for rapid, 20 on-site detection of pecan RKN, M. partityla. The LAMP assay will facilitate accurate pecan 21 nematode diagnosis in the field and contribute to the management of the pathogen. 3 1 diagnosis 3 Introduction 4 Pecan (Carya illinoinensis) is an important nut crop in North America. A variety of diseases caused 5 by bacteria, fungi, virus, and nematode can attack these trees, and if not properly managed, they 6 can cause economic damage to pecans. Root-knot nematodes (RKNs) of the genus Meloidogyne 7 are economically important plant-parasitic nematodes, which cause significant damage to pecan 8 production. Three species of RKNs, Meloidogyne incognita, M. arenaria and M. partityla have, 9 have been reported as pathogenic to pecan [1-3]. Among these species M. partityla is the dominant 10 RKN parasitizing pecan which has a greater incidence in southern United States [2-6]. M. partityla 11 was originally found infecting pecan in South Africa, and was likely introduced into the United 12 States during importation of infected pecan seedlings [7]. Due to the complexity and lengthy 13 process of diagnosis and species identification, it is difficult to conduct extensive surveys, which 14are required for determining the pecan RKN distribution. Therefore, a quick diagnosis approach 15 will be helpful to know the latest status of the pecan RKNs in s...

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Section: Discussionsupporting

confidence: 82%

Rapid detection of Pecan Root-Knot Nematode,Meloidogyne partityla, in laboratory and field conditions using loop-mediated isothermal amplification

Waliullah

Bell

Stackhouse

et al. 2020

Preprint

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“…Due to their huge economic and trade impacts, it is crucial to distinguish these species accurately using diagnostic tools. Morphological identification, based on a few characters of the second-stage juvenile (J2) and of the perineal area of the cyst, has been quite successful but always carries some uncertainty (Bačić et al, 2013;Seesao et al, 2016;Tirchi et al, 2016). Therefore, due to the variability of the main morphological features and the overlapping of standard diagnostic parameters in these two species, a confirmation through molecular methods is recommended.…”

mentioning

confidence: 99%

Morphological and molecular identification of the potato cyst nematodes Globodera rostochiensis and G. pallida in Portuguese potato fields

Camacho

Nóbrega

Lima

et al. 2017

Nematol

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The potato cyst nematodes (PCN) Globodera rostochiensis and G. pallida pose one of the greatest threats to potato crops worldwide and are subject to strict quarantine regulations in many countries. The identification of these Globodera species based on morphology may be ambiguous due to the variability of the main morphological features and the overlapping of the standard parameters in these two species; thus, confirmation via molecular methods is recommended. Multiplex PCR with speciesspecific primers (ITS5/PITSp4 + PITSr3) allows both species to be distinguished. However, despite the development of molecular identification methods, the morphological approach remains useful as a complementary diagnostic technique. In this work, we report results of morphological and molecular analyses that were carried out in two Globodera species from Portuguese potato fields. The average morphometric values of 40 cysts and 40 second-stage juveniles were generally within the expected ranges for G. pallida and G. rostochiensis with some variations noted. Molecular analysis with multiplex PCR confirmed the morphometric identification. The present results confirmed the occurrence of two potato cyst nematode species, G. rostochiensis and G. pallida. Surprisingly, the analysis of soils from Portuguese potato fields detected a greater number of samples infested with G. pallida, which is contrary to expectation as G. rostochiensis has been considered the most widespread species in Portugal. The distinction between the two species is therefore essential in order to detect their presence in the country with a view to re-evaluating the control measures implemented so far and adopting more effective practices.

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“…As nucleotide sequences vary among species, restriction sites differ in their locations along the genome, resulting in fragments of different sizes. Finally, restriction products are separated by gel electrophoresis (SEESSAO et al, 2017). This technique allows the identification of M. hapla, M. incognita and M. arenaria (HAN et al, 2004).…”

Section: Restriction Fragment Length Polymorphism (Rflp)mentioning

confidence: 99%

Diagnostic methods for identification of root-knot nematodes species from Brazil

et al. 2018

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The accurate identification of root-knot nematode (RKN) species (Meloidogyne spp.) is essential for implementing management strategies. Methods based on the morphology of adults, isozymes phenotypes and DNA analysis can be used for the diagnosis of RKN. Traditionally, RKN species are identified by the analysis of the perineal patterns and esterase phenotypes. For both procedures, mature females are required. Over the last few decades, accurate and rapid molecular techniques have been validated for RKN diagnosis, including eggs, juveniles and adults as DNA sources. Here, we emphasized the methods used for diagnosis of RKN, including emerging molecular techniques, focusing on the major species reported in Brazil.

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A review of methods for nematode identification

Cited by 65 publications

References 116 publications

Rapid detection of Pecan Root-Knot Nematode,Meloidogyne partityla, in laboratory and field conditions using loop-mediated isothermal amplification

Rapid detection of Pecan Root-Knot Nematode,Meloidogyne partityla, in laboratory and field conditions using loop-mediated isothermal amplification

Morphological and molecular identification of the potato cyst nematodes Globodera rostochiensis and G. pallida in Portuguese potato fields

Diagnostic methods for identification of root-knot nematodes species from Brazil

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