Molecular Identification of the Fall Armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) Using PCR-RFLP

Lewter, Jennifer A.; Szalanski, Allen L.

doi:10.3954/1523-5475-24.2.51

Cited by 13 publications

(6 citation statements)

References 12 publications

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“…Furthermore, the morphological characterization of the eggs of S. eridania was performed by Valverde (2007). However, when morphological identiÞcation is impossible, molecular diagnostic protocols have been recommended to distinguish among species (Lewter and Szalanski 2007). Currently, DNA extraction can be performed within a few minutes, followed by am-pliÞcation and restriction analysis to obtain diagnostic DNA proÞles in one or two working days.…”

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confidence: 99%

Morphological and Molecular Characterization of the Eggs of Some Noctuid Species Associated with Soybean in Brazil

Sosa-Gómez

Yano

Specht

et al. 2013

Annals of the Entomological Society of America

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The most important species of Lepidoptera that attack soybean, in descending order of importance, are Chrysodeixis includens (Walker), Anticarsia gemmatalis Hü bner, Spodoptera cosmioides (Walker), Spodoptera eridania (Stoll), and Spodoptera albula (Walker). In addition, Rachiplusia nu (Guené e) is one of the most common Plusiinae in the southern region of Brazil, encompassing the states of Paraná, Santa Catarina, and Rio Grande do Sul. The correct identiÞcation of these species is essential for choosing the appropriate control measures, as they differ in their susceptibility to insecticides and biological control agents. To distinguish Lepidoptera species commonly found in Brazil, the eggshells of all these species were morphologically characterized by scanning electron microscopy. Diagnostic characters for differentiating species include shape of rosette petals, number of primary cells, number of secondary cells, number of ribs, and rosette diameter. Also, a molecular diagnostic method using polymerase chain reaction and restriction fragment length polymorphism (PCRÐRFLP) analysis was developed to distinguish noctuid species commonly found in Brazil. A 658-bp region of the mitochondrial DNA cytochrome oxidase subunit I gene was ampliÞed using PCR and then sequenced. The Þve Lepidoptera species were distinguished by restriction enzymes Bpm I and MboI. RFLPs produced by Bpm I endonuclease were useful to discriminate species from within Spodoptera (S. cosmioides, S. eridania, and S. albula) and R. nu from C. includens. However, Bpm I did not digest the amplicons from S. eridania and C. includens, which were discriminated by RFLP patterns produced by the restriction enzyme MboI. PCR-RFLP can be performed in a short period, and it is useful to distinguish the most important Brazilian Lepidoptera soybean pests.

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confidence: 99%

Morphological and Molecular Characterization of the Eggs of Some Noctuid Species Associated with Soybean in Brazil

Sosa-Gómez

Yano

Specht

et al. 2013

Annals of the Entomological Society of America

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“…Because FAW can cause such devastation in a short period of time, there is a global need for a rapid molecular diagnostic test to assist with early and accurate incursion responses. Some of the current methods used to identify FAW include restriction fragment length polymorphism (RFLP) 24 ; PCR and Sanger sequencing of both COI 25 and TPI 20 , 26 gene regions; species-specific multiplex PCR primers 27 ; and real-time PCR assays 28 . These techniques, while effective at diagnosing the target insect, are time-consuming and are often expensive, requiring highly specialized laboratory facilities and expert staff.…”

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confidence: 99%

A diagnostic LAMP assay for rapid identification of an invasive plant pest, fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)

Agarwal

Rako

Schutze

et al. 2022

Sci Rep

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Fall armyworm (FAW), Spodoptera frugiperda (Lepidoptera: Noctuidae), is a highly polyphagous invasive plant pest that has expanded its global geographic distribution, including recently into much of Australia. Rapid diagnostic tests are required for identification of FAW to assist subsequent management and control. We developed a new loop-mediated isothermal amplification (LAMP) assay based on the mitochondrial cytochrome c oxidase subunit I (COI) gene for accurate and timely diagnosis of FAW in the field. The specificity of the new assay was tested against a broad panel of twenty non-target noctuids, including eight other Spodoptera species. Only S. frugiperda samples produced amplification within 20 min, with an anneal derivative temperature of 78.3 ± 0.3 °C. A gBlock dsDNA fragment was developed and trialled as a synthetic positive control, with a different anneal derivative of 81 °C. The new FAW LAMP assay was able to detect FAW DNA down to 2.4 pg, similar to an existing laboratory-based real-time PCR assay. We also trialled the new FAW assay with a colorimetric master mix and found it could successfully amplify positive FAW samples in half the time compared to an existing FAW colorimetric LAMP assay. Given the high sensitivity and rapid amplification time, we recommend the use of this newly developed FAW LAMP assay in a portable real-time fluorometer for in-field diagnosis of FAW.

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“…This method amplifies a particular gene, then uses carefully selected restriction enzymes to cut at target sites, resulting in species-specific bands when the digested polymerase chain reaction product is viewed on an agarose gel (Hoy 2013). Comparative band lengths created with restriction enzymes have successfully been used as a rapid diagnostic technique in agricultural studies (Brunner et al 2002;Lewter and Szalanski 2007;Arneodo et al 2015;Hua et al 2018), including those identifying closely related Diptera species such as the members of the Calliphoridae (Ratcliffe et al 2003), Tephritidae (Armstrong et al 1997;Muraji and Nakahara 2002), and Drosophilidae families (Kim et al 2014).…”

Section: Introductionmentioning

confidence: 99%

Identification of morphologically challenging Delia (Diptera: Anthomyiidae) species from field vegetable crops using polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP)

Mlynarek

Sim

2021

Can Entomol

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Root feeding by the larvae of multiple Delia species can lead to economic loss in many agricultural crops. Field vegetables are subject to infestations by a species complex composed of Delia radicum (Linnaeus) (Diptera: Anthomyiidae), a pest in brassica crops (Brassicaceae), Delia antiqua (Meigen), believed to cause the majority of crop damage in onions, and the generalists Delia florilega (Zetterstedt), Delia platura (Meigen), and Botanophila fugax (Meigen) (Diptera: Anthomyiidae). Correct species identification is necessary to implement field management strategies, but these species are challenging to identify morphologically. We propose a polymerase chain reaction–restriction fragment length polymorphism method as a molecular tool to distinguish between five species of Delia and between two genetic lines of D. platura. The mitochondrial DNA cytochrome c oxidase subunit 1 barcode fragment is targeted, then the polymerase chain reaction product digested with four different restriction enzymes (AccI, BsrI, MlyI, and StyI). The BsrI enzyme distinguishes the two genetic lines of D. platura and D. florilega. The MlyI enzyme identifies B. fugax from the Delia species. Combining BsrI, StyI, AccI, and MlyI into double digestion reactions allows for rapid diagnostics among the species tested. Our method was validated using DNA from specimens collected in eastern Canada. This method provides tools in ecological and environmental studies where these species are of interest.

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Molecular Identification of the Fall Armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) Using PCR-RFLP

Cited by 13 publications

References 12 publications

Morphological and Molecular Characterization of the Eggs of Some Noctuid Species Associated with Soybean in Brazil

Morphological and Molecular Characterization of the Eggs of Some Noctuid Species Associated with Soybean in Brazil

A diagnostic LAMP assay for rapid identification of an invasive plant pest, fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae)

Identification of morphologically challenging Delia (Diptera: Anthomyiidae) species from field vegetable crops using polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP)

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