A qPCR-based method for detecting parasitism of<i>Fopius arisanus</i>(Sonan) in oriental fruit flies,<i>Bactrocera dorsalis</i>(Hendel)

Liang, Guanghong; Heller, Wade P.; Chang, Chiou Ling; Chen, Jia Hua; Zhang, Fei Ping; Geib, Scott M.

doi:10.1002/ps.3976

Cited by 6 publications

(6 citation statements)

References 51 publications

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“…Liang et al . 51 obtained similar results using qPCR, with an egg detection threshold of 0.25 ng for parasitoid DNA [( Fopius arisanus (Sonan)] in 40 ng of host DNA [ Bactrocera dorsalis (Hendel)], for a dilution ratio of ~0.006. In addition, our detection threshold appeared sensitive enough to detect all the developmental stages of our parasitoids, including embryonic stages.…”

Section: Discussionmentioning

confidence: 58%

“…Host larva mortality remains the main limitation for an accurate and reproducible estimation of parasitism rates in the rearing laboratory 52,53 . In this case, DNA metabarcoding is useful for estimating parasitism rates because it can be used to detect parasitoids in the host body, whether the larval parasitoid is alive or dead 51 . For example, we can take the case of S. sahelensis , which was detected by both methods (DNA metabarcoding and rearing), and its detection threshold (~0.001) was low enough to detect all parasitoid stages.…”

Section: Discussionmentioning

confidence: 99%

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Deciphering host-parasitoid interactions and parasitism rates of crop pests using DNA metabarcoding

Sow

Brévault

Benoit

et al. 2019

Sci Rep

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An accurate estimation of parasitism rates and diversity of parasitoids of crop insect pests is a prerequisite for exploring processes leading to efficient natural biocontrol. Traditional methods such as rearing have been often limited by taxonomic identification, insect mortality and intensive work, but the advent of high-throughput sequencing (HTS) techniques, such as DNA metabarcoding, is increasingly seen as a reliable and powerful alternative approach. Little has been done to explore the benefits of such an approach for estimating parasitism rates and parasitoid diversity in an agricultural context. In this study, we compared the composition of parasitoid species and parasitism rates between rearing and DNA metabarcoding of host eggs and larvae of the millet head miner, Heliocheilus albipunctella De Joannis (Lepidoptera, Noctuidae), collected from millet fields in Senegal. We first assessed the detection threshold for the main ten endoparasitoids, by sequencing PCR products obtained from artificial dilution gradients of the parasitoid DNAs in the host moth. We then assessed the potential of DNA metabarcoding for diagnosing parasitism rates in samples collected from the field. Under controlled conditions, our results showed that relatively small quantities of parasitoid DNA (0.07 ng) were successfully detected within an eight-fold larger quantity of host DNA. Parasitoid diversity and parasitism rate estimates were always higher for DNA metabarcoding than for host rearing. Furthermore, metabarcoding detected multi-parasitism, cryptic parasitoid species and differences in parasitism rates between two different sampling sites. Metabarcoding shows promise for gaining a clearer understanding of the importance and complexity of host-parasitoid interactions in agro-ecosystems, with a view to improving pest biocontrol strategies.

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Section: Discussionmentioning

confidence: 58%

Section: Discussionmentioning

confidence: 99%

Deciphering host-parasitoid interactions and parasitism rates of crop pests using DNA metabarcoding

Sow

Brévault

Benoit

et al. 2019

Sci Rep

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“…We did not find other studies that tried to identify ghost parasitoids in host carcasses weeks after host death, nor did we find studies using MCA to detect and identify parasitoids. The closest method to ours to detect and identify parasitoids was the use of qPCR, in which detection was inferred using the quantification cycle threshold (Cq) method for presence/absence and the specificity assured by the use of species-specific probes (Liang et al, 2015(Liang et al, , 2018 TA B L E 3 Percentage of parasitoids detected by MCA in host carcasses after parasitoid emergence in relation to the total number of parasitoids observed by rearing primer or probe specificity is critically important to ensure that only the desired amplicon is amplified. These authors used the melting curves to confirm the absence of non-specific PCR products.…”

Section: Discussionmentioning

confidence: 99%

“…In the case of using species‐specific probes, there is no need to use species‐specific primers (e.g. Liang et al., 2015).…”

Section: Discussionmentioning

confidence: 99%

Melting curve analysis for detection and identification of ghost parasitoids in host carcasses a month after host death

Paula

Andow

2021

Methods Ecol Evol

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1. Incidence of parasitism is often underestimated because 'ghost' parasitoids (dead unemerged parasitoids or those that have emerged leaving the host carcasses) are difficult to detect and identify. This study demonstrates that the use of melting curve analysis (MCA) of host carcasses can detect and identify DNA of ghost parasitoids even a month after host death. 2. The coccinellid hosts Cycloneda sanguinea, Eriopis connexa, Harmonia axyridis and Hippodamia convergens were sampled from cole crops in 2017 and 2018 in the Midwest of Brazil, and reared and observed daily for parasitoid emergence. Dead coccinellids were held for 30 days after death before storage and only host carcasses with parasitoid emergence observed were analysed. Species-specific primers were designed for the identification of the parasitoid species that emerged during host rearing: Dinocampus coccinellae, Homalotylus mirabilis, Ho. terminalis, Strongygaster triangulifera and Phalacrotophora sp. The melting temperatures (T m ) of their amplicons were used as positive controls in MCA post-amplification in qPCR.3. Detection of parasitoid DNA in host carcasses was possible for D. coccinellae, Ho. mirabilis and Phalacrotophora sp. with a limit of detection (LOD) for all the parasitoids <1 pg of DNA, except for Phalacroptopthora sp. (LOD = 1.9 ng). Parasitoids were detected in 31 out of 70 host carcasses (44.3%) with detection ranging from 0% to 71%, depending on the parasitoid species.4. Synthesis and applications. This work demonstrates that MCA could be used to detect and rapidly identify the DNA of some parasitoid species (e.g. Ho. mirabilis, D. coccinellae and Phalacrotophora sp.) in host carcasses up to a month after parasitoid emergence with high sensitivity and specificity. MCA can be used on any field-collected host and, for some parasitoid species, it should lead to more accurate estimates of the incidence of parasitism. Our approach can be easily adapted and applied to study other parasitoid-host interactions.

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“…In an agricultural ecosystem, insect parasitoids play very important roles in the trophic networks and attract much research for their potential in biological control (Godfray 1994), especially as some species have largely been reared artificially and then released in the fields to suppress the pest populations (Liang et al 2015). Each species kills and lives at the expense of host as a result of its development after ovipositing its egg within or on the surface of the host (Eggleton and Gaston 1990;Rachel et al 2016).…”

Section: Introductionmentioning

confidence: 99%

A new species of Glyptapanteles Ashmead (Hymenoptera, Braconidae, Microgastrinae) within Macrobrochis gigas (Lepidoptera, Arctiidae, Lithosiidae) in Fujian, China

Tang

Dong

et al. 2020

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The south-east coastal area of Fujian, China, belongs to the Oriental Realm, and is characterized by a high insect species richness. In this work, a new species of Hymenopteran parasitoid, Glyptapanteles gigas Liang & Song, sp. nov. found in Jinjiang within hosts of caterpillars Macrobrochis gigas (Lepidoptera: Arctiidae), is described and illustrated, with differences from similar species. Additionally, we presumed that both parasitoid and host species play very important role in the coevolution and tritrophic interaction between plants, phytophagous insects, and their parasitoids, because these insects probably broke the sporangia and made contributions to their colonization, or some spores were spread for long distances by adult moths after their emergence, or some parasitoids were attracted by the eggs and larvae of these caterpillars, which was also thought to be helpful to spread of spores.

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A qPCR-based method for detecting parasitism ofFopius arisanus(Sonan) in oriental fruit flies,Bactrocera dorsalis(Hendel)

Abstract: This method is a rapid, sensitive and specific technique to determine the parasitism rate of F. arisanus across all life stages of B. dorsalis, which will be useful to predict parasitoid output from mass rearing and evaluate the outcome of pest suppression after mass release in the field.

Cited by 6 publications

References 51 publications

Deciphering host-parasitoid interactions and parasitism rates of crop pests using DNA metabarcoding

Deciphering host-parasitoid interactions and parasitism rates of crop pests using DNA metabarcoding

Melting curve analysis for detection and identification of ghost parasitoids in host carcasses a month after host death

A new species of Glyptapanteles Ashmead (Hymenoptera, Braconidae, Microgastrinae) within Macrobrochis gigas (Lepidoptera, Arctiidae, Lithosiidae) in Fujian, China

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