Identification and characterization of multiple dsRNases from a lepidopteran insect, the tobacco cutworm, Spodoptera litura (Lepidoptera: Noctuidae)

Peng, Yajun; Wang, Kangxu; Zhu, Guan-Heng; Han, Qunxin; Chen, Jiasheng; Elzaki, Mohammed Esmail Abdalla; Sheng, Cheng-Wang; Zhao, Chunqing; Palli, Subba Reddy; Han, Zhaojun

doi:10.1016/j.pestbp.2019.09.011

Cited by 44 publications

(60 citation statements)

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“…These results suggest bioinformatic analyses (e.g., 21 base pair matches) alone cannot predict potential dsRNA sensitivity to target species (and insensitivity to non-target species). Other factors including refractory genes, presence of high levels of dsRNase, and exposure to low environmental concentrations may prevent RNAi-mediated effects [ 23 , 35 , 36 ].…”

Section: Discussionmentioning

confidence: 99%

“…These three studies employed dsRNA-treated artificial diets rather than treated-host plant leaves. Of note, tobacco cutworm larvae that fed on cabbage leaves had greater dsRNA-degrading activity than larvae that were reared on an artificial diet [ 36 ]. These authors suggest that artificial diet could potentially influence dsRNase expression, dsRNA stability, and RNAi efficiency.…”

Section: Discussionmentioning

confidence: 99%

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Evaluating toxicity of Varroa mite (Varroa destructor)-active dsRNA to monarch butterfly (Danaus plexippus) larvae

et al. 2021

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Varroa mites (Varroa destructor) are parasitic mites that, combined with other factors, are contributing to high levels of honey bee (Apis mellifera) colony losses. A Varroa-active dsRNA was recently developed to control Varroa mites within honey bee brood cells. This dsRNA has 372 base pairs that are homologous to a sequence region within the Varroa mite calmodulin gene (cam). The Varroa-active dsRNA also shares a 21-base pair match with monarch butterfly (Danaus plexippus) calmodulin mRNA, raising the possibility of non-target effects if there is environmental exposure. We chronically exposed the entire monarch larval stage to common (Asclepias syriaca) and tropical (Asclepias curassavica) milkweed leaves treated with concentrations of Varroa-active dsRNA that are one- and ten-fold higher than those used to treat honey bee hives. This corresponded to concentrations of 0.025–0.041 and 0.211–0.282 mg/g leaf, respectively. Potassium arsenate and a previously designed monarch-active dsRNA with a 100% base pair match to the monarch v-ATPase A mRNA (leaf concentration was 0.020–0.034 mg/g) were used as positive controls. The Varroa mite and monarch-active dsRNA’s did not cause significant differences in larval mortality, larval or pupal development, pupal weights, or adult eclosion rates when compared to negative controls. Irrespective of control or dsRNA treatment, larvae that consumed approximately 7500 to 10,500-mg milkweed leaf within 10 to 12 days had the highest pupal weights. The lack of mortality and sublethal effects following dietary exposure to dsRNA with 21-base pair and 100% base pair match to mRNAs that correspond to regulatory genes suggest monarch mRNA may be refractory to silencing by dsRNA or monarch dsRNase may degrade dsRNA to a concentration that is insufficient to silence mRNA signaling.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Evaluating toxicity of Varroa mite (Varroa destructor)-active dsRNA to monarch butterfly (Danaus plexippus) larvae

et al. 2021

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“…Foi rodado novamente uma busca, porém com o comando "tblastx", utilizando os mesmos parâmetros e, dessa vez, foram encontrados 5 contigs potenciais para a REase de T. (GUAN et al, 2018a(GUAN et al, , 2019. Porém, a identificação de múltiplas dsRNases atuando junto na degradação de moléculas de dsRNA em um inseto recalcitrante ao RNAi já foi demonstrada anteriormente (PENG et al, 2020). As dsRNases são assim denominadas pela sua atividade de degradação de moléculas de dsRNA, se relacionando com a eficiência da técnica de…”

Section: Resultsunclassified

“…Em trabalho posterior, foi demonstrado que um homólogo da REase em Helicoverpa armigera teve sua expressão aumentada na presença de dsRNA, corroborando os dados anteriormente demonstrados em Ostrinia furnacalis (GUAN et al, 2018a(GUAN et al, , 2019. Além disso, outras dsRNases têm sido reportadas em outras espécies e ordens de insetos e têm sido apontadas como possível mecanismo associado à baixa eficiência no silenciamento gênico desencadeado pela molécula de dsRNA (SPIT et al, 2017;PRENTICE et al, 2019;PENG et al, 2020 (URBANEJA et al, 2013). Os adultos são mariposas pequenas, com comprimento de 10-11 mm, apresentam coloração cinza-prateada, antenas filiformes e palpos labiais recurvados (COELHO;FRANÇA, 1987;HAJI et al, 1988).…”

Section: Introductionunclassified

“…). Em um estudo com o objetivo de avaliar a contribuição de mais de uma dsRNase na redução da eficiência do silenciamento gênico por RNAi em Spodoptera litura, foram encontrados seis genes que codificam endonucleases não específicas de DNA/RNA (DNA/RNA non-specific endonuclease) em que três eram especificamente expressas no intestino médio da fase larval desse inseto(PENG et al, 2020). Nesse caso, foi demonstrada a atividade de dsRNase para 4 das 5 proteínas testadas, onde a maior atividade de degradação foi observada no terceiro ínstar de lagartas de S. litura.…”

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Identificação de ribonuclease potencialmente associada a degradação de dupla fita de RNA (dsRNA) em >i<Tuta absoluta>/i< (Meyrick) e a eficiência no silenciamento gênico induzido pelo RNA de interferência (RNAi)

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Identification and functional analysis of dsRNases in spotted‐wing drosophila, Drosophila suzukii

Yoon

Ahn

Flinn

et al. 2021

Arch Insect Biochem Physiol

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RNAi efficiency in insects is different from species to species; some species in Coleoptera are relatively more amenable to RNA interference (RNAi) than other species.One of the major factors is the presence of dsRNAdegrading enzymes, called dsRNases, in saliva, gut, or hemolymph in insects, which degrade the double-stranded RNA (dsRNA) introduced, resulting in the low efficacy of RNAi. In this study, we report a dsRNA-degrading activity in the gut homogenates from the spotted-wing drosophila, Drosophila suzukii, by ex vivo assay. Then, we identified two Drosophila suzukii dsRNase genes, named DrosudsRNase1 and DrosudsRNase2. In silico analysis shows that the gene structures are similar to dsRNases found in other insects.When dsRNases expressed in Sf9 cells were compared for their dsRNA degrading activities, dsRNase1 was more vital than dsRNase2. Both dsRNases were expressed highly and exclusively in the gut compared to the rest of body. Also, they were highly expressed during larval and adult stages but not in embryonic and pupal stages, suggesting the dsRNases protect foreign RNA molecules received during the feeding periods. DsRNase1 was expressed at a higher level in adults, whereas dsRNase2 showed more expression in early larvae. Our study on the tissue and development-This article has been contributed to by US Government employees and their work is in the public domain in the USA.

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Identification and characterization of multiple dsRNases from a lepidopteran insect, the tobacco cutworm, Spodoptera litura (Lepidoptera: Noctuidae)

Cited by 44 publications

References 43 publications

Evaluating toxicity of Varroa mite (Varroa destructor)-active dsRNA to monarch butterfly (Danaus plexippus) larvae

Evaluating toxicity of Varroa mite (Varroa destructor)-active dsRNA to monarch butterfly (Danaus plexippus) larvae

Identificação de ribonuclease potencialmente associada a degradação de dupla fita de RNA (dsRNA) em >i<Tuta absoluta>/i< (Meyrick) e a eficiência no silenciamento gênico induzido pelo RNA de interferência (RNAi)

Identification and functional analysis of dsRNases in spotted‐wing drosophila, Drosophila suzukii

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