A TaqMan qPCR method for detecting kdr resistance in Aphis gossypii demonstrates improved sensitivity compared to conventional PCR–RFLP

Suann, Monica; Bogema, Daniel R.; Mansfield, Sarah; Barchia, I.; Herron, Grant A

doi:10.1007/s10340-015-0651-5

Cited by 7 publications

(5 citation statements)

References 28 publications

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“…Point mutations in VGSC genes have been identified in many insect species (Dong et al, 2014). In our samples, we did not observe the presence of the kdr (L1014F) mutation previously found in Australian cotton aphids (Marshall et al, 2012;Suann et al, 2015). Interestingly, in addition to the known super-kdr (M918L) mutation, we discovered another substitution at position 918 (M918V) in two of the three examined cotton aphid populations collected in Xinjiang.…”

Section: Discussioncontrasting

confidence: 65%

“…Molecular diagnostic tests are powerful tools for detecting resistant genotypes rapidly. Molecular-based methodology was already developed for detecting the kdr-associated L1014F mutation (Marshall et al, 2012;Suann et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

“…Specifically, M918L (super-kdr) was commonly observed in cotton aphids collected across China in 2011-2013 (Chen et al, 2017) and in Cameroon (Carletto et al, 2010), while the L1014F (kdr) was found to be distributed in Australia (Marshall et al, 2012;Suann et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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First report of voltage‐gated sodium channel M918V and molecular diagnostics of nicotinic acetylcholine receptor R81T in the cotton aphid

Munkhbayar

Liu

et al. 2020

J Applied Entomology

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The cotton aphid, Aphis gossypii Glover, is one of the most important agricultural insect pests. Pyrethroid and neonicotinoid insecticides have generally shown excellent control of A. gossypii, but many populations of this pest have developed resistance against these classes of insecticides. The success of insecticide resistance management strategies requires detailed knowledge of both phenotype and genotype of the target insect pest. In this study, we attempted to understand the molecular status of insecticide resistance in cotton aphid populations in Xinjiang Uygur Autonomous Region of China, the major cotton planting region of China. In addition to the previously reported M918L mutation, we discovered another substitution (M918V) in the voltage-gated sodium channel (VGSC). Moreover, we developed a molecular assay that could be used to detect precisely the R81T mutation in the nicotinic acetylcholine receptor (nAChR). This survey revealed that 918L was the predominant VGSC allele with a frequency ranging from 50.0% to 56.7%. Notably, appreciable frequencies (between 10% and 40%) of the resistance 81T allele of the nAChR gene were detected in three investigated populations. The prevalent co-occurrence of both VGSC 918L/V and nAchR 81T indicates a worrisome situation of multiple resistance to both pyrethroids and neonicotinoids.

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Section: Discussioncontrasting

confidence: 65%

Section: Discussionmentioning

confidence: 99%

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First report of voltage‐gated sodium channel M918V and molecular diagnostics of nicotinic acetylcholine receptor R81T in the cotton aphid

Munkhbayar

Liu

et al. 2020

J Applied Entomology

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“…The mRNA expression of glycogen phosphorylase ( HaGP ) and glycogen synthase ( HaGS ) were detected via qRT-PCR ( Suann et al, 2015 ). As a result, HaGP increased while HaGS expression decreased during the starvation period ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Effects of starvation on the carbohydrate metabolism in Harmonia axyridis (Pallas)

Shi

Wang

et al. 2017

Biology Open

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Trehalose plays an important role in energy storage, metabolism, and protection from extreme environmental conditions in insects. Trehalose is the main blood sugar in insects, and it can be rapidly used as an energy source in times of need. To elucidate the mechanisms of the starvation response, we observed the effects of starvation on trehalose and glycogen, trehalase activity, and the relative gene expression of genes in the trehalose and glycogen metabolic pathways in the invasive beetle Harmonia axyridis. Our results show that trehalose levels and the activities of two types of trehalases decreased significantly in the first 8 h of starvation, while the relative expression of HaTreh1-1 increased. While trehalose remained nearly constant at a relatively high level from 8 to 24 h, glycogen levels decreased significantly from 8 h to 24 h of starvation. Likewise, glycogen phosphorylase (HaGP) expression was significantly higher at 12 to 24 h starvation than the first 8 h, while the expression of glycogen synthase (HaGS) was relatively stable. Furthermore, trehalose decreased significantly from 24 h starvation to 72 h starvation, while trehalase activities and the relative expression of some HaTreh genes generally increased toward the end of the starvation period. The expression of trehalose-6-phosphate synthase (HaTPS) increased significantly, supporting the increase in trehalose synthesis. These results show that trehalose plays a key role in the energy provided during the starvation process through the molecular and biochemical regulation of trehalose and glycogen metabolism.

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“…One microgram of total RNA was used as a template to synthesize first-strand cDNA using a PrimeScript RT with gDNA Eraser kit (TaKaRa, Japan). The expression of HaTreh1 - 5 , HaTreh2 - like , and HaTreh2 was estimated by qRT-PCR (Suann et al 2015) using a Bio-Rad CFX96™ system (Bio-Rad, USA) and SsFast™ EvaGreen Supermix (Bio-Rad, USA). Primers were designed to determine the expression of the genes, HaTreh1 - 5 , HaTreh2 - like, and HaTreh2 (Table 1).…”

Section: Methodsmentioning

confidence: 99%

Three novel trehalase genes from Harmonia axyridis (Coleoptera: Coccinellidae): cloning and regulation in response to rapid cold and re-warming

et al. 2019

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Trehalose is the main blood sugar in insects. To study the function of trehalase during exposure to low temperatures, three other novel cDNAs of trehalase were cloned from Harmonia axyridis by transcriptome sequencing and rapid amplification of cDNA ends. One of the cloned cDNAs encoded a soluble trehalase, the second trehalase cDNA encoded a transmembrane-like domain, and the third cDNA encoded a membrane-bound protein. Therefore, these cDNAs were, respectively, named HaTreh1 - 5 , HaTreh2 - like , and HaTreh2 . HaTreh1 - 5 , HaTreh2 - like , and HaTreh2 cDNAs encoded proteins containing 586, 553, and 633 amino acids with predicted masses of approximately 69.47, 63.46, and 73.66 kDa, and pIs of 9.20, 5.52, and 6.31, respectively. All three novel trehalases contained signal motifs “PGGINKESYYLDSY”, “QWDYPNAWPP”, and a highly conserved glycine-rich (GGGGEY) region. The expression levels of HaTreh1 - 5 and HaTreh2 mRNAs were high during adult stages, whereas HaTreh2 - like was expressed in low amounts in the fourth larval stage. The results showed that the activity of membrane-bound trehalases decreased from 25 to 10 °C and from 5 to − 5 °C during cooling. The results also revealed a decreasing trend in expression of the three HaTreh mRNAs during the cooling treatment, and an initial decrease followed by an increase during the process of re-warming. Electronic supplementary material The online version of this article (10.1007/s13205-019-1839-9) contains supplementary material, which is available to authorized users.

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A TaqMan qPCR method for detecting kdr resistance in Aphis gossypii demonstrates improved sensitivity compared to conventional PCR–RFLP

Cited by 7 publications

References 28 publications

First report of voltage‐gated sodium channel M918V and molecular diagnostics of nicotinic acetylcholine receptor R81T in the cotton aphid

First report of voltage‐gated sodium channel M918V and molecular diagnostics of nicotinic acetylcholine receptor R81T in the cotton aphid

Effects of starvation on the carbohydrate metabolism in Harmonia axyridis (Pallas)

Three novel trehalase genes from Harmonia axyridis (Coleoptera: Coccinellidae): cloning and regulation in response to rapid cold and re-warming

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