CRISPR/Cas9-mediated knockout of both the PxABCC2 and PxABCC3 genes confers high-level resistance to Bacillus thuringiensis Cry1Ac toxin in the diamondback moth, Plutella xylostella (L.)

2019 ◽  
Vol 107 ◽  
pp. 31-38 ◽  
Author(s):  
Zhaojiang Guo ◽  
Dan Sun ◽  
Shi Kang ◽  
Junlei Zhou ◽  
Lijun Gong ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Plutella Xylostella ◽  
Diamondback Moth ◽  
Cry1ac Toxin ◽  
High Level ◽  
Level Resistance

Insecticide Resistance Monitoring of the Diamondback Moth (Lepidoptera: Plutellidae) Populations in China

2021 ◽  
Author(s):  
Jing Wang ◽  
Xiaobin Zheng ◽  
Jiangjiang Yuan ◽  
Shuaiyu Wang ◽  
Baoyun Xu ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Insecticide Resistance ◽  
Plutella Xylostella ◽  
Yunnan Province ◽  
Diamondback Moth ◽  
Resistance Management ◽  
Resistance Monitoring ◽  
Crop Pest ◽  
High Level ◽  
Selection Of

Abstract The diamondback moth, Plutella xylostella L., is a worldwide crop pest that is difficult to control because of its ability to develop resistance to many insecticides. To provide a reference for resistance management of P. xylostella in China, the present study used a leaf-dip bioassay to monitor the resistance of P. xylostella to nine insecticides in eight regions of China. The results showed that P. xylostella had developed a high level of resistance to beta-cypermethrin (resistance ratio [RR] > 112), and moderate (RR < 40) to high levels of resistance to indoxacarb, abamectin, and chlorfluazuron. For chlorantraniliprole, RRs > 100 were found in Midu (Yunnan Province) and Jinghai (Tianjin). In most regions, the resistance to spinetoram and chlorfenapyr and Bacillus thuringiensis (Bt) was low. No resistance was detected to diafenthiuron. Overall, P. xylostella resistance to insecticides was higher in Midu than in other regions. The data in this study should help guide the selection of insecticides for management of P. xylostella in China.

High-Level Resistance to Bacillus thuringiensis Toxin Cry1Ac and Cadherin Genotype in Pink Bollworm

2006 ◽  
Vol 99 (6) ◽  
pp. 2125-2131 ◽  
Author(s):  
Bruce E. Tabashnik ◽  
Robert W. Biggs ◽  
Jeffrey A. Fabrick ◽  
Aaron J. Gassmann ◽  
Timothy J. Dennehy ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Pink Bollworm ◽  
Bacillus Thuringiensis Toxin ◽  
High Level ◽  
Level Resistance

Towards CRISPR/Cas9-based gene drive in the diamondback moth Plutella xylostella

2021 ◽  
Author(s):  
Xuejiao Xu ◽  
Tim Harvey-Samuel ◽  
Hamid Anees Siddiqui ◽  
Joshua Ang ◽  
Michelle E Anderson ◽  
...  
Keyword(s):  
Genetic Control ◽  
Plutella Xylostella ◽  
Control Strategies ◽  
Diamondback Moth ◽  
Regulatory Elements ◽  
Strategy Development ◽  
Gene Drive ◽  
Global Agriculture ◽  
High Level ◽  
Gene Drives

Promising to provide powerful genetic control tools, gene drives have been constructed in multiple dipterans, yeast and mice, for the purposes of population elimination or modification. However, it remains unclear whether these techniques can be applied to lepidopterans. Here, we used endogenous regulatory elements to drive Cas9 and sgRNA expression in the diamondback moth, (Plutella xylostella), and test the first split-drive system in a lepidopteran. The diamondback moth is an economically important global agriculture pest of cruciferous crops and has developed severe resistance to various insecticides, making it a prime candidate for such novel control strategy development. A very high level of somatic editing was observed in Cas9/sgRNA transheterozygotes, although no significant homing was revealed in the subsequent generation. Although heritable, Cas9-medated germline cleavage, as well as maternal and paternal Cas9 deposition was observed, rates were far lower than for somatic cleavage events, indicating robust somatic but limited germline activity of Cas9/sgRNA under the control of selected regulatory elements. Our results provide valuable experience, paving the way for future construction of gene drive-based genetic control strategies in DBM or other lepidopterans.

scholarly journals CRISPR-Mediated Knockout of the ABCC2 Gene in Ostrinia furnacalis Confers High-Level Resistance to the Bacillus thuringiensis Cry1Fa Toxin

Toxins ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 246 ◽  
Author(s):  
Xingliang Wang ◽  
Yanjun Xu ◽  
Jianlei Huang ◽  
Wenzhong Jin ◽  
Yihua Yang ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Deletion Mutation ◽  
Ostrinia Furnacalis ◽  
Corn Borer ◽  
Resistance Evolution ◽  
Loop Region ◽  
Truncating Mutation ◽  
High Level ◽  
Level Resistance

The adoption of transgenic crops expressing Bacillus thuringiensis (Bt) insecticidal crystalline (Cry) proteins has reduced insecticide application, increased yields, and contributed to food safety worldwide. However, the efficacy of transgenic Bt crops is put at risk by the adaptive resistance evolution of target pests. Previous studies indicate that resistance to Bacillus thuringiensis Cry1A and Cry1F toxins was genetically linked with mutations of ATP-binding cassette (ABC) transporter subfamily C gene ABCC2 in at least seven lepidopteran insects. Several strains selected in the laboratory of the Asian corn borer, Ostrinia furnacalis, a destructive pest of corn in Asian Western Pacific countries, developed high levels of resistance to Cry1A and Cry1F toxins. The causality between the O. furnacalis ABCC2 (OfABCC2) gene and resistance to Cry1A and Cry1F toxins remains unknown. Here, we successfully generated a homozygous strain (OfC2-KO) of O. furnacalis with an 8-bp deletion mutation of ABCC2 by the CRISPR/Cas9 approach. The 8-bp deletion mutation results in a frame shift in the open reading frame of transcripts, which produced a predicted protein truncated in the TM4-TM5 loop region. The knockout strain OfC2-KO showed much more than a 300-fold resistance to Cry1Fa, and low levels of resistance to Cry1Ab and Cry1Ac (<10-fold), but no significant effects on the toxicities of Cry1Aa and two chemical insecticides (abamectin and chlorantraniliprole), compared to the background NJ-S strain. Furthermore, we found that the Cry1Fa resistance was autosomal, recessive, and significantly linked with the 8-bp deletion mutation of OfABCC2 in the OfC2-KO strain. In conclusion, in vivo functional investigation demonstrates the causality of the OfABCC2 truncating mutation with high-level resistance to the Cry1Fa toxin in O. furnacalis. Our results suggest that the OfABCC2 protein might be a functional receptor for Cry1Fa and reinforces the association of this gene to the mode of action of the Cry1Fa toxin.

Enhancement of δ-Endotoxin Activity by Toxin-Free Spore of Bacillus thuringiensis against the Diamondback Moth, Plutella xylostella

1994 ◽  
Vol 63 (1) ◽  
pp. 111-112 ◽  
Author(s):  
Minoru Miyasono ◽  
Shyuichiro Inagaki ◽  
Makiko Yamamoto ◽  
Katsuaki Ohba ◽  
Takeo Ishiguro ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Plutella Xylostella ◽  
Diamondback Moth ◽  
Endotoxin Activity ◽  
Free Spore

scholarly journals Insecticidal Effects of Organotin(IV) Compounds on Plutella Xylostella (L.) Larvae. II. Inhibitory Potencies Against Acetylcholinesterase and Evidence for Synergism in Tests With Bacillus Thuringiensis(BER.) and Malathion

1994 ◽  
Vol 1 (1) ◽  
pp. 1-17 ◽  
Author(s):  
Nazni W. Ahmad ◽  
Tay Siew Huang ◽  
S. Balabaskaran ◽  
K. M. Lo ◽  
V. G. Kumar Das
Keyword(s):  
Bacillus Thuringiensis ◽  
Plutella Xylostella ◽  
Resistant Strain ◽  
Synergistic Effects ◽  
Diamondback Moth ◽  
Ache Inhibition ◽  
Microbial Insecticide ◽  
In Vitro Inhibition ◽  
Sublethal Concentrations

Features of pesticide synergism and acetylcholinesterase (AChE) inhibition (in vitro) were studied using a selected range of organotin compounds against the early 4th instar larvae of a highly resistant strain of the diamondback moth (DBM), Plutella xylostella, a major universal pest of cruciferous vegetables.Fourteen triorganotin compounds were evaluated for their ability to enhance the toxicity of the microbial insecticide, Bacillus thuringiensis (BT) and of the commercial insecticide, Malathion to Plutella xylostella larvae. Supplemental synergism was observed with triphenyl- and tricyclopentyltin hydroxides in combinations with Bacillus thuringiensis. Increased synergism was observed with an increase in the number of cyclopentyl groups on tin in the mixed series, CypnPh3-n SnX, where X = OH, and 1-(1,2,4-triazolyl). The combination of (p-chlorophenyl)diphenyltin N,N-dimethyldithiocarbamate at LD10 and LD25 concentrations with sublethal concentrations of Malathion as well as of tricyclohexyltin methanesulphonate at the 0.01% (w/v) concentration with Malathion exerted strong synergistic effects (supplemental synergism) with toxicity index (T.I) values of 7.2, 19.8 and 10.1, respectively.Studies on the in vitro inhibition of acetylcholinesterase prepared from the DBM larvae showed that while most of the triorganotin Compounds tested were without effect on the enzyme, compounds containing the thiocarbamylacetate or the dithiocarbamylacetate moieties demonstrated appreciable levels of inhibition, being comparable in efficacy to commercial grades of Malathion and Methomyl.

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