scholarly journals Genetic Knockouts Indicate That the ABCC2 Protein in the Bollworm Helicoverpa zea Is Not a Major Receptor for the Cry1Ac Insecticidal Protein

Genes ◽  
2021 ◽  
Vol 12 (10) ◽  
pp. 1522
Author(s):  
Omaththage P. Perera ◽  
Nathan S. Little ◽  
Heba Abdelgaffar ◽  
Juan Luis Jurat-Fuentes ◽  
Gadi V. P. Reddy

Members of the insect ATP binding cassette transporter subfamily C2 (ABCC2) in several moth species are known as receptors for the Cry1Ac insecticidal protein from Bacillus thuringiensis (Bt). Mutations that abolish the functional domains of ABCC2 are known to cause resistance to Cry1Ac, although the reported levels of resistance vary widely depending on insect species. In this study, the function of the ABCC2 gene as a putative Cry1Ac receptor in Helicoverpa zea, a major pest of over 300 crops, was evaluated using CRISPR/Cas9 to progressively eliminate different functional ABCC2 domains. Results from bioassays with edited insect lines support that mutations in ABCC2 were associated with Cry1Ac resistance ratios (RR) ranging from 7.3- to 39.8-fold. No significant differences in susceptibility to Cry1Ac were detected between H. zea with partial or complete ABCC2 knockout, although the highest levels of tolerance were observed when knocking out half of ABCC2. Based on >500–1000-fold RRs reported in similar studies for closely related moth species, the low RRs observed in H. zea knockouts support that ABCC2 is not a major Cry1Ac receptor in this insect.

Author(s):  
Omaththage P. Perera ◽  
Nathan S LIttle ◽  
Heba Abdelgaffar ◽  
Juan Luis Jurat-Fuentes ◽  
Gadi V. P. Reddy

Members of the insect ATP binding cassette transporter subfamily C2 (ABCC2) in several moth species are known as receptors for the Cry1Ac insecticidal protein from Bacillus thuringiensis (Bt). Mutations that abolish the functional domains of ABCC2 are known to cause resistance to Cry1Ac, although the reported levels of resistance vary widely depending on insect species. In this study, the function of the ABCC2 gene as putative Cry1Ac receptor in Helicoverpa zea, a major pest of over 300 crops, was evaluated using CRISPR/Cas9 to progressively eliminate different functional ABCC2 domains. Results from bioassays with edited insect lines support that muta-tions in ABCC2 was associated with Cry1Ac resistance ratios (RR) ranging from 7.3- to 39.8-fold. No significant differences in susceptibility to Cry1Ac were detected between H. zea with partial or complete ABCC2 knockout, although highest levels of tolerance were observed when knocking out half of ABCC2. Based on >500-1,000-fold RRs reported in similar studies for closely related moth species, the low RRs observed in H. zea knockouts support that ABCC2 is not a major Cry1Ac receptor in this insect.


1996 ◽  
Vol 31 (3) ◽  
pp. 340-346 ◽  
Author(s):  
Steven R. Sims ◽  
Jay C. Pershing ◽  
Barbara J. Reich

Twelve independently transformed lines of transgenic corn (Zea mays L.) expressing the CryIA(b) insecticidal protein from Bacillus thuringiensis var. kurstaki were field tested to evaluate their resistance to the corn earworm, Helicoverpa zea (Boddie). Silks of the primary (=top) ears of transgenic [CryIA(b) positive] and isoline control plants [no CryIA(b) protein] were artificially infested with first-instar H. zea larvae and the length of ear penetration was measured after 19 d. Eight of the 12 lines had significantly less ear damage than their respective isoline controls; 3 transgenic lines reduced H. zea feeding damage by > 75% and stunted surviving H. zea larvae. Concentration of the CryIA(b) protein (μg/g fresh weight) in silks of the transgenic lines, determined using ELISA, ranged from 0.0 to 1.28 μg/g. Within transgenic lines, there was a weak (P < 0.06) negative relationship between the concentration of CryIA(b) protein in fresh silks and the length of H. zea ear penetration.


1998 ◽  
Vol 33 (2) ◽  
pp. 129-135 ◽  
Author(s):  
A. O. Ameen ◽  
J. R. Fuxa ◽  
A. R. Richter

Interactions between formulations of the aizawai and kurstaki subspecies of Bacillus thuringiensis Berliner were evaluated by bioassay in Heliothis virescens (F.) and Helicoverpa zea (Boddie). In preliminary experiments, a formulation of subspecies aizawai, Xentari AS®, had significantly (P < 0.05) higher median lethal concentrations (LC50s) in both insect species than formulations based on subspecies kurstaki. Helicoverpa zea was significantly (P < 0.05) more susceptible than H. virescens to one formulation of subspecies kurstaki (Dipel ES®), but the two insects did not differ in susceptibility to Xentari AS® or to a second formulation of subspecies kurstaki (Dipel 6AF®). In H. virescens, Xentari AS® was additive with Dipel 6AF® and significantly (P< 0.05) antagonistic with Dipel ES® and with a third formulation of subspecies kurstaki, Dipel 48A®. In H. zea, Xentari AS® was significantly antagonistic with all three formulations of subspecies kurstaki. This suggests that certain toxin combinations from B. thuringiensis subspecies might not be effective for managing H. virescens and H. zea populations.


1998 ◽  
Vol 33 (4) ◽  
pp. 365-377 ◽  
Author(s):  
R. G. Luttrell ◽  
Abbas Ali ◽  
S. Y. Young ◽  
Kathy Knighten

Diet incorporation and spray chamber assays compared the activity of three commercial formulations of Bacillus thuringiensis Berliner against Heliothis virescens (F.), Helicoverpa zea (Boddie), Pseudoplusia includens (Walker), and Spodoptera exigua (Hübner). The commercial products evaluated were Condor OF® (Ecogen Inc., Langhorne, PA), Dipel ES® (Abbott Laboratories, North Chicago, IL), and Javelin WG® (Sandoz Crop Protection Corp., Des Plaines, IL). Variation in results of the diet incorporation studies illustrated the importance of including a standard reference formulation in the design of the studies and suggested that some standardization of insect strains used in such assays may be important also. Overall results of the diet incorporation studies were similar to those obtained in the spray chamber studies where insects were assayed on sprayed cotton and soybean. Dose-mortality regressions were developed for each B. thuringiensis formulation applied against each insect species on cotton and soybean over a 28-day observation period. These regressions may be helpful in the development of recommendations for the use of B. thuringiensis on cotton and soybean. They may also have value in establishing base-line levels of susceptibility for future resistance monitoring efforts. Median lethal concentrations (LC50s) tended to be slightly higher on cotton than on soybean. The most susceptible insect species was H. virescens, and the least susceptible was S. exigua. LC50s for H. zea and P. includens were similar and intermediate between those of H. virescens and S. exigua. Javelin WG tended to exhibit the highest activity per unit weight of commercial product, but the relative differences among the commercial formulations were influenced by insect species.


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