Development and characterization of diamondback moth resistance to transgenic broccoli expressing high levels of Cry1C

J. Z. Zhao, H. L. Collins, J. D. Tang, J. Cao, E. D. Earle, Richard T. Roush, S. Herrero, B. Escriche, J. Ferre, A. M. Shelton

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

A field-collected colony of the diamondback moth, Plutella xylostella, had 31-fold resistance to Cry1C protoxin of Bacillus thuringiensis. After 24 generations of selection with Cry1C protoxin and transgenic broccoli expressing a Cry1C protein, the resistance that developed was high enough that neonates of the resistant strain could complete their entire life cycle on transgenic broccoli expressing high levels of Cry1C. After 26 generations of selection, the resistance ratios of this strain to Cry1C protoxin were 12, 400- and 63, 100-fold, respectively, for the neonates and second instars by a leaf dip assay. The resistance remained stable until generation 38 (G38) under continuous selection but decreased to 235-fold at G38 when selection ceased at G28. The Cry1C resistance in this strain was seen to be inherited as an autosomal and incompletely recessive factor or factors when evaluated using a leaf dip assay and recessive when evaluated using Cry1C transgenic broccoli. Saturable binding of 125I-Cry1C was found with brush border membrane vesicles (BBMV) from both susceptible and Cry1C-resistant strains. Significant differences in Cry1C binding to BBMV from the two strains were detected. BBMV from the resistant strain had about sevenfold-lower affinity for Cry1C and threefold-higher binding site concentration than BBMV from the susceptible strain. The overall Cry1C binding affinity was just 2.5-fold higher for BBMV from the susceptible strain than it was for BBMV from the resistant strain. These results suggest that reduced binding is not the major mechanism of resistance to Cry1C.

Original languageEnglish (US)
Pages (from-to)3784-3789
Number of pages6
JournalApplied and environmental microbiology
Volume66
Issue number9
DOIs
StatePublished - Sep 25 2000

Fingerprint

Moths
Brassica
Plutella xylostella
broccoli
Microvilli
vesicle
moth
brush border membrane vesicles
genetically modified organisms
membrane
Membranes
fold
neonate
dip
assay
neonates
Life Cycle Stages
border
life cycle
Binding Sites

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

Cite this

Zhao, J. Z. ; Collins, H. L. ; Tang, J. D. ; Cao, J. ; Earle, E. D. ; Roush, Richard T. ; Herrero, S. ; Escriche, B. ; Ferre, J. ; Shelton, A. M. / Development and characterization of diamondback moth resistance to transgenic broccoli expressing high levels of Cry1C. In: Applied and environmental microbiology. 2000 ; Vol. 66, No. 9. pp. 3784-3789.
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Zhao, JZ, Collins, HL, Tang, JD, Cao, J, Earle, ED, Roush, RT, Herrero, S, Escriche, B, Ferre, J & Shelton, AM 2000, 'Development and characterization of diamondback moth resistance to transgenic broccoli expressing high levels of Cry1C', Applied and environmental microbiology, vol. 66, no. 9, pp. 3784-3789. https://doi.org/10.1128/AEM.66.9.3784-3789.2000

Development and characterization of diamondback moth resistance to transgenic broccoli expressing high levels of Cry1C. / Zhao, J. Z.; Collins, H. L.; Tang, J. D.; Cao, J.; Earle, E. D.; Roush, Richard T.; Herrero, S.; Escriche, B.; Ferre, J.; Shelton, A. M.

In: Applied and environmental microbiology, Vol. 66, No. 9, 25.09.2000, p. 3784-3789.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Development and characterization of diamondback moth resistance to transgenic broccoli expressing high levels of Cry1C

AU - Zhao, J. Z.

AU - Collins, H. L.

AU - Tang, J. D.

AU - Cao, J.

AU - Earle, E. D.

AU - Roush, Richard T.

AU - Herrero, S.

AU - Escriche, B.

AU - Ferre, J.

AU - Shelton, A. M.

PY - 2000/9/25

Y1 - 2000/9/25

N2 - A field-collected colony of the diamondback moth, Plutella xylostella, had 31-fold resistance to Cry1C protoxin of Bacillus thuringiensis. After 24 generations of selection with Cry1C protoxin and transgenic broccoli expressing a Cry1C protein, the resistance that developed was high enough that neonates of the resistant strain could complete their entire life cycle on transgenic broccoli expressing high levels of Cry1C. After 26 generations of selection, the resistance ratios of this strain to Cry1C protoxin were 12, 400- and 63, 100-fold, respectively, for the neonates and second instars by a leaf dip assay. The resistance remained stable until generation 38 (G38) under continuous selection but decreased to 235-fold at G38 when selection ceased at G28. The Cry1C resistance in this strain was seen to be inherited as an autosomal and incompletely recessive factor or factors when evaluated using a leaf dip assay and recessive when evaluated using Cry1C transgenic broccoli. Saturable binding of 125I-Cry1C was found with brush border membrane vesicles (BBMV) from both susceptible and Cry1C-resistant strains. Significant differences in Cry1C binding to BBMV from the two strains were detected. BBMV from the resistant strain had about sevenfold-lower affinity for Cry1C and threefold-higher binding site concentration than BBMV from the susceptible strain. The overall Cry1C binding affinity was just 2.5-fold higher for BBMV from the susceptible strain than it was for BBMV from the resistant strain. These results suggest that reduced binding is not the major mechanism of resistance to Cry1C.

AB - A field-collected colony of the diamondback moth, Plutella xylostella, had 31-fold resistance to Cry1C protoxin of Bacillus thuringiensis. After 24 generations of selection with Cry1C protoxin and transgenic broccoli expressing a Cry1C protein, the resistance that developed was high enough that neonates of the resistant strain could complete their entire life cycle on transgenic broccoli expressing high levels of Cry1C. After 26 generations of selection, the resistance ratios of this strain to Cry1C protoxin were 12, 400- and 63, 100-fold, respectively, for the neonates and second instars by a leaf dip assay. The resistance remained stable until generation 38 (G38) under continuous selection but decreased to 235-fold at G38 when selection ceased at G28. The Cry1C resistance in this strain was seen to be inherited as an autosomal and incompletely recessive factor or factors when evaluated using a leaf dip assay and recessive when evaluated using Cry1C transgenic broccoli. Saturable binding of 125I-Cry1C was found with brush border membrane vesicles (BBMV) from both susceptible and Cry1C-resistant strains. Significant differences in Cry1C binding to BBMV from the two strains were detected. BBMV from the resistant strain had about sevenfold-lower affinity for Cry1C and threefold-higher binding site concentration than BBMV from the susceptible strain. The overall Cry1C binding affinity was just 2.5-fold higher for BBMV from the susceptible strain than it was for BBMV from the resistant strain. These results suggest that reduced binding is not the major mechanism of resistance to Cry1C.

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EP - 3789

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

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