The compatibility of a nucleopolyhedrosis virus control with resistance management for Bacillus thuringiensis: Co-infection and cross-resistance studies with the diamondback moth, Plutella xylostella

doi:10.1016/j.jip.2006.06.003

Journal of Invertebrate Pathology

Volume 93, Issue 2, October 2006, Pages 114-120

https://doi.org/10.1016/j.jip.2006.06.003 Get rights and content

Abstract

The use of genetically modified crops expressing Bacillus thuringiensis (Bt) toxins can lead to the reduction in application of broad-spectrum pesticides and an increased opportunity for supplementary biological control. Bt microbial sprays are also used by organic growers or as part of integrated pest management programs that rely on the use of natural enemies. In both applications the evolution of resistance to Bt toxins is a potential problem. Natural enemies (pathogens or insects) acting in combination with toxins can accelerate or decelerate the evolution of resistance to Bt. In the present study we investigated whether the use of a nucleopolyhedrovirus (AcMNPV) could potentially affect the evolution of resistance to the Bt toxin Cry1Ac in Plutella xylostella. At low toxin doses there was evidence for antagonistic interactions between AcMNPV and Cry1Ac resistant and susceptible insects. However, this antagonism was much stronger and more widespread for susceptible larvae; interactions were generally not distinguishable from additive for resistant larvae. Selection for resistance to Cry1Ac in two populations of P. xylostella with differing resistance mechanisms did not produce any correlated changes in resistance to AcMNPV. Stronger antagonistic interactions between Bt and AcMNPV on susceptible rather than resistant larvae can decrease the relative fitness between Bt-resistant and susceptible larvae. These interactions and the lack of cross-resistance between virus and toxin suggest that the use of NPV is compatible with resistance management to Bt products.

Introduction

The demand for environmentally friendly control methods and an increasing problem of resistance to synthetic insecticides has led to the increased use of Bacillus thuringiensis (Bt) microbial sprays. While the percentage share of the insecticide market accounted for by Bt microbial sprays is small this type of biological control is a key product in forestry, in Integrated Pest Management (IPM) and for organic growers (Lacey et al., 2001). Bt has also been successfully exploited for its production of insect-specific Cry toxins. Genes encoding these toxins have been incorporated into genetically modified (GM) crops such as cotton and maize (Shelton et al., 2002). The consequences of the application of insect resistant GM crops are several; one notable advantage has been the reduction in use of broad-spectrum insecticides (Pyke and Fitt, 1998, Pray et al., 2002, Carpenter et al., 2004, Hails and Raymond, 2004), a pattern accompanied by some evidence for increasing reliance on natural enemies and biological control (EPA, 2001).

A reduction in the application of broad-spectrum pesticides increases the potential for IPM (van Lenteren, 2000) and can allow the build-up of natural enemies. In this context, the use of narrow-spectrum microbial products that do not interfere with a conserved natural enemy complex can be of added value in pest management. Nucleopolyhedroviruses (NPVs) are one such microbial insecticide. NPV products are commercially available for heliothine pests in the US and Australia and have been successfully used globally in various agricultural and horticultural settings (Hunter-Fujita et al., 1998) and in the development of IPM in cotton (Mensah, 2002). While NPVs have limitations as insecticides, especially in their cost of production, they can, through the build-up of infectious particles and secondary cycles of infection in pest populations, be an effective means of long-term population control (Reed and Springett, 1971, Entwistle et al., 1983, Dwyer and Elkinton, 1993, Fuxa and Richter, 1994, Young, 1998, Bonsall, 2004). This, in turn can lead to increased spray intervals and reduced costs for growers (Moscardi, 1999).

However, concerns have been expressed about the compatibility of microbial and natural enemy control with Bt resistance management. In particular, parasitoid attack and the action of a fungal pathogen were predicted to increase the rate of adaptation of Heliothis virescens to GM tobacco (Johnson and Gould, 1992, Johnson et al., 1997a, Johnson et al., 1997b, Gould, 1998). Nevertheless, parasitoid attack on Bt-resistant Plutella xylostella was not predicted to affect the relative fitness or evolution of resistance on GM oil seed rape (Schuler et al., 1999, Schuler et al., 2004). Given the importance of resistance management for the sustainability of Bt products and the potential of many target insects to evolve resistance to Bt toxins (Ferré and Van Rie, 2002, Janmaat and Myers, 2003) it would be valuable to know whether other invertebrate pathogens might have similarly deleterious consequences for the evolution of resistance to Bt. In this study we investigated how the combined use of a nucleopolyhedrovirus (AcMNPV) and a Bt toxin, Cry1Ac, might affect the evolution of resistance to Bt. More specifically, we tested how co-exposure to NPV and toxin would interact and whether these interactions would vary between Cry1Ac resistant and susceptible insects. Secondly, we tested whether selection for resistance to a Bt toxin would produce correlated changes in susceptibility to an NPV.

Section snippets

General methods

Two populations of P. xylostella from Malaysia, Serd4 and Karak, derived from field populations with high levels of field-evolved resistance to B. t. kurstaki (Btk) and Cry1Ac, its most important constituent toxin, were used in this study (Sayyed and Wright, 2001, Sayyed et al., 2004). These populations have distinct and independently evolved mechanisms of resistance to Cry1Ac (Sayyed et al., 2001, Sayyed and Wright, 2001). Insects were fed on Chinese cabbage, Brassica pekinensis var. “One Kilo

Co-infection with AcMNPV and Cry1Ac

Comparison of toxin-only LC50s from resistant and susceptible Karak strains indicates that relative resistance to Cry1Ac was 23-fold at the time of this experiment (Fig. 1, Fig. 2). This is compatible with estimates of relative resistance (20 to 30-fold) from bio-assays immediately prior and after this experiment (data not shown). For Cry1Ac resistant insects the analysis of covariance found no evidence for interactions between toxin and virus (Table 1a, Fig. 3). While there was evidence of

Discussion

There was no clearly no impact of selection for Bt resistance on susceptibility to AcMNPV in two independent populations of P. xylostella. Thus, neither the loss of binding resistance mechanisms in the Serd4 and Karak population, nor the reduced toxin activation and an un-described third Bt resistance mechanism in Serd4 (Sayyed et al., 2001, Sayyed et al., 2004, Sayyed and Wright, 2001) appear to have any effects on the susceptibility of larvae to NPV infection. Although there are few other

Acknowledgments

We thank the Biotechnology and Biological Sciences Research Council (Grant D15960) for funding this study. Dr. Rosie Hails assisted in the development of this project. We also thank Dan Rose and Angeliki Martinou for technical assistance.

References (48)

M.M. Bradford
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
Anal. Biochem.
(1976)
E.K. Engelhard et al.
Developmental resistance in 4th-instar Trichoplusia ni orally inoculated with Autographa-californica nuclear polyhedrosis virus
Virology
(1995)
J.A. Fuxa
Interactions of the microsporidium Vairimorpha necatrix with a bacterium, virus and fungus in Heliothis zea
J. Invertebr. Pathol.
(1979)
J.R. Fuxa et al.
Response of nuclear polyhedrosis virus-resistant Spodoptera frugiperda larvae to other pathogens and to chemical insecticides
J. Invertebr. Pathol.
(1990)
L.A. Lacey et al.
Insect pathogens as biological control agents: Do they have a future?
Biol. Control
(2001)
T.H. Schuler et al.
Effects of Bt plants on the development and survival of the parasitoid Cotesia plutellae (Hymenoptera : Braconidae) in susceptible and Bt-resistant larvae of the diamondback moth, Plutella xylostella (Lepidoptera : Plutellidae)
J. Insect Physiol.
(2004)
A.G. Spies et al.
Effect of sublethal Bacillus thuringiensis crystal endotoxin treatment on the larval midgut of a moth, Manduca—SEM Study
Tissue Cell
(1985)
J.C. van Lenteren
A greenhouse without pesticides: Fact or fantasy?
Crop Prot.
(2000)
W. Abbott
A method of computing the effectiveness of insecticide
J. Econ. Entomol.
(1925)
M.R. Bell et al.
Heliothis virescens and Heliothis zea (Lepidoptera, Noctuidae)—dosage effects of feeding mixtures of Bacillus thuringiensis and a nuclear polyhedrosis-virus on mortality and growth
Environ. Entomol.
(1986)

M.B. Bonsall

The impact of diseases and pathogens on insect population dynamics

Physiol. Entomol.

(2004)

Carpenter, J.E., Sankula, S., Silvers, C.S., Gianessi, L.P., 2004. Insecticidal Bacillus thurgingiensis plants versus...

D. Collett

Modelling Binary Data

(1991)

M.J. Crawley

Statistical Computing, an Introduction to Data Analysis using S-Plus

(2002)

G. Dwyer et al.

Using simple models to predict virus epizootics in gypsy moth populations

J. Anim. Ecol.

(1993)

P.F. Entwistle et al.

Epizootiology of nuclear polyhedrosis virus (Baculoviridae) in European spruce sawfly (Gilpinia hercyniae)—spread of disease from small epicentres in comparison with spread of disease in other hosts

J. Appl. Ecol.

(1983)

Environmental Protection Agency, 2001. Bt-plant incorporated protectants October 15 2001; Biopesticides Registration...

R.R. Farrar et al.

Activity of the nucleopolyhedrovirus of the fall armyworm (Lepidoptera:Noctuidae) on foliage of transgenic sweet corn expressing a CryIA (b) toxin

Environ. Entomol.

(2004)

J. Ferré et al.

Biochemistry and genetics of insect resistance to Bacillus thuringiensis

Annu. Rev. Entomol.

(2002)

J.R. Fuxa et al.

Persistence and distribution of wild-type and recombinant nucleopolyhedroviruses in soil

Microb. Ecol.

(2001)

J.R. Fuxa et al.

Distance and rate of spread of Anticarsia gemmatalis (Lepidoptera, Noctuidae) nuclear polyhedrosis-virus released into soybean

Environ. Entomol.

(1994)

T.R. Glare et al.

Bacillus thuringiensis: Biology, Ecology and Safety

(2000)

F. Gould

Sustainability of transgenic insecticidal cultivars: integrating pest genetics and ecology

Annu. Rev. Entomol.

(1998)

R.S. Hails et al.

Insect resistant transgenic plants and their environmental impact

Cited by (31)

HearNPV susceptibility in Helicoverpa armigera and Helicoverpa punctigera strains resistant to Bt toxins Cry1Ac, Cry2Ab, and Vip3Aa
2021, Journal of Invertebrate Pathology
Citation Excerpt :
The focus of this work was to explore patterns of relative efficacy of HearNPV across strains within each Helicoverpa species to inform future laboratory and field experiments and stimulate thinking about a potential IRM fit for this product. The lack of any cross-resistance between HearNPV and other insecticides is consistent with previous studies in other insect species (Ignoffo and Roush, 1986; Fuxa and Richter, 1990; Raymond et al., 2006; Bentivenha et al., 2018). Bentivenha et al. (2018) showed that Spodoptera frugiperda larvae resistant to Bt or 5 additional insecticides were equally sensitive to SfMNPV relative to an insecticide-susceptible strain.
Genetically engineered crops expressing insecticidal toxins from Bacillus thuringiensis (Bt) have improved the management of targeted lepidopteran pests and reduced the use of insecticide sprays. These benefits explain an increasing adoption of Bt crops worldwide, intensifying the selection pressure on target species and the risk of resistance. Nucleopolyhedroviruses (NPVs) are effective bioinsecticides against numerous important lepidopteran pests. If Bt-resistant insects are shown to be susceptible to NPVs then these bioinsecticides could be a valuable component of Insecticide Resistance Management (IRM) strategies for Bt crops. We assessed the effectiveness of a Helicoverpa nucleopolyhedrovirus (HearNPV) against several different Bt-resistant strains. Utilising a droplet feeding bioassay we confirmed susceptibility to HearNPV in Helicoverpa punctigera and Helicoverpa armigera larvae resistant to the Bt toxins Cry1Ac, Cry2Ab, and Vip3A. Dual resistant H. punctigera, (Cry1Ac/Cry2Ab, and Cry2Ab/Vip3A) and dual resistant H. armigera (Cry2Ab/Vip3A) were also susceptible to HearNPV. Regardless of their specific resistance profile, Bt-resistant larvae displayed statistically similar lethal concentration (LC₅₀) and lethal time (LT₅₀) responses to HearNPV when compared to Bt-sensitive control insects. These results indicate that Bt-resistant H. armigera and H. punctigera are not cross-resistant to HearNPV. Consequently, the use of HearNPV against these pests may be a valuable tool to an IRM strategy for controlling Bt-resistant populations.
ABCC2 participates in the resistance of Plutella xylostella to chemical insecticides
2020, Pesticide Biochemistry and Physiology
Citation Excerpt :
For example, the fufenozide-resistant strain showed high resistance to dibenzoylhydrazines and benzoylphenylureas, and low cross-resistance to avermectin (Sun et al., 2012); the indoxacarb-resistant strain showed cross-resistance to metaflumizone, beta-cypermethrin, and chlorfenapyr (Zhang et al., 2017), and the field-evolved pyridalyl-resistant strain showed high resistance to all insecticides tested (including avermectin, emamectin benzoate, chlorantraniliprole, fipronil, and chlorfenapyr) (Yin et al., 2019). Interestingly, although a great deal of cross-resistance between different Bt toxins had been previously reported (e.g. Cry1C-resistant P. xylostella strains are cross-resistant to Cry1A, Cry1F, and other toxins) (Liu et al., 2001; Sayyed and Wright, 2001; Zhao et al., 2001; Sayyed et al., 2008), few reports have shown that Bt-resistant strains can exert cross-resistance with chemical insecticides and there is no cross-resistance between Bt toxins and the bioinsecticide AcMNPV (Raymond et al., 2006). According to this study, ABCC2 expression in P. xylostella is closely related to its metabolic resistance.
The ABCC2 protein of Plutella xylostella is an important target of Cry1A toxins from Bacillus thuringiensis (Bt), but whether this protein is involved in the resistance of P. xylostella to other insecticides remains unclear. In this study, the abcc2 gene of P. xylostella was cloned and the expression levels of Pxabcc2 in susceptible and resistant strains were investigated. ABCC2 was found to be expressed 3.2–6.7-fold higher in the resistant strain than in the susceptible strain; in the surviving P. xylostella, ABCC2 levels were significantly higher when treated with indoxacarb, avermectin, and beta-cypermethrin. We constructed a stable ABCC2-expressing HEK-293 cell line to reveal the contribution of ABCC2 to insecticide resistance. The avermectin and chlorfenapyr sensitivities of the stably-transfected cell line were significantly lower than those of the control cells. The intracellular avermectin concentration was significantly lower in the stably-transfected cell line than in the control cells after four hours of exposure. This study shows that up-regulated ABCC2 expression is related to insecticide resistance in P. xylostella. Moreover, we used RNA interference technology to reduce ABCC2 levels in P. xylostella. Down-regulating ABCC2 expression did not significantly affect avermectin or chlorfenapyr resistance in P. xylostella. We speculate that increased ABCC2 expression can enhance metabolic resistance in P. xylostella. This study also provides new insights into cross-resistance between B. thuringiensis toxins and chemical insecticides.
Microbial biopesticides for invertebrate pests and their markets in the United States
2019, Journal of Invertebrate Pathology
Citation Excerpt :
Primary targets include the cabbage looper, Trichoplusia ni, corn earworm, Heliothis zea, and beet armyworm, Spodoptera exigua. These products are highly specific and can be compatible with parasitoids and other biological control methods (Raymond et al., 2006). Another product (AcNPV) for cabbage looper, Trichoplusia ni, is pending (Silvana Niedermann, pers.
Microbial pesticides based on bacteria, fungi and viruses or their bioactive compounds have long been developed as alternatives for synthetic pesticides to control invertebrate pests. However, concern for environmental and human health from excessive reliance on chemical pesticides, changes in residue standards, and increased demand for organically grown produce has contributed to a considerable growth in their use in recent years. There are currently 356 registered biopesticide active ingredients in the U.S., including 57 species and/or strains of microbes or their derivatives, labelled for use against pestiferous insects, mites and nematodes. Strains of Bacillus thuringiensis for Lepidoptera remain the most popular products, but newer bacterial strains and their metabolites have been developed against a wider range of arthropods for use on fruit, vegetable and ornamental crops. Currently, ten fungal species/strains are registered against thrips, whiteflies, aphids, or other sucking pests and plant parasitic nematodes in greenhouse, nursery and field crops, while five nucleopolyhedroviruses and three granuloviruses are registered for Lepidoptera in field and greenhouse grown vegetables and ornamentals, tree fruit and nuts, forestry, and stored products. Many of these products are organic listed and most have 4 h or less reentry and no pre-harvest restrictions. Investment by multinational companies, advances in screening, industrial fermentation and storage of new microorganisms, are increasing the market share for microbials. Here, we summarize the market for microbial-based pesticides labelled for invertebrates in the U.S. We cover current uses and recent advances that further advance their use in additional markets in the coming decades.
No cross-resistance between ChinNPV and chemical insecticides in Chrysodeixis includens (Lepidoptera: Noctuidae)
2019, Journal of Invertebrate Pathology
Chrysodeixis includens nucleopolyhedrovirus (ChinNPV: Baculoviridae: Alphabaculovirus) is an active ingredient of a biological-based insecticide (Chrysogen®) recommended against soybean looper (SBL), Chrysodeixis includens (Walker, [1858]), in soybean in Brazil. We investigated if SBL strains resistant to chemical insecticides are cross-resistant to the baculovirus ChinNPV. In droplet feeding bioassays, SBL strains resistant to lambda-cyhalothrin and teflubenzuron showed equivalent susceptibility to ChinNPV as heterozygous and susceptible strains, indicating no cross-resistance between ChinNPV and chemical insecticides in SBL. Therefore, the ChinNPV is a valuable new “mode-of-action” tool for SBL resistance management in Brazil.
Biocontrol of Insect Pests
2016, Ecofriendly Pest Management for Food Security
Immediately after World War II, the success of dichlorodiphenyltrichloroethane (DDT) and 2,4-dichlorophenoxyacetic acid (2,4-D) emphasized the development of synthetic pesticides and their extensive use in agriculture. However, their deleterious effects on the environment and in agriculture soon proposed the need for corrective measures to attain sustainable agriculture and environmental safety. Biocontrol eventually attained global preference over synthetic pesticides for being an effective means to suppress the population of insect pests. During the past two decades, steady progress has been made worldwide to safeguard crops from insect pests through the use and manipulation of biocontrol agents/natural enemies. Still, there is an increasing demand to search more natural enemies (viz., predators, parasites, and pathogens) and assess their efficacy against various agricultural pests. Despite a growing demand for biocontrol agents, major populations of the world, especially those of developing nations, are still unaware about the different biocontrol methods. Moreover, only recently have scientists and practitioners attempted to assign economic value to biocontrol. In the present chapter, various aspects of biocontrol methods have been discussed that would hopefully make the readers aware about the ways by which they are being implemented in integrated pest management systems.
Resistance to Bacillus thuringiensis in the cabbage looper (Trichoplusia ni) increases susceptibility to a nucleopolyhedrovirus
2010, Journal of Invertebrate Pathology
Cabbage loopers, Trichoplusia ni, are pests in many agricultural settings including vegetable greenhouses in British Columbia (Canada), where microbial insecticides based on Bacillus thuringiensis (Bt) toxins are commonly used. Frequent use of these insecticides has led to resistance in some populations. An alternative microbial control is the multiple nucleopolyhedrovirus of the alfalfa looper (Autographa californica), AcMNPV which occurs naturally, but at low frequencies in T. ni populations. Bioassays show that T. ni resistant to Bt were twice as susceptible to AcMNPV as were individuals from the Bt-susceptible strain and AcMNPV could be complementary in a resistance management program for T. ni.

View all citing articles on Scopus

View full text

The compatibility of a nucleopolyhedrosis virus control with resistance management for Bacillus thuringiensis: Co-infection and cross-resistance studies with the diamondback moth, Plutella xylostella

Abstract

Introduction

Section snippets

General methods

Co-infection with AcMNPV and Cry1Ac

Discussion

Acknowledgments

Anal. Biochem.

Virology

J. Invertebr. Pathol.

J. Invertebr. Pathol.

Biol. Control

J. Insect Physiol.

Tissue Cell

Crop Prot.

A method of computing the effectiveness of insecticide

J. Econ. Entomol.

Heliothis virescens and Heliothis zea (Lepidoptera, Noctuidae)—dosage effects of feeding mixtures of Bacillus thuringiensis and a nuclear polyhedrosis-virus on mortality and growth

Environ. Entomol.

The impact of diseases and pathogens on insect population dynamics

Physiol. Entomol.

Modelling Binary Data

Statistical Computing, an Introduction to Data Analysis using S-Plus

Using simple models to predict virus epizootics in gypsy moth populations

J. Anim. Ecol.

Epizootiology of nuclear polyhedrosis virus (Baculoviridae) in European spruce sawfly (Gilpinia hercyniae)—spread of disease from small epicentres in comparison with spread of disease in other hosts

J. Appl. Ecol.

Activity of the nucleopolyhedrovirus of the fall armyworm (Lepidoptera:Noctuidae) on foliage of transgenic sweet corn expressing a CryIA (b) toxin

Environ. Entomol.

Biochemistry and genetics of insect resistance to Bacillus thuringiensis

Annu. Rev. Entomol.

Persistence and distribution of wild-type and recombinant nucleopolyhedroviruses in soil

Microb. Ecol.

Distance and rate of spread of Anticarsia gemmatalis (Lepidoptera, Noctuidae) nuclear polyhedrosis-virus released into soybean

Environ. Entomol.

Bacillus thuringiensis: Biology, Ecology and Safety

Sustainability of transgenic insecticidal cultivars: integrating pest genetics and ecology

Annu. Rev. Entomol.

Insect resistant transgenic plants and their environmental impact