Engineered Repressible Lethality for Controlling the Pink Bollworm, a Lepidopteran Pest of Cotton

Morrison, Neil I.; Simmons, Gregory S.; Fu, Guoliang; O’Connell, Sinead; Walker, Adam; Dafa'alla, Tarig; Walters, Michelle; Claus, John; Tang, Guolei; Jin, Li; Marubbi, Thea; Epton, Matthew J; Harris, Claire; Staten, R. T.; Miller, E.; Miller, Thomas A.; Alphey, Luke

doi:10.1371/journal.pone.0050922

Cited by 30 publications

(22 citation statements)

References 43 publications

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“…In this autoregulatory tTAV system, tTAV serves as both the transactivator and the effector, and high-level expression of tTAV from a proposed positive feedback configuration provides a lethal effect in B. mori, as was seen previously for the dipterans, Ceratitis capitata, Bactrocera oleae, and Aedes aegypti (17,25,26), and the lepidopterans, pink bollworm, and diamondback moth Plutella xylostella (29). The incorporation of a sex-specific, alternative-splicing module from the pink bollworm ortholog of doublesex makes the effect sex-specific (13).…”

Section: Discussionmentioning

confidence: 76%

“…The nonautonomous piggyBac vector allows germ-line transformation of insects via microinjection with separately supplied piggyBac transposase. The fluorescent marker is DsRed2 under the control of a baculovirus-derived promoter, Hr5ie1; this provides strong red fluorescence in a range of insect species (25,29,33,34). The conditional female-specific gene is designed to derive its lethal effect via hyperaccumulation of tTAV from a positive feedback loop, leading to transcriptional suppression (25,26).…”

Section: Resultsmentioning

confidence: 99%

“…As a result, female lethality is induced during embryonic and early larval stages caused by mass accumulation of tTAV in the absence of dietary tetracycline. Interestingly, embryonic lethality was not observed in transgenic strains of other pest insects carrying molecular constructs of similar design (13,17,25,26,29); the difference may result from the relatively slow development of silkworm allowing more time for accumulation of the toxic protein. Female lethality can be repressed when animals are provided with dietary tetracycline, and females and males survive to adult in approximately equal proportions.…”

Section: Discussionmentioning

confidence: 99%

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Transgene-based, female-specific lethality system for genetic sexing of the silkworm, Bombyx mori

Tan

Jin

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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120

140

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Transgene-based genetic sexing methods are being developed for insects of agricultural and public health importance. Male-only rearing has long been sought in sericulture because males show superior economic characteristics, such as better fitness, lower food consumption, and higher silk yield. Here we report the establishment of a transgene-based genetic sexing system for the silkworm, Bombyx mori. We developed a construct in which a positive feedback loop regulated by sex-specific alternative splicing leads to high-level expression of the tetracycline-repressible transactivator in females only. Transgenic animals show female-specific lethality during embryonic and early larval stages, leading to male-only cocoons. This transgene-based female-specific lethal system not only has wide application in sericulture, but also has great potential in lepidopteran pest control.Lepidoptera | doublesex T he mulberry silkworm, Bombyx mori, is a completely domesticated insect and is the foundation of sericulture, an endeavor of great economic importance. It is believed that sericulture originated in China and has been conducted there for more than 5,000 y (1). Male-only rearing techniques for B. mori are desirable because males show higher resistance to disease, lower food consumption, and better silk quality (2). To this end, several B. mori strains for male-only rearing have been developed by classical genetics. In the last century, Strunnikov (3) established a sex-linked, balanced-lethal system using radiation-induced chromosome translocations. However, conventional approaches involving selective breeding or irradiation for developing male-only silkworm strains are time and labor consuming. Thus, novel approaches are desired to improve modern silkworm breeding. In recent years, advances in B. mori genetic manipulation, notably genetic transformation, have been successfully established and applied extensively in gene function analysis and the production of bioreactors (1, 4-7). These technologies provide a potential basis for improvements in sericulture including the development of a male-only rearing system.Transgene-based genetic sexing systems have been developed in Drosophila melanogaster (8, 9) and several medically and agriculturally important insect species (10-13) as part of a series of genetics-based improvements and alternatives to the sterile insect technique (14). Systems based on sex-specific lethality for improving silk production are preferable to those using differential expression of a marker gene, for example fluorescence, which then would require manual or automated examination of each individual as part of the sorting process. Molecular designs developed for Diptera should be able to provide genetic sexing in B. mori, as the ability to transfer systems from one species to another is a key advantage of transgenic approaches over classical genetic methods (14-16). Furthermore, despite early reports to the contrary, more recent work has established clearly that transgenic strains can be developed with good...

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Section: Discussionmentioning

confidence: 76%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Transgene-based, female-specific lethality system for genetic sexing of the silkworm, Bombyx mori

Tan

Jin

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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120

140

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“…As per Morrison et al, (2012), a transgenic pink bollworm had survivability of 100 per cent under permissive condition but without permissive condition mortality was also 100 in different strains of transgenic pink bollworm…”

Section: Transgenesis In Pest Controlmentioning

confidence: 96%

Area Wide Pest Management: Concept and Approaches

Dalal¹,

Rathee²,

Singh³

2017

Int.J.Curr.Microbiol.App.Sci

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“…One variation of the RIDL system leads to the death of both sexes in the absence of tetracycline ('bi-sex lethality'). This was successfully demonstrated in C. capitata [Gong et al, 2005], A. aegypti [Phuc et al, 2007], and the lepidopteran P. gossypiella [Morrison et al, 2012]. However, in most cases the elimination of the female cohort from the release generation is considered highly advantageous for sterile-insect programmes [Papathanos et al, 2009].…”

Section: Use Of Sex Determination Alternative Splicing For Insect Pesmentioning

confidence: 99%

Practical Applications of Insects' Sexual Development for Pest Control

Koukidou

Alphey

2014

Sex Dev

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Elucidation of the sex differentiation pathway in insects offers an opportunity to understand key aspects of evolutionary developmental biology. In addition, it provides the understanding necessary to manipulate insects in order to develop new synthetic genetics-based tools for the control of pest insects. Considerable progress has been made in this, especially in improvements to the sterile insect technique (SIT). Large scale sex separation is considered highly desirable or essential for most SIT targets. This separation can be provided by genetic methods based on sex-specific gene expression. Investigation of sex determination by many groups has provided molecular components and methods for this. Though the primary sex determination signal varies considerably, key regulatory genes and mechanisms remain surprisingly similar. In most cases studied so far, a primary signal is transmitted to a basal gene at the bottom of the hierarchy (dsx) through an alternative splicing cascade; dsx is itself differentially spliced in males and females. A sex-specific alternative splicing system therefore offers an attractive route to achieve female-specific expression. Experience has shown that alternative splicing modules can be developed with cross-species function; modularity and standardisation and re-use of parts are key principles of synthetic biology. Both female-killing and sex reversal (XX females to phenotypic males) can in principle also be used as efficient alternatives to sterilisation in SIT-like methods. Sexual maturity is yet another area where understanding of sexual development may be applied to insect control programmes. Further detailed understanding of this crucial aspect of insect biology will undoubtedly continue to underpin innovative practical applications.

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Engineered Repressible Lethality for Controlling the Pink Bollworm, a Lepidopteran Pest of Cotton

Cited by 30 publications

References 43 publications

Transgene-based, female-specific lethality system for genetic sexing of the silkworm, Bombyx mori

Transgene-based, female-specific lethality system for genetic sexing of the silkworm, Bombyx mori

Area Wide Pest Management: Concept and Approaches

Practical Applications of Insects' Sexual Development for Pest Control

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