Establishment of an<i>RTA-Bddsx</i>hybrid system for female-specific splicing that can affect the sex ratio of<i>Bactrocera dorsalis</i>(Hendel) after embryonic injection

Huang, Chieh-Chen; Huang, Chia Chia; Dai, Shu‐Mei; Chang, Cheng‐Nan

doi:10.1002/ps.3990

Cited by 3 publications

(12 citation statements)

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“…In this study, a transgenic approach was used to test whether RTA‐Bddsx expressed in vivo can lead to female‐specific splicing, as observed in vitro , and show a female‐specific lethal effect in a transient expression assay . The actin 5C promoter ( Ac p) of B. dorsalis was chosen to drive the RTA‐Bddsx construct CF21‐26, which has been demonstrated to produce active RTA in females after alternative splicing to remove intron 3 of Bddsx . Because Ac p is a housekeeping gene promoter, we expected broad expression and toxic effects in multiple tissues.…”

Section: Resultsmentioning

confidence: 99%

“…The well‐established Tet‐Off system, which can be stringently regulated by tetracycline, matches our requirements. A Tet‐Off system with sex‐specific splicing of the tra gene has been applied successfully in the sexing of Anastrepha suspense , Ceratitis capitata and Lucilia cuprina , and can be easily adapted to B. dorsalis , as previously described …”

Section: Resultsmentioning

confidence: 99%

“…Several lines of evidence support the concept of using the sex‐differential toxicity of RTA to achieve female‐specific lethality in vitro . Furthermore, transient expression of RTA‐Bddsx genes was performed through embryonic injection, and we observed that the percentages of females were significantly reduced in association with Acp‐CF26‐21, for which female‐specific splicing was observed in vitro . The results suggested that the RTA‐Bddsx system shows great potential for B. dorsalis control.…”

Section: Introductionmentioning

confidence: 84%

“…In contrast to classical genetic sexing methods, several female‐lethal systems have been developed using the unique alternative splicing properties of sex determination genes. Both the transformer and doublesex genes have been applied successfully in female‐specific lethal systems …”

Section: Introductionmentioning

confidence: 99%

“…Both the transformer and doublesex genes have been applied successfully in female-specific lethal systems. 19,20,38,39 Ricin is a natural plant toxin belonging to the class II ribosome-inactivating protein (RIP) family. It consists of an A chain and a B chain (RTA and RTB) linked by a disulfide bond.…”

Section: Introductionmentioning

confidence: 99%

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Introduction of the RTA‐Bddsx gene induces female‐specific lethal effects in transformed Bactrocera dorsalis (Hendel)

Huang

Dai

Chang

2015

Pest Management Science

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 84%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Introduction of the RTA‐Bddsx gene induces female‐specific lethal effects in transformed Bactrocera dorsalis (Hendel)

Huang

Dai

Chang

2015

Pest Management Science

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Introduction of a cold sensitivity‐conferring mutation into the RTA‐Bddsx hybrid system of Bactrocera dorsalis for establishment of a thermally controllable homozygous line

Dai

Huang

Chang

2021

Pest Management Science

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BACKGROUND: For efficient control of the economically important fruit pest Bactrocera dorsalis, a hybrid system combining ricin toxicity and sex-related alternative splicing of the doublesex gene has been developed. This system exhibits the expected female-specific lethal effect; however, the transgenic females do not survive, making it difficult to raise stable homozygous lines. Since modification of ricin toxin A chain (RTA) through a single-residue change (Gly 212 > Arg 212 ) leads to cold-sensitive posttranslational repression of its toxicity, we utilized this unique property to obtain RTA-Bddsx females that survive at low temperature for line maintenance.RESULTS: In transient expression experiments using embryonic injection, two groups treated with RTAcs-derived DNA (LERQcs and RTAcs) exhibited temperature-dependent effects. The toxicity was higher at 29°C than at 18°C. The proportion of males was close to 50% at 18°C in all the tested groups except LERQcs-treated flies, which exhibited a high proportion of males (over 70%) at 29°C. The results indicate the cold-sensitive responses of RTA and further suggest a female-specific lethal effect. Subsequently, 14 putative RTAcs-Bddsx transgenic Ds-Red + G 1 males were identified, and female-specific lethal effects were observed in Ds-Red + G 2 and G 3 lines under cultivation at 29°C but not at 18°C. The male ratio can be increased to up to 95% in G 3 line 001, indicating that RTAcs functions well in B. dorsalis. CONCLUSION:The improved RTAcs-Bddsx system with conditional toxicity represents a novel and promising step toward the practical control of B. dorsalis.

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Genetically Engineered Ricin SuppressesBactrocera dorsalis(Diptera: Tephritidae) based on Demographic Analysis of Group-Reared Life Table

et al. 2016

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The oriental fruit fly, Bactrocera dorsalis (Hendel), reduces the quantity and quality of many host fruits through the process of oviposition and larval feeding, and this insect has been considered a major insect pest in several Asian countries for decades. Using an earlier-developed, female-specific system that combines the toxicity of the ricin A chain (RTA) and the alternative RNA splicing property of doublesex (Bddsx), we show that transgenic male flies harboring the RTA-Bddsx transgene unevenly repress the pest population through inheritable effects. In age-stage, two-sex life-table analyses, high larval mortality and a delay in pupation were observed after introducing the transgene. The high male to female ratio in DsRed flies demonstrates the lethal effect of ricin on females. The fitness of both the DsRed- and DsRed-transformed females was reduced as shown in the decrease of the net reproductive rate (R), intrinsic rate (r), and finite rate (λ) values compared with the wild-type populations. The integrity of the RTA-Bddsx transgene remained in more than 80% of DsRed males after ten generations, supporting the stable inheritance of the transgene. All of the data from this study support the proposed RTA-Bddsx SIT approach, which provides a species-specific and environmentally friendly method of suppressing, rather than eradiating, B. dorsalis.

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Establishment of anRTA-Bddsxhybrid system for female-specific splicing that can affect the sex ratio ofBactrocera dorsalis(Hendel) after embryonic injection

Abstract: Our results indicate that the RTA-Bddsx hybrid system offers a novel and promising approach for oriental fruit fly control.

Cited by 3 publications

References 58 publications

Introduction of the RTA‐Bddsx gene induces female‐specific lethal effects in transformed Bactrocera dorsalis (Hendel)

Introduction of the RTA‐Bddsx gene induces female‐specific lethal effects in transformed Bactrocera dorsalis (Hendel)

Introduction of a cold sensitivity‐conferring mutation into the RTA‐Bddsx hybrid system of Bactrocera dorsalis for establishment of a thermally controllable homozygous line

Genetically Engineered Ricin SuppressesBactrocera dorsalis(Diptera: Tephritidae) based on Demographic Analysis of Group-Reared Life Table

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