Introduction of the <i><scp>RTA</scp>‐Bddsx</i> gene induces female‐specific lethal effects in transformed <i>Bactrocera dorsalis</i> (Hendel)

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

doi:10.1002/ps.4094

Cited by 5 publications

(15 citation statements)

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“…1B). The appearance of transformed females indicates that the actin 5C promoter-RTA-Bddsx transgene lost its function during chromosomal rearrangement (Huang et al 2016b). The transformed females constituted 9.51% of the total transformed F 4 population.…”

Section: Resultsmentioning

confidence: 99%

“…Life Table: High Larval Mortality, a Delay in Pupation, and a High Male/Female Ratio After Introduction of the RTA-Bddsx Transgene Eight transgenic F 3 males from high-male-percentage (94.6-100%) F 2 flies were selected and mated with wild-type females, and a parallel wild-type experiment was simultaneously conducted as a control (Huang et al 2016b). A total of 1,378 eggs were collected from the transformed population (Table 1), and the expression of the DsRed gene, a fluorescent marker, was used as an indicator of external DNA fragment insertion.…”

Section: Resultsmentioning

confidence: 99%

“…In this study, a laboratory strain of B. dorsalis, which has been maintained in the laboratory for over 13 years, was used as a source of the wild-type flies. Eight transgenic F 3 males from LERQ-1-2(F2) M08, LERQ-1-7(F2) M10, and LERQ-1-8(F2) M02 revealed high male percentages (94.6-100%) and were randomly selected and mated with wild-type females (Huang et al 2016b). The flies were maintained at 28 C under a photoperiod of 12:12 (L:D) h. Adult flies were housed in cages for mating and egg-laying; hatched larvae were cultured through periodic transfer to fresh food.…”

Section: Fruit Fly Strainsmentioning

confidence: 99%

“…Genomic DNA Isolation and Analysis of the Integrity of the RTA-Bddsx Transgene Genomic DNA was isolated from the heads of the transformed fruit flies according to a previously reported method (Huang et al 2016b). The primers PB-5587F (5 0 -CCCGTAATGCAGAAGAA GACCATG-3 0 ) and PB-6522R (5 0 -CGCAGCGTGACCGCTACA CTTG-3 0 ) were used to examine the integrity of the actin 5C promoter and the RTA-Bddsx gene, and PCR amplification was performed as follows: 94 C for 2 min; 35 cycles of 94 C for 45 s, 56 C for 1 min, and 72 C for 2.5 min; and 72 C for 10 min.…”

Section: Life Table Analysismentioning

confidence: 99%

“…In previous studies, we combined the female-specific alternative splicing property of Bddsx with the toxicity of RTA to develop a hybrid system for female-specific lethality in vitro and in vivo (Huang et al 2016a). Subsequently, an RTA-Bddsx transgene driven by the actin 5C promoter of B. dorsalis in the piggyBac vector was used to establish transgenic flies (Huang et al 2016b).…”

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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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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Fruit Fly Strainsmentioning

confidence: 99%

Section: Life Table Analysismentioning

confidence: 99%

mentioning

confidence: 99%

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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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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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Development of a piggyBac transgenic system in Bactrocera dorsalis and its potential for research on olfactory molecular targets

Zhang,

Wang,

Yuan

et al. 2024

Journal of Integrative Agriculture

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

Abstract: This transgenic study provides direct evidence for the female-specific lethal effects of RTA-Bddsx in B. dorsalis and offers a novel and promising approach for the control of B. dorsalis in the future. © 2015 Society of Chemical Industry.

Cited by 5 publications

References 57 publications

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

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

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

Development of a piggyBac transgenic system in Bactrocera dorsalis and its potential for research on olfactory molecular targets

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