Development of a Stage- and Species-Specific RNAi System for Molecular Insights in <i>Trichogramma</i> Wasps
<i>Trichogramma</i> wasps, egg parasitoids widely used to control lepidopteran pests, have long eluded in-depth molecular mechanistic studies due to their minute size and genetic tool scarcity. While previous RNAi efforts were restricted to <i>T. dendrolimi</i>, we developed...
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2025-06-01
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| author | Zelong Yang Yan Lu Zhuo Jiang Xilin Jiao Han Lin Wanning Jiang Wenmei Du Xue Zhang Zhao Peng Junjie Zhang Xiao Wang Ying Hu |
| author_facet | Zelong Yang Yan Lu Zhuo Jiang Xilin Jiao Han Lin Wanning Jiang Wenmei Du Xue Zhang Zhao Peng Junjie Zhang Xiao Wang Ying Hu |
| author_sort | Zelong Yang |
| collection | DOAJ |
| description | <i>Trichogramma</i> wasps, egg parasitoids widely used to control lepidopteran pests, have long eluded in-depth molecular mechanistic studies due to their minute size and genetic tool scarcity. While previous RNAi efforts were restricted to <i>T. dendrolimi</i>, we developed the first cross-species RNAi system for both <i>T. dendrolimi</i> and the previously intractable <i>T. ostriniae</i>. Temporal expression profiling identified <i>white</i> and <i>laccase 2</i> as stage-specific RNAi targets, peaking during prepupal/pupal stages, which were tested across species and developmental stages using microinjection and soaking dsRNA delivery methods. Survival analysis prioritized soaking for <i>T. dendrolimi</i> prepupae/pupae, while microinjection was essential for <i>T. ostriniae</i> to bypass prepupal mortality during soaking. Concentration-dependent RNAi targeting the <i>white</i> gene achieved 85.61% transcript reduction in <i>T. dendrolimi</i> via soaking and 89.36% in <i>T. ostriniae</i> via microinjection at 2000 ng/μL, correlating with 64.06% and 32.09% white-eyed pupae, causing a significant reduction in eye pigments. For the <i>laccase 2</i> gene, soaking at 2000 ng/μL induced 88.35% transcript reduction in <i>T. dendrolimi</i> and 73.31% in <i>T. ostriniae</i>, leading to incomplete cuticle tanning and sclerotization. This study resolves the long-standing challenge of genetic manipulation in <i>Trichogramma</i> wasps, providing a universally applicable framework to decipher parasitoid–host interactions at the molecular scale, which is useful for sustainable pest management strategies. |
| format | Article |
| id | doaj-art-8c79aaabc05f40c8b759c808ed51048f |
| institution | Kabale University |
| issn | 2075-4450 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Insects |
| spelling | doaj-art-8c79aaabc05f40c8b759c808ed51048f2025-08-20T03:58:30ZengMDPI AGInsects2075-44502025-06-0116767310.3390/insects16070673Development of a Stage- and Species-Specific RNAi System for Molecular Insights in <i>Trichogramma</i> WaspsZelong Yang0Yan Lu1Zhuo Jiang2Xilin Jiao3Han Lin4Wanning Jiang5Wenmei Du6Xue Zhang7Zhao Peng8Junjie Zhang9Xiao Wang10Ying Hu11Engineering Research Center of Natural Enemies, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, ChinaEngineering Research Center of Natural Enemies, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, ChinaEngineering Research Center of Natural Enemies, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, ChinaEngineering Research Center of Natural Enemies, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, ChinaEngineering Research Center of Natural Enemies, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, ChinaEngineering Research Center of Natural Enemies, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, ChinaEngineering Research Center of Natural Enemies, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, ChinaEngineering Research Center of Natural Enemies, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, ChinaDepartment of Plant Pathology, College of Plant Protection, Jilin Agricultural University, Changchun 130118, ChinaEngineering Research Center of Natural Enemies, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, ChinaEngineering Research Center of Natural Enemies, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, ChinaEngineering Research Center of Natural Enemies, Institute of Biological Control, Jilin Agricultural University, Changchun 130118, China<i>Trichogramma</i> wasps, egg parasitoids widely used to control lepidopteran pests, have long eluded in-depth molecular mechanistic studies due to their minute size and genetic tool scarcity. While previous RNAi efforts were restricted to <i>T. dendrolimi</i>, we developed the first cross-species RNAi system for both <i>T. dendrolimi</i> and the previously intractable <i>T. ostriniae</i>. Temporal expression profiling identified <i>white</i> and <i>laccase 2</i> as stage-specific RNAi targets, peaking during prepupal/pupal stages, which were tested across species and developmental stages using microinjection and soaking dsRNA delivery methods. Survival analysis prioritized soaking for <i>T. dendrolimi</i> prepupae/pupae, while microinjection was essential for <i>T. ostriniae</i> to bypass prepupal mortality during soaking. Concentration-dependent RNAi targeting the <i>white</i> gene achieved 85.61% transcript reduction in <i>T. dendrolimi</i> via soaking and 89.36% in <i>T. ostriniae</i> via microinjection at 2000 ng/μL, correlating with 64.06% and 32.09% white-eyed pupae, causing a significant reduction in eye pigments. For the <i>laccase 2</i> gene, soaking at 2000 ng/μL induced 88.35% transcript reduction in <i>T. dendrolimi</i> and 73.31% in <i>T. ostriniae</i>, leading to incomplete cuticle tanning and sclerotization. This study resolves the long-standing challenge of genetic manipulation in <i>Trichogramma</i> wasps, providing a universally applicable framework to decipher parasitoid–host interactions at the molecular scale, which is useful for sustainable pest management strategies.https://www.mdpi.com/2075-4450/16/7/673<i>Trichogramma</i> waspsRNA interferencesoakingmicroinjectionspecies-specific optimization |
| spellingShingle | Zelong Yang Yan Lu Zhuo Jiang Xilin Jiao Han Lin Wanning Jiang Wenmei Du Xue Zhang Zhao Peng Junjie Zhang Xiao Wang Ying Hu Development of a Stage- and Species-Specific RNAi System for Molecular Insights in <i>Trichogramma</i> Wasps Insects <i>Trichogramma</i> wasps RNA interference soaking microinjection species-specific optimization |
| title | Development of a Stage- and Species-Specific RNAi System for Molecular Insights in <i>Trichogramma</i> Wasps |
| title_full | Development of a Stage- and Species-Specific RNAi System for Molecular Insights in <i>Trichogramma</i> Wasps |
| title_fullStr | Development of a Stage- and Species-Specific RNAi System for Molecular Insights in <i>Trichogramma</i> Wasps |
| title_full_unstemmed | Development of a Stage- and Species-Specific RNAi System for Molecular Insights in <i>Trichogramma</i> Wasps |
| title_short | Development of a Stage- and Species-Specific RNAi System for Molecular Insights in <i>Trichogramma</i> Wasps |
| title_sort | development of a stage and species specific rnai system for molecular insights in i trichogramma i wasps |
| topic | <i>Trichogramma</i> wasps RNA interference soaking microinjection species-specific optimization |
| url | https://www.mdpi.com/2075-4450/16/7/673 |
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