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Transgenic potato lines expressing hairpin RNAi construct of molting-associated EcR gene exhibit enhanced resistance against Colorado potato beetle (Leptinotarsa decemlineata, Say).
Transgenic Res. 2019 02; 28(1):151-164.TR

Abstract

Most of the commercialized insect resistant transgenic crops express cry gene(s) isolated from Bacillus thuringiensis; however, intensive cultivation of Bt crops over almost two decades has been questioned regarding its sustainability and durability in pest management. The present study focused on silencing of highly specific molting-associated Ecdysone receptor (EcR) gene of Colorado potato beetle (CPB) using RNA interference (RNAi) approach. The partial cDNA of EcR gene of CPB was amplified using specific primers in sense and anti-sense orientations, and cloned in pRNAi-GG vector flanked by an intronic sequence (pdk). Leaf and internodal explants of Agria and Lady Olympia potato cultivars were infected with Agrobacterium strain LBA4404 harboring constructs under the control of CaMV 35S promoter. Standard molecular analysis of primary transformants showed proper integration of T-DNA in plant genome. The transgenic plants of both cultivars were evaluated for their efficacy against first, second and third instar CPB larvae. The leaf biotoxicity assays revealed 15-80% of CPB mortality. A significantly lower fold-change (0.87-4.14×) in larval weight was observed in insects fed on transgenic plants compared to the ones fed on control plants (1.87-6.53×). Furthermore, CPB larvae fed on transgenic plants exhibited reduced EcR transcripts, indicating the functionality of dsRNA EcR in silencing EcR gene expression. This study is an excellent example of the integration of an alternative, effective and reliable method to cope with potato insect pests that incur significant losses to potato production in the world.

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Authors+Show Affiliations

Hussain T
Department of Agricultural Genetic Engineering, Faculty of Agricultural Sciences and Technologies, Nigde Omer Halisdemir University, 51240, Nigde, Turkey.
Aksoy E
Department of Agricultural Genetic Engineering, Faculty of Agricultural Sciences and Technologies, Nigde Omer Halisdemir University, 51240, Nigde, Turkey.
Çalışkan ME
Department of Agricultural Genetic Engineering, Faculty of Agricultural Sciences and Technologies, Nigde Omer Halisdemir University, 51240, Nigde, Turkey.
Bakhsh A
Department of Agricultural Genetic Engineering, Faculty of Agricultural Sciences and Technologies, Nigde Omer Halisdemir University, 51240, Nigde, Turkey. abthebest@gmail.com.

MeSH

AnimalsBacillus thuringiensisColeopteraInsect ProteinsLarvaMoltingPest ControlPlant LeavesPlants, Genetically ModifiedRNA InterferenceReceptors, SteroidSolanum tuberosum

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

30607744

Citation

Hussain, Tahira, et al. "Transgenic Potato Lines Expressing Hairpin RNAi Construct of Molting-associated EcR Gene Exhibit Enhanced Resistance Against Colorado Potato Beetle (Leptinotarsa Decemlineata, Say)." Transgenic Research, vol. 28, no. 1, 2019, pp. 151-164.
Hussain T, Aksoy E, Çalışkan ME, et al. Transgenic potato lines expressing hairpin RNAi construct of molting-associated EcR gene exhibit enhanced resistance against Colorado potato beetle (Leptinotarsa decemlineata, Say). Transgenic Res. 2019;28(1):151-164.
Hussain, T., Aksoy, E., Çalışkan, M. E., & Bakhsh, A. (2019). Transgenic potato lines expressing hairpin RNAi construct of molting-associated EcR gene exhibit enhanced resistance against Colorado potato beetle (Leptinotarsa decemlineata, Say). Transgenic Research, 28(1), 151-164. https://doi.org/10.1007/s11248-018-0109-7
Hussain T, et al. Transgenic Potato Lines Expressing Hairpin RNAi Construct of Molting-associated EcR Gene Exhibit Enhanced Resistance Against Colorado Potato Beetle (Leptinotarsa Decemlineata, Say). Transgenic Res. 2019;28(1):151-164. PubMed PMID: 30607744.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Transgenic potato lines expressing hairpin RNAi construct of molting-associated EcR gene exhibit enhanced resistance against Colorado potato beetle (Leptinotarsa decemlineata, Say). AU - Hussain,Tahira, AU - Aksoy,Emre, AU - Çalışkan,Mehmet Emin, AU - Bakhsh,Allah, Y1 - 2019/01/03/ PY - 2018/08/27/received PY - 2018/12/11/accepted PY - 2019/1/5/pubmed PY - 2019/3/30/medline PY - 2019/1/5/entrez KW - Ecdysone receptor KW - Insect resistance KW - Molting KW - Transgenic technology SP - 151 EP - 164 JF - Transgenic research JO - Transgenic Res VL - 28 IS - 1 N2 - Most of the commercialized insect resistant transgenic crops express cry gene(s) isolated from Bacillus thuringiensis; however, intensive cultivation of Bt crops over almost two decades has been questioned regarding its sustainability and durability in pest management. The present study focused on silencing of highly specific molting-associated Ecdysone receptor (EcR) gene of Colorado potato beetle (CPB) using RNA interference (RNAi) approach. The partial cDNA of EcR gene of CPB was amplified using specific primers in sense and anti-sense orientations, and cloned in pRNAi-GG vector flanked by an intronic sequence (pdk). Leaf and internodal explants of Agria and Lady Olympia potato cultivars were infected with Agrobacterium strain LBA4404 harboring constructs under the control of CaMV 35S promoter. Standard molecular analysis of primary transformants showed proper integration of T-DNA in plant genome. The transgenic plants of both cultivars were evaluated for their efficacy against first, second and third instar CPB larvae. The leaf biotoxicity assays revealed 15-80% of CPB mortality. A significantly lower fold-change (0.87-4.14×) in larval weight was observed in insects fed on transgenic plants compared to the ones fed on control plants (1.87-6.53×). Furthermore, CPB larvae fed on transgenic plants exhibited reduced EcR transcripts, indicating the functionality of dsRNA EcR in silencing EcR gene expression. This study is an excellent example of the integration of an alternative, effective and reliable method to cope with potato insect pests that incur significant losses to potato production in the world. SN - 1573-9368 UR - https://www.unboundmedicine.com/medline/citation/30607744/Transgenic_potato_lines_expressing_hairpin_RNAi_construct_of_molting_associated_EcR_gene_exhibit_enhanced_resistance_against_Colorado_potato_beetle__Leptinotarsa_decemlineata_Say__ DB - PRIME DP - Unbound Medicine ER -