Evaluation of dsRNAs targeting different genes in control of Colorado potato beetle (Leptinotarsa decemlineata)
Küçük Resim Yok
Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Niğde Ömer Halisdemir Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
RNA interferans (RNAi) tekniği genlerin seçici bir şekilde inhibe edilmesini sağlayan bir genetik araç olarak, bitki zararlıların kontrolünde umut vadetmektedir. Bu çalışmada, RNAi'nin patates böceği (Leptinotarsa decemlineata Say) üzerindeki etkilerini çift zincirli RNA'lar (dsRNA'lar), L4440 vektörü ve Escherichia coli HT115 suşu kullanılarak üretilen Vacuolar ATPase (V-ATPase), Ultraspiracle Protein (USP) ve Ecdysone receptor (EcR) mRNA'sını kullanılarak araştırılmıştır. Kullanılan RNAi'nin L. decemlineata ölüm oranında artma, larva ağırlığında ve besin tüketiminde azalma ve yumurta bırakma sayısında düşme gibi sonuçlar göstermiştir. RNAi uygulamaları azaltılmış mRNA seviyeleri ile RNAi'nin etkinliğini doğruladığı bulunmuştur. Ayrıca, dsUSP ve dsEcR'yi birlikte kullanımı bunların tek başına uygulanmalarına kıyasla, larvalarda %90 oranında ölüme yol açarken, dişilerin bıraktığı yumurta sayısında %88 azalmaya neden olduğu saptanmıştır. Sonuç olarak, bu çalışma, RNAi teknolojisinin L. decemlineata kontrolünde kullanılma potansiyeline sahip olduğunu göstermekte olup, bu zararlının popülasyonunu azaltmada kullanılabileceğini göstermektedir. Anahtar Kelimeler: RNA interferans, besleme biyotest, mortalite, mRNA seviyesi, sinerjizm
RNA interference (RNAi) serves as a versatile genetic tool capable of selectively inhibiting the expression of specific genes, providing a promising avenue for preventing crops from being infested by insect pests and diseases. In this study, RNAi was employed to investigate its effects on the Colorado Potato Beetle (Leptinotarsa decemlineata Say). The double-stranded RNAs (dsRNAs) were synthesized using L4440 vector in Escherichia coli HT115 strain to silence the L. decemlineata Vacuolar ATPase (V-ATPase), Ultraspiracle protein (USP), and Ecdysone receptor (EcR) mRNAs which are essential for a range of physiological and developmental processes. The results revealed that RNAi increased mortality rates, significant effects on larval weight gain, reduced foliage consumption, and suppression of egg-laying behavior in L. decemlineata. The targeted downregulation of gene expression, as evidenced by reduced mRNA levels, confirmed the efficacy of RNAi. Additionally, combining dsUSP and dsEcR showed a synergistic effect, demonstrating the greatest mortality rate recorded at 90% in larvae, with 88% reduction in the number of laid eggs compared to their individual applications in feeding bioassays. This study shows that employing RNAi technology has the potential to effectively control this devastating potato pest by providing a targeted approach to suppress its population. Keywords: RNA interference, feeding bioassay, mortality, mRNA level, synergism
RNA interference (RNAi) serves as a versatile genetic tool capable of selectively inhibiting the expression of specific genes, providing a promising avenue for preventing crops from being infested by insect pests and diseases. In this study, RNAi was employed to investigate its effects on the Colorado Potato Beetle (Leptinotarsa decemlineata Say). The double-stranded RNAs (dsRNAs) were synthesized using L4440 vector in Escherichia coli HT115 strain to silence the L. decemlineata Vacuolar ATPase (V-ATPase), Ultraspiracle protein (USP), and Ecdysone receptor (EcR) mRNAs which are essential for a range of physiological and developmental processes. The results revealed that RNAi increased mortality rates, significant effects on larval weight gain, reduced foliage consumption, and suppression of egg-laying behavior in L. decemlineata. The targeted downregulation of gene expression, as evidenced by reduced mRNA levels, confirmed the efficacy of RNAi. Additionally, combining dsUSP and dsEcR showed a synergistic effect, demonstrating the greatest mortality rate recorded at 90% in larvae, with 88% reduction in the number of laid eggs compared to their individual applications in feeding bioassays. This study shows that employing RNAi technology has the potential to effectively control this devastating potato pest by providing a targeted approach to suppress its population. Keywords: RNA interference, feeding bioassay, mortality, mRNA level, synergism
Açıklama
Fen Bilimleri Enstitüsü, Tarım Bilimleri ve Teknolojileri Ana Bilim Dalı
Anahtar Kelimeler
Ziraat, Agriculture
