Bento Flavia Mm, Marques Rodrigo N, Campana Felippe B, Demétrio Clarice Gb, Leandro Roseli A, Parra José Roberto P, Figueira Antonio
Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, Piracicaba, Brazil.
Escola Superior de Agricultura "Luiz de Queiroz" (ESALQ), Universidade de São Paulo, Piracicaba, Brazil.
Pest Manag Sci. 2020 Jan;76(1):287-295. doi: 10.1002/ps.5513. Epub 2019 Jul 16.
RNA interference (RNAi) has been evaluated in several insect pests as a novel strategy to be included in integrated pest management. Lepidopterans are recognized to be recalcitrant to gene silencing by RNAi. As such, double-stranded RNA (dsRNA) delivery needs to be adjusted to assure its stability until it reaches the target gene transcript for silencing. Gene silencing by RNAi offers the potential to be used in the control of Tuta absoluta (Meyrick), one of the main insect pests of tomato (Solanum lycopersicum) worldwide. Here, we tested the delivery of dsRNA expressed in Escherichia coli HT115(DE3) and supplied to larvae in an artificial diet by screening target genes for silencing. We tested six target genes: juvenile hormone inducible protein (JHP); juvenile hormone epoxide hydrolase protein (JHEH); ecdysteroid 25-hydroxylase (PHM); chitin synthase A (CHI); carboxylesterase (COE); and arginine kinase (AK).
Based on larval mortality, the duration of the larval stage in days, pupal weight, and the accumulation of the target gene transcript, we demonstrated the efficacy of bacterial dsRNA delivery for the functional effects on larval development. Providing dsRNA targeted to JHP, CHI, COE and AK by bacteria led to a significant decrease in transcript accumulation and an increase in larval mortality.
Bacteria expressing dsRNA targeting essential T. absoluta genes supplied in artificial diet are efficient to screen RNAi target-genes. The oral delivery of dsRNA by bacteria is a novel potential alternative for the control of T. absoluta based on RNAi. © 2019 Society of Chemical Industry.
RNA干扰(RNAi)已在多种害虫中进行评估,作为综合害虫管理中一种新的策略。鳞翅目昆虫被认为对RNAi介导的基因沉默具有抗性。因此,双链RNA(dsRNA)的递送需要进行调整,以确保其在到达靶基因转录本进行沉默之前的稳定性。RNAi介导的基因沉默具有用于防治番茄潜叶蛾(Tuta absoluta, Meyrick)的潜力,番茄潜叶蛾是全球番茄(Solanum lycopersicum)的主要害虫之一。在此,我们通过筛选用于沉默的靶基因,测试了在大肠杆菌HT115(DE3)中表达并通过人工饲料提供给幼虫的dsRNA的递送情况。我们测试了六个靶基因:保幼激素诱导蛋白(JHP);保幼激素环氧水解酶蛋白(JHEH);蜕皮甾体25-羟化酶(PHM);几丁质合成酶A(CHI);羧酸酯酶(COE);以及精氨酸激酶(AK)。
基于幼虫死亡率、幼虫期天数、蛹重以及靶基因转录本的积累情况,我们证明了细菌递送dsRNA对幼虫发育具有功能影响的有效性。通过细菌提供靶向JHP、CHI、COE和AK的dsRNA导致转录本积累显著减少,幼虫死亡率增加。
在人工饲料中提供表达靶向番茄潜叶蛾必需基因的dsRNA的细菌,对于筛选RNAi靶基因是有效的。基于RNAi,通过细菌口服递送dsRNA是防治番茄潜叶蛾的一种新的潜在替代方法。© 2019化学工业协会。