Vatanparast M, Kazzazi M, Mirzaie-Asl A, Hosseininaveh V
Department of Plant Protection, College of Agriculture, Bu-Ali Sina University, Hamedan, Iran.
Department of Biotechnology, College of Agriculture, Bu-Ali Sina University, Hamedan, Iran.
Bull Entomol Res. 2017 Dec;107(6):777-790. doi: 10.1017/S0007485317000293. Epub 2017 May 9.
Helicoverpa armigera is a significant agricultural pest and particularly notorious for its resistance to many types of common insecticides. RNA interference (RNAi) is a mechanism of post-transcriptional gene silencing and trigged by double-strand RNA (dsRNA), has become a widely used reverse genetics and potent tool for insect pest control. In this study, the effect of ingestion and injection delivery methods of dsRNA related two important enzyme genes, α-amylase (HaAMY48, Ha-AMY49) and juvenile hormone esterase (Ha-JHE), were examined on growth and development of H. armigera. After 24, 48, 72 and 96 h of feeding bioassay, significant down regulation was observed about; 56, 68, 78, 80.75% for HaAMY48, 60, 70, 86.5 and 96.75%, for Ha-AMY49 and 14, 27.5, 23 and 31.7% for Ha-JHE, respectively. The results for injection assay was 61.5, 71.5, 74 and 95.8% for Ha-AMY48; 70, 88, 91.5 and 97.7% for Ha-AMY49 and 22, 61, 75 and 74% for Ha-JHE after 24, 48 and 72 h of last injecting, respectively. Larvae that treated with dsRNA, fed or injected, lost more than half of their weight. 50% mortality in treated larvae was observed in the case injection bioassay with dsHa-JHE and 59% of larvae that fed of dsRNA-treated cubes survived. DsHa-AMY48 and 49 have significant mortality, but mixing of them is more effective in both bioassays. Injection bioassay has a potent inhibitory effect on α-amylase-specific activity about more than 87% in treated larvae with mix of dsHa-AMY48 and 49. Adult malformation percent was evaluated for feeding (28, 35.5 and 43% for Ha-AMY48, 49 and Ha-JHE, respectively) and injection bioassay (23, 42 and 29% for Ha-AMY48, 49 and Ha-JHE, respectively). All these finding suggest that Ha-AMY48, Ha-AMY49 and Ha-JHE can be new candidates to scheming effective dsRNAs pesticide for H. armigera control.
棉铃虫是一种重要的农业害虫，尤其以对多种常见杀虫剂具有抗性而臭名昭著。RNA干扰（RNAi）是一种转录后基因沉默机制，由双链RNA（dsRNA）触发，已成为一种广泛应用的反向遗传学和控制害虫的有力工具。在本研究中，研究了与两个重要酶基因α-淀粉酶（HaAMY48、Ha-AMY49）和保幼激素酯酶（Ha-JHE）相关的dsRNA的摄入和注射递送方法对棉铃虫生长发育的影响。在喂食生物测定24、48、72和96小时后，观察到HaAMY48分别显著下调约56%、68%、78%、80.75%；Ha-AMY49分别下调60%、70%、86.5%和96.75%；Ha-JHE分别下调14%、27.5%、23%和31.7%。注射测定的结果是，在最后一次注射24、48和72小时后，Ha-AMY48分别为61.5%、71.5%、74%和95.8%；Ha-AMY49分别为70%、88%、91.5%和97.7%；Ha-JHE分别为22%、61%、75%和74%。用dsRNA处理（喂食或注射）的幼虫体重减轻了一半以上。在用dsHa-JHE进行注射生物测定的情况下，观察到处理后的幼虫死亡率为50%，而喂食dsRNA处理过的方块的幼虫中有59%存活。DsHa-AMY48和49具有显著的致死率，但在两种生物测定中，将它们混合使用效果更佳。在用dsHa-AMY48和49混合处理的幼虫中，注射生物测定对α-淀粉酶比活性具有超过87%的强效抑制作用。对成虫畸形率进行了评估，喂食生物测定中（Ha-AMY48、49和Ha-JHE分别为28%、35.5%和43%），注射生物测定中（Ha-AMY48、49和Ha-JHE分别为23%、42%和29%）。所有这些发现表明Ha-AMY48、Ha-AMY49和Ha-JHE可成为设计有效dsRNAs杀虫剂以控制棉铃虫的新候选基因。
