Firmino Alexandre Augusto Pereira, Pinheiro Daniele Heloísa, Moreira-Pinto Clidia Eduarda, Antonino José Dijair, Macedo Leonardo Lima Pepino, Martins-de-Sa Diogo, Arraes Fabrício Barbosa Monteiro, Coelho Roberta Ramos, Fonseca Fernando Campos de Assis, Silva Maria Cristina Mattar, Engler Janice de Almeida, Silva Marília Santos, Lourenço-Tessutti Isabela Tristan, Terra Walter Ribeiro, Grossi-de-Sa Maria Fátima
Embrapa Genetic Resources and Biotechnology, Brasília, Brazil.
Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany.
Front Physiol. 2020 Nov 16;11:591569. doi: 10.3389/fphys.2020.591569. eCollection 2020.
The cotton boll weevil, , is the most economically important pest of cotton in Brazil. Pest management programs focused on are based mostly on the use of chemical insecticides, which may cause serious ecological impacts. Furthermore, has developed resistance to some insecticides after their long-term use. Therefore, alternative control approaches that are more sustainable and have reduced environmental impacts are highly desirable to protect cotton crops from this destructive pest. RNA interference (RNAi) is a valuable reverse genetics tool for the investigation of gene function and has been explored for the development of strategies to control agricultural insect pests. This study aimed to evaluate the biological role of the () gene in and its potential as an RNAi target for the control of this insect pest. We found that is expressed throughout the development of with significantly higher expression in pupal and adult developmental stages. In addition, the immunolocalization of the AgraLac2 protein in third-instar larvae using specific antibodies revealed that AgraLac2 is distributed throughout the epithelial tissue, the cuticle and the tracheal system. We also verified that the knockdown of in resulted in an altered cuticle tanning process, molting defects and arrested development. Remarkably, insects injected with ds exhibited defects in cuticle hardening and pigmentation. As a consequence, the development of ds-treated insects was compromised, and in cases of severe phenotypic defects, the insects subsequently died. On the contrary, insects subjected to control treatments did not show any visible phenotypic defects in cuticle formation and successfully molted to the pupal and adult stages. Taken together, our data indicate that is involved in the cuticle tanning process in and may be a promising target for the development of RNAi-based technologies.
棉铃象甲是巴西棉花在经济上最重要的害虫。针对棉铃象甲的害虫管理计划主要基于化学杀虫剂的使用,这可能会造成严重的生态影响。此外,长期使用某些杀虫剂后,棉铃象甲已对它们产生了抗性。因此,非常需要更具可持续性且对环境影响较小的替代控制方法,以保护棉花作物免受这种毁灭性害虫的侵害。RNA干扰(RNAi)是一种用于研究基因功能的有价值的反向遗传学工具,并已被探索用于开发控制农业害虫的策略。本研究旨在评估AgraLac2基因在棉铃象甲中的生物学作用及其作为控制这种害虫的RNAi靶标的潜力。我们发现AgraLac2在棉铃象甲的整个发育过程中都有表达,在蛹期和成虫发育阶段表达显著更高。此外,使用特异性抗体对三龄幼虫中AgraLac2蛋白进行免疫定位显示,AgraLac2分布于整个上皮组织、表皮和气管系统。我们还证实,在棉铃象甲中敲低AgraLac2会导致表皮鞣化过程改变、蜕皮缺陷和发育停滞。值得注意的是,注射dsRNA的昆虫在表皮硬化和色素沉着方面出现缺陷。因此,经dsRNA处理的昆虫的发育受到损害,在严重表型缺陷的情况下,昆虫随后死亡。相反,接受对照处理的昆虫在表皮形成方面未表现出任何明显的表型缺陷,并成功蜕皮至蛹期和成虫期。综上所述,我们的数据表明AgraLac2参与了棉铃象甲的表皮鞣化过程,可能是基于RNAi技术开发的一个有前景的靶标。