Noriega Daniel D, Arraes Fabricio B M, Antonino José Dijair, Macedo Leonardo L P, Fonseca Fernando C A, Togawa Roberto C, Grynberg Priscila, Silva Maria C M, Negrisoli Aldomario S, Grossi-de-Sa Maria F
Embrapa Genetic Resources and Biotechnology, Brasília, Brazil.
Department of Cellular Biology, University of Brasília, Brasília, Brazil.
Front Physiol. 2020 Oct 28;11:588450. doi: 10.3389/fphys.2020.588450. eCollection 2020.
The sugarcane giant borer (SGB), , is a pest that has strong economic relevance for sugarcane producers. Due to the endophytic behavior of the larva, current methods of management are inefficient. A promising biotechnological management option has been proposed based on RNA interference (RNAi), a process that uses molecules of double-stranded RNA (dsRNA) to specifically knock down essential genes and reduce insect survival. The selection of suitable target genes is often supported by omic sciences. Studies have shown that genes related to feeding adaptation processes are good candidates to be targeted by RNAi for pest management. Among those genes, esterases are highlighted because of their impact on insect development. In this study, the objective was to evaluate the transcriptome responses of the SGB's gut in order to provide curated data of genes that could be used for pest management by RNAi in future studies. Further, we validated the function of an esterase-coding gene and its potential as a target for RNAi-based control. We sequenced the gut transcriptome of SGB larvae by Illumina HiSeq and evaluated its gene expression profiles in response to different diets (sugarcane stalk and artificial diet). We obtained differentially expressed genes (DEGs) involved in detoxification, digestion, and transport, which suggest a generalist mechanism of adaptation in SGB larvae. Among the DEGs, was identified and characterized a candidate juvenile hormone esterase gene (). We knocked down the gene by oral delivery of dsRNA molecules and evaluated gene expression in the gut. The survival and nutritional parameters of the larvae were measured along the developmental cycle of treated insects. We found that the gene acts as a regulator of feeding behavior. The knockdown of triggered a forced starvation state in late larval instars that significantly reduced the fitness of the larvae. However, the mechanism of action of this gene remains unclear, and the correlation between the expression of and the levels of juvenile hormone (JH) metabolites in the hemolymph of the SGB must be assessed in future research.
甘蔗大螟(SGB)对甘蔗种植者具有重大经济影响,是一种害虫。由于其幼虫的内生习性,目前的防治方法效率低下。基于RNA干扰(RNAi)提出了一种有前景的生物技术防治方案,该过程利用双链RNA(dsRNA)分子特异性敲除关键基因并降低昆虫存活率。合适靶基因的选择通常得到组学科学的支持。研究表明,与取食适应过程相关的基因是RNAi用于害虫防治的良好候选靶标。其中,酯酶因其对昆虫发育的影响而备受关注。本研究旨在评估甘蔗大螟肠道的转录组反应,以提供可用于未来RNAi害虫防治研究的基因数据。此外,我们验证了一个酯酶编码基因的功能及其作为RNAi防治靶标的潜力。我们通过Illumina HiSeq对甘蔗大螟幼虫的肠道转录组进行测序,并评估其在不同食物(甘蔗茎和人工饲料)条件下的基因表达谱。我们获得了参与解毒、消化和运输的差异表达基因(DEGs),这表明甘蔗大螟幼虫具有普遍的适应机制。在这些DEGs中,鉴定并表征了一个候选保幼激素酯酶基因()。我们通过口服dsRNA分子敲低该基因,并评估肠道中的基因表达。在处理昆虫的发育周期中测量幼虫的存活和营养参数。我们发现该基因作为取食行为的调节因子。敲低该基因会在末龄幼虫中引发强制饥饿状态,显著降低幼虫的适合度。然而,该基因的作用机制尚不清楚,未来研究必须评估该基因的表达与甘蔗大螟血淋巴中保幼激素(JH)代谢物水平之间的相关性。