South African National Bioinformatics Institute, South African Medical Research Council Bioinformatics Unit, The University of the Western Cape, Bellville, Western Cape, South Africa.
AgriProtein Technologies (Pty) Limited, Philippi, Western Cape, South Africa.
PLoS One. 2022 Mar 17;17(3):e0265492. doi: 10.1371/journal.pone.0265492. eCollection 2022.
The growing demands on protein producers and the dwindling available resources have made Hermetia illucens (the black soldier fly, BSF) an economically important species. Insights into the genome of this insect will better allow for robust breeding protocols, and more efficient production to be used as a replacement of animal feed protein. The use of microRNA as a method to understand how gene regulation allows insect species to adapt to changes in their environment, has been established in multiple species. The baseline and life stage expression levels established in this study, allow for insight into the development and sex-linked microRNA regulation in BSF. To accomplish this, microRNA was extracted and sequenced from 15 different libraries with each life stage in triplicate. Of the total 192 microRNAs found, 168 were orthologous to known arthropod microRNAs and 24 microRNAs were unique to BSF. Twenty-six of the 168 microRNAs conserved across arthropods had a statistically significant (p < 0.05) differential expression between Egg to Larval stages. The development from larva to pupa was characterized by 16 statistically significant differentially expressed microRNA. Seven and 9 microRNA were detected as statistically significant between pupa to adult female and pupa to adult male, respectively. All life stages had a nearly equal split between up and down regulated microRNAs. Ten of the unique 24 miRNA were detected exclusively in one life stage. The egg life stage expressed five microRNA (hil-miR-m, hil-miR-p, hil-miR-r, hil-miR-s, and hil-miR-u) not seen in any other life stages. The female adult and pupa life stages expressed one miRNA each hil-miR-h and hil-miR-ac respectively. Both male and female adult life stages expressed hil-miR-a, hil-miR-b, and hil-miR-y. There were no unique microRNAs found only in the larva stage. Twenty-two microRNAs with 56 experimentally validated target genes in the closely related Drosophila melanogaster were identified. Thus, the microRNA found display the unique evolution of BSF, along with the life stages and potential genes to target for robust mass rearing. Understanding of the microRNA expression in BSF will further their use in the crucial search for alternative and sustainable protein sources.
对蛋白质生产者的需求不断增长，而可用资源却日益减少，这使得家蝇（黑蝇，BSF）成为一种具有重要经济意义的物种。深入了解这种昆虫的基因组，将有助于制定稳健的繁殖方案，并提高其作为动物饲料蛋白替代品的生产效率。在多个物种中，已经证实了 microRNA 作为一种了解基因调控如何使昆虫物种适应环境变化的方法。本研究建立了 BSF 发育和性连锁 microRNA 调控的基线和生命阶段表达水平。为此，从 15 个不同的三重复本的生命阶段文库中提取并测序了 microRNA。总共发现了 192 个 microRNA，其中 168 个与已知的节肢动物 microRNA 同源，24 个 microRNA 是 BSF 特有的。在节肢动物中保守的 168 个 microRNA 中有 26 个在卵到幼虫阶段的表达有统计学意义（p < 0.05）。从幼虫到蛹的发育阶段有 16 个 microRNA 的表达有统计学意义。在蛹到雌成虫和蛹到雄成虫之间，分别检测到 7 个和 9 个 microRNA 有统计学意义。所有生命阶段的上调和下调 microRNA 几乎相等。在 24 个独特的 microRNA 中，有 10 个仅在一个生命阶段被检测到。卵期表达了 5 个 microRNA（hil-miR-m、hil-miR-p、hil-miR-r、hil-miR-s 和 hil-miR-u），在其他生命阶段均未检测到。雌成虫和蛹期分别表达了 hil-miR-h 和 hil-miR-ac 各一个 microRNA。雄成虫和雌成虫都表达了 hil-miR-a、hil-miR-b 和 hil-miR-y。幼虫阶段没有发现独特的 microRNA。在密切相关的黑腹果蝇中，鉴定出了 22 个具有 56 个实验验证靶基因的 microRNA。因此，所发现的 microRNA 显示了 BSF 的独特进化，以及用于大规模繁殖的潜在靶基因。对 BSF 中 microRNA 表达的理解将进一步促进其在替代和可持续蛋白质来源的关键研究中的应用。
