Zhang Jing, Blessing Danso, Wu Chenyu, Liu Na, Li Juan, Qin Sheng, Li Muwang
School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang, Jiangsu, China.
The Sericultural Research Institute, Chinese Academy of Agricultural Science, Zhenjiang, Jiangsu, China.
PLoS One. 2017 Jun 15;12(6):e0179560. doi: 10.1371/journal.pone.0179560. eCollection 2017.
Wings of Bombyx mori (B. mori) develop from the primordium, and different B. mori strains have different wing types. In order to identify the key factors influencing B. mori wing development, we chose strains P50 and U11, which are typical for normal wing and minute wing phenotypes, respectively. We dissected the wing disc on the 1st-day of wandering stage (P50D1 and U11D1), 2nd-day of wandering stage (P50D2 and U11D2), and 3rd-day of wandering stage (P50D3 and U11D3). Subsequently, RNA-sequencing (RNA-Seq) was performed on both strains in order to construct their gene expression profiles. P50 exhibited 628 genes differentially expressed to U11, 324 up-regulated genes, and 304 down-regulated genes. Five enriched gene ontology (GO) terms were identified by GO enrichment analysis based on these differentially expressed genes (DEGs). KEGG enrichment analysis results showed that the DEGs were enriched in five pathways; of these, we identified three pathways related to the development of wings. The three pathways include amino sugar and nucleotide sugar metabolism pathway, proteasome signaling pathway, and the Hippo signaling pathway. The representative genes in the enrichment pathways were further verified by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). The RNA-Seq and qRT-PCR results were largely consistent with each other. Our results also revealed that the significantly different genes obtained in our study might be involved in the development of the size of B. mori wings. In addition, several KEGG enriched pathways might be involved in the regulation of the pathways of wing formation. These results provide a basis for further research of wing development in B. mori.
家蚕的翅膀由原基发育而来，不同的家蚕品系具有不同的翅膀类型。为了确定影响家蚕翅膀发育的关键因素，我们选择了P50和U11品系，它们分别是正常翅膀和微小翅膀表型的典型代表。我们在化蛹前1天（P50D1和U11D1）、化蛹前2天（P50D2和U11D2）和化蛹前3天（P50D3和U11D3）解剖了翅芽。随后，对这两个品系进行了RNA测序（RNA-Seq），以构建它们的基因表达谱。P50与U11相比，有628个基因差异表达，其中324个基因上调，304个基因下调。基于这些差异表达基因（DEGs），通过GO富集分析确定了五个富集的基因本体（GO）术语。KEGG富集分析结果表明，这些DEGs富集在五条通路中；其中，我们确定了三条与翅膀发育相关的通路。这三条通路包括氨基糖和核苷酸糖代谢通路、蛋白酶体信号通路和Hippo信号通路。通过定量实时逆转录聚合酶链反应（qRT-PCR）进一步验证了富集通路中的代表性基因。RNA-Seq和qRT-PCR结果基本一致。我们的结果还表明，我们研究中获得的显著差异基因可能参与了家蚕翅膀大小的发育。此外，几个KEGG富集通路可能参与了翅膀形成通路的调控。这些结果为进一步研究家蚕翅膀发育提供了依据。
