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Title: | 球背象鼻蟲鱗片基因表現 Gene expression during scale development in Pachyrhynchus weevils |
Authors: | 陳奕臻 Yi-Chen Chen |
Advisor: | 曾惠芸 Hui-Yun Tseng |
Keyword: | 球背象鼻蟲,RNA-seq,次世代定序,qPCR,鱗片發育, Pachyrhynchus weevils,NGS,RNA-seq,qPCR,scale development, |
Publication Year : | 2023 |
Degree: | 碩士 |
Abstract: | 生物身上多樣且鮮豔的色彩常具備不同的功能,其中球背象鼻蟲屬(Pachyrhynchus)多樣化的色彩與斑紋常與性擇與警戒色訊號有關。球背象鼻蟲鮮豔的色彩為翅鞘鱗片內光子晶體結構所造成,且為遺傳所調控,推測可能有一些特定的基因涉及鱗片顏色調節機制,但相關調控基因仍然未知。本篇研究利用三種球背象鼻蟲(斷紋球背象鼻蟲、大圓斑球背象鼻蟲與球斑球背象鼻蟲)進行分析,首先以斷紋球背象鼻蟲進行次世代定序,篩選出可能參與鱗片顏色調控的基因及調節的相關路徑,並利用qPCR 檢測候選基因於球背象鼻蟲表現情形。候選基因從次世代定序結果中的差異表現量基因(differentially expressed genes, DEGs)篩選,並設計qPCR引子。定序結果顯示羽化後6小時及24小時分別可辨識3172、2890個DEGs,且隨著羽化時間增加,表現量上調的DEGs數量上升。在heatmap分析的結果中,篩選的候選基因分成翅鞘及鱗片兩個類群,並偵測到actin (gene) 、tyrosine (gene) 、cuticle (gene) 等特定基因表現量可以區分翅鞘或鱗片的差異。以qPCR比較球背象鼻蟲羽化後6小時及24小時基因表現量可以發現不同物種有不同的情況; calcium (pathway) 在大圓斑球背象鼻蟲羽化後24小時的表現量有明顯降低,而球斑球背象鼻蟲則是wnt (pathway) 表現量顯著降低。比較羽化6小時種間的差異可以看出actin (pathway) 在斷紋球背象鼻蟲的表現量明顯低於球斑球背象鼻蟲;而斷紋球背象鼻蟲tyrosine (pathway) 的表現量顯著高於大圓斑球背象鼻蟲。此外,羽化後24小時tyrosine (pathway) 在大圓斑球背象鼻蟲的表現量比球斑球背象鼻蟲高。總而言之,4個基因tyrosine (gene)、wnt (pathway)、actin (gene)、calcium (pathway) 在不同種不同羽化時間的表現量呈現差異。另外有4個基因出現在特定物種或是時間點,如lectin (gene)表現只有在斷紋球背象鼻蟲被偵測到;chitin (gene)表現於羽化後24小時的大圓斑球背象鼻蟲及球斑球背象鼻蟲;球斑球背象鼻蟲與斷紋球象鼻蟲檢測到ras (pathway)表現;而hedgehog (gene)僅在球斑球背象鼻蟲檢測到。綜合上述,本研究結果顯示這些特定基因可能於球背象鼻蟲鱗片發育的成色機制中扮演其中一角,並在羽化後不同時間點或是物種間的差異會受到不同基因不同程度的調控。 The diverse and colorful patterns found in organism serve various functions. In Pachyrhynchus weevils, these colors and patterns function as signals for sexual selection and aposematism, which was composed of 3D photonic architectures. The study of photonic structure through hybridization suggests the involvement of specific genes in color regulation. However, the genes responsible for regulating scale color in Pachyrhynchus weevils have remained unknown. In this study, RNA-seq was conducted in P. nobilis to identify DEGs (differentially expressed genes), and candidate genes potentially involved in scale regulation or related pathway were selected from the DEGs. The expression of candidate genes was then detected using qPCR in three Pachyrhynchus species (P. nobilis, P. sarcitis, and P. orbifer). At 6 hours and 24 hours after emergence, 3172 and 2890 DEGs were identified in NGS data, respectively, with the number of up-regulated DEGs increasing over time. In the heatmap, the candidate genes were clustered into elytra and scale groups. Certain genes, such as actin (gene), tyrosine (gene) and cuticle (gene), exhibited high expression in both elytra and scale tissues. In the qPCR comparison between 6 hours and 24 hours after emergence, the expression of calcium (pathway) decreased at 24 hours in P. sarcitis, and the expression level of wnt (pathway) significantly decreased at 24 hours in P. orbifer. When comparing among species at 6 hours, the expression of actin (gene) in P. orbifer was significantly higher than that in P. nobilis, and the expression of tyrosine (pathway) was significantly higher in P. nobilis than in P. sarcitis. However, at 24 hours, P. sarcitis exhibited higher expression in tyrosine (pathway) than in P. orbifer. Overall, the expression of four genes, tyrosine (pathway), wnt (pathway), actin (gene), and calcium (pathway), differed among Pachyrhynchus weevils. Additionally, the expression of four genes varied between different species or at specific time points. For instance, lectin (gene) was only detected in P. nobilis, ras (pathway) was detected in P. nobilis and P. orbifer, the expression of chitin (gene) was found in P. sarcitis and P. orbifer at 24 hours after emergence and hedgehog (gene) was detected in P. orbifer. Based on the above findings, this study suggests that these specific genes may play roles in color regulation during scale development in Pachyrhynchus weevils, with varying expression at different time points after emergence or among species. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/90733 |
DOI: | 10.6342/NTU202302538 |
Fulltext Rights: | 同意授權(限校園內公開) |
Appears in Collections: | 昆蟲學系 |
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