本文選題：中華鋸齒米蝦 + 轉(zhuǎn)錄組�。� 參考：《集美大學(xué)》2017年碩士論文

【摘要】：中華鋸齒米蝦(Neocaridina denticulata sinensis)廣泛分布于亞洲淡水水域,因其體色豐富多樣,自2003年開始成為一種重要的觀賞蝦。但是其體色的分子遺傳基礎(chǔ)尚未知。在觀賞中華鋸齒米蝦中,紅、黃和透明3種純色品系的體色遺傳最穩(wěn)定。因此,本文通過比較紅、黃和透明3種純色米蝦皮膚組織的轉(zhuǎn)錄組,獲得了大量體色差異表達(dá)基因,并利用qPCR在純色品系和琉璃蝦中對重要候選基因進(jìn)行驗(yàn)證。希望為揭示米蝦體色遺傳機(jī)制提供資料。主要研究結(jié)果如下:1.一共獲得了268,629,208個clean reads,經(jīng)TRINITY軟件拼接后獲得45,434條Unigenes。Unigene的平均長度為837 bp,N50為1551 bp。與NCBI nr、Swiss-prot、KOG和Kegg數(shù)據(jù)庫比對后,共有19,992條Unigenes獲得注釋。2.通過對3種純色品系米蝦皮膚轉(zhuǎn)錄組的基因表達(dá)量分析,共得到14,915個差異表達(dá)Unigene。核糖體蛋白基因、actin、calmodulin、EF1α和AK等基因可能通過色素遷移及色素體狀態(tài)維持這一細(xì)胞學(xué)過程引起米蝦體色差異。紅色品系的鳥嘌呤合成通路高表達(dá),表明米蝦紅色素的主要成分可能是蝶啶類色素。黃色品系沒有顯著高表達(dá)色素合成通路,但是甲殼藍(lán)蛋白基因在黃色品系米蝦中高表達(dá),表明米蝦黃色素可能與類胡蘿卜素有關(guān)。3.通過與已知果蠅、家蠶和赤擬谷盜體色基因的比對發(fā)現(xiàn)了大量可能參與色素沉積的基因,其中包括黑色素、眼色素、蝶呤等合成酶系及其調(diào)節(jié)基因并從中篩選出25條與體色相關(guān)的候選基因。利用實(shí)時熒光定量PCR獲得25條候選基因在紅、黃和透明3種純色品系及在2種琉璃蝦的表達(dá)模式。3種純色品系的RT-qPCR結(jié)果與轉(zhuǎn)錄組RPKM值的高低趨勢吻合。4.原位雜交實(shí)驗(yàn)結(jié)果表明,ebony、GC-I、SPR和XDH基因在米蝦的復(fù)眼期幼體和仔蝦階段均有表達(dá),而在無節(jié)幼體階段沒有檢測到表達(dá)信號。在色素沉積過程中,米蝦這4個基因可能具有與其昆蟲的同源基因類似的作用。轉(zhuǎn)錄組獲得這些數(shù)據(jù)為米蝦體色相關(guān)基因的發(fā)掘和體色遺傳機(jī)制的解析提供豐富的序列資源。
[Abstract]:Neocaridina denticulata sinensis is widely distributed in the freshwater waters of Asia. Because of its rich variety, it has become an important ornamental shrimp since 2003. But the molecular genetic basis of its body color is unknown. Among the ornamental Chinese sawtooth prawns, the color of red, yellow and transparent 3 species of pure color is the most stable. In this paper, a large number of differential expression genes were obtained by comparing the red, yellow and transparent 3 kinds of skin tissue of pure color prawns. The important candidate genes were verified by qPCR in the pure color and glazed shrimp. We hope to provide information for revealing the genetic mechanism of the body color of the prawns. The main results are as follows: 1. a total of 268629208 Clean reads, after TRINITY software splicing, the average length of 45434 Unigenes.Unigene is 837 BP, N50 is 1551 bp., and NCBI NR, Swiss-prot, KOG, and Kegg databases are compared, and a total of 19992 notes are obtained by analyzing the gene expression of the 3 species of pure color shrimp skin transcriptome, and a total of 14915 difference tables are obtained. The genes of Unigene. ribosomal protein, actin, calmodulin, EF1 a, and AK may cause the difference in the body color of the shrimp through the pigment migration and the status of the pigments. The guanine synthesis pathway in the red line is highly expressed, indicating that the main component of the red pigment in the shrimp may be the pellid pigment. The yellow line is not significantly higher. The expression of the pigment synthesis pathway, but the chitin gene is highly expressed in the Yellow strain of the shrimp, suggesting that the yellow pigment may be associated with the carotenoid.3. by comparing with the known fruit fly, silkworm, and hemilyx body color genes, a large number of genes that may be involved in pigment deposition, including melanin, eye pigments, and sphenotrexate, are found. 25 candidate genes related to body color were screened from the enzyme line and its regulatory gene. The RT-qPCR results of 25 candidate genes in red, yellow and transparent and the expression pattern of 2 kinds of glazed shrimp were obtained by real-time fluorescent quantitative PCR, and the RPKM value of the RPKM value of the transcriptional group was consistent with the.4. in situ hybridization experimental knot. The results showed that ebony, GC-I, SPR and XDH genes were expressed in the larval and shrimp stage of the shrimp, but the expression signal was not detected in the naupli stage. In the process of pigment deposition, the 4 genes of the shrimp may have similar effects on the homologous genes of the insects. The analysis of genetic mechanism of excavation and body color provides abundant sequence resources.
【學(xué)位授予單位】：集美大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S917.4

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