中华锯齿米虾色素沉积相关基因的发掘与证明

发布时间：2018-07-06 19:31

本文选题：中华锯齿米虾 + 转录组　；参考：《集美大学》2017年硕士论文

【摘要】：中华锯齿米虾(Neocaridina denticulata sinensis)广泛分布于亚洲淡水水域,因其体色丰富多样,自2003年开始成为一种重要的观赏虾。但是其体色的分子遗传基础尚未知。在观赏中华锯齿米虾中,红、黄和透明3种纯色品系的体色遗传最稳定。因此,本文通过比较红、黄和透明3种纯色米虾皮肤组织的转录组,获得了大量体色差异表达基因,并利用qPCR在纯色品系和琉璃虾中对重要候选基因进行验证。希望为揭示米虾体色遗传机制提供资料。主要研究结果如下:1.一共获得了268,629,208个clean reads,经TRINITY软件拼接后获得45,434条Unigenes。Unigene的平均长度为837 bp,N50为1551 bp。与NCBI nr、Swiss-prot、KOG和Kegg数据库比对后,共有19,992条Unigenes获得注释。2.通过对3种纯色品系米虾皮肤转录组的基因表达量分析,共得到14,915个差异表达Unigene。核糖体蛋白基因、actin、calmodulin、EF1α和AK等基因可能通过色素迁移及色素体状态维持这一细胞学过程引起米虾体色差异。红色品系的鸟嘌呤合成通路高表达,表明米虾红色素的主要成分可能是蝶啶类色素。黄色品系没有显著高表达色素合成通路,但是甲壳蓝蛋白基因在黄色品系米虾中高表达,表明米虾黄色素可能与类胡萝卜素有关。3.通过与已知果蝇、家蚕和赤拟谷盗体色基因的比对发现了大量可能参与色素沉积的基因,其中包括黑色素、眼色素、蝶呤等合成酶系及其调节基因并从中筛选出25条与体色相关的候选基因。利用实时荧光定量PCR获得25条候选基因在红、黄和透明3种纯色品系及在2种琉璃虾的表达模式。3种纯色品系的RT-qPCR结果与转录组RPKM值的高低趋势吻合。4.原位杂交实验结果表明,ebony、GC-I、SPR和XDH基因在米虾的复眼期幼体和仔虾阶段均有表达,而在无节幼体阶段没有检测到表达信号。在色素沉积过程中,米虾这4个基因可能具有与其昆虫的同源基因类似的作用。转录组获得这些数据为米虾体色相关基因的发掘和体色遗传机制的解析提供丰富的序列资源。
[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.
【学位授予单位】：集美大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S917.4

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