四带小鲃的长鳍表型及其转录组分析
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Q 785;S 917.4

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观赏鱼斗鱼遗传改良技术开发(D-8006-19-0166)


Phenotypic and transcriptomic analysis of long fins of tiger barb (Puntius tetrazona)
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    摘要:

    为了了解野生型和长鳍突变型四带小鲃的体长、尾鳍长、尾鳍分节数目和分节长度,实验利用转录组学方法和t 检验统计方法对上述指标进行了分析。结果显示,6月龄野生型 (n=30)与突变型 (n=30)成鱼体长之间没有显著差异,但二者尾鳍长、尾鳍分节数目和分节长度3个指标间存在显著差异。同时,对不同体长野生型(n=80)和突变型(n=80)四带小鲃尾鳍长度、分节数目和分节长度进行了线性回归分析。结果显示,相对于野生型,突变型四带小鲃尾鳍长度增加速率更快 (野生型:k=0.28,突变型:k=0.62),尾鳍分节数目增加频率基本一致 (野生型:k=0.79,突变型:k=0.82),尾鳍分节长度更长 (野生型:k=0.000 4,突变型:k=0.006 8)。野生型和突变型成鱼尾鳍组织的转录组分析结果显示,转录组从头组装共获得56 271个unigene,野生型组 (n=3)和突变型组 (n=3)之间共有1 304个差异表达基因,其中971个基因上调,333个基因下调。差异表达基因 KEGG通路显著富集到细胞周期和DNA复制等与细胞增殖相关的信号通路,其中细胞周期蛋白基因 (cycAcycBcycE)和细胞周期依赖性激酶基因 (cdk1)等显著上调。最后,对ckap2lki-67、cdc20、prc1、urgcpcdk1共6个基因的表达水平进行了实时荧光定量PCR (qRT-PCR)分析,其结果与转录组分析结果基本一致。研究表明,长鳍四带小鲃突变基因引起细胞周期相关基因表达量上升,造成尾鳍分节片段的细胞增殖速率加快,尾鳍分节长度增加,呈现为长尾鳍表型。

    Abstract:

    Puntius tetrazona is a small tropical ornamental fish with high economic value. Through the method of t-test, the body length (BL), caudal fin length (CL), segment number (SN) and segment length (SL) of caudal fin of P. tetrazona (wild type, WT) and (long fin, LF) were statistically analyzed. The results showed that there was no significant difference in BL between the WT (n=30) and LF (n=30) (P>0.05), but there were significant differences in CL (P<0.000 1), SN (P<0.000 1) and SL (P<0.000 1). At the same time, linear regression analyses were performed on the caudal fin length, segment number and length of caudal fin of WT (n=80) and LF (n=80) P. tetrazona with different body length. The results showed that compared to WT, the caudal fin of LF P. tetrazona grew more rapidly (WT: k=0.28, LF: k=0.62, P<0.000 1), the increasing frequency of the segment number of caudal fin was similar (WT: k=0.79, LF: k=0.82, P=0.006), and the segment length of caudal fin was much greater (WT: k=0.000 4, LF: k=0.006 8, P<0.000 1). Transcriptome analysis of caudal fin of WT and LF adults showed that 56 271 unigenes were obtained from de novo transcriptome assembly. There were 1 304 differentially expressed genes between WT group (n=3) and LF group (n=3), of which 971 genes were up-regulated and 333 genes were down-regulated. The KEGG pathway enrichment analysis showed that the differently expressed genes were significantly enriched into cell cycle and DNA replication signal pathways related to cell proliferation (P<0.05), in which cyclin (cycA, cycB, and cycE) and cyclin-dependent kinase 1 genes were significantly up-regulated. Finally, we determined the expression level of cytoskeleton associated protein 2-like, proliferation marker protein Ki-67, cell division cycle protein 20, protein regulator of cytokinesis 1, up-regulator of cell proliferation and cyclin-dependent kinase 1 by qRT-PCR. The results of qRT-PCR were consistent with those of RNA-seq. The above results showed that a mutated gene in LF P. tetrazona caused the up-regulated expression of cell cycle related genes, which accelerated the cell proliferation rate of caudal fin and resulted in increase of caudal fin segment length, showing the phenotype with a long caudal fin.

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杨继辉,燕男,崔文耀,熊加奉,黄嵘,任建峰.四带小鲃的长鳍表型及其转录组分析[J].水产学报,2022,46(8):1324~1333

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  • 收稿日期:2021-04-11
  • 最后修改日期:2021-07-05
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  • 在线发布日期: 2022-08-16
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