不同日龄白羽王鸽胸肌中lncRNA和mRNA功能分析

Functional Analysis of lncRNA and mRNA in the Pectoralis Muscle of White King Pigeons at Different Days of Age

【目的】分析不同日龄乳鸽胸肌发育的差异长链非编码RNA(lncRNA)和mRNA,确定影响乳鸽生长发育的关键候选基因,探究乳鸽生长发育的分子机制,为促进乳鸽生长发育的研究提供理论基础。【方法】乳鸽于1(D1)、7(D7)、14(D14)和25(D25)日龄分别称重,随后在D7和D25时进行屠宰,利用RNA-Seq技术对所选日龄白羽王鸽胸肌的lncRNA进行测序并预测其靶基因,构建lncRNA-mRNA互作网络,对筛选的差异表达lncRNA进行GO功能和KEGG通路富集分析,并利用实时荧光定量PCR验证测序结果的可靠性。【结果】乳鸽于7~14日龄日增重最快,25日龄乳鸽体重到达上市日龄。从构建的6个文库中共获得56421个lncRNAs和30208个mRNAs,其中2335个lncRNAs差异表达,靶向1553个mRNAs。lncRNA-mRNA调控网络显示,lncRNA与mRNA间存在相互调控。GO与KEGG分析结果显示,差异基因与GTpase活性等相关,差异lncRNAs及其靶基因主要富集在MAPK信号通路、骨骼肌组织发育、TGF-β信号通路、Notch信号通路等信号通路,关键lncRNAs可能通过这些信号通路调控乳鸽胸肌发育。其中LTCONS_00012834、LTCONS_00032767、LTCONS_00045211和LTCONS_00002003分别靶向XIRP 1、PITX 3、BMPR 1 B、MYLK 4基因,发挥关键作用。实时荧光定量结果与高通量测序结果趋势一致,说明测序数据的可靠性。【结论】本研究共获得了2335个差异表达lncRNAs和1553个mRNAs,4个差异表达lncRNAs:LTCONS_00032767、LTCONS_00012834、LTCONS_00002003、LTCONS_00045211分别靶向与骨骼肌发育相关基因PITX 3、XIRP 1、MYLK 4和BMPR 1 B,且这些基因受到多个lncRNA的调控。结果为阐明乳鸽胸肌生长发育的分子机制以及白羽王鸽的分子选育提供了理论基础。

【Objective】Differential lncRNA and mRNA of pectoralis muscle development in pigeons of different day-olds were analyzed to identify the key candidate genes affecting the growth and development of pigeons,explore the molecular mechanism of growth and development of pigeons,and provide a theoretical basis for the research on the promotion of the growth and development of pigeons.【Method】White King pigeons were weighed at 1(D1),7(D7),14(D14)and 25(D25)days of age,followed by slaughtering at D7 and D25.lncRNAs from the pectoralis muscles of the selected day-old White King pigeons were sequenced to predict the target genes using RNA-Seq technology,construct the lncRNA-mRNA interactions network,and the screened differentially expressed lncRNAs were analyzed by GO and KEGG enrichment,and the reliability of sequencing results was verified by Real-time quantitative PCR.【Result】Pigeons gained the fastest daily weight at the stage from 7 to 14 days of age,and the weight of breast pigeons reached the market age at 25 days of age.A total of 56421 lncRNAs and 30208 mRNAs were obtained from the six libraries constructed,of which 2335 lncRNAs were expressed differentially and 1553 mRNAs were targeted in the two groups.The lncRNA-mRNA regulatory network showed that there was mutual regulation between lncRNA and mRNA.The results of KEGG and GO analyses showed that the differential genes were associated with GTpase activity,etc.The differential lncRNAs and their target genes were mainly enriched in signaling pathways such as MAPK signaling pathway,skeletal muscle tissue development,TGF-βsignaling pathway,Notch signaling pathway and other signaling pathways,and the key lncRNAs might regulate the pectoralis muscle development of the breast muscle of the pigeon through these signaling pathways.Among them,LTCONS_00012834,LTCONS_00032767,LTCONS_00045211,and LTCONS_00002003 targeted XIRP 1,PITX 3,BMPR 1 B,and MYLK 4,respectively,and played key roles.Real-time quantitative PCR results were consistent with the high-throughput sequencing results,indicating the reliability of the sequencing data.【Conclusion】A total of 2335 differentially expressed lncRNAs and 1553 mRNAs were obtained in this study,and four differentially expressed lncRNAs:LTCONS_00032767,LTCONS_00012834,LTCONS_00002003,and LTCONS_00045211 targeted genes related to skeletal muscle development PITX 3,XIRP 1,MYLK 4,and BM PR1 B,respectively,and these genes were regulated by multiple lncRNAs.The theoretical basis was provided for elucidating the molecular mechanism of pectoralis muscle growth and development in pigeons as well as the molecular selection and breeding of White King pigeons.