蜜蜂卵巢激活和产卵过程差异表达的编码RNA与非编码RNA的筛选和鉴定
发布时间：2018-07-29 18:31
【摘要】：蜜蜂是重要的授粉昆虫,其生产的蜂蜜、王浆等蜂产品对人类机体具有保健作用。蜜蜂产卵性状是蜜蜂蜂群发展的基础,是影响蜂群性状表现的重要基础因素。因此,研究蜜蜂卵巢激活和产卵过程、寻找对蜜蜂卵巢激活和产卵有显著作用的基因,对于了解蜜蜂产卵的遗传机理和提高产卵有重要意义。本研究利用高通量测序的方法,对蜜蜂蜂王卵巢RNA的表达变化规律进行了研究。本研究选取处女蜂王、正常产卵蜂王、产卵抑制蜂王、产卵恢复蜂王作为研究对象(n=3/组),利用转录组测序的方法筛选获得了卵巢激活过程、产卵抑制过程和产卵恢复过程差异表达的lncRNA、mRNA、miRNA和 circRNA: (1)卵巢激活过程,224 个 lncRNA、3218 个 mRNA、39个 miRNA 和 228 个 circRNA 表达上调;516 个 lncRNA、2263 个 mRNA、42个miRNA和272个circRNA表达下调;(2 )产卵抑制过程,57个lncRNA、266 个 mRNA、9 个 miRNA 和 273 个 circRNA 表达上调;31 个 lncRNA、72个mRNA、4个miRNA和227个circRNA表达下调;(3)产卵恢复过程,40 个 lncRNA、256 个 mRNA、2 个 miRNA 和 236 个 circRNA 表达上调;60 个 lncRNA、241 个 mRNA、2 个 miRNA 和 264 个 circRNA表达下调。对筛选到的差异表达RNA进行功能富集分析,发现在卵巢激活和产卵过程中显著富集的生物学过程主要有系统发育、生物调节和神经系统的发育。有多个生物学通路与蜜蜂卵巢激活和产卵密切相关,包括Wnt、hippo、TGF-beta、notch、MAPK、FoxO 和 mTOR 信号通路等。通过将获得的差异表达mRNA和lncRNA的物理位置与已知的蜜蜂卵巢大小相关QTL区间进行比对,得到了 73个候选基因和14个候选lncRNA,它们是与蜜蜂卵巢大小和产卵紧密相关的候选基因。接下来,我们对差异circRNA的靶基因构建了 gene-act-network,鉴定了网络中的关键基因:ECR、E75、E74、IR-B和RACK1。另外,构建了 lncRNA-miRNA-mRNA 和 circRNA-miRNA-mRNA 网络,发现一些RNA在网络中承担“桥梁”的功能,连接不同的基因和RNA。对网络中发挥关键作用的基因,CCDC93和Hsc70-3,进行了讨论。本研究筛选鉴定了蜜蜂蜂王卵巢激活和产卵过程中差异表达的lncRNA、mRNA、miRNA和circRNA,数据分析表明多种RNA在蜜蜂产卵活动中发挥重要作用。研究为进一步探明蜜蜂产卵的遗传机理奠定了基础。
[Abstract]:Bee is an important pollination insect. Honey, royal jelly and other bee products have health care effect on human body. Bee spawning character is the basis of honeybee colony development and an important basic factor affecting the performance of bee colony traits. Therefore, it is important to study the process of honeybee ovary activation and oviposition, and to find the genes that play a significant role in the activation and oviposition of honeybee ovaries, which is of great significance to understand the genetic mechanism of honeybee oviposition and to improve its oviposition. In this study, high-throughput sequencing was used to study the expression of RNA in the ovary of bee queen. In this study, the virgin queen, the normal spawning queen, the oviposition inhibition queen and the oviposition recovery queen were selected as the study subjects. The ovarian activation process was obtained by transcriptional sequencing method. IncRNA-mRNA-miRNAs and circRNAs differentially expressed during oviposition inhibition and oviposition recovery: (1) 3218 RNAs of 22ncRNAs in ovarian activation process, 2263 mRNAs expressed in 39 miRNA and 228 circRNA, and (2) down-regulated expression of 42 miRNA and 272 circRNA in oviposition; (2) oviposition. During the inhibition process, the expression of 57 lncRNAs, 266 mRNAs, 9 miRNA and 273 circRNA up-regulated 31 miRNA, 72 mRNAs, 4 miRNA and 227 circRNA down-regulated; (3) during oviposition recovery, 40 LNcRNAs, 256 RNAs, 2 miRNA, and 236 circRNA, up-regulated the expression of 60 LNcRNAs. The expression of 2 miRNA and 264 circRNA were down-regulated. The results of functional enrichment analysis of differentially expressed RNA showed that the biological processes in the process of ovarian activation and oviposition were mainly phylogeny, biological regulation and nervous system development. There are several biological pathways closely related to ovarian activation and oviposition of honeybees, including the Wnttpapo TGF-beta MAPK, FoxO and mTOR signaling pathways. By comparing the physical positions of differentially expressed mRNA and lncRNA with known QTL regions of honeybee ovary size, 73 candidate genes and 14 candidate LNcRNAs were obtained, which were closely related to ovarian size and oviposition of honeybee. Next, we constructed gene-act-network for the target genes of differential circRNA, and identified the key genes in the network, namely: ECRN E75, E74, IR-B and RACK1. In addition, the lncRNA-miRNA-mRNA and circRNA-miRNA-mRNA networks are constructed, and some RNA functions as a "bridge" in the network, connecting different genes and RNAs. The genes CCDC93 and Hsc70-3, which play a key role in the network, are discussed. In this study, we screened and identified the differentially expressed RNA miRNAs and circRNAs in the process of ovarian activation and oviposition of honeybee queen bee. The data analysis showed that many kinds of RNA played an important role in honeybee oviposition. The study laid a foundation for the further study of the genetic mechanism of honeybee spawning.
【学位授予单位】：中国农业科学院
【学位级别】：博士后
【学位授予年份】：2017
【分类号】：S891
本文编号：2153620
[Abstract]:Bee is an important pollination insect. Honey, royal jelly and other bee products have health care effect on human body. Bee spawning character is the basis of honeybee colony development and an important basic factor affecting the performance of bee colony traits. Therefore, it is important to study the process of honeybee ovary activation and oviposition, and to find the genes that play a significant role in the activation and oviposition of honeybee ovaries, which is of great significance to understand the genetic mechanism of honeybee oviposition and to improve its oviposition. In this study, high-throughput sequencing was used to study the expression of RNA in the ovary of bee queen. In this study, the virgin queen, the normal spawning queen, the oviposition inhibition queen and the oviposition recovery queen were selected as the study subjects. The ovarian activation process was obtained by transcriptional sequencing method. IncRNA-mRNA-miRNAs and circRNAs differentially expressed during oviposition inhibition and oviposition recovery: (1) 3218 RNAs of 22ncRNAs in ovarian activation process, 2263 mRNAs expressed in 39 miRNA and 228 circRNA, and (2) down-regulated expression of 42 miRNA and 272 circRNA in oviposition; (2) oviposition. During the inhibition process, the expression of 57 lncRNAs, 266 mRNAs, 9 miRNA and 273 circRNA up-regulated 31 miRNA, 72 mRNAs, 4 miRNA and 227 circRNA down-regulated; (3) during oviposition recovery, 40 LNcRNAs, 256 RNAs, 2 miRNA, and 236 circRNA, up-regulated the expression of 60 LNcRNAs. The expression of 2 miRNA and 264 circRNA were down-regulated. The results of functional enrichment analysis of differentially expressed RNA showed that the biological processes in the process of ovarian activation and oviposition were mainly phylogeny, biological regulation and nervous system development. There are several biological pathways closely related to ovarian activation and oviposition of honeybees, including the Wnttpapo TGF-beta MAPK, FoxO and mTOR signaling pathways. By comparing the physical positions of differentially expressed mRNA and lncRNA with known QTL regions of honeybee ovary size, 73 candidate genes and 14 candidate LNcRNAs were obtained, which were closely related to ovarian size and oviposition of honeybee. Next, we constructed gene-act-network for the target genes of differential circRNA, and identified the key genes in the network, namely: ECRN E75, E74, IR-B and RACK1. In addition, the lncRNA-miRNA-mRNA and circRNA-miRNA-mRNA networks are constructed, and some RNA functions as a "bridge" in the network, connecting different genes and RNAs. The genes CCDC93 and Hsc70-3, which play a key role in the network, are discussed. In this study, we screened and identified the differentially expressed RNA miRNAs and circRNAs in the process of ovarian activation and oviposition of honeybee queen bee. The data analysis showed that many kinds of RNA played an important role in honeybee oviposition. The study laid a foundation for the further study of the genetic mechanism of honeybee spawning.
【学位授予单位】：中国农业科学院
【学位级别】：博士后
【学位授予年份】：2017
【分类号】：S891
【参考文献】
相关期刊论文 前1条
1 孙晓阳,王雁玲;Wnt信号通路与哺乳动物生殖[J];生物化学与生物物理进展;2003年02期
，本文编号：2153620
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