【摘要】：蜜蜂是重要的授粉昆蟲,其生產(chǎn)的蜂蜜、王漿等蜂產(chǎn)品對人類機(jī)體具有保健作用。蜜蜂產(chǎn)卵性狀是蜜蜂蜂群發(fā)展的基礎(chǔ),是影響蜂群性狀表現(xiàn)的重要基礎(chǔ)因素。因此,研究蜜蜂卵巢激活和產(chǎn)卵過程、尋找對蜜蜂卵巢激活和產(chǎn)卵有顯著作用的基因,對于了解蜜蜂產(chǎn)卵的遺傳機(jī)理和提高產(chǎn)卵有重要意義。本研究利用高通量測序的方法,對蜜蜂蜂王卵巢RNA的表達(dá)變化規(guī)律進(jìn)行了研究。本研究選取處女蜂王、正常產(chǎn)卵蜂王、產(chǎn)卵抑制蜂王、產(chǎn)卵恢復(fù)蜂王作為研究對象(n=3/組),利用轉(zhuǎn)錄組測序的方法篩選獲得了卵巢激活過程、產(chǎn)卵抑制過程和產(chǎn)卵恢復(fù)過程差異表達(dá)的lncRNA、mRNA、miRNA和 circRNA: (1)卵巢激活過程,224 個 lncRNA、3218 個 mRNA、39個 miRNA 和 228 個 circRNA 表達(dá)上調(diào);516 個 lncRNA、2263 個 mRNA、42個miRNA和272個circRNA表達(dá)下調(diào);(2 )產(chǎn)卵抑制過程,57個lncRNA、266 個 mRNA、9 個 miRNA 和 273 個 circRNA 表達(dá)上調(diào);31 個 lncRNA、72個mRNA、4個miRNA和227個circRNA表達(dá)下調(diào);(3)產(chǎn)卵恢復(fù)過程,40 個 lncRNA、256 個 mRNA、2 個 miRNA 和 236 個 circRNA 表達(dá)上調(diào);60 個 lncRNA、241 個 mRNA、2 個 miRNA 和 264 個 circRNA表達(dá)下調(diào)。對篩選到的差異表達(dá)RNA進(jìn)行功能富集分析,發(fā)現(xiàn)在卵巢激活和產(chǎn)卵過程中顯著富集的生物學(xué)過程主要有系統(tǒng)發(fā)育、生物調(diào)節(jié)和神經(jīng)系統(tǒng)的發(fā)育。有多個生物學(xué)通路與蜜蜂卵巢激活和產(chǎn)卵密切相關(guān),包括Wnt、hippo、TGF-beta、notch、MAPK、FoxO 和 mTOR 信號通路等。通過將獲得的差異表達(dá)mRNA和lncRNA的物理位置與已知的蜜蜂卵巢大小相關(guān)QTL區(qū)間進(jìn)行比對,得到了 73個候選基因和14個候選lncRNA,它們是與蜜蜂卵巢大小和產(chǎn)卵緊密相關(guān)的候選基因。接下來,我們對差異circRNA的靶基因構(gòu)建了 gene-act-network,鑒定了網(wǎng)絡(luò)中的關(guān)鍵基因:ECR、E75、E74、IR-B和RACK1。另外,構(gòu)建了 lncRNA-miRNA-mRNA 和 circRNA-miRNA-mRNA 網(wǎng)絡(luò),發(fā)現(xiàn)一些RNA在網(wǎng)絡(luò)中承擔(dān)“橋梁”的功能,連接不同的基因和RNA。對網(wǎng)絡(luò)中發(fā)揮關(guān)鍵作用的基因,CCDC93和Hsc70-3,進(jìn)行了討論。本研究篩選鑒定了蜜蜂蜂王卵巢激活和產(chǎn)卵過程中差異表達(dá)的lncRNA、mRNA、miRNA和circRNA,數(shù)據(jù)分析表明多種RNA在蜜蜂產(chǎn)卵活動中發(fā)揮重要作用。研究為進(jìn)一步探明蜜蜂產(chǎn)卵的遺傳機(jī)理奠定了基礎(chǔ)。
[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.
【學(xué)位授予單位】：中國農(nóng)業(yè)科學(xué)院
【學(xué)位級別】：博士后
【學(xué)位授予年份】：2017
【分類號】：S891

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相關(guān)期刊論文 前1條

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本文編號：2153620