本文選題：意大利蜜蜂 + 中華蜜蜂�。� 參考：《福建農(nóng)林大學(xué)》2017年碩士論文

【摘要】：白堊病是一種最具代表性的蜜蜂真菌病,該病由蜜蜂球囊菌(Ascosphaeraapis,簡(jiǎn)稱球囊菌)特異性侵染蜜蜂幼蟲而導(dǎo)致,嚴(yán)重危害養(yǎng)蜂生產(chǎn)。意大利蜜蜂(意蜂)幼蟲極易被球囊菌侵染而罹患白堊病,而中華蜜蜂(中蜂)幼蟲的球囊菌抗性較強(qiáng)。前期球囊菌的研究主要涉及分類鑒定、培養(yǎng)方法、病原檢測(cè)、流行病學(xué)、侵染過程以及免疫防御等方面,由于參考基因組信息的缺失,球囊菌的分子研究進(jìn)展緩慢。相關(guān)研究主要集中在球囊菌侵染意蜂幼蟲的諸多方面,有關(guān)球囊菌侵染中蜂幼蟲的研究非常滯后。目前,白堊病的組學(xué)研究極其有限,有關(guān)蜜蜂幼蟲響應(yīng)球囊菌脅迫的轉(zhuǎn)錄組學(xué)研究未見報(bào)道。因此,本研究用107孢子/mL的球囊菌純化孢子飼喂感染意蜂及中蜂幼蟲,利用RNA-seq技術(shù)對(duì)對(duì)照組和處理組蜜蜂幼蟲腸道進(jìn)行深度測(cè)序,首先組裝并注釋中蜂幼蟲腸道的參考轉(zhuǎn)錄組,進(jìn)而對(duì)兩種蜜蜂幼蟲腸道響應(yīng)球囊菌脅迫的免疫應(yīng)答進(jìn)行深入分析,獲得的主要結(jié)果如下:(1)首次通過形態(tài)學(xué)和分子生物學(xué)手段證實(shí)從自然蜂群中的中蜂白堊狀幼蟲尸體上分離的病原為球囊菌,且中蜂幼蟲可在實(shí)驗(yàn)室條件下通過飼喂接種球囊菌而發(fā)生白堊病。(2)成功組裝并注釋中蜂幼蟲腸道參考轉(zhuǎn)錄組,可為中蜂及其幼蟲的分子生物學(xué)及組學(xué)研究提供重要的參考信息;基于此轉(zhuǎn)錄組數(shù)據(jù)開發(fā)出21個(gè)中蜂幼蟲的SSR分子標(biāo)記,可應(yīng)用于中蜂及其幼蟲的基因圖譜構(gòu)建、基因多樣性分析以及基因定位等研究,同時(shí)說明利用轉(zhuǎn)錄組數(shù)據(jù)大規(guī)模開發(fā)非模式生物的SSR是一種高效快速的可行途徑。(3)通過中蜂幼蟲腸道在球囊菌脅迫過程中的差異表達(dá)基因(DEGs)分析和趨勢(shì)分析,發(fā)現(xiàn)富集在細(xì)胞免疫通路的DEGs中表現(xiàn)為上調(diào)趨勢(shì)的基因數(shù)遠(yuǎn)多于下調(diào)趨勢(shì),富集在體液免疫通路的DEGs均表現(xiàn)為上調(diào)趨勢(shì),表明宿主的細(xì)胞和體液免疫通路均被球囊菌顯著激活,全面解析了中蜂幼蟲腸道響應(yīng)球囊菌脅迫的免疫應(yīng)答。(4)通過意蜂幼蟲在球囊菌脅迫過程中的DEGs分析和趨勢(shì)分析,發(fā)現(xiàn)富集在細(xì)胞免疫通路上的部分DEGs表現(xiàn)為上調(diào)趨勢(shì),而更多DEGs表現(xiàn)為下調(diào)趨勢(shì),富集在體液免疫通路上的絕大多數(shù)DEGs表現(xiàn)為上調(diào)趨勢(shì),表明宿主的細(xì)胞免疫通路被部分激活,而體液免疫通路被顯著激活,全面解析了意蜂幼蟲腸道響應(yīng)球囊菌脅迫的免疫應(yīng)答。
[Abstract]:Chalky disease is one of the most representative fungal diseases of bees. It is caused by the specific infection of bee larvae by Ascosphaeraapis, which seriously endangers the production of beekeeping. The larvae of Italian honeybee (Italian bee) are susceptible to chalkiness, but the larvae of Chinese bee (Chinese bee) are more resistant to Balloon. The previous studies of balloon bacteria mainly involved in classification and identification, culture methods, pathogen detection, epidemiology, infection process and immune defense, etc. Due to the lack of reference genome information, the molecular research progress of the balloon bacteria was slow. The related studies mainly focused on many aspects of the infecting of Italian wasp larvae by Balloon bacteria, and the study on the larvae of Spinel wasps infected by Balloonella spp was very late. At present, the study of chalky disease is very limited, and there is no report on the transcriptome of bee larvae responding to the stress of Balloon. In this study, the purified spores of 107spore / mL spore were fed to infected bee and Chinese bee larvae. The intestinal tract of the control and treated groups were further sequenced by RNA-seq technique. First, the reference transcription group of bee larval intestine was assembled and annotated. Then the immune responses of the intestinal tract of two honeybee larvae in response to the stress of Balloon bacteria were analyzed in depth. The main results obtained are as follows: (1) for the first time, the pathogen isolated from the corpses of the Chinese bee chalkiness larvae in the natural bee colony was proved to be a balloon bacterium by morphological and molecular biological means. And the Chinese bee larvae can be successfully assembled and annotated by inoculation of the chalkiness disease by inoculating Balloonella spp. under laboratory conditions, which can provide important reference information for the molecular biology and stratigraphic study of the Chinese bee larvae and their larvae. Based on the transcriptional data, the SSR markers of 21 larvae were developed, which can be applied to the construction of gene map, gene diversity analysis and gene mapping of Chinese wasp and its larvae. At the same time, using transcriptome data to develop SSR of non-model organisms on a large scale is a feasible and efficient and rapid approach. It can be used to analyze the differentially expressed genes in the intestinal tract of wasp larvae during Balloon stress. It was found that the number of up-regulation genes in DEGs enriched in cellular immune pathway was much more than that in down-regulation, and that DEGs enriched in humoral immune pathway was up-regulated, indicating that the cellular and humoral immune pathways of host were significantly activated by Balloon. The immune response of the intestinal tract of Chinese bee larvae to Balloon stress was analyzed. Through the DEGs analysis and trend analysis, it was found that some of the DEGs enriched in the cellular immune pathway showed an up-regulation trend, through the DEGs analysis and the trend analysis of the sputum larvae during the stress process of the sputum. However, more DEGs showed down-regulation, and most of DEGs enriched in humoral immune pathway were up-regulated, indicating that the cellular immune pathway of host was partially activated, while humoral immune pathway was significantly activated. The immune response of Italian bee larvae to balloon stress was analyzed.
【學(xué)位授予單位】：福建農(nóng)林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S895

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本文編號(hào)：1895177