本文選題：白蛉 + 生態(tài)習(xí)性　； 參考：《第二軍醫(yī)大學(xué)》2017年碩士論文

【摘要】：白蛉是一類體積較小的吸血昆蟲,隸屬雙翅目(Diptera)、毛蠓科(Psychodidae),在我國分布廣泛,基于形態(tài)特征分類已記錄白蛉40余種,部分蛉種可傳播利什曼病、巴爾通體病和白蛉熱等疾病,具有非常重要的醫(yī)學(xué)意義。對(duì)白蛉的準(zhǔn)確鑒別,了解白蛉的生態(tài)習(xí)性,深入闡明傳病蛉種的生物學(xué)特點(diǎn)尤為重要。本課題的研究策略:首先,在我國河南、四川和海南等地?zé)粽T和人工采集白蛉,調(diào)查當(dāng)?shù)匕昨鹊纳鷳B(tài)習(xí)性,分子鑒定白蛉的血源動(dòng)物,依據(jù)海南海口全年的燈誘采集標(biāo)本,初步分析闡明其季節(jié)消長規(guī)律。其次,檢視白蛉的形態(tài)特征,鑒定種類;擴(kuò)增對(duì)應(yīng)樣本的mt DNA-COI、Cytb和rDNA-18S基因片段,測(cè)定和分析序列,結(jié)合形態(tài)鑒定的結(jié)果,分析分子特征在種內(nèi)與種間的差異,篩選適合的DNA條形碼用于白蛉種類鑒別。再者,應(yīng)用高通量測(cè)序技術(shù),對(duì)現(xiàn)場(chǎng)采集的中華白蛉成蟲分為雌、雄的頭部、胸腹部4組樣本,分別進(jìn)行轉(zhuǎn)錄組測(cè)序、功能注釋與差異表達(dá)基因分析。獲得以下主要結(jié)果:1、我國四川和河南兩地的中華白蛉的生態(tài)習(xí)性存在一定差異。采自四川的白蛉孳生地為山洞和土洞,近家棲,河南的白蛉孳生地為人居的土窯洞,家棲。分子鑒定的血源動(dòng)物包括:豬、兔、雞、狗、牛和人等,四川的白蛉主要吸血對(duì)象是豬,而河南是雞,兩地均檢測(cè)到人和狗,其中人的比例分別為4.54%(四川)和41.67%(河南)。2、2015年8月-2016年9月在海南海口全年共捕獲377只白蛉,種群密度總體不高,高峰呈雙峰,為6月和11月。3、本研究共檢視和記錄了2屬7種白蛉的形態(tài)鑒別特征,包括:白蛉屬的中華白蛉和施氏白蛉,司蛉屬的貝氏司蛉、應(yīng)氏司蛉、歌樂山司蛉、鱗喙司蛉和鱗胸司蛉。獲得對(duì)應(yīng)蛉種的mt DNA-COI和Cytb序列,以及rDNA-18S序列(除歌樂山司蛉)。綜合分析結(jié)果顯示,mtDNA-COI、Cytb和rDNA-18S均可作為DNA條形碼鑒別上述白蛉種類。4、貝氏白蛉復(fù)合體存在多個(gè)隱種,我國的貝氏司蛉與泰國的貝氏司蛉關(guān)系密切,與貝氏司蛉A和B種的mtDNA-COI序列差別大,暫定為貝氏司蛉C種。5、高通量測(cè)序獲得中華白蛉的轉(zhuǎn)錄組序列,共133,835,342條原始reads,大于Q20的reads數(shù)128,090,788條,占原始數(shù)據(jù)95.71%,共31.7kM。組裝獲得32,628條unigene,分別注釋到Nr、Swiss-Prot、KEGG、COG和GO數(shù)據(jù)庫。篩選獲得4374個(gè)具有顯著性表達(dá)差異的基因,經(jīng)qRT-PCR驗(yàn)證10個(gè)基因片段表達(dá)量和轉(zhuǎn)錄組測(cè)序結(jié)果保存一致。中華白蛉的頭部和胸腹部的差異表達(dá)基因與其功能關(guān)系密切,不同性別間的差別基因也是與其攝食和生殖功能相關(guān)。
[Abstract]:Lacewing is a small blood-sucking insect belonging to Diptera Diptera. It is widely distributed in China. More than 40 species have been recorded on the basis of morphological classification. Some of them can spread leishmaniasis, Bartonella disease and phlegm fever. Has very important medical significance. It is very important to identify the laceflies accurately, understand their ecological habits and elucidate the biological characteristics of disease transmission species. The research strategies of this subject are as follows: first of all, in Henan, Sichuan and Hainan provinces in China, the ecological habits of the local sandflies were investigated, and the blood source animals were identified by molecular analysis, and the samples were collected according to the annual lamp trap in Haikou, Hainan Province. A preliminary analysis was made to illustrate the seasonal variation and variation. Secondly, the morphological characteristics of the sandfly were examined to identify the species, the mt DNA-COII Cytb and rDNA-18S gene fragments of the corresponding samples were amplified, the sequences were determined and analyzed, and the differences of molecular characteristics between species and species were analyzed by combining the results of morphological identification. The suitable DNA bar code was selected for the species identification of sandflies. Furthermore, high-throughput sequencing technique was used to analyze four groups of female, male head, chest and abdomen samples of adult sandfly collected in the field, respectively, for transcriptional sequencing, functional annotation and differentially expressed gene analysis. The main results are as follows: 1. There are some differences in ecological habits between Sichuan and Henan provinces in China. The breeding ground of sandfly collected from Sichuan is cave and soil cave, near home habitat, and Henan lacewing breeding place is soil cave of human settlement. Molecular identification of the origin of blood animals include: pigs, rabbits, chickens, dogs, cattle and people, Sichuan sandfly is the main blood sucking pigs, while Henan is chicken, both human and dog were detected. The proportion of people was 4.54 and 41.67, respectively. From August 2015 to September 2016, 377 sandflies were captured in Haikou, Hainan Province. The population density was not high, and the peak was double peak. For June and Nov. 3, this study examined and recorded the morphological characteristics of 2 genera and 7 species of sandflies, including: the Chinese and Scheffler sandflies of the genus Phlewing, the Bessler, the Spinelian, and the Geles of the genus, respectively. The squame-beak and the squamothorax. The mt DNA-COI and Cytb sequences and rDNA-18S sequences of the corresponding species were obtained. The results of comprehensive analysis showed that both mtDNA-COICytb and rDNA-18S could be used as DNA bar codes to identify the species of the above mentioned species. There were many codon species in the complex, and there was a close relationship between China and Thailand. The mtDNA-COI sequences of the species A and B were different from those of species A and B, and were tentatively classified as C. 5. The transcriptome sequences were obtained by high-throughput sequencing. A total of 133835342 primordial READSs and 128090788 reads sequences larger than Q20 were obtained, accounting for 95.71 and 31.7kMof the original data. 32628 Unigenees were assembled and annotated into the NrSwiss-ProtKEGG COG and go databases, respectively. 4374 genes with significant difference in expression were screened and confirmed by qRT-PCR. The expression levels of 10 gene fragments were consistent with the results of transcriptome sequencing. The differentially expressed genes in head, chest and abdomen were closely related to their functions, and the difference genes between genders were also related to their feeding and reproductive functions.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R384.1

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