發(fā)布時(shí)間：2018-04-19 08:29

  本文選題：蜜蜂 + 幼蟲(chóng)��； 參考：《江西農(nóng)業(yè)大學(xué)》2016年博士論文

【摘要】：蜜蜂是一種典型的真社會(huì)性昆蟲(chóng),它的獨(dú)特生物學(xué)特性一直受廣大科學(xué)研究者的關(guān)注。本研究以意大利蜜蜂(Apis mellifera ligustica,簡(jiǎn)稱(chēng)意蜂)為實(shí)驗(yàn)材料,利用Needle trap技術(shù)與氣質(zhì)聯(lián)用技術(shù)分析鑒定出9種工蜂幼蟲(chóng)信息素。發(fā)現(xiàn)E-β-羅勒烯為工蜂幼蟲(chóng)的饑餓信息素,在饑餓組幼蟲(chóng)中含量顯著高于飼喂組及純食物組,且2日齡幼蟲(chóng)組顯著高于4日齡幼蟲(chóng)組。同時(shí),行為學(xué)實(shí)驗(yàn)結(jié)果表明添加E-β-羅勒烯可引起哺育蜂的探頭哺育行為,進(jìn)一步證實(shí)了E-β-羅勒烯為工蜂幼蟲(chóng)饑餓信息素。利用RNA-Seq技術(shù)分析鑒定出了E-β-羅勒烯在幼蟲(chóng)體內(nèi)的生物合成通路,并且發(fā)現(xiàn)參與該通路的3個(gè)基因在2d幼蟲(chóng)中表達(dá)顯著高于4d幼蟲(chóng),并在饑餓30分鐘時(shí)表達(dá)量達(dá)到最高峰。利用上述相同的Needle trap與氣質(zhì)聯(lián)用技術(shù),分析鑒定出了蜂王幼蟲(chóng)與雄蜂幼蟲(chóng)的饑餓信息素。實(shí)驗(yàn)結(jié)果表明,蜂王與雄蜂幼蟲(chóng)也以E-β-羅勒烯作為其饑餓信息素。各饑餓幼蟲(chóng)組所含E-β-羅勒烯均顯著高于飼喂幼蟲(chóng)組與食物組。2日齡蜂王、雄蜂與工蜂幼蟲(chóng)饑餓組E-β-羅勒烯含量均差異不顯著,但4日齡蜂王幼蟲(chóng)饑餓組E-β-羅勒烯含量顯著低于雄蜂與工蜂幼蟲(chóng)組。同時(shí),發(fā)現(xiàn)蜂王幼蟲(chóng)含有一種雄蜂與工蜂幼蟲(chóng)沒(méi)有的幼蟲(chóng)信息素----2-庚酮。RNA-Seq結(jié)果表明,蜂王與雄蜂幼蟲(chóng)也具有與工蜂幼蟲(chóng)相同的E-β-羅勒烯生物合成通路與相關(guān)基因。使用RNA-Seq測(cè)序技術(shù)檢測(cè)了意蜂三型蜂2日齡與4日齡幼蟲(chóng)的mRNA,分析比較了三型蜂幼蟲(chóng)在發(fā)育過(guò)程中的基因表達(dá)差異。各三型蜂幼蟲(chóng)組檢測(cè)到的總基因個(gè)數(shù)沒(méi)有差異,但各組之間存在大量差異表達(dá)基因。2日齡雄蜂幼蟲(chóng)與蜂王幼蟲(chóng)對(duì)比組差異表達(dá)基因數(shù)為475個(gè),高于雄蜂與工蜂幼蟲(chóng)對(duì)比組(197個(gè))及蜂王與工蜂幼蟲(chóng)對(duì)比組(121個(gè));4日齡蜂王與雄蜂幼蟲(chóng)對(duì)比組差異基因達(dá)687個(gè),工蜂與雄蜂幼蟲(chóng)對(duì)比組為604個(gè),而蜂王與工蜂幼蟲(chóng)對(duì)比組僅為475個(gè),表明雄蜂幼蟲(chóng)與其他兩種雌性蜂之間的差異高于兩種雌性蜂之間的差異。而4日齡各對(duì)比組的差異基因數(shù)明顯高于2日齡對(duì)比組,表明隨著日齡的增加三型蜂幼蟲(chóng)的發(fā)育更為分化。其次,2d與4d工蜂幼蟲(chóng)之間差異表達(dá)基因?yàn)?190個(gè),2d與4d蜂王幼蟲(chóng)對(duì)比組為1181個(gè),而雄蜂幼蟲(chóng)對(duì)比組僅為598個(gè),表明蜜蜂單雙倍體遺傳背景在其發(fā)育上的調(diào)控模式存在差異。基因共表達(dá)聚類(lèi)分析結(jié)果表明2日齡工蜂與雄蜂幼蟲(chóng)先聚類(lèi)在一起,再與2日齡蜂王聚類(lèi),4日齡則兩種雌性蜂聚類(lèi)更近。雌性蜜蜂幼蟲(chóng)與雄蜂幼蟲(chóng)相比較,發(fā)現(xiàn)大量差異表達(dá)基因參與了蜜蜂激素合成分泌、蜂毒的合成、眼睛和飛行肌肉的生長(zhǎng)發(fā)育、眼睛發(fā)育、卵巢成熟、以及一些調(diào)控基因表達(dá)的轉(zhuǎn)錄因子與信號(hào)通路等,這與雌性蜂與雄蜂在形態(tài)、生理與行為上的特征相符。
[Abstract]:Honeybee is a typical true-social insect, its unique biological characteristics have been concerned by many scientific researchers.In this study, 9 species of pheromones of worker bee larvae were identified by means of Needle trap and GC-MS using Apis mellifera ligustica (Italian bee) as the experimental material.It was found that E- 尾 -basilerene was the pheromone of worker bee larvae, and the content of E- 尾 -basilerene in the starved larvae was significantly higher than that in the feeding and pure food groups, and that in the 2-day-old larvae was significantly higher than that in the 4-day-old larvae.At the same time, the results of behavioral experiments showed that the addition of E- 尾 -basilerene could induce the feeding behavior of nursing bees, which further confirmed that E- 尾 -basilerene was a hunger pheromone of worker bee larvae.The biosynthesis pathway of E- 尾 -basilerene in larva was identified by RNA-Seq technique. It was found that the three genes involved in the biosynthesis pathway were significantly higher in 2 d larva than in 4 d larva, and reached the highest level at 30 minutes of starvation.By using the same Needle trap and GC-MS techniques, the hunger pheromones of the queen bee larvae and the male larvae were analyzed and identified.The results showed that E- 尾-basilerene was also used as the hunger pheromone of the queen bee and the larvae of the male bee.The content of E- 尾-basilerene in the starved larva group was significantly higher than that in the feeding larval group and the feeding group. The content of E- 尾-basilerene in the starved group of male bee and worker bee was not significantly different from that in the feeding group and the food group.However, the content of E- 尾-basilerene in the starvation group was significantly lower than that in the male and worker larvae groups.At the same time, it was found that the queen bee larva contained a pheromone 2-heptanone, RNA-Seq, which was not found in the male bee and worker bee larvae. The results showed that the queen bee larva and the male bee larva also had the same E- 尾 -basilerene biosynthesis pathway and related genes as the worker bee larvae.The RNA-Seq sequencing technique was used to detect the mRNAs of the 2-day-old and 4-day-old larvae of the Italian bee, and the difference of gene expression during the development of the larvae was analyzed and compared.There was no difference in the number of total genes detected in each larvae group, but there were a large number of differentially expressed genes between the groups. The number of differentially expressed genes between the two groups was 475.There were 687 differentially expressed genes in the control group (197) higher than those in the control group (197) and the control group (121) and 604 (P < 0.05) respectively, and the difference was significantly higher than that in the control group (n = 197) and the control group (n = 121) and the control group (n = 604) respectively, which were higher than those in the control group (n = 197).The difference between male bee larva and other two female wasps was higher than that between two female wasps, but the comparison group between queen bee and worker bee larvae was 475, which indicated that the difference between male bee larva and other two kinds of female wasps was higher than that between them.The number of differentially expressed genes in the 4-day-old control group was significantly higher than that in the 2-day-old control group, indicating that the development of the larvae of the three-type wasp was more differentiated with the increase of the age of 4 days.Secondly, there were different expression bases between worker bee larvae (2d and 4d), because 1181 were in the contrast group of 1190 larva and 1181 in the control group, and only 598 in the control group, which indicated that there were differences in the developmental regulation pattern of the haploidy genetic background of honeybee.The results of gene coexpression cluster analysis showed that the two species of female wasps were clustered closer to the 2-day-old worker bee and the male bee larva, and then to the 2-day-old bee king cluster at 4 days old.A large number of differentially expressed genes were found to be involved in the synthesis and secretion of bee hormones, the synthesis of bee venom, the growth and development of eyes and flying muscles, the development of eyes, and the maturation of ovaries.Some transcriptional factors and signaling pathways regulating gene expression were found to be consistent with the morphological, physiological and behavioral characteristics of female and male bees.
【學(xué)位授予單位】：江西農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：S891
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本文編號(hào)：1772342