本文選題：滯育 + 棉鈴蟲(chóng)�。� 參考：《中山大學(xué)》2016年博士論文

【摘要】：自然環(huán)境并非總是適合于生物的生長(zhǎng)發(fā)育,面對(duì)惡劣環(huán)境時(shí),昆蟲(chóng)往往會(huì)選擇進(jìn)入滯育狀態(tài)以躲過(guò)不利環(huán)境的影響,待到環(huán)境條件適合再重新啟動(dòng)發(fā)育。因此,滯育是昆蟲(chóng)賴以生存的重要策略。滯育的主要特征是個(gè)體代謝顯著下調(diào),抗逆性增強(qiáng)以及發(fā)育極其緩慢,由此可以看出滯育是整體變化,十分復(fù)雜,涉及昆蟲(chóng)體內(nèi)許多生理生化調(diào)控。一般認(rèn)為激素是調(diào)節(jié)滯育的關(guān)鍵因素,且已經(jīng)得到大量的證據(jù)證實(shí),其中蛹滯育是由于促前胸腺激素-蛻皮激素信號(hào)下調(diào)導(dǎo)致的。雖然滯育過(guò)程生理生化變化已經(jīng)有大量的研究,但是這些生理生化背后的分子機(jī)制卻尚不十分清楚,因此本研究以蛹滯育的棉鈴蟲(chóng)為研究對(duì)象,試圖闡明棉鈴蟲(chóng)滯育過(guò)程中代謝的變化原因以及胰島素受體對(duì)滯育的影響。為了調(diào)查棉鈴蟲(chóng)滯育過(guò)程中代謝下調(diào)的機(jī)制,克隆了糖酵解途徑的第一個(gè)關(guān)鍵基因己糖激酶(HK),并調(diào)查了它在棉鈴蟲(chóng)發(fā)育或者滯育個(gè)體腦中表達(dá)和酶活。結(jié)果發(fā)現(xiàn)HK mRNA和蛋白在發(fā)育型蛹腦中表達(dá)明顯高于滯育型蛹腦,且其活性也與表達(dá)水平一致,在發(fā)育型蛹腦中活性較高。下調(diào)HK表達(dá)活性可以引起代謝降低以及ROS增加,從而降低細(xì)胞活性并延遲蛹的發(fā)育。暗示著HK是一個(gè)調(diào)節(jié)昆蟲(chóng)發(fā)育的重要因子。隨后我們鑒定了三個(gè)轉(zhuǎn)錄因子CREB、c-Myc和POU特異性結(jié)合到HK啟動(dòng)子上調(diào)節(jié)HK活性,但有趣的是POU和c-Myc是特異性調(diào)節(jié)HK表達(dá)的轉(zhuǎn)錄因子,而CREB卻是非特異性的。另外,POU和c-Myc能夠響應(yīng)上游激素信號(hào)蛻皮激素的調(diào)節(jié)。在滯育個(gè)體中低水平的蛻皮激素導(dǎo)致低水平的POU和c-Myc表達(dá),從而抑制了HK的表達(dá)水平,導(dǎo)致代謝的下調(diào)和ROS的積累,抑制蛹繼續(xù)發(fā)育而進(jìn)入滯育狀態(tài)。細(xì)胞能量代謝的主要場(chǎng)所是線粒體,以前的報(bào)道中發(fā)現(xiàn)滯育型棉鈴蟲(chóng)蛹腦中COX活性下調(diào),暗示著線粒體活性也可能處于較低水平,但是具體機(jī)制并不十分清楚。因此,首先調(diào)查了線粒體DNA含量以及COX活性,發(fā)現(xiàn)線粒體活性在滯育型蛹腦中的確受到抑制。隨后的調(diào)查結(jié)果顯示,HIF-1α在滯育型蛹腦中表達(dá)較發(fā)育型蛹腦增高,且HIF-1α高表達(dá)能夠抑制線粒體活性。HIF-1α抑制線粒體活性是通過(guò)蛋白酶體降解c-Myc蛋白發(fā)揮的作用。線粒體轉(zhuǎn)錄因子A(TFAM)是一個(gè)調(diào)節(jié)線粒體轉(zhuǎn)錄和線粒體DNA復(fù)制的重要轉(zhuǎn)錄因子,c-Myc能夠直接結(jié)合到TFAM啟動(dòng)子上并激活其表達(dá),而TFAM活性的缺失是導(dǎo)致線粒體活性下調(diào)的重要原因,證實(shí)了HIF-1α-c-Myc-TFAM信號(hào)參與到調(diào)節(jié)昆蟲(chóng)滯育過(guò)程中線粒體的活性。胰島素(insulin)信號(hào)通路是調(diào)節(jié)發(fā)育的重要途徑,有許多研究發(fā)現(xiàn)insulin信號(hào)參與到昆蟲(chóng)滯育的調(diào)節(jié)并起到重要作用。為了研究insulin信號(hào)通路在棉鈴蟲(chóng)滯育中的作用,克隆并調(diào)查了胰島素受體(InR)在棉鈴蟲(chóng)蛹腦中的表達(dá)。結(jié)果發(fā)現(xiàn)InR在滯育型蛹腦中高表達(dá),且P-InR水平在滯育型蛹腦中明顯高于發(fā)育型蛹腦;insulin通過(guò)P-InR調(diào)節(jié)P-ERK表達(dá);ROS能夠激活P-InR,P-AKT和P-ERK;且ROS對(duì)P-AKT和P-ERK的激活是通過(guò)P-InR調(diào)控的。結(jié)合我們已有的一些結(jié)果,發(fā)現(xiàn)了ROS-P-InR-P-ERK信號(hào)能夠參與到棉鈴蟲(chóng)滯育過(guò)程中蛋白酶體的調(diào)控,降解控制發(fā)育的相關(guān)蛋白,使蛹順利進(jìn)入滯育。
[Abstract]:Natural environment is not always suitable for biological growth and development. In the face of bad environment, insects often choose diapause to avoid the effect of adverse environment and adapt to environmental conditions to restart and develop. Therefore, diapause is an important strategy for the survival of insects. It can be seen that diapause is a whole, very complex and involves many physiological and biochemical regulation in the body of insects. It is generally believed that hormone is the key factor regulating diapause, and a large amount of evidence has been obtained, in which the diapause of pupae is caused by the downregulation of the pre thymus hormone - ecdysone. The physiological and biochemical changes in diapause have been studied, but the molecular mechanisms behind these physiological and biochemical factors are still not very clear. Therefore, this study aims to elucidate the causes of metabolic changes and the effect of insulin receptor on diapause during the diapause of the cotton bollworm. In order to investigate the stagnation of cotton bollworm. The mechanism of down regulation of metabolism, the first key gene of hexokinase (HK), the key gene of glycolysis, was cloned, and the expression and enzyme activity in the individual brain of the cotton bollworm and diapause were investigated. The results showed that the expression of HK mRNA and protein in the developmental pupa brain was obviously higher than that of diapause pupa brain, and its activity was consistent with the expression level. The activity of the pupae in the pupae is high. Down regulation of HK expression activity can cause the decrease of metabolism and the increase of ROS, thus reducing cell activity and delayed the development of pupae. It suggests that HK is an important factor regulating the development of insects. Then we identified three transcription factors CREB, c-Myc and POU specific binding to the HK promoter to regulate HK activity, but it is interesting. It is that POU and c-Myc are the transcription factors that specifically regulate the expression of HK, while CREB is nonspecific. In addition, POU and c-Myc can respond to the regulation of the upstream hormone signal ecdysone. Low levels of ecdysone in diapause individuals lead to low levels of POU and c-Myc expression, thus inhibiting the expression level of HK, leading to metabolic downregulation and ROS. Accumulation, inhibition of the continued development of pupae and diapause. The main site of cell energy metabolism is mitochondria. In previous reports, the COX activity in the diapause type Helicoverpa armigera was downregulated in the pupa brain, suggesting that mitochondrial activity may also be at a lower level, but the specific mechanism is not very clear. Therefore, the content of mitochondrial DNA and C are investigated first. OX activity was found to be suppressed in the diapause pupa brain. The results of the subsequent investigation showed that the expression of HIF-1 alpha in diapause pupa brain was higher than that of the developmental pupae, and the high expression of HIF-1 a could inhibit the mitochondrial activity of the mitochondrial activity by inhibiting the mitochondrial activity by protease degradation of the c-Myc protein. Transcriptional factor A (TFAM) is an important transcription factor regulating mitochondrial transcription and mitochondrial DNA replication. C-Myc can directly bind to TFAM promoter and activate its expression, and the absence of TFAM activity is an important cause of mitochondrial activity downregulation. It is confirmed that the HIF-1 alpha -c-Myc-TFAM signal is involved in the regulation of mitochondria in the process of insect diapause. Activity. Insulin (insulin) signaling pathway is an important way to regulate development. Many studies have found that insulin signals play an important role in regulating and regulating insect diapause. In order to study the role of insulin signaling pathway in the diapause of cotton bollworm, the expression of insulin receptor (InR) in the chrysalis of cotton bollworm was cloned and investigated. The results found that In R was highly expressed in the diapause pupa brain, and the level of P-InR was significantly higher in the diapause pupa brain than in the developmental pupa brain; insulin was used to regulate P-ERK expression through P-InR; ROS could activate P-InR, P-AKT and P-ERK; and ROS to P-AKT and P-ERK activation was regulated. The regulation of proteasome during the diapause of Helicoverpa armigera leads to the degradation of related proteins in development and the smooth passage of pupae into diapause.
【學(xué)位授予單位】：中山大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q963

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 徐衛(wèi)華;;昆蟲(chóng)滯育研究進(jìn)展[J];昆蟲(chóng)知識(shí);2008年04期

2 黎萬(wàn)順;陳斌;何正波;;蔥蠅非滯育、冬滯育和夏滯育蛹發(fā)育和形態(tài)特征比較[J];昆蟲(chóng)學(xué)報(bào);2012年07期

3 王宗舜;昆蟲(chóng)滯育與激素調(diào)節(jié)[J];昆蟲(chóng)知識(shí);1985年04期

4 薛瑞德;蚊蟲(chóng)的滯育[J];昆蟲(chóng)知識(shí);1990年05期

5 仵均祥,李長(zhǎng)青,李怡萍,成衛(wèi)寧;小麥吸漿蟲(chóng)滯育研究進(jìn)展[J];昆蟲(chóng)知識(shí);2004年06期

6 劉流;賀莉芳;劉暉;晏容;萬(wàn)啟惠;;滯育對(duì)家蠅幼蟲(chóng)蛋白質(zhì)和核酸的影響[J];中國(guó)媒介生物學(xué)及控制雜志;2009年02期

7 黎萬(wàn)順;馮國(guó)忠;陳斌;何正波;;蔥蠅冬滯育蛹的全長(zhǎng)cDNA文庫(kù)的構(gòu)建[J];昆蟲(chóng)知識(shí);2010年01期

8 黎萬(wàn)順;陳斌;馮國(guó)忠;李廷景;;蔥蠅非滯育蛹的全長(zhǎng)cDNA文庫(kù)構(gòu)建[J];重慶師范大學(xué)學(xué)報(bào)(自然科學(xué)版);2010年01期

9 肖海軍;魏兆軍;薛芳森;;昆蟲(chóng)滯育關(guān)聯(lián)熱休克蛋白的研究進(jìn)展[J];昆蟲(chóng)學(xué)報(bào);2011年09期

10 徐晶晶;陳斌;郝友進(jìn);司鳳玲;何正波;;昆蟲(chóng)滯育相關(guān)激素調(diào)節(jié)的研究進(jìn)展[J];重慶師范大學(xué)學(xué)報(bào)(自然科學(xué)版);2012年04期

相關(guān)會(huì)議論文 前10條

1 黃少虹;江幸福;羅禮智;;昆蟲(chóng)滯育發(fā)生與調(diào)控的生理生態(tài)機(jī)制[A];植物保護(hù)與現(xiàn)代農(nóng)業(yè)——中國(guó)植物保護(hù)學(xué)會(huì)2007年學(xué)術(shù)年會(huì)論文集[C];2007年

2 肖海軍;何海敏;朱杏芬;薛芳森;;低溫和光周期對(duì)黑紋粉蝶滯育解除的影響[A];植�？萍紕�(chuàng)新與病蟲(chóng)防控專業(yè)化——中國(guó)植物保護(hù)學(xué)會(huì)2011年學(xué)術(shù)年會(huì)論文集[C];2011年

3 李秉，

本文編號(hào)：2053578