本文選題：家蠶 + 耐氟��； 參考：《西南大學(xué)》2017年碩士論文

【摘要】：本研究以鱗翅目模式昆蟲家蠶中的敏感品種734和耐氟品種T6為材料,應(yīng)用差異顯示技術(shù)對(duì)家蠶耐氟性相關(guān)基因進(jìn)行篩選,并分析這些差異基因的功能。氟化物通過污染桑葉造成家蠶大量死亡,嚴(yán)重危害蠶桑業(yè)的發(fā)展,同時(shí)人類及其他生物也深受其害。闡明家蠶耐氟中毒的機(jī)理,為設(shè)計(jì)相關(guān)藥物防御及治療氟中毒提供新的方向。其主要結(jié)論如下:1.統(tǒng)計(jì)分析家蠶734品種和T6品種5齡期浸葉法添食200mg/kg氟化鈉后的生理生化改變。在200mg/kg的氟化鈉濃度下,T6品種能正常生長,734品種表現(xiàn)明顯氟中毒癥狀,T6耐氟品種死亡率比敏感品種734低,體重增長率與敏感品種734差異顯著。提示T6品種可能存在耐氟基因。2.通過mRNA差異顯示技術(shù),在家蠶耐氟品種T6和敏感品種734脂肪體中篩選參與解氟代謝相關(guān)的基因,獲得了一個(gè)位于線粒體膜上與轉(zhuǎn)運(yùn)磷酸根離子至線粒體內(nèi)與ADP合成ATP相關(guān)的磷酸根轉(zhuǎn)運(yùn)蛋白Bmptp基因,半定量結(jié)果顯示Bmptp基因在T6耐氟品種所受氟化物的影響明顯比734品種低,說明氟化物可能通過影響B(tài)mptp基因表達(dá)從而影響ATP合成,導(dǎo)致家蠶供能不足,引起中毒癥狀。3.利用氟離子選擇電極法測量家蠶耐氟品種T6和敏感品種734脂肪體的氟濃度,結(jié)果顯示T6能通過氟代謝使體內(nèi)氟濃度維持在安全閥值內(nèi),而734則累積氟,提示脂肪體可能參與了氟代謝。4.差異顯示實(shí)驗(yàn)過程中獲得了另一個(gè)在家蠶耐氟品種T6高表達(dá)而在敏感品種734低表達(dá)的基因Bmcyp333a2。RT-PCR結(jié)果表明,其在T6品種脂肪體中表達(dá)量相對(duì)較高,在各組織都有表達(dá)而家蠶添氟后氟離子逐漸遍布累積于各組織,推測其可能與家蠶耐氟有關(guān)。
[Abstract]:In this study, sensitive varieties 734 and T6 of Bombyx mori of Lepidoptera model insect were used as materials. The genes related to fluoride tolerance of Bombyx mori were screened by differential display technique, and the function of these genes was analyzed. Fluoride caused a large number of deaths of silkworm by contaminating mulberry leaves, seriously endangering the development of sericulture, and at the same time, human beings and other living things suffered greatly. To elucidate the mechanism of fluorosis tolerance of silkworm, and to provide a new direction for the design of drug defense and the treatment of fluorosis. The main conclusions are as follows: 1. The physiological and biochemical changes of silkworm (Bombyx mori) 734 and T6 treated with sodium 200mg/kg fluoride at 5 age stage were analyzed statistically. Under the concentration of sodium fluoride of 200mg/kg, the death rate of T6 variety with fluoride tolerance was lower than that of sensitive variety 734, and the growth rate of body weight was significantly different from that of sensitive variety 734. These results suggest that fluorine tolerance gene. 2. MRNA differential display technique was used to screen the genes involved in defluorination metabolism in the fluorine-tolerant silkworm T6 and the sensitive 734 fat body. A phosphate radical transporter (Bmptp) gene related to the synthesis of ATP by ADP was obtained in mitochondria membrane and transported to mitochondria. The semi-quantitative results showed that the effect of Bmptp gene on fluoride was significantly lower in T6 fluorine-tolerant cultivars than in 734 varieties. It is suggested that fluoride may affect the synthesis of ATP by affecting the expression of Bmptp gene, resulting in insufficient energy supply and toxic symptoms. 3. Fluorine concentration in fat body of silkworm (Bombyx mori) resistant variety T6 and sensitive variety 734 was measured by fluorine ion selective electrode method. The results showed that T6 could maintain the concentration of fluoride in the body at the safety threshold through fluorine metabolism, while 734 could accumulate fluorine. It suggests that adipose body may be involved in fluorine metabolism. 4. During the differential display experiment, another Bmcyp333a2.RT-PCR gene was obtained, which was highly expressed in fluorine-tolerant silkworm (Bombyx mori) and low in sensitive variety 734. The results showed that the gene expressed in the fat body of T6 was relatively high. The fluorine ions were accumulated in all tissues after fluorine addition, which may be related to the fluoride tolerance of silkworm (Bombyx mori).
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S884.95

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