家蚕ptp和cyp333a2基因的鉴定及其耐氟性功能研究

发布时间：2018-05-23 12:01

本文选题：家蚕 + 耐氟　；参考：《西南大学》2017年硕士论文

【摘要】：本研究以鳞翅目模式昆虫家蚕中的敏感品种734和耐氟品种T6为材料,应用差异显示技术对家蚕耐氟性相关基因进行筛选,并分析这些差异基因的功能。氟化物通过污染桑叶造成家蚕大量死亡,严重危害蚕桑业的发展,同时人类及其他生物也深受其害。阐明家蚕耐氟中毒的机理,为设计相关药物防御及治疗氟中毒提供新的方向。其主要结论如下:1.统计分析家蚕734品种和T6品种5龄期浸叶法添食200mg/kg氟化钠后的生理生化改变。在200mg/kg的氟化钠浓度下,T6品种能正常生长,734品种表现明显氟中毒症状,T6耐氟品种死亡率比敏感品种734低,体重增长率与敏感品种734差异显著。提示T6品种可能存在耐氟基因。2.通过mRNA差异显示技术,在家蚕耐氟品种T6和敏感品种734脂肪体中筛选参与解氟代谢相关的基因,获得了一个位于线粒体膜上与转运磷酸根离子至线粒体内与ADP合成ATP相关的磷酸根转运蛋白Bmptp基因,半定量结果显示Bmptp基因在T6耐氟品种所受氟化物的影响明显比734品种低,说明氟化物可能通过影响Bmptp基因表达从而影响ATP合成,导致家蚕供能不足,引起中毒症状。3.利用氟离子选择电极法测量家蚕耐氟品种T6和敏感品种734脂肪体的氟浓度,结果显示T6能通过氟代谢使体内氟浓度维持在安全阀值内,而734则累积氟,提示脂肪体可能参与了氟代谢。4.差异显示实验过程中获得了另一个在家蚕耐氟品种T6高表达而在敏感品种734低表达的基因Bmcyp333a2。RT-PCR结果表明,其在T6品种脂肪体中表达量相对较高,在各组织都有表达而家蚕添氟后氟离子逐渐遍布累积于各组织,推测其可能与家蚕耐氟有关。
[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).
【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S884.95

【参考文献】