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题名: 环境内分泌干扰物对土壤跳虫的繁殖毒性及分子标志物筛选
作者: 张倩倩1
学位类别: 硕士
答辩日期: 2017-05
授予单位: 中国科学院大学
授予地点: 北京
导师: 乔敏
关键词: 土壤跳虫,五氯酚,十溴联苯醚,毒性,基因表达 ; soil collembola, PCP, BDE209, toxicity, gene expression
其他题名: Reproductive toxicity and molecular biomarkers of endocrine disrupting chemicals on soil Collembola
学位专业: 生物工程
中文摘要: 五氯酚( PCP)及其钠盐常被用作杀虫剂和木材防腐剂等。十溴联苯醚 (BDE209)是一种溴代阻燃剂,被广泛用于电子、化工和纺织等领域。PCP和 BDE-209均被广泛地使用,且两者在环境条件下都难以降解,因此引起了严重的 环境问题。本研究选择 PCP和 BDE209这两种典型内分泌干扰物为研究对象, 以跳虫 Folsomia candida为实验动物,研究了 PCP和 BDE209暴露下跳虫的繁殖 情况、蜕皮行为以及基因表达。这对诊断该类物质土壤污染和开展土壤生态风险 评价,具有重要意义。主要研究内容及结论如下: (1)多代繁殖实验表明跳虫亲代在幼虫发育至成虫期暴露于 PCP会对其繁 殖子一代产生抑制,子一代如果继续在卵至幼虫期暴露于 PCP会对其繁殖子二 代产生抑制。对于 BDE209,只有在亲代和子一代连续暴露条件下对子一代繁殖 量产生抑制,但对子二代无影响。 (2)利用显微镜在亚致死剂量下对跳虫长期暴露于 PCP和 BDE209时的蜕 皮情况观察发现,PCP能显著延迟跳虫的产卵和蜕皮,而本实验条件下,BDE209 对跳虫的蜕皮和产卵无显著影响。 (3)利用 qPCR技术,研究了不同浓度 PCP和 BDE209暴露下,跳虫 HSP70、 CYP450和 GST基因以及蜕皮相关基因表达的变化。本实验条件下,PCP对跳 虫HSP70和 CYP450基因的表达无显著影响。PCP能显著诱导GST基因Fcc00494 的表达,显著抑制几丁质酶基因 Fcc00881和几丁质结合域基因 Fcc01306的表达。 Fcc00494、Fcc00881和 Fcc01306有望作为指示土壤 PCP污染的分子生物标志物。 这为开展土壤 PCP污染的快速诊断和生态风险评价提供了支持。BDE209对所选 基因的表达无显著影响。 (4)利用基因芯片,研究了跳虫在 BDE209暴露下的基因表达谱的变化, 并比较了 BDE209和 PCP暴露下跳虫基因表达谱的变化。GO富集分析表明, BDE209诱导了胚胎发育和细胞周期等相关基因的表达,产生氧化胁迫。BDE209 和 PCP都能诱导细胞周期相关基因的表达。PCP和 BDE209都能抑制跳虫的繁 殖,PCP抑制跳虫繁殖的原因之一可能是 PCP干扰了跳虫的蜕皮,而 BDE209 抑制跳虫繁殖的原因之一可能是 BDE209干扰了跳虫卵的形成。
英文摘要: Pentachlorophenol (PCP) is commonly used as wood preservative, pesticide, etc. Decabromodiphenyl ether (BDE209) is a kind of brominated flame retardants and is commonly used in electronic industry, chemical industry, textile industry, etc. PCP and BDE209 are widely used and have resulted in serious environmental issues because of their persistence. In this paper, the two endocrine disrupting chemicals, PCP and BDE209 were selected as target compounds. The reproduction, molt and gene expression of Folsomia candida under exposure to PCP and BDE209 were studied. This is important for the diagnosis and ecological risk assessment of soil pollution by these chemicals. The main results are as follows: (1) The results of the multigenerational reproduction tests showed that the quantities of the first filial generation were reduced after F. candida parental generation exposed to PCP during the juvenile stage to the adult stage. When the first filial generation continually exposed to PCP from the egg stage to the juvenile stage, the quantities of the second filial generation were reduced. Only when the parental generation and the first filial generation continually exposed to BDE209, the quantities of the first filial generation were reduced but the second filial generation was not affected. (2) The molting of F. candida under long term exposure to sublethal dose of PCP and BDE209 were observed by microscope. The results showed that the molting and oviposition of F. candida was significantly delayed by PCP and BDE209 did not significantly affect the molting and oviposition of F. candida in our experiment. (3) The expression of HSP70 genes, CYP450 genes, GST genes and molting-related genes of F. candida exposed to different concentration of PCP and BDE209 were detected by qPCR. PCP did not cause significant expression of HSP70 and CYP450 gene. PCP significantly induced the expression of the GST gene Fcc00494 and repressed the expression of the chitinase gene Fcc00881 as well as the chitin binding domain gene Fcc001306. Fcc00494, Fcc00881 and Fcc001306 are potential biomarkers indicating PCP pollution in soil. BDE209 cause no significant changes of these genes and thus these genes are not suitable biomarkers for indicating BDE209 pollution in soil. (4) The gene expression profile of F. candida exposed to BDE209 was analyzed by microarray approach. The gene expression profile of F. candida exposed to BDE209 and PCP were compared. GO enrichment analysis of the differently expressed genes showed that BDE209 upregulated genes involved in embryo development and cell cycle and induced oxidative stress. Both BDE209 and PCP upregulated genes involved in cell cycle. Both BDE209 and PCP can inhibit the reproduction of F. candida. One of the possible reasons why PCP inhibits reproduction is that PCP disrupts the molting of F. candida while for BDE209, disrupting egg formation may be one of the reasons.
内容类型: 学位论文
URI标识: http://ir.rcees.ac.cn/handle/311016/38746
Appears in Collections:中澳联合土壤环境研究室_学位论文

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作者单位: 1.中国科学院生态环境研究中心

Recommended Citation:
张倩倩. 环境内分泌干扰物对土壤跳虫的繁殖毒性及分子标志物筛选[D]. 北京. 中国科学院大学. 2017.
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