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摘要：沃尔巴克氏体是一种在自然界分布中最为广泛的共生菌。已知赤拟谷盗体内不感染沃尔巴克氏体，杂拟谷盗体内感染此内生菌，为了搞清沃尔巴克氏体在这两个近缘种的种间传播情况，我们首先利用PCR扩增技术检测两种拟谷盗体内共生菌沃尔巴氏体的存在情况，然后将两种拟谷盗混合饲养和杂交，用来检查沃尔巴克氏体能否能够进行种间的水平传递和垂直传递。结果表明，混合饲养后，赤拟谷盗及其与杂拟谷盗的正反杂交后代体内均没有检测到沃尔巴克氏体的存在，未发现沃尔巴克氏体在杂拟谷盗和赤拟谷盗种间的水平和垂直传播。本研究为今后利用沃尔巴克氏体对拟谷盗进行生物防治提供了理论依据。33494
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毕业论文关键词：赤拟谷盗；杂拟谷盗；沃尔巴克氏体；种间传播；PCR扩增
Study on Interspecific Transmission of Symbiotic Bacteria Wolbachia in Tribolium castoneum and T. confusum
Abstract：Wolbachia is a kind of widespread symbiotic bacteria. It is known that Wolbachia does not exist in Tribolium castoneum （Herbst）,but exists in Tribolium confusum (Jac. du Val.), To clarify the interspecific horizontal and vertical transmission of Wolbachia in the two closely related flour beetles T. castaneum and T. confusum, we first detected them in both beetle species by PCR amplification and electrophoresis separation of ftsZ target gene fragment; then, after same-sex or opposite-sex mixed feeding for them, we determined the infection status of Wolbachia in the red flour beetles T. castaneum and their reciprocal hybrids. The results showed that, the confused flour beetles T. confusum were infected with Wolbachia, but the red flour beetles T. castaneum were not. Although the two beetle species T. castaneum and T. confusum can mate and produce hybrid offsprings, the red flour beetles T. castaneum and their reciprocal hybrids were not infected with Wolbachia. This suggests that interspecific horizontal and vertical transmissions of Wolbachia do not occur in the two closely related flour beetles T. castaneum and T. confusum, and the red flour beetles T. castaneum in vivo may contain an internal factor inhibiting the normal reproduction and survival of Wolbachia. This study provides the theoretical basis for potential bio-control of Tribilium flour beetles using Wolbachia in the future.
Key words：T. castoneum；T. confusum；Wolbachia；interspecific transmission；PCR amplification
目录
引言 1
1 材料与方法 2
1.1 供试昆虫 2
1.2 仪器设备和试剂 2
1.2.1 主要仪器 2
1.2.2 主要试剂 3
1.2.2.1 DNA提取缓冲液的配制 3
1.2.2.2 琼脂糖凝胶电泳所用试剂的配制 3
1.3 试验方法 3
1.3.1 赤拟谷盗与杂拟谷盗体内沃尔巴克氏体的PCR检测 3
1.3.1.1 总DNA的提取 3
1.3.1.2 PCR 扩增、测序 3
1.3.2 自然条件下沃尔巴克氏体在赤拟谷盗与杂拟谷盗种间的水平传播 3
1.3.3 沃尔巴克氏体在赤拟谷盗与杂拟谷盗种间的垂直传播 4
2 结果与分析 4
2.1 赤拟谷盗和杂拟谷盗体内沃尔巴克氏体的基因检测 4
2.2 自然条件下沃尔巴克氏体在同性赤拟谷盗与杂拟谷盗种间的水平传播 5
2.3 自然条件下沃尔巴克氏体在异性赤拟谷盗与杂拟谷盗种间的水平传播 5
2.4 自然条件下沃尔巴克氏体在赤拟谷盗与杂拟谷盗种间的垂直传播 6
3 讨论 7
致谢 8
参考文献 8
引言