| 摘要: |
| 【目的】 研究紫外线UV-B对蚜虫种下分化的影响，筛选、鉴别UV-B诱导条件下差异表达的基因，阐明蚜虫对UV的耐受和反应机理以及分子变异机制，为蚜虫UV B胁迫条件下的分子生态遗传与进化研究提供理论依据。【方法】 以UV-B胁迫桃蚜种群cDNA为试验方(Tester), 以正常条件下桃蚜种群cDNA 为驱动方(Driver)，应用抑制差减杂交（SSH）技术，研究了UV-B胁迫下桃蚜特异基因的表达,并随机选取100个EST克隆测序，分析差异基因的功能。【结果】 蚜虫体内参与紫外胁迫的相关基因有6类，其中胁迫诱导相关基因占总基因的10.64%，核酸、脂肪酸代谢相关基因占14.89%，转录相关基因占9.57%，结构蛋白和蛋白合成相关基因占9.57%，共生菌相关基因占8.51%，未知功能基因占19.15%，未知基因占27.66%。半定量RT-PCR扩增结果显示，热休克蛋白70、辅酶 NADH、氧化还原酶、表皮蛋白在处理和对照之间的差异均达极显著水平(P＜0.01),而看家基因在对照和处理间无显著差异。【结论】 蚜虫种群对紫外线胁迫的反应不仅依赖于抗氧化作用,还是一个多种抗性途径和多基因协同作用的复杂体系。 |
| 关键词: 桃蚜 紫外线胁迫 分子生态遗传 SSH差减文库 |
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| 基金项目:国家自然科学基金项目(30470268) |
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| Preliminary study on molecular ecology of gene expression of Myzus persicae under UV-B stress |
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| Abstract: |
| 【Objective】 In order to probe into the molecular variation mechanism,UV-B effect on aphid differential was researched.【Method】 A molecular analysis of gene expression differences that occur between the UV-B induced aphids and control aphids was performed using the Suppression subtractive hybridization (SSH) method.Non treated Myzus persicae was used as the driver and cDNA from M.persicae exposed to UV-B was for tester.100 EST clones which were randomly selected were sequenced and analyzed.【Result】 These ESTs were categorized into six groups encoding proteins involved in stress related proteins (10.64%),nuclear acid and fatty acid metabolism (14.89%),transcription (9.57%),structure protein and protein synthesis (9.57%),Wolbachia endosymbiont (8.51%),putative protein (19.15%) and unknown proteins (27.66%).The semi quantitative RT-PCR analysis of the over expressed genes showed that difference was marked (P＜0.01) between the two aphid populations in HSK70,NADH ubiquinone oxidoreductase,oxidreductase activity,cuticle protein.【Conclusion】 Functional classification suggested that the response to UV B stress not only depends on the antioxygen system,but also may be a complex system with multiple genes co-regulation,which provides the start points to reveal the mechanisms of aphid populations stress tolerance to UV-B. |
| Key words: Myzus persicae UV-B stress molecular ecological genetics suppression substraction hybridization |
