摘要复眼是赤拟谷盗和杂拟谷盗的主要视觉器官，视蛋白是存在于复眼内光感受细胞膜的受体，经过长期的进化和适应，拟谷盗的视蛋白进化为长波（LW）视蛋白和紫外线敏感（UV）视蛋白两类。为了明确赤拟谷盗与杂拟谷盗复眼外部形态和视蛋白基因的进化，本研究首先在体视显微镜下对赤拟谷盗和杂拟谷盗复眼的外部形态进行了比较观察，然后运用RACE技术克隆出赤拟谷盗与杂拟谷盗的LW视蛋白和UV视蛋白的基因全长，对它们进行种间序列比对分析，并构建相对应的系统发育树。结果表明，赤拟谷盗与杂拟谷盗复眼的外部形态存在明显不同，赤拟谷盗的复眼比杂拟谷盗大。赤拟谷盗和杂拟谷盗的LW视蛋白基因片段长度为1360bp，种间存在8个变异位点，系统发育树显示赤拟谷盗与杂拟谷盗LW和UV视蛋白基因聚为一支，表明赤拟谷盗和杂拟谷盗的LW和UV视蛋白基因同源性高，种间亲缘关系近。赤拟谷盗与杂拟谷盗视蛋白基因的克隆与序列分析进化，为深入研究视蛋白在储粮害虫趋光行为中所发挥的作用奠定基础，不仅在理论上具有重要的意义，在实际的应用中也有潜在价值，可能为储粮害虫的生物绿色防治提供新的思路和方法。50711

该论文有图2幅，表9个，参考文献15篇。

毕业论文关键词：赤拟谷盗  杂拟谷盗  复眼  视蛋白基因  RACE技术  

Comparative analysis of LW opsin gene sequence between Tribolium castaneum and Tribolium confusum

Abstract

Compound eye is the main vision organ of insect which plays a significant role in its foraging, finding mate, learning and memory, etc. Insect persity is very rich, and by long-term light adaptation, the visual pigments of Tribolium castaneum and Tribolium confusum evolve into two categories: long-wavelenth (LW) opsin and opsin and ultra-violet (UV) opsin. In this study, to clarify the evolution of compound eyes and opsin genes in T. castaneum and T. confusum, the stereo microscope examination of compound eyes was first made in T. castaneum and T. confusum. Then, the full length of LW and UV gene was cloned by rapid amplification of cDNA ends (RACE). Finally, the sequences were aligned, and phylogenetic tree was constructed. The results showed that there were significant differences in the morphology of compound eyes of T. castaneum and T. confusum. The LW opsin gene was 1360bp in length, and there were 8 mutation sites in LW opsin genes between species. Phylogeny incorporating other species revealed that, LW opsin genes of T. castaneum and T. confusum were highly homologous, and T. castaneum were more closely related to T. confusum. The evolutionary analyses of compound eye and opsin gene in T. castaneum and T. confusum lay the foundation for further study on the role of opsin in phototaxis behavior of stored-grain pests. It not only has important significance in theory, but also are there potential values in practical application, which may provide new ideas for the biological control of stored grain pests.

Key words：Tribolium castaneum  Tribolium confusum  compound eye    opsin gene  RACE

目录

摘要 I

Abstract II

目录 III

1绪论 1

2材料与方法 2

2.1供试昆虫 2

2.2体视显微镜下的观察 2

2.3赤拟谷盗和杂拟谷盗头部RNA的提取 2

2.4 cDNA第一条链的合成 3

2.5 TDT加尾反应 3

2.6 LW视蛋白基因的克隆