【作者】 文菊华；

【导师】 颜亨梅；

【作者基本信息】 湖南师范大学 ， 动物学， 2004， 硕士

【摘要】 本研究应用同工酶技术和RAPD分子标记技术分别探讨了蜘蛛目狼蛛科中部分属种间的亲缘关系，得出了如下结论。 1．在用同工酶电泳技术对狼蛛科3属6种间亲缘关系的研究中，发现了： (1)同种个体不同部位EST同工酶因其所含组织、器官不同而存在差异(0.25＜P＜0.75)，可能和不同部位所执行的代谢活动功能以及蜘蛛消化道在体躯中的位置有关；另外同种不同地域及海拔高度的狼蛛EST同工酶也有差别(0.25＜P＜0.75)，这种差异与地理异质性有关，蜘蛛生存环境的自然生态因子对同工酶的活性有一定的影响。 (2)同种蜘蛛不同个体的EST同工酶存在微小差异(0.90＜P＜0.95)，这种差别在种内相对较稳定；同种蜘蛛不同性别个体也存在差异(0.75＜P＜0.90)，雄性个体的酯酶表达更为充分且雌雄性别上的差异大于个体差异。 (3)同种个体不同发育时期的EST同工酶因其在不同发育阶段的生理特点而存在较大差异(0.25＜P＜0.75)；同种不同饥饿时间处理后个体EST同工酶无论是在酶带的条数及酶的活性上差异都较明显(0.10＜P＜0.25)，通过本次实验得出，在对蜘蛛进行同工酶研究时，最佳的饥饿处理时间在10—25天之间。 (4)酯酶同工酶在狼蛛科低级阶元中具有稳定的种属特异性，对这一类群的研究具有可行性，因而可以作为属内不同种之间(0.75(P(0.90)、不同属不同种之间(0.25(P(0.75)的分类指标之一，同时对这一类群分类地位的探索具有一定价值。 2.本研究采用了与前人不同的蜘蛛总DNA提取方法一一一毛TAB法，成功地运用这一简单、经济、高效的方法提取到了能完全满足RAPD一PCR扩增需要的DNA样品。改进后的蜘蛛类总DNA提取方法简便、快速、对设备和试剂要求都不高，而收率和纯度都可满足实验的要求，特别是对DNA完整性要求较高的RAPD技术。 3.应用RAPD技术对常见的7种狼蛛的遗传多样性进行了分析，所得结论与现行的形态学分类相吻合，这说明RAPD技术是近缘属种分类鉴定的一种有效辅助手段，可以为蜘蛛系统学的研究提供新的分子水平上的信息，有助于建立客观的分类体系，值得进一步探索研究。更多还原

【Abstract】 In this thesis, by applying the technology of isoenzyme and random amplified polymorphic DNA(RAPD) markers, we respectively study the genetic relationship between some species in wolf spider family. The corollaries are as follows:1. In the research about the genetic relationship to the sixth species, the third genera in the wolf spider family by using the technology of isoenzyme electric-pulse, we find:(l)The differences (0. 25<P<0. 75) exist in different parts of EST isoenzyme patterns in the same individual due to the differences in its tissues, segments, and relating to the metabolism of its different parts and the spider’ s digesting channel’ s position in its body. In addition, there are differences (0. 25<P<0.75) in wolf spider esterase isoenzyme patterns in different geographic region and elevation, which may relate to geographical heterogeneity. The ecological factors of the living environment of spiders may exert certain effect on activation of isoenzyme.(2) The slight differences (0.90<P<0.95) in esterase isoenzyme patterns of different individual spiders of the same genus are relatively stable within the genus; There are also differences (0.75<P<0.90) in sexually different individualspiders of the same genus, with male’ s fuller expression in esterase. Moreover, the differences in sex are wider than those in individuals.(3) There are relatively wide differences in esterase isoenzyme patterns of the same individual in different growing periods, due to the physiological features in different growing periods; After the individual esterase isoenzymes of the same genus are tested in different hungry time, there exist relatively apparent differences (0. 10<P<0.25) either on the number of esterase patterns or on the activation of esterase. We conclude by this experiment that the best hungry time is 10-20 days.(4)The esterase isoenzyme in Lycosidae family has a stable polymorphosis, which is of feasibility to the research of this category so that we can take it as the indentification of Lycosidae family between the different species of the same genus(0.75<P<0.90), and between the different species in different genera(0. 25<P<0.75), and at the same time it is of some value to the research about the classifying status of this category.2. In this research, a new, simple, economical, and high-efficiency method for spiders’ genomic DNAextraction-CTAB-is applied successfully to extract the needed DNA samples, which are enough to satisfy the requirement of enlargement of RAPD-PCR. The improved method of spiders’ genomic DNA extraction is succinct and expeditious, with relatively low requirements to the facilities and reagents. Its production and. purity can meet with the requirements of the experiment, especially of the technology of RAPD, which has a high requirement for the integrity of DNA.3. The genetic diversity of 7 species of common wolf spiders is analyzed by applying the RAPD technology, and the result is suitable for the present morphological taxonomy, which shows that the RAPD technology is an efficient complementary method for the classification and identification of closely related species and genera. It deserves a further study and exploration because it is helpful for the establishment of a subject classification system by providing information for the research of spider systematics at new molecular level.更多还原