【作者】 牛红星；

【导师】 刘敬泽；

【作者基本信息】 河北师范大学 ， 动物学， 2008， 博士

【摘要】 从2003年10至2007年12月,对河南省翼手类进行了较为全面的研究。1研究了河南省翼手类的种类、数量、分布及生态习性,评估了蝙蝠栖息地的保护现状并提出了保护性建议。2采用“取骨髓-空气干燥法”和“取骨髓-蒸气固定法”对河南省5种蝙蝠科蝙蝠和2种菊头蝠科蝙蝠进行了核型研究,初步探讨了核型的演化;对不同产地的10种蝙蝠进行了RAPD分析,确定了它们的亲缘关系。3采用聚丙烯酰胺凝胶垂直平板电泳技术,对九种蝙蝠(八种组织:肺脏、心脏、肝脏、肾脏、肌肉、胃、小肠、舌)的同工酶(EST、LDH、MDH、ME)表达情况、电泳图谱进行了比较研究。4利用线粒体细胞色素B基因,研究了河南省翼手类的分子系统发育,构建了系统发育树,探讨了它们之间的亲缘关系,并对亚洲长翼蝠Miniopterus schreibersii和马铁菊头蝠Rhinolophus ferrumequinum进行了种群研究。主要结果如下:1.共观察到蝙蝠130000余只,采集标本628号,隶属3科10属26种,占全国总种数的20.8%。其中古北界种4种,占15.4%;东洋界种11种,占42.3%;广布种11种,占42.3%,河南省的翼手类以东洋界种和广布种为主。2.河南省的翼手类动物主要分布在山区,山区的种类多,数量集中,平原地区种类少,数量大,但分布不集中。3.栖息蝙蝠种类和数量最多的洞穴是西峡云华蝙蝠洞和米坪洞、南召的仙人洞、济源的黄楝树引水渠、商城的乌龙洞及桐柏回龙的熊洞,因其为多种蝙蝠提供繁殖和冬眠地而具有较高保护价值,但所有的栖息地保护现状均不容乐观,蝙蝠的生存正在受到严重的威胁,急需加强保护。4.根据翼手类的组成、分布、数量、区系特征及自然地理条件,将河南省翼手类动物地理划分为5个动物地理省:I、豫西北太行山地动物地理省;II、豫西伏牛山地丘陵台地动物地理省;III、豫西伏南山地丘陵盆地动物地理省;IV、豫南桐柏-大别山地森林动物地理省;V、豫东黄淮海平原动物地理省。5.蝙蝠科5种蝙蝠和菊头蝠科2种蝙蝠的核型研究结果表明:大棕蝠Eptesicus serotinus的染色体数为2n=50,N.F=48,其核型与南蝠Ia io的核型比较接近;亚洲长翼蝠Miniopterus schreibersii的染色体数为2n=46,N.F=50;毛腿鼠耳蝠Myotis fimbriatus的染色体数为2n=44,N.F=52;东亚蝙蝠Vespertilio superans的染色体数为2n=38,N.F=48;东亚伏翼Pipistrellus abramus的染色体数为2n=26,N.F=44;中菊头蝠Rhinolophus affinis染色体数为2n=62;马铁菊头蝠Rhinolophus ferrumequinum染色体数为2n=58, N.F=60。其中大棕蝠、马铁菊头蝠的核型为首次报道。6.蝙蝠科7种蝙蝠RAPD分析结果表明:7种蝙蝠分为四大类群,亚洲长翼蝠为一个类群,白腹管鼻蝠构成另一个类群,3种鼠耳蝠、大棕蝠为一个类群,东亚伏翼为第四类群。鼠耳蝠较为原始,与其关系最近的是大棕蝠,关系最远的是长翼蝠。菊头幅科3种蝙蝠RAPD分析结果表明:角菊头蝠属于比较原始的类群,它与中菊头蝠的亲缘关系近于其与马铁菊头蝠的亲缘关系。不同产地的同种蝙蝠出现了不同程度的分化,其遗传多态性明显高于同一产地同种蝙蝠的遗传多态性,同一区域的同种蝙蝠个体之间也存在着分化,而同一冬眠群内的蝙蝠之间几乎没有分化。7.同工酶研究结果表明:在八种组织中,EST、LDH、MDH和ME同工酶表达上有明显的差异。EST同工酶在肾脏、肝脏和小肠中占优势,表现为多型性,活性高;LDH同工酶和大多数哺乳动物一样,具有组织特异性,在胸肌、肾脏等耗氧型组织中,以B亚基为主,在肝组织中,以A亚基为主;MDH同工酶具有组织特异性,s-MDH和m-MDH酶带在八种组织中的分布差异表明蝙蝠主要以线粒体型酶为主,这与翼手目动物适应飞行、消耗能量大有着密切联系;在八种组织中肌肉的ME同工酶表现的酶带最多,活性也较强。同属蝙蝠的同工酶谱型差异较小,仅表现为酶带的强弱稍有不同及某些亚带出现的多少和位置,不同属蝙蝠差异稍大。8.基于Cytb基因的系统发育树表明:菊头蝠科与蹄蝠科、蝙蝠科的关系都较近,而蹄蝠科与蝙蝠科的关系较远;在蝙蝠科内,管鼻蝠属与鼠耳蝠属的关系比较近,长翼蝠属与其他属的关系最远。河南省的长翼蝠与中国海南的长翼蝠应分属两个不同的亚种,而河南省的长翼蝠与日本的长翼蝠差异较小应属于一个亚种。长翼蝠的种群研究显示,在不同地点之间的长翼蝠遗传距离差异非常小,这表明河南省的地理环境不足以对蝙蝠形成阻隔,在各个蝙蝠种群间存在广泛的基因交流。菊头蝠科的系统发育树表明:应将吉林的马铁菊头蝠定名为马铁菊头蝠日本亚种(Rhinolophus ferrumequinum nippon),将云南样本鉴定为马铁菊头蝠尼泊尔亚种(Rhinolophus ferrumequinum tragatus),而河南样本可能为马铁菊头蝠新亚种。将菊头蝠科6种蝙蝠聚为两个独立的分支即角菊头蝠、单角菊头蝠、菲菊头蝠聚为一支,马铁菊头蝠、大耳菊头蝠、中菊头蝠和皮氏菊头蝠聚为另一支,分歧时间大约为2.9-7百万年前。更多还原

【Abstract】 A comprehensive research on Henan province bats was performed from October 2003 to December 2007. The research included: 1.Analyzing the species, relative abundance, distributions, living styles and behavior of bats, evaluating their habitats and giving some advice on protection. 2. Bone marrow cells from five species of Vespertilionidae bats and two species of Rhinolophidae bats that widely distributed in Henan Province, were prepared by means of air-dried method or steam-fixed method, then the karyotypes and their karyotype evolution were analyzed. Phylogenetic relationships among the seven species of Vespertilionidae bats and three species of Rhinolophidae bats from different locations were investigated by RAPD assay; 3. The four isozymes including esterase (EST), lactate dehydrogenase (LDH), malate dehydrogenase (MDH) and malic enzyme (ME) from different organ were systematically compared with each other for the expression pattern and zymogram pattern by using discontinuous vertical polyacrylamide gel electrophoresis (PAGE), aiming at providing clues for bat research; 4. The phylogenetic relatives of Henan bats and the populations on M. schreibersii and R. ferrumequinum were analyzed based on Cytb gene DNA sequence. The main results were as follows:1. Approximately 130000 bats were recorded and 628 specimens were collected, which represented 26 species, 10 genus, 3 families, and occupied 20.8% of Chinese bat. Among them, 4 palaearctic realm species (15.4%), 11 Oriental realm species (42.3%) and 11 wide spread species (42.3%).2. Most of bats distributed in mountains where the species was diversity, and the quantity was centralized. While in plain, the quantity was large and the distribution was not centralized.3. The caves with the highest diversity and largest amount of bats inhabited in were Yunhua and Miping cave in Xixia, Xianren cave in Nanzhao, Huanglianshu cave in Jiyuan, Wulong cave in Shang cheng and Xiong cave in Huilong, Tongbai. Though high value for bats reproduction and hibernation, those caves were not effectively protected and the bats were being subjected to danger of extinction.4. According to the Chiropter composition, distribution, quantity, fauna feature and physical geography conditions, Henan bats were divided into five zoological geographic province: (1) North-west Taihang mountainous zoological geographic province; (2) West Funiu mountainous and mesa zoological geographic province; (3) West Funiu mountainous and basin zoological geographic province; (4) South Tongbai-Dabie mountainous and forest zoological geographic province; (5) East Huanghuai plain zoological geographic province.5. The karyotypes studies of the five Vespertilionidae bats and the two Rhinolophidae bats showed a diploid chromosome number of 2n=50, N.F (fundamental number) =48 in Eptesicus serotirus, with karyotype similar to that of La io., 2n=46 and N.F=50 in Miniopterus schreibersi; 2n=44 and N.F=52 in Myotis fimbriatus; 2n=38 and N.F=48 in Vespertilio superans 2n=26 and N.F=44 in Pipistrellus abramus, 2n=62 and N.F=60 in Rhinolophus affinis, 2n=58 and N.F=60 in Rhinolophus ferrumequinum respectively. Of the 7 species bats, the karyotypes of E. serotirus and R. ferrumequinum were reported for the first time.6. RAPD analysis of 7 species of Vespertilionidae bats suggested that they were divided into four groups: M. schreibersi, P. abramus and Murina leucogaster belonged to I, II andⅢgroup, respectively, while E. serotirus and three species of Myotis belonged to the fourth group.The Myotis was an relative primitive group with close relationship with E.serotirus and distant relationship with M. schreibersii. RAPD analysis of 3 Rhinolophus bats also showed that Rhinolophus cornutus was an older group, which had nearer relationship with R. affinis than with R. ferrumequinum. To some extent, the divergence occurred among the bats of same species but different locations. Their genetic polymorphism was obviously higher than that of bats of same species and same locations. The divergence also existed among individual bats of same species and same locations. However, almost no divergence was found among the bats within the same hibernation group.7. The expression patterns of four isozymes in eight tissues were obviously different. EST exhibited polytypism and higher activities in kidney, liver and small intestine. LDH showed tissue specificity, with the same expression pattern as that of most of mammalian in which the B-subunit was abundant in oxygen consumption tissue, such as chest muscle and kidney, while the A-subunit was abundant in liver. MDH showed tissue specificity either. Based on differential staining intensity and the bands distribution of s-MDH and m-MDH from eight tissues in electrophoresis gel, it could be concluded that m-MDH (mitochondria MDH) was dominant in bats, and this was closely related with flying adaptation and energy consumed of Chiroptera. Of all the eight tissues, ME in muscle displayed diversity and the strongest activity. Isozymes electrophoretograms from congeneric bats showed a little difference in the number, location and intensity of bands. But the difference was big when comparing electrophoretograms from different genus bats.8. The phylogenic trees based on Cytb gene suggested that the relationship between family Vespertilionidae and Hipposideridae or Rhinolophidae was closer, but the relationship between family Hipposideridae and Vespertilionidae was further. Within the family Vespertilionidae the relationship between genus Murina and Myotis was closer but genus Miniopterus was further to the others. Henan M. schreibersii and Hannai M. schreibersii belonged to two different subspecies, while Nara specimen belonged to another subspecies. Among the five sampling sites over 600 km, there was no distinct gene differentiation. In the M. schreibersii population level, the value of haplotype proportion, frequency and diversity were high, which indicated that there was widely gene communication among these sites.更多还原