西藏马鹿（Cervus wallichii）分子生态学与营养生态学研究

发布时间：2018-04-05 20:46

  本文选题：西藏马鹿　切入点：分子生态学　出处：《东北林业大学》2016年博士论文

【摘要】：目前,西藏马鹿(Cervus wallichii)是我国特有、濒危、珍贵鹿类动物,主要分布于西藏山南地区桑日县境内。该物种曾一度被国际组织认定为已经灭绝的物种,上个世纪90年代被中外科学家在野外调查时重新发现。本研究于2013年和2014年(8月初-9月末)连续两年秋季,采用非损伤取样法——粪便样品,对西藏马鹿的遗传多样性、种群性比及数量、食物组成、食物选择、营养成分测定以及生理健康评价等内容进行了研究,获得如下主要成果：1、从野外采集的199份西藏马鹿粪便样品中,利用非损伤取样法成功提取出了87份DNA分子片段。对12个多态性高,无紧密连锁关系的微卫星位点进行了PCR分析,并进行了个体识别,最终识别出的87份粪便样品分属于50只西藏马鹿个体。对这50只个体性别鉴定结果为：雌：雄≈1：0.79,表明西藏马鹿种群的性别比例雌性个体数量多于雄性个体数量,有利于西藏马鹿种群的发展。2、运用R软件Capwire包中的TIRM和ECM两种模型进行模拟得到的结果分别别为：TIRM模块得到西藏马鹿数量为87只,97.5%的判别率下的置信区间为87～90只；ECM模块得到的西藏马鹿数量为65只,97.5%的判别率下的置信区间为65～67只。利用该软件包的似然比测试得到的P值不显著偏离0,说明调查区域内的不同的个体的捕捉率不同,因此以模块TIRM结果为种群数量估计值。本次粪便采样覆盖面积(即调查抽样面积)为264.5km2,在97.5%的判别率下马鹿种群数量置信区间为87～90只,密度为0.329～0.340只/km2。本次实地调查确认：西藏马鹿目前仅分布于面积2680.6 km2的西藏桑日马鹿自然保护区内。西藏马鹿实际适栖面积为650.4 km2,其余2030.2km2为白唇鹿(C. albirostris)的主要分布区。故2013～2014年秋季西藏马鹿种群总数量约为214～221只。3、通过50只个体基因型数据统计指出,西藏马鹿种群平均等位基因数为7.58±0.18个,有效等位基因数平均为4.91±0.16个,单个微卫星座位的PIC在0.39～0.90之间,平均值为0.67±0.013,12个座位中只有位点T123为中度多态性位点,其他11个位点均为高度多态性位点。期望杂合度变化范围为0.45～0.91,平均为0.72±0.01；平均表现杂合度为0.52±0.11。遗传分析数据显示,目前西藏马鹿的遗传多样性较高,种群不会由于自身遗传因素而导致衰退。4、通过粪便显微切片技术分析得出西藏马鹿秋季采食的植物共有37种,隶属于18个科,26属,采食比率较高的依次为菊科(21.62%)、豆科(13.51%)、禾本科(13.51%)、蔷薇科(8.11%)；对西藏马鹿所食植物中的水分(W)、粗蛋白(CP)、粗脂肪EE)、能量(E),纤维素(C)和单宁(T)等物质含量测定结果：水分(62.13%)、粗蛋白(11.06%)、粗脂肪(8.88%)、能量(17812.53J/g)、纤维素(22.86%)、单宁(0.14%)。5、选择食物组成、能量、纤维素和蛋白质4个主要营养成分为因变量,建立响应曲面模型,分析结果显示西藏马鹿偏向蛋白质和能量高的食物,对蛋白质高纤维素含量低,能量高纤维素含量低的植物有一定的选择性,但是并未出现明显的回避,分析认为西藏马鹿秋季取食策略为食物营养均衡型策略。此外,还通过对西藏马鹿粪便样品中氮含量与秋季主要采食的20种植物的平均蛋白含量的测定结果,初步探讨了鹿粪氮含量与可食植物蛋白质含量之间的关系。6、采用沉淀法和漂浮法对西藏马鹿粪便样品中寄生虫含量进行了测定；利用酶联免疫法对粪便样品中的免疫球蛋白进行了测定。结果显示西藏马鹿粪便中寄生虫感染率低,免疫球蛋白较圈养梅花鹿含量低,说明西藏马鹿生理健康状态良好。综上所述,尽管我国西藏马鹿种群遗传多样性较高,营养状态良好,但鉴于目前分布面积狭窄,种群数量稀少,当地有关部门应尽快加大该物种目前的唯一分布地——西藏桑日马鹿自然保护区的建设投资和保护管理力度,以利于这一特有、濒危、珍贵鹿科动物种群数量的迅速恢复。
[Abstract]:At present, Tibet red deer (Cervus wallichii) is endemic to China, endangered, rare deer animal, mainly in the territory of SANGRI County Shannan Prefecture of Tibet. This species has been identified as the international organization of extinct species, the last century in 90s by the Chinese and foreign scientists to found in field investigation. This research in 2013 and 2014 (-9 at the beginning of August at the end of the month) for two consecutive years in autumn, using noninvasive sampling, faecal samples, genetic diversity of Tibet red deer, the number of population sex ratio and food composition, food selection, determination of nutrition and physical health evaluation content of the study, obtained the main results as follows: 1, from the 199 Tibet red deer feces the samples collected in the field, using noninvasive sampling successfully extracted 87 DNA fragments. 12 highly polymorphic microsatellite loci without linkage analysis of PCR and For the individual identification, 87 fecal samples eventually identified belong to 50 of the 50 Tibet wapiti individuals. Only individual gender identification results: female: male = 1:0.79, showed that the sex ratio of Tibet wapiti population female more than male individual quantity, is conducive to the development of Tibet.2 horse deer populations, the use of R Capwire software package TIRM and ECM two models of the results obtained by simulation respectively: TIRM module to get Tibet wapiti number 87, 97.5% confidence interval discriminant rate under 87~90; the number of Tibet wapiti ECM module was obtained 65, 97.5% confidence interval discriminant rate under 65~67 the likelihood ratio test. By using software package P values did not deviate significantly from 0, that in the area of different individual capture rate is different, so the module of the TIRM results for population estimates. The stool samples cover The area (i.e. sampling area) for 264.5km2, in the deer population dismount discrimination rate confidence interval 87~90 97.5%, density of 0.329 ~ 0.340 /km2., the survey confirmed: Tibet red deer is currently only distributed in the area of 2680.6 km2 of the Tibet nature reserve. Red deer, Tibet red deer habitat suitable actual area is 650.4 km2, the rest of the 2030.2km2 (C. albirostris) for Bai Chunlu the main distribution area. The 2013~2014 autumn Tibet wapiti population the total number of about 214~221.3, the 50 individual genotype data statistics show that the Tibet red deer population, the average number of alleles was 7.58 + 0.18, the average effective number of alleles was 4.91 + 0.16 a single, microsatellite loci of PIC in 0.39 ~ 0.90, an average of only 0.67 sites of T123 + 0.013,12 seats for site of moderate polymorphisms, the other 11 loci were highly polymorphic loci. 鏈涙潅鍚堝害鍙樺寲鑼冨洿涓，

本文编号：1716391