發(fā)布時(shí)間：2018-02-01 00:50

  本文關(guān)鍵詞： 微衛(wèi)星 林麝 遺傳多樣性 磁珠富集法　出處：《西北農(nóng)林科技大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：林麝(Moschus berezovskii)是我國(guó)重要的經(jīng)濟(jì)動(dòng)物,因其所產(chǎn)麝香有極高的經(jīng)濟(jì)價(jià)值而被大量捕殺,導(dǎo)致種群數(shù)量銳減,2002年被列為我國(guó)Ⅰ級(jí)重點(diǎn)保護(hù)的珍稀瀕危野生動(dòng)物。目前,開(kāi)展人工養(yǎng)麝是保護(hù)林麝野生資源的重要手段之一,但圈養(yǎng)林麝又帶來(lái)了新的問(wèn)題,遺傳相關(guān)問(wèn)題尤為突出。因林麝保護(hù)級(jí)別高,分布區(qū)域有限,樣品獲取較難,目前有關(guān)林麝的研究報(bào)道較少。為豐富林麝遺傳學(xué)有關(guān)數(shù)據(jù),同時(shí)為麝場(chǎng)的養(yǎng)殖管理提供資料,本文開(kāi)展了以下研究:運(yùn)用磁珠富集法構(gòu)建林麝微衛(wèi)星富集文庫(kù),從文庫(kù)中篩選多態(tài)微衛(wèi)星位點(diǎn);用8個(gè)林麝多態(tài)微衛(wèi)星位點(diǎn)分析陜西某麝場(chǎng)圈養(yǎng)林麝群體的遺傳多樣性,并對(duì)所采40個(gè)林麝樣進(jìn)行UPGMA聚類(lèi)分析。所得主要結(jié)果如下:1.采用磁珠富集法構(gòu)建林麝(AC)n微衛(wèi)星文庫(kù),獲得16個(gè)微衛(wèi)星位點(diǎn),其中完全型微衛(wèi)星8個(gè),占50%,不完全型微衛(wèi)星7個(gè),占43.75%,復(fù)合型微衛(wèi)星1個(gè),占6.25%。2.設(shè)計(jì)14個(gè)林麝微衛(wèi)星位點(diǎn)引物,經(jīng)篩選發(fā)現(xiàn)3個(gè)多態(tài)的林麝微衛(wèi)星位點(diǎn),可用于林麝群體遺傳多樣性分析;1個(gè)林麝和馬麝的種間鑒別位點(diǎn)。3.圈養(yǎng)林麝種群的遺傳多樣性分析結(jié)果顯示,8個(gè)林麝微衛(wèi)星位點(diǎn)的多態(tài)信息含量在0.679~0.870之間,平均多態(tài)信息含量為0.798;共檢測(cè)到68個(gè)等位基因,平均等位基因數(shù)為8.5,平均有效等位基因數(shù)為5.8770;圈養(yǎng)林麝種群的平均觀測(cè)雜合度為0.5428,平均期望雜合度為0.8337,平均Shannon’s信息指數(shù)為1.8971�？ǚ綑z驗(yàn)結(jié)果顯示,除Mber76C位點(diǎn)外,其余7個(gè)微衛(wèi)星位點(diǎn)偏離Hardy-Weinberg平衡,其中2個(gè)位點(diǎn)雜合子過(guò)剩,5個(gè)位點(diǎn)雜合子缺失。8個(gè)微衛(wèi)星位點(diǎn)共檢測(cè)到14個(gè)稀有等位基因,占等位基因總數(shù)的20.59%。4.40個(gè)圈養(yǎng)林麝樣品的聚類(lèi)分析結(jié)果表明,群體的相似系數(shù)在0.72~0.97之間,整個(gè)群體可分為9類(lèi),同一類(lèi)群個(gè)體數(shù)最多為22個(gè),最少為1個(gè)。本研究豐富了林麝微衛(wèi)星標(biāo)記數(shù)據(jù),分析發(fā)現(xiàn)陜西麝場(chǎng)圈養(yǎng)林麝群體的遺傳多樣水平較高,但為了麝場(chǎng)長(zhǎng)遠(yuǎn)的發(fā)展,仍需不斷提高麝場(chǎng)的養(yǎng)殖管理水平,建立完善的群體遺傳譜系,實(shí)時(shí)進(jìn)行群體遺傳監(jiān)測(cè),避免種群退化的風(fēng)險(xiǎn)。
[Abstract]:Moschus berezovskii (Moschus berezovskii) is an important economic animal in China. Because of its high economic value, Moschus berezovskii has been killed in large numbers, resulting in a sharp decrease in population. In 2002, it was listed as a rare and endangered wild animal in China. At present, artificial breeding of musk deer is one of the important means to protect the wild resources of forest musk deer, but the captive forest musk deer has brought new problems. Because of the high protection level and limited distribution area, it is difficult to obtain samples, so there are few reports about forest musk deer. It is to enrich the genetic data of forest musk deer. At the same time, the following studies were carried out for the culture management of musk deer farm: the microsatellite enrichment library of forest musk deer was constructed by magnetic bead enrichment method, and the polymorphic microsatellite loci were screened from the library; The genetic diversity of captive forest musk deer populations in a musk deer farm in Shaanxi Province was analyzed by using 8 loci of microsatellite polymorphism of forest musk deer. The main results were as follows: 1. The microsatellite library of ACN was constructed by magnetic bead enrichment method, and 16 microsatellite loci were obtained. There were 8 complete microsatellites (50 cents), 7 incomplete microsatellites (43.75) and 1 compound microsatellite (6.25.2.). Fourteen primers were designed for the microsatellite loci of forest musk deer. Three polymorphic microsatellite loci were identified, which can be used to analyze the genetic diversity of forest musk deer populations. The results of genetic diversity analysis showed that the polymorphic information content of microsatellite loci of 8 forest musk deer was between 0.679 and 0.870. The average polymorphic information content was 0.798; 68 alleles were detected, the average number of alleles was 8.5 and the number of effective alleles was 5.8770; The average observed heterozygosity, expected heterozygosity and Shannon's information index were 0.5428, 0.8337 and 1.8971 respectively. The chi-square test results showed that the observed heterozygosity was 0.5428, the average expected heterozygosity was 0.8337, and the average Shannon's information index was 1.8971. Except for Mber76C, the other 7 microsatellite sites deviated from the Hardy-Weinberg equilibrium, and 2 of them had excess heterozygotes. A total of 14 rare alleles were detected in 8 microsatellite loci, accounting for 20.59% of the total alleles, and the cluster analysis of 4.40 captive forest musk deer samples showed that. The similarity coefficient of the population was between 0.72 and 0.97, the whole population could be divided into 9 categories, the number of individuals in the same group was 22 at most, and at least one. This study enriched the microsatellite marker data of forest musk deer. It was found that the genetic diversity of the population in the captive forest of Shaanxi musk deer farm was high, but in order to develop the musk deer farm for a long time, it was still necessary to improve the management level of the musk deer farm and establish a complete genetic pedigree of the population. Population genetic monitoring is carried out in real time to avoid the risk of population degradation.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S865.41
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本文編號(hào)：1480461