分子标记在家畜遗传多样性研究中的应用

家畜遗传多样性是生物多样性的重要组成部分,与人类未来的生存与发展密切相关。中国拥有丰富的地方家畜品种资源,但由于保种不当和盲目引进外来品种进行杂交、资金缺乏等原因,许多品种已濒临灭绝。如何保存利用好我国优良的家畜遗传资源已成为越来越严峻的问题。随着生物学技术的不断发展与完善,许多分子标记技术已经应用于畜禽遗传资源的研究。简要介绍了在家畜遗传多样性研究中应用最广的几种分子标记及其特性,讨论了在家畜遗传多样性的研究中如何选用适当的分子标记。     

发育工程与家畜改良

二十世纪七十年代后胚胎移植几乎引起一场家畜繁殖技术的革命,并波及到胚胎学、发育生物学、免疫学、遗传学等基础学科。随之产生了一些新尖的技术与之相互配合,形成了熔各种知识和技术手段于一炉,瞄准应用的研究领域——发育工程。本文集中讨论胚胎移植、冷冻、性别鉴定、卵母细胞体外受精、受精卵核移植和嵌合体及转基因动物制作等发育工程技术的兴起及其对家畜遗传改良所产生的积极影响。     

中国6个国家级保护鸭种群遗传多样性分析

为探讨国家级保护鸭种群的遗传多样性和遗传分化关系,利用17个微卫星标记,对我国6个国家级保护鸭品种资源的遗传多样性进行了分析,统计了平均遗传杂合度(H)、多态信息含量(PIC)、基因多样度(FST)、平均遗传分化系数(GST)和遗传距离等。结果表明,各鸭品种的杂合度较高,除两个群体表现为显著的杂合子缺失外,其他群体均处于Hard-Weinberg平衡状态。6个鸭种群间平均FST值为17.0%,平均遗传分化系数GST为14.7%,各品种平均杂合度为0.706～0.604,平均多态信息含量(PIC)为0.561～0.663。本研究说明我国6个国家级保护鸭品种资源各品种内和品种间的遗传变异大,遗传多样性丰富。     

中国地方黄牛的遗传多样性*

中国黄牛起源复杂,我国地方黄牛群体不同品种在毛色、形态外貌、细胞遗传学、血液蛋白座位分析均表现出多样性。计算我国黄牛群体6个毛色座位平均杂合度和6个血液蛋白座位平均杂合度分别为0.3144和0.4873,表明我国地方黄牛群体的遗传多样性非常丰富。计算我国黄牛群体的6个毛色座位和6个血液蛋白座位的基因分化系数分别为0.3404和0.095,表明我国黄牛群体毛色差异中有34.04％是由品种间的差异造成的。血液蛋白的多态性有9.5％是由品种间的差异造成的。我国黄牛群体的遗传多样性主要来自品种内的遗传多样性。保存我国黄牛品种资源多样性不仅要从整个中国黄牛群体上考虑,而且要针对每个品种（或类群）进行保种。     

红花品种资源的同工酶遗传多样性及分类研究

本研究采用同工酶遗传标记对89份红花品种的遗传多样性进行了评价和分类研究。在4种酶,即酯酶(EST)、乙醇脱氢酶(ADH)、酸性磷酸酶(APC)和莽草酸脱氢酶(SKDH)的分析中,共解释出8个同工酶基因位点,包括25个等位基因。在8个同工酶位点即ESTl、EST2、ADH1、ADH2、APCI、APC2、APC3和SKDH1中,7个呈现多态性,占87．5％,只有ADH2在所有材料中出现一致性。在25个等位基因中,15个表现出多样性,占60％。这些等位基因的平均分布频率在东亚材料中最高,为0．013。分布频率最低的是中东和土耳其的材料,只有0．009。同工酶位点杂合性平均为0．54,但位点间有很大差异,从0．16～0．87不等。利用聚类分析对89份红花品种进行了分类,形成4个组群。分类结果表明,印度材料多样性丰富,各组群均有分布。土耳其的材料与中东国家的材料关系密切;未知来源材料可能主要来自印度、土耳其和中东国家。     

家畜试验中的一种相关扰动回归模型

本文根据数量遗传学原理,从家畜群体中同一家系内的个体之间存在的遗传相关性出发,建立了扰动项相关的回归模型,运用回归理论讨论一类相关样本的统计推断及其应用。     

SSR标记揭示的云南地方稻品种遗传多样性及其保育意义

为了探索水稻(Oryza sativa L.)地方品种的遗传多样性及其有效保育方法,对采自云南省17个村寨的82个水稻地方品种和3个国际常用的典型籼稻和粳稻品种进行了微卫星(SSR)分子标记的分析.利用19对SSR引物在85个水稻品种中共扩增出了83个基因型,其分子量变异在100～500 bp之间.基于各品种SSR基因型遗传相似系数聚类分析而获得的UPGMA树状图表明各水稻品种之间存在较大的遗传多样性,其相似系数变异在0.15～0.90之间.但这些地方品种的遗传多样性并非呈均等的地理分布.这85个水稻品种在相似系数为0.52之处分为二组,其中一组包括几乎所有的籼稻品种,而另一组包括全部的粳稻品种,表明SSR标记能很好揭示水稻籼-粳分化.同时,有些来自不同采集地的同名品种表现出一定的遗传差异,说明同名异物的现象存在.云南水稻地方品种具有丰富的遗传多样性,对其有效保育十分重要和迫切,但只有根据遗传多样性的水平和分布特点,采用正确的保育对策和取样方法才能确保对云南水稻地方品种的有效保育.结果进一步表明,选用适当的微卫星引物,可以为准确鉴定籼稻和粳稻品种及研究其进化规律提供有效的分子标记方法,并有利于有目标的水稻遗传资源保育和育种创新.     

桂花品种资源的遗传多样性分析

根据桂花（Osmanthus fragrans）品种的形态特征,结合AFLP分子标记,对部分桂花栽培品种进行了遗传多样性分析。结果表明,桂花品种之间存在着较为丰富的遗传多样性,AFLP分子标记检测到的多态性条带占总扩增条带的57.46%。根据桂花品种的主要性状特征,利用数值分类法对其进行分类,并用UPGMA法对AFLP结果进行聚类分析,结果均显示桂林地区的桂花品种存在明显的区域性,花色可以作为重要的分类标准,同时对桂花品种的分类系统进行了探讨。     

桂花品种资源的遗传多样性分析

根据桂花(Osmanthus fragrans)品种的形态特征, 结合AFLP分子标记, 对部分桂花栽培品种进行了遗传多样性分析。结果表明, 桂花品种之间存在着较为丰富的遗传多样性, AFLP分子标记检测到的多态性条带占总扩增条带的57.46%。根据桂花品种的主要性状特征, 利用数值分类法对其进行分类, 并用UPGMA法对AFLP结果进行聚类分析, 结果均显示桂林地区的桂花品种存在明显的区域性, 花色可以作为重要的分类标准, 同时对桂花品种的分类系统进行了探讨。     

家马的驯化起源与遗传演化特征

人类文明发展历史中, 家马(Equus ferus caballus)曾是推动文化交流、促进人类社会发展的主要动力。关于家马何时、何地被驯化以及在此过程中其遗传演化如何被人类影响等一直备受关注。近年来随着遗传学技术的发展, 人们对该问题有了更为深入的理解。本文回顾了近二十年来相关研究所取得的成果, 探讨了家马的驯化起源中心和驯化过程中的遗传演化特征, 并对未来的研究方向以及遗传资源保护提出了建议。分子标记遗传学和考古学研究认为家马可能来自多个驯化起源地种群, 然而最近的古DNA研究结果表明, 现代家马的驯化起源可能比之前人们所猜测的更加复杂, 古代博泰马被认为是最早被驯化的家马, 然而最近被证实并不是现代家马的直系祖先。如此复杂的驯化问题可能从多学科的层次才能解析清楚。人类社会活动直接或间接影响了家马的演化历程, 特别是工业革命以来家马的遗传基础发生了巨大变化, 其遗传多样性开始急剧衰退, 不少地方品种正逐渐走向衰落甚至灭绝。为确保农业生态安全不受威胁, 建议加强家马遗传资源保护与动物遗传学和文化地理之间的联系研究。     

Individual assignment using microsatellite DNA reveals unambiguous breed identification in the domestic dog

Modern individual clustering methods utilising hypervariable nuclear microsatellite DNA polymorphisms are being increasingly applied in the field of population genetics. This study explores the efficiency of the clustering methods in identifying the breeds of origin of 250 domestic dog (Canis familiaris) individuals based on 10 microsatellite loci. An allele sharing distance (DAS) matrix and the corresponding neighbour-joining tree of individuals revealed monophyletic assemblages that corresponded perfectly with the breeds of origin of the dogs. Individual assignment tests using a Bayesian statistical approach, an allele frequency based method, and a DCE genetic distance based method were all extremely powerful. Most strikingly, the Bayesian method provided 100% assignment success of individuals into their correct breeds of origin and 100% exclusion success of individuals from all alternate reference populations with a high level of statistical confidence (P < 0.0001). A Bayesian Markov Chain Monte Carlo clustering approach revealed clear distinction of individuals into groups according to their breeds of origin, with a near-zero level of 'genetic admixture' among breeds. The results demonstrate that an FST of 0.18, mean expected gene diversity of 0.6 across 10 loci, and approximately 50 individuals per reference population suffice to provide maximum individual assignment success in C. familiaris. This refutes the traditional view that DNA based dog breed identification is not feasible at the individual level of resolution.     

边际多样性方法及其在绵羊品种保护中的应用

本文介绍了边际多样性方法,并利用微卫星DNA遗传距离、灭绝概率等结果分析了中国北方11个绵羊品种的品种贡献率、边际多样性及保护潜力。结果表明,苏尼特羊(Sunite sheep)的边际多样性最高（－0.2008）,其次为滩羊(Tan sheep)(－0.1932)、兰州大尾羊(Lanzhou large tailed sheep)(－0.1843);岷县黑裘皮羊(Minxian black fur sheep)最低（－0.1268）。保护潜力最大的品种为兰州大尾羊（0.1419）,其次为同羊(Tong sheep)（0.1017）,最小的为小尾寒羊(Small tailed Han sheep)（0.0365）。根据边际多样性方法,作者认为确定需要保护的品种应该依据最大保护潜力而不是品种的濒危程度。11个绵羊品种的保护顺序依次是兰州大尾羊、同羊、汉中羊(Hanzhong sheep)、甘家羊(Ganjia sheep)、欧拉羊(Oula sheep)、岷县黑裘皮羊、乔科羊(Qiaoke sheep)、苏尼特羊、乌珠穆沁羊(Ujumqin sheep)和小尾寒羊。     

Is That Dog a Pit Bull? A Cross-Country Comparison of Perceptions of Shelter Workers Regarding Breed Identification

Bull breeds are commonly kept as companion animals, but the pit bull terrier is restricted by breed-specific legislation (BSL) in parts of the United States and throughout the United Kingdom. Shelter workers must decide which breed(s) a dog is. This decision may influence the dog's fate, particularly in places with BSL. In this study, shelter workers in the United States and United Kingdom were shown pictures of 20 dogs and were asked what breed each dog was, how they determined each dog's breed, whether each dog was a pit bull, and what they expected the fate of each dog to be. There was much variation in responses both between and within the United States and United Kingdom. UK participants frequently labeled dogs commonly considered by U.S. participants to be pit bulls as Staffordshire bull terriers. UK participants were more likely to say their shelters would euthanize dogs deemed to be pit bulls. Most participants noted using dogs' physical features to determine breed, and 41% affected by BSL indicated they would knowingly mislabel a dog of a restricted breed, presumably to increase the dog's adoption chances.     

Genetic variation (protein markers and microsatellites) in endangered Catalonian donkeys

Genetic variation of the endangered Catalonian donkey breed (Equus asinus) has been analysed at 19 loci including seven protein loci and 12 microsatellite loci isolated from the domestic horse, in 98 individuals of both sexes. Only four protein markers and three microsatellites were polymorphic. Allele frequencies of the analysed loci showed close agreement with Hardy–Weinberg proportions, with the exception of the MPZ002 locus (P<0.01). The within-population inbreeding estimate was not significantly different from zero (as measured by F_IS-statistic). The cumulative-exclusion probability for all polymorphic loci was 82.9%, this value still being very low so that these markers could efficiently be utilised for verification of parentage.     

Animal genetic resources and domestic animal diversity in Nepal

Domestic livestock in Nepal belong to 17 species, including five Bovidae (and interspecific hybrids of two of them), seven Aves, two Equidae (not including mules), pig (Sus scrofa), rabbit (Oryctolagus cuniculus) and elephant (Loxodonta africana). Cattle (Bos taurus and Bos indicus) are the most numerous quadruped, followed by goats (Capra hircus), buffalo (Bubalus bubalis) and sheep (Ovis aries). Domestic fowl (Gallus domesticus) are the most common poultry, followed by pigeons (Columba livia) and ducks (Anas platyrhyncos). Government policy is to upgrade and replace local types with improved stock. This policy has had limited success through lack of funding, inconsistent long-term objectives and an inability to pursue programmes in traditional farming systems. The purity of native types is being lost in some areas due to informal programmes by farmers themselves. Except for one type of goat there are no conservation programmes for native breeds. Wild relatives of livestock, including arnee (Bubalus arnee), gaur (Bibos gaurus), wild boar (Sus scrofa), jungle fowl (Gallus gallus) and rock dove (Columba livia), are being infiltrated by genes of domestic animals. Little has been done to characterize Nepal's livestock resources yet continuous attempts have been made to replace them. Conservation needs are to: (i) identify all genetic resources comprising the gene pool of species used as domestic livestock; (ii) develop and use the associated diversity to increase production, achieve sustainable systems and meet demands for particular products; (iii) monitor resources represented by small numbers or that are being displaced; and (iv) preserve resources not in current demand. A start has been made on this by Nepal's membership of a regional network.     

织金瘰螈生物学特性及保护措施

织金瘰螈Paramesotriton zhijinensis是2008年才发现的中国特有珍稀两栖动物.发现于贵州省织金县,它是世界上最稀少的两栖动物之一,在世界其它各地至今还没有被发现,也是被关注和研究最少的濒危动物之一.由于生境的逐渐丧失、环境的恶化、人为的严重干扰等诸多因素导致其种群数量急剧减少.以现在的种群数量和消亡速度来看,如不善加保护,它很可能不久就会灭绝.本文对织金瘰螈的分布区域、栖息环境、体型特征、生活习性、食性及种群资源状况进行了简单分析,希望能为该物种的资源保护提供重要的理论依据.     

Genetic distances within and across cattle breeds as indicated by biallelic AFLP markers

We tested the use of biallelic Amplified fragment length polymorphism (AFLP) polymorphisms for the estimation of relative genetic distances of cattle individuals within or across breeds. An allele permutation procedure was developed to estimate the stochastic variation of the genetic distance that is inherent to a given dataset. In a panel of 47 Holstein-Friesian cattle analysed with 248 polymorphic markers, the average genetic distance of bulls selected for breeding was slightly lower than the distance of the cows. The observed standard deviation (SD) of the distance indicated genetic subdivision, which for the bulls was explained by variation in the additive relationship derived from herdbook data. Animals from three different breeds, the highly selected Holstein-Friesian, the Italian Brown and the historic Maremmana, were compared on the basis of 106 polymorphic markers. No breed-specific fragments were observed. The mean pair-wise genetic distance within breeds was 85% of the value across breeds, but principal coordinates analysis clustered the animals according to their breed of origin. Calculation of distances between the breeds indicated a relatively divergent position of the Maremmana, relative to the two other breeds. However, biallelic markers indicate that the process of breed formation had only a limited effect on the diversity at marker loci.