我国主要地方绵羊品种随机扩增多态DNA研究

对蒙古羊、湖羊、滩羊、小尾寒羊、乌珠穆沁羊、藏绵羊、阿勒泰羊7个地方绵羊品种和无角陶赛特羊、德国美利奴羊、萨福克羊3个引入品种基因组DNA进行了RAPD分析。结果表明：（1）RAPD可作为一种有效的标记用于绵羊品种之间遗传亲缘关系的分析。（2）在所使用的43种随机引物中,有35种引物扩增出多态谱带,多态频率为66.24%,说明RAPD技术用于研究绵羊核DNA的遗传变异具有较高的检出率和灵敏度。（3）总群体平均遗传多样性指数（HSP）为0.9139,说明绵羊群体具有较为丰富的遗传多样性。（4）我国地方绵羊品种间的分子聚类关系与其所处的地理位置、考古学结果,以及细胞遗传学研究结果基本一致,引入品种间的分子聚类关系也与其育成史基本一致。 Abstract:The genetic polymorphism and relationship of 7 indigenous sheep breeds of China and 3 imported sheep breeds were studied using random amplified polymorphic DNA (RAPD).The results indicated that the RAPD was an effective marker for the analysis of genetic relationship among sheep breeds.Among 43 arbitrary primers,35 were polymorphic.The percentage of polymorphic markers was 66.24%,which indicated that the RAPD had higher efficiency of polymorphism detection and sensitivity in studying the genetic variation among sheep breeds.The average index of genetic polymorphism for whole population (Hsp) was 0.9139,which showed that the genetic polymorphism was abundant between sheep populations.The genetic relationship between different indigenous sheep breeds in China was in accord with their localities,the results from archeology and cytogenetics and the genetic relationship between imported sheep breeds was in accord with their breeding history.     

RAPD analysis of natural populations of Acanthopanax brachypus

Random amplified Polymorphic DNA(RAPD) analysis is a new technology of molecular marking which has proved very powerful in detecting genetic diversity at the level of population.The genomic DNAs used in our experiment were extracted from fresh leaves taken from 59 individuals sampled from three natural populations in Yan an,Shanxi Province.Through more than 2,000 PCRs,deep-going RAPD analysis was carried out on DNA samples from 49 inviduals.The percentage of polymorphic RAPD loci found in these three populations were respectively 27.2%,18.6% and 5.4%;the average genetic distances within population,0.055,0.026 and 0.008;the average genetic distances between populations (I-II),(I-III) and (II-III),0.105,0.096 and 0.060.The genetic diversity of A.brachypus within and between populations was found,for the first time,to be rather poor,thus revealing innate factors as the cause contributing to its endangered status.In addition,our work also provides basic materials for elucidating the underlying cause of its endangerment and for its protection biology.     

Random amplified polymorphic DNA analysis of eel genome

Eel family is a huge one, in which many kinds of eels especially some migratory eels, bear strong resemblance to each other, and are therefore difficult to be identified. In this study 29 random primers were used to make RAPD analysis for Japaneses eel (Anguilla japonica), European eel (Anguilla anguilla) and Pike eel (Muraenesox cinereus).And totally 299 fragments were counted.Shared or specific fragments were counted and genetic similarity or genetic distance were calculated.The genetic similarity between Japanese eel and Pike eel is 0.68 and the genetic distance between them is 0.32;those between European eel and Pike eel are 0.72 and 0.28 respectively,and between Japanese eel and European eel are 0.74 and 0.25 respectively.The method has been shown to be suitable to molecular identification of eels.It provides an alternative approach to determine the relationship between species.     

Use of Random Amplified Polymorphic DNA Analysis for Economically Important Food Crops

The objective of this review is to summarize numerous studies on the use of the random amplified polymorphic DNA （RAPD） technique on rice, corn, wheat, sorghum, barley, rye, and oats to examine its feasibility and validity for assessment of genetic variation, population genetics, mapping, linkage and marker assisted selection, phylogenetic analysis, and the detection of somaclonal variation. Also we discuss the advantages and limitations of RAPD. Molecular markers have entered the scene of genetic improvement in different fields of agricultural research. The simplicity of the RAPD technique made it ideal for genetic mapping, plant and animal breeding programs, and DNA fingerprinting, with particular utility in the field of population genetics.     

用DNA指纹图技术分析Wistar大鼠的遗传距离

采用JL-02多位点探针和Southern杂交对Wistar大鼠基因组进行了DNA指纹分析,并对国内最具有典型的6个地区9个Wistar大鼠群体内和群体间进行了DNA指纹分析比较。结果表明,DNA指纹图较好地反映了封闭群动物的遗传本质,具有良好的多态性。同一群体不同个体间Wistar大鼠遗传距离主要为0.2~0.6,将不同群体的Wistar大鼠DNA指纹图带进行分析表明,所有群体间的DNA指纹图的遗传距离在0.2~0.7。 Abstract:DNA fingerprinting of Wistar rat were studied with JL-02 Mulilocus probe and Southern hybridization,and comparing with different individuals of nine groups Wistar rat from six national classical area and different groups.It was indicated that DNA fingerprinting could reflect genetic material of outbred strain rat,and were more polymorphic.The genetic distances of Wistar rat were distributed for 0.2~0.6 among different individuals within same groups,and the genetic distances of Wistar rat were distributed for 0.2~0.7 among different groups．     

用整合微流控芯片系统分析黄山松RAPD片段多态性

随机扩增多态DNA（randomly amplified polymorphic DNA,RAPD）标记可快速提供连锁信息,特别是在针叶树单倍体的大配子体中RAPD基因型也能得以确定［3］。在对多态性的RAPD片段进行分析时,传统的平板凝胶电泳分析法因人工操作,其有效的鉴别受到一定程度的限制,只能得到一些有关RAPD片段半定量信息。我们将整合微流控芯片系统作为一种新的工具运用于黄山松的多态性RAPD片段分析中。发现基于芯片的检测方法比琼脂糖凝胶电泳方法灵敏度更高,所需的样品量要少得多,所需的时间仅为其1/4。并且能自动对DNA片段进行定性、定量分析。它是一种高效、灵敏、迅速、重复性好的检测新技术。 Abstract:Randomly amplified polymorphic DNA (RAPD) markers quickly provide linkage information［1~2］,especially in conifers where haploid megagametophytes can be used for genotyping［3］.Traditionally use of slab gel electrophresis results in qualitative data that can be manually manipulated to gain semiquantitative information about the polymorphic RAPD fragments.We have proposed the use of an integrated microfluidic chip-based system as a new tool in the analysis of polymorphic RAPD fragments.The chip-based method was found to be very sensitive,requiring much less sample and only quarter the time compared to the agarose gel method.The automated data analysis sizes and quantitates the DNA fragments,thus yielding a more thorough,reproducible,sensitive,and rapid analysis.     

利用RAPD标记分析鸵鸟种群的遗传变异

为探讨分子遗传标记在鸵鸟(Struthiocamelus)育种中的应用,利用RAPD技术分析了我国养殖的三个主要鸵鸟品种内和品种间的遗传变异.蓝颈鸵鸟、澳洲灰和非洲黑三个品种在品种内和品种间显示出低水平的变异.品种间的遗传距离较好地说明了三个品种的育种改良过程,同时预示出用澳洲灰与其他品种鸵鸟进行杂交育种的可行性.选择的20条在鸡中能稳定扩增的RAPD引物其75%能在鸵鸟基因组中扩增出产物,显示出两种禽类DNA具有同源性,为今后利用鸡的微卫星引物对鸵鸟进行基因组扫描(genomescan)分析奠定基础。 Abstract:To evaluate the molecular genetic markers on ostrich (Struthio camelus)breeding,we have analyzed the genetic variability of three main ostrich breeds by using RAPD.The studies show that the Blue,Australia Grey and African Black have low genetic variability.The genetic distance between breeds clearly illustrates the breeding history,at the same time,it also showes the possibility that cross breeding can be applied to Australia Grey and the breeds.75 percent selected RAPD primers working well in chicken could stably amplify products in ostriches,which implied DNA identity between chicken and ostrich.The work provided a foundation for genomic scanning.     

PCR扩增近交系大鼠微卫星位点DNA多态性的研究

本实验选取大鼠7条染色体上的微卫星位点合成了10对引物,利用聚合酶链反应(PCR)扩增技术对国内北京和哈尔滨等4家单位提供的6个品系(SHR、SHRSP、LEW、RCS、WKY和F344)的8个近交系大鼠群体进行了DNA多态性分析的研究.结果表明:9个微卫星位点具有显著多态性;不同品系个体之间具有多态性;同一群体不同个体之间除SHR(哈)的SMST位点和WKY(哈)的AGT位点出现一定的差异外,其他均没有差异;不同地区同一品系的不同个体之间也存在一定的差异.该方法能有效地对近交系与杂交系、品系与品系、品系与亚系加以区分.因此,本实验为开展近交系大鼠遗传作图、基因定位和为实验动物的遗传背景监测提供可靠的信息,为大鼠遗传基因的研究提供了一个快捷简便、特异准确的方法。 Abstract:In the experiment,It were selected that 20 primers were assigned on 7 chromosomes of inbred rat.DNA polymorphism of 8 colonies from 6 inbred rat strains(SHR、SHRSP、WKY、LEW、RCS、F344)were studied in national Beijing and Harbin using PCR-analyzed microsatellites.The results indicated that there were remarkable polymorphism in 9 microsatellite loci;There is a polymorphism among the various rat strains;and no polymorphism in the same rat strains except SMST locus of SHR(Harbin)and AGT locus of WKY(Harbin)and there is a little difference among the same inbred rat strains in different areas.The method of PCR-analyzed microsatellites can be used for distinguishing between inbred and outbred、different strains、strain and substrain.and it provides a lot of information for genetic mapping,gene location and heredity probe of the inbred rat strains,and a speedy、convenient method for genetic research of the inbred rat.     

水稻一多拷贝微卫星DNA多态性分析

用一个多拷贝微卫星 DNA标记分析了238份栽培水稻的遗传多样性及遗传多样性从农家品种到现代栽培品种的动态变化。共检测出16种长度变异类型和32种表现型,但长度变异类型数、表现型数和表现型多样性水平在农家品种和现代栽培品种间没有显著差别。该标记在遗传资源评价和DNA指纹图谱建设中有重要的应用价值,也是研究环境或人工对多拷贝基因选择作用的一个良好模型。 Abstract:The genetic diversity of 238 cultivated rice and its difference between landraces and modern cultivars have been assayed using a multiple copy microsatellite DNA marker.Although 16 length variants and 32 phenotypes were found in all accessions,no significant differences in number of length variants,number of phenotypes and phenotypic diversity were detected between landraces and modern cultivars.This marker is of great value in evaluating genetic germplasm and constructing DNA fingerprints of rice and provides an elite model for tracing effectiveness of natural and artificial selection on multiple copy genes.     

Genetic diversity analysis by RAPD in Cathaya argyrophylla Chun et Kuang

Genetic diversity level of Cathaya argyrophylla was confirmed by random amplified polymorphic DNA (RAPD) markers. Seventy five samples (individuals), collected from Hunan and Sichuan provinces of China were used in the study. 21 10-mer oligonucleotide primers detected 106 sites, and 34 (32% ) of them were polymor-phic. The level of genetic variation in C. argyrophylla was lower than those of other conifers, and was considered to be associated with the complexity of habitats. The percentages of polymorphic sites (PPS) in the Hunan and Sichuan pop-ulations were 18% and 25% respectively. 7.99% of genetic variation existed between the two populations; this value was higher than the mean value (6.8%) among populations in conifers displayed by allozyme. Some subpopulations of C. argyrophylla were greatly differentiated because of site mutation and genetic drift. The highest value of genetic dif-ference between subpopulations amounted to 16. 23% . In addition, a concept of diversity coefficient(DC), a value us     

山羊经济性状标记辅助选择的遗传效应分析

本研究以辽宁绒山羊、柴达木绒山羊和柴达木山羊3个群体共147只山羊为研究对象,运用PAGE和RAPD技术对山羊的体重、绒产量和绒细度3个性状进行了与标记基因关系的遗传分析,结果表明：EsD2-2型、LAPBB型和PA-32-2型分别为体重、绒产量和绒细度性状的优势标记基因型;可利用标记辅助预测的方法充分利用多基因座标记基因间的互作效应;在体重上,寻找到有显著选择效应的RAPD条带11个,在绒产量和绒细度上分别为9和6个;在多目标性状选择中,CY0818/A0型和OPW19/C1型为体重和绒产量的双重优势RAPD标记,CY0818/G1型为体重和绒细度的双重优势RAPD标记。Abstract: The genetic relationships between economic traits and genetic markers were studied in 147 goats including Chaidamu goat (CS), Chaidamu Cashmere goat (CRS) and Liaoning Cashmere goat (LRS) in Qinghai province, China. CRS was the population of CS×LRS crossbred. The results showed as follows: the selection reaction of these blood protein polymorphisic loci were great, such as EsD, LAP and PA-3; and EsD2-2, LAPBB and PA-32-2 were the superior marker genotypes on body weight ,Cashmere yield and Cashmere fineness respectively by Least Square method. The interaction between marker genotypes at double loci was found frequently, and their ratio between interaction variance component and genetic variance was higher. With the method of marker assisted prediction( MAP), some interaction effect could be used effectively in the crossbreed population. On the aspect of random amplified polymorphic DNA (RAPD), the number of the superior RAPD marker bands were 11 on body weight trait, 9 and 6 RAPD marker bands on Cashmere yield and Cashmere fineness. For multi-goal traits, CY0818/A0 type and OPW19/C1 type were superior RAPD markers of body weight and Cashmere yield, CY0818/G1 type was superior one of body weight and Cashmere fineness.     

分子系统学研究中分子位点数与遗传差异信息可靠性的关系

用桑蚕(包括不同生态地理品种59个及野桑蚕)和桑属植物(包括12个种和2个变种)两类截然不同的材料,探讨了分子系统学研究中RAPD分子位点数与遗传差异信息可靠性间的关系。发现:(1)所分析的位点数多少与所能提供系统信息的量及可靠性之间有明显的关系;(2)当位点数在20以下时,得到的遗传差异的结果极不可靠,随着位点数的增加所提供的信息量及可靠性增加。当位点数超过70个时,所提供的信息可靠性趋于稳定;(3)对桑蚕和桑属植物两种截然不同的材料的分析,均得相似结果。由此推论:在用RAPD进行生物系统学研究中,以分析70个左右位点数为好;这一结论受研究对象的影响小,在其它类似的研究中或许具有一定的参考价值。 Abstract:We studied the correlation between the number of RAPD-loci investigated and the reliability of the information on genetic variation analysis using quite different two-kind materials,silkworms(including 59 geographic varieties of Bombyx mori belonging to different ecotype,as well as Bombyx mandarina M.)and mulberries(including 12 species and 2 varieties).The results showed as follows:(1)There was obvious correlation between the numbers of RAPD-loci investigated and the reliability of the information on genetic variation analysis.(2)The data were not suitable for the genetic variation analysis when the number of RAPD-loci in vestigated was smaller than 20.The more RAPD loci were analyzed,the more accurate genetic diversity would be identified when the number of the locus was varied from 20 to 70.Furthermore,the stable information to be available is shown when the number exceeded 70.(3)The similar regularities mentioned above were observed in both silkworm and mulberry.In order to get reliable information of genetic variation in molecular phylogenetic studies,at least 70 RPD loci would be investigated.And the results obtained in this study might be referential for AFLP and RFLP analysis.     

小麦异交群体选择分化的RAPD分析

用RAPD分子标记技术及数量遗传分析手段对小麦异交群体趋异选择4代得到的各子群体的遗传关系进行了分析,并对两种结果进行了相关分析。采用11个引物扩增出134个位点,RAPD分析结果表明,群体具有丰富的遗传变异,整个群体的多态位点百分率达0.8134,杂合度达0.3006;子群体间遗传分化较小,相对基因分化系数Gst为0.2310,固定指数平均0.2120,标准遗传距离为0.1044±0.048,表明遗传变异多数来自群体之内。而数量遗传分析发现,选择性状进展都与预期方向相符,主成分遗传距离大部分达显著水平,说明选择使群体发生了相当程度的分化。对两种遗传距离进行聚类分析表明,二种信息间关系各类群成员多不一致。其原因可能在于：数量性状是人工选择的目标,RAPD是基因组随机序列的扩增,二者之间的相关与引物选择有关。 Abstract：The genetic relations of some subpopulations in the fourth generation of selection in outbreeding population of winter wheat were examined by RAPD techniques and quantitative analysis. The correlations of the two results were examined.For RAPD analysis, 11 primers were adopted, and 134 sites were amplified. The results showed that abundant genetic variations existed in the populations. In the whole population, the percentage of polymorphic sites (P) is 0.8134, and the averaged heterozygosity is 0.3006. However,genetic differentiation among subpopulations is small. The relative gene differentiation (Gst) is 0.2310,fixed index averages 0.2120, and the standard genetic distance between subpopulations is 0.1044±0.048.All above shows that most of the genetic variations existed within populations.The cluster results from RAPD analysis and principal component distances showed that similarity relations were only found in small part of the subpopulations. The reason for the results may be that the genes concerned for quantitative traits are selected directly,while the genes involved in RAPD analysis are only random samples of the whole genomes.     

山羊经济性状标记辅助选择的遗传效应分析 Genetic Effect of the Marker Assisted Selection on Economic Traits of Goats

本研究以辽宁绒山羊、柴达木绒山羊和柴达木山羊3个群体共147只山羊为研究对象，运用PAGE和RAPD技术对山羊的体重、绒产量和绒细度3个性状进行了与标记基因关系的遗传分析，结果表明：EsD2-2型、LAPBB型和PA-32-2型分别为体重、绒产量和绒细度性状的优势标记基因型；可利用标记辅助预测的方法充分利用多基因座标记基因间的互作效应；在体重上，寻找到有显著选择效应的RAPD条带11个，在绒产量和绒细度上分别为9和6个；在多目标性状选择中，CY0818/A0型和OPW19/C1型为体重和绒产量的双重优势RAPD标记，CY0818/G1型为体重和绒细度的双重优势RAPD标记。Abstract: The genetic relationships between economic traits and genetic markers were studied in 147 goats including Chaidamu goat (CS), Chaidamu Cashmere goat (CRS) and Liaoning Cashmere goat (LRS) in Qinghai province, China. CRS was the population of CS×LRS crossbred. The results showed as follows: the selection reaction of these blood protein polymorphisic loci were great, such as EsD, LAP and PA-3; and EsD2-2, LAPBB and PA-32-2 were the superior marker genotypes on body weight ,Cashmere yield and Cashmere fineness respectively by Least Square method. The interaction between marker genotypes at double loci was found frequently, and their ratio between interaction variance component and genetic variance was higher. With the method of marker assisted prediction( MAP), some interaction effect could be used effectively in the crossbreed population. On the aspect of random amplified polymorphic DNA (RAPD), the number of the superior RAPD marker bands were 11 on body weight trait, 9 and 6 RAPD marker bands on Cashmere yield and Cashmere fineness. For multi-goal traits, CY0818/A0 type and OPW19/C1 type were superior RAPD markers of body weight and Cashmere yield, CY0818/G1 type was superior one of body weight and Cashmere fineness.     

Molecular markers of nuclear restoration gene Rf1 in sunflower using bulked segregant analysis-RAPD

Restoration of cytoplasmic male sterility (CMS) in sunflower was demonstrated to be controlled by polygenes by analysing 982 effective crosses among 109 self-crossed lines and 16 CMS lines. Two self-crossed lines and one CMS line with distinct genotypes were applied to creation of segregating populations for DNA bulks of the target gene Rfl. Bulked DNA was prepared in order to investigate single gene Rfl and its gene marker among polygenic characters at the same genetic background. Using 80 10-mer operon primers, 620 RAPD reactions were carried out between fertile and sterile DNA bulks. In about 800 loci, primary results showed that 8 were related to the restoration genes. Furthermore. 2 were confirmed as RAPD markers for gene Rfl by examining 9 maintenance and 7 restoration lines. This method is the improvement for bulked segregant analysis[1] with which markers of single gene of target can be identified rapidly among polygenic characters.     

Recycling Isolation of Plant DNA,A Novel Method

DNA is one of the most basic and essential genetic materials in the field of molecular biology.To date,isolation of sufficient and good-quality DNA is still a challenge for many plant species,though various DNA extraction methods have been published.In the present paper,a recycling DNA extraction method was proposed.The key step of this method was that a single plant tissue sample was recycled for DNA extraction for up to four times,and correspondingly four DNA precipitations(termed as the 1st,2nd,3rd and 4th DNA sample, respectively) were conducted.This recycling step was integrated into the conventional CTAB DNA extraction method to establish a recycling CTAB method.This modified CTAB method was tested in eight plant species,wheat,sorghum,barley,corn,rice,Brachypodium distachyon,Miscanthus sinensis and tung tree.The results showed that high-yield and good-quality DNA samples could be obtained by using this new method in all the eight plant species.The DNA samples were good templates for PCR amplification of both ISSR and SSR markers.The recycling method can be used in multiple plant species and can be integrated with multiple conventional DNA isolation methods,and thus is an effective and universal DNA isolation method.     

采用RAPD标记探讨瓣蕊唐松草(毛茛科唐松草属)的遗传多样性及其地理分布格局(英文)

Random amplified polymerphic DNA(RAPD)method was applied to assessg enetic variation and population structure of Thahctrum petalotdeum L(Ranunoulaceae),Two hundred and forty-six individuals from 11 populations of the species were investigated by RAPD profiles Twenty selected RAPD primers generated 125 bands．in which 120 were polymorphic Ther esults revealed a high level of genetic variation(ercentage of polymorphIc bands(PPB was 96％．Nei’s gene diversity(りwas 03502 and shannon’s information index(I) was 0．5199 at the species level) The differentiation among the populations was high(Gst=0．3511)in this species．Result of analyzing of molecularvariance(AMOVA)showedthat38．88％of genetic variance was found among the populations Positive correlation withr r=01945(P=00002)was found between genetic distance and geographic distance amongpo pulations Two populations distributed in the drainage basin of YanELz River affined genedcally and formed one clada and the rest nine populations formed the other clade in both unweighted pair-group method using arithmetic average(UPGMA)trees made by two different method different methods. It was yen/clear that these two populations were very special, andmust be closely related in history, despite the fact that they now share quite weak link to the restpopulations through gene communication.     

Identification and Assessing the Cultivars of Laminaria Lamx. （Phaeophyceae） with Molecular Markers

Molecular markers were used to identify and assess cultivars ofLaminaria Lamx. and to delineate their phylogenetic relationships. Random amplified polymorphic DNA (RAPD) analysis was used for detection. After screening, 11 primers were selected and they yielded 133 bands in all, of which approximately 99.2% were polymorphic. The genetic distances between gametophytes ranged from 0.412 to 0.956.Two clusters were formed with the unweighted pair group method with arithmetic mean (UPGMA) dendrogram based on the simple matching coefficient. All cultivars ofLaminariajaponica Aresch. used for breeding in China fell into one cluster. L.japonica from Japan, L. saccharina (L.) Lam., and L. angustata Kjellm.formed the other cluster and showed higher genetic variation than L. japonica from China. Nuclear ribosomal DNA (rDNA) sequences, including internal transcribed spacers (ITS1 and ITS2) were studied and aligned. The nucleotides of the sequences ranged from 634 to 668, with a total of 692 positions including ITS1, ITS2, and the 5.8S coding region. The phylogenetic tree obtained by the neighbor-joining method favored, to some extent, the results revealed by RAPD analysis. The present study indicates that RAPD and ITS analyses could be used to identify and assess Laminaria germplasm and to distinguish some species and, even intraspecies, in Laminaria.     

DNA分子标记在番茄遗传育种研究中的应用

本文综述了DNA分子标记在番茄遗传图谱构建、番茄种质资源研究与品种纯度的鉴定、番茄基因分子标记研究及番茄基因图位克隆方面的应用研究进展。 Abstract:This paper reviewed the recent advance of the application of DNA molecular marker in various aspects of tomato breeding including genetic map construction,germplasm research and purity control of cultivars,identification markers linked to important genes and map-based gene cloning.