口腔扁平苔藓患者的CIITA基因单倍型分析

目的:分析CIITA基因中单核苷酸多态性(Single Nucleotide Polymorphism,SNP)的单倍型与口腔扁平苔藓(Oral Lichen Planus,OLP)之间的关系,并探讨计算单倍型的新方法。方法:在得到42名患者与86名对照的CIITA基因中15个SNP位点的信息后,通过Haploview和SHEsis软件以及本研究组设计的频数计数法进行单倍型分析。组间比较使用χ2检验,并以P=0.05作为统计检验标准。结果:在本研究群体中,CIITA基因上的15个SNP位点能够形成两个单体域(haploblock),其中由位点rs6498124,rs11647384和rs4774所组成的单倍型GAC在三种单倍型分析方法中均显示出显著的统计学差异(Haploview:Chi2=6.127,P=0.013;SHEsis:Chi2=6.469 P=0.011;频数计数法:Chi2=5.460,P=0.019)。结论:在由CIITA基因的SNP位点rs6498124,rs11647384,rs4774组成的单体域中,单倍型GAC对OLP的患病具有潜在的保护性。频数计数法显示,rs4774位点本身的保护性及其与同一单体域内另外两个位点之间的完全连锁不平衡关系是单体型GAC显示出保护性的主要依据。提示单倍型对疾病易感性的解释在于其中的特定SNP位点等位基因型,及其该位点与其它相关位点间的连锁不平衡关系。     

角倍蚜细胞色素氧化酶(CO Ⅰ)基因的多样性分析

角倍蚜是五倍子蚜的重要种类之一,其干雌寄生于盐肤木上,在完全封闭的虫瘿内生活,瘿内为高单宁、高湿度和高CO2的特殊小生境,而越冬若蚜寄生于提灯藓类上.本文对角倍蚜2个虫态的线粒体细胞色素C氧化酶Ⅰ(CO Ⅰ)基因序列进行分析,发现这两个虫态具有不同程度的遗传分化,其中干雌的单倍型多样性和核苷酸多样性分别为0.308和0.101,而越冬若蚜为0.176和0.047.角倍蚜CO Ⅰ基因序列共有5个单突变变异位点,包含6种单倍型,其中Hap_1、Hap_4为广布单倍型,Hap_2、Hap_3和Hap_5为干雌的私有单倍型,Hap_6为越冬若蚜的私有单倍型.Network分析单倍型间的遗传距离得知单倍型Hap_1与Hap_2、Hap_3、Hap_4、Hap_5、Hap_6的距离分别为472、495、286、520和194.Hap_5距Hap_1最远,是最晚分化出的单倍型,其次为Hap_3和Hap_2;Hap_6离Hap_1最近,是最早从Hap_1分化出来的单倍型.故推测越冬若蚜分化时间早于干雌.     

三角帆蚌GPX基因结构特征及抗性相关SNP的筛选

根据三角帆蚌(Hyriopsis cumingii)谷胱甘肽过氧化物酶(Glutathione peroxidase,GPX)基因cDNA序列,通过PCR和基因组步移法,从三角帆蚌基因组DNA中扩增出GPX基因全长及其5′调控区。序列分析表明,该基因序列全长6 708 bp,含有2个外显子,1个内含子。5′调控区为992 bp,含有启动子的核心序列TATA盒以及其他一些转录调控元件,如AP1、C/EBP、CdxA。2个外显子长度分别为273 bp和991 bp,内含子长度为4 491 bp。通过直接测序法在三角帆蚌抗性群体和易感群体中筛选GPX基因的SNPs,并研究这些多态性位点与抗逆性状的相关性。共获得了16个SNP位点,其中启动子区的A-99G位点、A-86C位点、A-49C位点,内含子区的A2841T位点、C2847T位点、G3146C位点、A3150G位点以及G4645T位点共8个SNP位点的基因型频率和等位基因频率在抗性和易感群体中均存在显著性差异(P<0.05)。连锁不平衡分析结果显示GPX基因A-86C位点、A-49C位点、C2847T位点、A3150G位点与G4645T位点之间,以及A2841T位点与G3146C位点之间均存在强连锁不平衡。同时单倍型分析发现,在抗性群体中单倍型分别为ACTGT和TG的个体出现的频率显著高于易感群体中出现的频率。这些结果表明,GPX基因的部分SNPs可作为三角帆蚌抗病辅助育种的候选分子标记。     

斑点叉尾GHRH基因3个SNPs位点及其单倍型组合与生长性状的关联分析

研究旨在探讨生长激素释放激素基因(Growth hormone-releasing hormone,GHRH)对斑点叉尾(Ictalurus punctatus)生长性状的影响。采用DNA混池测序法筛选GHRH基因的单核苷酸多态性(Single nucleotide polymorphisms,SNPs)位点,使用SNa Pshot法将筛选到的SNPs多态性位点进行分型,并对这些位点进行连锁不平衡和单倍型分析。结果表明,在GHRH基因内含子区域共检测到4个SNPs位点,并成功地对3个位点进行了分型,3个位点间均不存在强连锁不平衡;3个SNPs位点在176尾斑点叉尾中形成了6种有效单倍型。关联分析表明SNP位点g.6301 GA的AA基因型的体质量显著性地高于AG和GG型(P0.05),比群体的平均体质量高14%。单倍型组合H1/H4和H1/H5个体的体质量和体长极显著性地高于其他单倍型组合(P0.01),体质量比群体平均体质量分别高30%和15%,体长比群体平均体长分别高7%和6%。研究为斑点叉尾生长性状分子标记辅助选育和QTL定位提供了参考依据。     

基质金属蛋白酶-9 基因单核苷酸多态性与肺结核病的相关性研究

目的:探讨基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)基因多态性(single nucleotide polymorphism,SNP)与肺结核的相关性。方法:对符合纳入及排除标准的肺结核病例组224例及健康对照组249例进行血样收集与临床资料采集。采用飞行时间质谱分析方法对MMP-9基因rs17576、rs2236416、rs3787268、rs3918254共4个多态性位点进行基因分型,数据统计分析采用SPSS20.0和Haplo View 4.0软件进行。结果:我们首次发现,在病例及对照组中,rs17576基因型频率分布存在统计学差异(X~2=7.822,P=0.020)。与对照组相比,病例组G等位基因频率显著高于对照组(X~2=7.335,P=0.007,OR=1.463,95%CI=1.110-1.927)。病例组rs17576基因型分布中,GG和AG基因型患者吸烟史显著高于AA基因型患者;GG和AG基因型患者卡介苗接种史显著低于AA基因型患者。连锁不平衡分析发现一个单倍型(rs17576-rs3918254)高度连锁(D'0.7;r~20.8)。在病例组及对照组中,G-C和A-C单倍型频率分布存在显著性差异,病例组中G-C单倍型频率显著高于对照组(P=0.022),对照组中A-C单倍型频率显著高于疾病组(P=0.024)。结论:MMP-9基因rs17576多态性位点可能与肺结核有关,携带有rs17576位点G等位基因的个体更易发生肺结核。携带G-C(rs17576-rs3918254)单倍型的个体更易患肺结核病,携带A-C(rs17576-rs3918254)单倍型的个体相对不易患肺结核病。     

新疆维吾尔族人群过氧化物酶体增殖物激活受体基因相关SNP位点的多态性

对过氧化物酶体增殖物激活受体基因(PPARG)的31个SNP位点进行群体遗传学分析,利用质谱检测技术检测PPARG基因的31个SNPs位点多态性,并根据质谱峰图判读样本目标位点基因型,统计分析31个SNP位点的基因型和等位基因的分布频率,利用x2检验确定筛选的SNP位点是否符合Hardy-Weinberg平衡定律。结果发现31个SNP位点中,23个位点的次等位基因分布频率MAF≥0.05,在新疆维吾尔族人群中具有多态性,其他8个SNP位点的MAF0.05,没显示多态性。基因型和等位基因频率在男女两组间均无统计学差异(P0.05),表明这些位点等位基因分布不存在性别差异。     

AGT基因单倍型与原发性高血压

选取血管紧张素原（angiotensinogen, AGT）基因启动子区 -217, -152, -20, -6, 内含子1 的+31, 第二外显子T174M（3889）和 T235M（4072）共7个位点,对497例的样本（高血压患者298例,血压正常对照199例） 用PCR-RFLP、和最大期望值（expectation maximization,EM）算法为基础的最大似然法(maximum likelihood estimate,MLE)检测和估算,本群体AGT基因A-6G,C+31T,T235M三位点两两存在完全连锁不平衡(D^,=1);G-217A和G-152A位点,G-152A和3889T位点平衡传递。存在7种单倍型,单倍型H2(-217: A, -152: G, -20: A, -6: G, +31: T, 174: T, 235: M) 在正常血压个体中的频率高于高血压组。研究结果提示AGT基因中H2单倍型可能与控制血压的保护性因素连锁不平衡。此外,本研究结果支持基因剂量效应可能存在于单倍型中,而不与单个位点直接关联。     

猪CD4基因多态性及生物信息学分析

本研究利用PCR-RFLP方法检测猪CD4基因部分外显子区的单核苷酸多态性,分析CD4基因SNPs之间的连锁不平衡程度,并采用生物信息学软件预测SNPs对CD4蛋白的潜在影响,为猪的抗病育种研究提供相应的分子标记。研究表明,大白猪CD4基因外显子4、6和7都存在一个突变位点,分别可使CD4蛋白T80S、P290L和M348I发生改变;每个SNP位点都有三种基因型,多态信息含量均处于中等多态(0.25PIC0.5)。三个SNPs呈强连锁不平衡状态(r~20.8),而且仅构成两种主要单倍型CTC和GCG型(两者的频率占99%以上)。经序列预测分析可知,两种CD4单倍型纯合子的理化性质和二级结构存在差异,但三个错义突变是否能影响猪CD4蛋白的功能需进一步研究。     

POPULATION GENETIC STRUCTURE OF ENDANGERED SPECIES PLATΥPHARODON EXTREMUS ( SCHIZOTHORACINAE, CYPRINIDAE )IN YELLOW RIVER, CHINA

极边扁咽齿鱼Platypharodon extremus是黄河上游的特有鱼类,近年来由于过度捕捞、环境变迁等因素,其资源量剧减,种群处于濒危状态.研究中采集了分布于黄河上游的3个种群(n= 107),基于线粒体DNA控制区695bp 序列,共检测到101个可变位点,占总分析位点的14.5％,其中58个为简约信息位点.3种群共界定了87个单倍型,种群平均单倍型多样性h= 0.995,核苷酸多样性π=0.0129.结果表明,极边扁咽齿鱼的种群遗传多样性水平较高.分子方差分析(AMOVA)表明,3群体间总遗传分化系数Fst= 0.0528,群体间尚未有显著的遗传分化.系统发生树显示3种群的单倍型混合分布,各进化枝之间分歧度很低,没有形成明显的单倍型组,也没有显示出单倍型与地理位置的对应关系.3个群体共享1个单倍型(Hap_ 31),推测它们来自于共同的祖先.歧点分布和Fu’Fs中性检测显示极边扁咽齿鱼并未经历种群扩张.     

本土绵羊品种GH与IGF-1基因多态性分析

生长性状是绵羊育种中关注的重要性状.GH和IGF-1基因已被证明是影响动物生长发育的重要候选基因.因此,了解中国本土绵羊GH和IGF-1基因的遗传多样性,将为提高本土绵羊的生产效率、制定遗传育种和品种改良措施奠定基础.本研究以本土绵羊品种呼伦贝尔羊、藏羊、湖羊、阿勒泰羊、小尾寒羊和滩羊为主要研究对象,以澳白羊为参考,通过Sanger测序检测了340只绵羊GH和IGF-1外显子单核苷酸多态性.研究发现,GH和IGF-1基因外显子区分别检测到11个和3个SNPs,呈品种特异性分布,在群体中处于低度至中度多态.连锁不平衡分析发现,GH基因的SNP1、SNP3、SNP6、SNP7和SNP8呈强连锁不平衡,构建的7种单倍型中,CGACAG是优势单倍型(52.6％),而IGF-1基因的3个SNPs之间连锁关系很弱.生物信息学分析发现,检测到的14个SNPs中GH的SNP2、SNP4、SNP8和IGF-1的SNP1、SNP3为新发现的多态位点,其中SNP4和SNP9为错义突变,可能会导致编码蛋白质二级结构与三级结构发生改变.研究结果表明,与其他本土品种和澳白羊相比,湖羊的GH和IGF-1基因具有较为丰富的遗传变异.已有研究证实GH的SNP6和SNP9以及IGF-1的SNP2不同基因型与绵羊的生长和胴体性状相关联,且在本土品种中均有多态,推测GH基因的SNP6、SNP9和IGF-1基因的SNP2可作为本土绵羊生长性状分子标记辅助选择的候选SNPs.     

Genetic divergence between North American ancestral soybean lines and introductions with resistance to soybean cyst nematode revealed by chloroplast haplotype

Domesticated soybean [Glycine max (L.) Merr.] is a major crop with an established ancestral relationship to wild soybean (Glycine soja Sieb. & Zucc.) native to Asia. Soybean genetic diversity can be assessed at different levels by identification of polymorphic alleles at genetic loci, in either the plastid or nuclear genomes. The objective of this study was to evaluate genetic diversity based on chloroplast haplotypes for soybean genotypes present in the USDA germplasm resource collection. Shared chloroplast haplotypes represent broad groups of genetic relatedness. Previous work categorized three-quarters of the cultivated soybeans from Asia into a single haplotype group. Our results confirmed the close relationship of North American soybean ancestors and G. max plant introductions previously identified as representing potential sources of soybean genetic variation with the finding that these genotypes belonged to a single chloroplast haplotype group. Genetic diversity was identified in soybean genotypes determined to have a high density of single nucleotide polymorphisms and in a screen of accessions with resistance to soybean cyst nematode. Characterization of soybean plant introduction lines into chloroplast haplotype group may be an important initial step in evaluating the appropriate use of particular soybean genotypes.     

Highly variable patterns of linkage disequilibrium in multiple soybean populations

Prospects for utilizing whole-genome association analysis in autogamous plant populations appear promising due to the reported high levels of linkage disequilibrium (LD). To determine the optimal strategies for implementing association analysis in soybean (Glycine max L. Merr.), we analyzed the structure of LD in three regions of the genome varying in length from 336 to 574 kb. This analysis was conducted in four distinct groups of soybean germplasm: 26 accessions of the wild ancestor of soybean (Glycine soja Seib. et Zucc.); 52 Asian G. max Landraces, the immediate results of domestication from G. soja; 17 Asian Landrace introductions that became the ancestors of North American (N. Am.) cultivars, and 25 Elite Cultivars from N. Am. In G. soja, LD did not extend past 100 kb; however, in the three cultivated G. max groups, LD extended from 90 to 574 kb, likely due to the impacts of domestication and increased self-fertilization. The three genomic regions were highly variable relative to the extent of LD within the three cultivated soybean populations. G. soja appears to be ideal for fine mapping of genes, but due to the highly variable levels of LD in the Landraces and the Elite Cultivars, whole-genome association analysis in soybean may be more difficult than first anticipated.     

Importance of genetic characterization and conservation of plant genetic resources: The breeding system and genetic diversity of wild soybean (Glycine soja)

Abstract Plant genetic resources play an important role in the improvement of cultivated plants. To characterize and evaluate the ecological and reproductive features of wild soybean ( Glycine soja Sieb. et Zucc.), which is the most probable ancestor of cultivated soybean ( G. max (L) Merr.), the breeding system and genetic diversity of G. soja were investigated. The extent of natural cross-pollination of G. soja was estimated in four populations along the Omono River in Akita Prefecture, Japan by examining allozyme variation. Although it has been previously believed that G. soja is autogamous, as is cultivated soybean, the mean multilocus outcrossing rate ( t m) estimate was 13%. These values are much higher than the outcrossing rate previously reported for both G. soja and G. max . Frequent visits by honeybees and carpenter bees to flowers were also observed, which supported this conjecture. Furthermore, to evaluate the genetic variation of G. soja as a genetic resource, the genetic structure of 447 populations over Japan were analyzed. Wild soybean populations had a higher degree of variation of isozyme loci. The G ST coefficient of gene differentian values among the sites within the district were particularly high, revealing that the isozyme genotype was greatly different among site populations and homogeneous within the sites. The genetic differentiation among nine districts was observed in the allele frequencies of a few loci, indicating that geographic isolation in the wild soybean population was effectively created through the distance between the districts. The difference in the allele frequency among the districts may be produced under genetic drift. Finally, the importance of the preservation of natural plant populations and the habitats of wild progenitors (i.e. the in situ conservation of plant genetic resources) was emphasized.     

Genetic Variation in an Inbred Plant: Variation in Tissue Cultures of Soybean [Glycine Max (L.) Merrill]

Although soybean [Glycine max (L.) Merrill] grows as an inbreeding, generally homozygous, plant, the germplasm of the species contains large amounts of genetic variation. Analysis of soybean DNA has indicated that variation of RFLP (restriction fragment length polymorphism) markers within the species usually entails only two alleles at any one locus and that mixtures of such dimorphic loci account for virtually all of the restriction fragment variation seen in soybean (G. max), and in its ancestors, G. soja and G. gracilis. We report here that tissue cultures prepared from root tissue of individual soybean plants develop RFLP allelic differences at various loci. However, these newly generated alleles are almost always the same as ones previously found and characterized in other varieties of cultivated soybean (cultivars). This repeated generation of particular alleles suggests that much of the genetic variation seen in soybean could be the consequence of specific, relatively frequently employed, recombinational events. Such a mechanism would allow inbred cultivars to generate genetic variation (in the form of alternative alleles) in a controlled manner, perhaps in response to stress.     

大豆种质资源SRAP分子标记中的引物筛选

以113个大豆栽培品种和20个野生品种为材料,从288对引物组合中筛选出12对多态性丰富、条带清晰、可重复性好的SRAP引物组合。用筛选出的12对引物组合对大豆品种进行PCR扩增,获得了带型丰富和清晰可辨的DNA的PAGE指纹图谱;共扩增出251条谱带,其中多态性条带220条,多态性谱带比率为87.6%,平均每个引物扩增出18.3条谱带。结果显示,所筛选出的12对引物组合可以有效的应用于大豆种质资源的SRAP分析。     

北京地区野生大豆种群SSR标记的遗传多样性评价

 使用40对SSR引物分析了北京地区野生大豆(Glycine soja)天然种群的遗传结构与遗传多样性。10个种群共检测到526个等位变异, 平均每对引物等位基因数为13.15个, 种群平均Shannon指数(I)为0.658, 群体平均位点预期杂合度(H_e)为0.369, 群体平均位点杂合度(H_o)为1.29 %。平均种群内遗传多样度(H_s)为0.362, 平均种群间遗传多样度(D_ST)为0.446, 基因分化程度(G_ST)为0.544。该研究显示, 中-西部生态区种群比北部和东部山区种群有较高的遗传多样性。在地理上, 环绕北京地区的太行山和燕山两大余脉区域野生大豆种群遗传分化表现出地理差异。可能是经过干旱选择而形成的有抗旱潜力的种群在遗传上表现单一化。期待该种群提供耐旱基因。     

The genetic diversity and population structure of wild soybean evaluated by chloroplast and nuclear gene sequences

Glycine soja, also called wild soybean, is the wild ancestor of domesticated soybean (Glycine max), and one of the world's major cultivated crops. Wild soybean is a valuable resource for the breeding of cultivated soybean and harbors useful genes or agronomic traits. To use and conserve this valuable resource, we conducted a study to evaluate the genetic diversity and population structure of wild soybean using the sequencing data of two nuclear loci (AF105221 and PhyB) and one chloroplast locus (trnQ-rps16) of more than 600 individuals representing 53 populations throughout the natural distribution range. The results showed that most of the variation was found within the populations and groups, but significant genetic differentiation was also detected among different eco-geographical groups. Correlations between genetic and geographical distance at all the loci were consistent with the isolation by distance gene flow model. G. soja exhibited the highest genetic diversity in middle and downstream of Yangzi River (MDYR) region, followed by North East China (NEC), and was the lowest in North West China (NWC). We concluded that both in situ and ex situ conservation strategies required for wild soybean populations, especially which are native to MDYR and NEC regions.     

Investigation of genetically modified soybean biosafety in the center of the origin and diversity of Russian Far East

Wild soybean (Glycine soja Sieb. & Zucc.) is the nearest relative of a soybean crop (Glycine max (L.) Merr.). Study of population genetic structure of wild-growing relatives ofgenetically modified (GM) plants in the centers of their origin is one of the main procedures before introduction of GM crops in these areas. We have studied genetic variability of nine wild growing soya populations of Primorye Territory using RAPD analysis. The level of G. soja genetic variability was considerably higher than that of G. max. We have analyzed phylogenetic relationships in the genus Glycine subgenus Soja using RAPD markers. Our data confirm validity of allocation G. gracilis in a rank of a species.     

Diversity of chloroplast DNA SSRs in wild and cultivated soybeans: evidence for multiple origins of cultivated soybean

Soybean [ Glycine max (L.) Merr.] is one of the major crops in the world and was domesticated from a wild progenitor, Glycine soja Sieb. & Zucc., in East Asia. In order to address the questions concerning the evolution and maternal lineage of soybean, we surveyed the variation in chloroplast DNA simple sequence repeats (cpSSR) of 326 wild and cultivated soybean accessions that were collected from various Asian countries. Twenty-three variants were detected at six cpSSRs in the accessions tested. All of the variants were found in wild soybean, whereas only 14 variants existed in the cultigen. Combining the variants at the six cpSSRs gave 52 haplotypes in the former and eight haplotypes in the latter. Both analyses indicated a considerably higher genetic diversity in the wild soybean. Around 75% of the cultivated accessions tested possessed a common haplotype (no. 49), which was detected in only seven wild accessions, six from southern Japan and one from southern China. The predominant haplotype in the cultigen may therefore have originated from a rare haplotype of the wild soybean that is presently distributed in the southern areas of Japan and China. The remaining seven haplotypes in the cultigen were distributed regionally, and except for three rare haplotypes, largely overlapped with the distributions of wild accessions with the same respective haplotypes. Our results strongly suggest that the cultivated soybeans with different cpDNA haplotypes originated independently in different regions from different wild gene pools and/or hybrid swarms between cultivated and wild forms.     

Population structure of the wild soybean (Glycine soja) in China: implications from microsatellite analyses

Background and Aims Wild soybean (Glycine soja), a native species of East Asia, is the closest wild relative of the cultivated soybean (G. max) and supplies valuable genetic resources for cultivar breeding. Analyses of the genetic variation and population structure of wild soybean are fundamental for effective conservation studies and utilization of this valuable genetic resource. Methods In this study, 40 wild soybean populations from China were genotyped with 20 microsatellites to investigate the natural population structure and genetic diversity. These results were integrated with previous microsatellite analyses for 231 representative individuals from East Asia to investigate the genetic relationships of wild soybeans from China. Key Results Analysis of molecular variance (AMOVA) revealed that 43·92 % of the molecular variance occurred within populations, although relatively low genetic diversity was detected for natural wild soybean populations. Most of the populations exhibited significant effects of a genetic bottleneck. Principal co-ordinate analysis, construction of a Neighbor-Joining tree and Bayesian clustering indicated two main genotypic clusters of wild soybean from China. The wild soybean populations, which are distributed in north-east and south China, separated by the Huang-Huai Valley, displayed similar genotypes, whereas those populations from the Huang-Huai Valley were different. Conclusions The previously unknown population structure of the natural populations of wild soybean distributed throughout China was determined. Two evolutionarily significant units were defined and further analysed by combining genetic diversity and structure analyses from Chinese populations with representative samples from Eastern Asia. The study suggests that during the glacial period there may have been an expansion route between south-east and north-east China, via the temperate forests in the East China Sea Land Bridge, which resulted in similar genotypes of wild soybean populations from these regions. Genetic diversity and bottleneck analysis supports that both extensive collection of germplasm resources and habitat management strategies should be undertaken for effective conservation studies of these important wild soybean resources.