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1.
两个人工雌核发育红白锦鲤群体的RAPD标记分析 总被引:23,自引:3,他引:20
采用PAPD方法,对两个人工雌核发育红白锦鲤群体进行了多态性及分子标记分析。结果表明,同一雌核发育群体具有基本一致的扩增产物,而不同雌核发育群体间的扩增产物则有较大不同;并从30个随机引物的扩增谱带中找到了7个引物(Opo-7,Opo-9,Opo-12,Opo-14,Opj-4,Opj-8和Opj-10)的扩增谱带可以作为两个不同雌核发育群体间的遗传标记。由UPGMA聚类法构建的分支系统树清晰地反映了两个雌核发育群体及其个体间的相互关系。 相似文献
2.
应用微卫星标记对雌核发育银鲫的遗传多样性初探 总被引:34,自引:5,他引:34
利用Crooijmans et al.(1997)分离的包含CA重复单元的普通鲤鱼(Cyprinus carpino.L)的8个微卫星DNA标记,对银鲫(Carassius auratus gibelio Bloch)的5个不同雌核发育系的24尾个体进行PCR扩增。分析电泳结果发现,除MFW28未能在银鲫中稳定地扩增出相应的同源序列,其余的7对引物扩增的重复性和稳定性都很好,随引物不同,各等位基因数为1-14个,大小在100-506bp。在MFW1、MFW4、MFW19、MFW20、MFW23和MFW246个微卫星的扩增图谱中,不同的雌核发育系扩增出各自独特的图谱,而同一系内的不同个体间具有高度的遗传同质性,但仍然在个别个体中检测到少量的多态片段。不同系间的扩增图谱呈现出高度的遗传异质性,共鉴定出23个可以用于有效区分5个不同雌核发育系的分子标记。这5个微卫星标记反映了银鲫5个雌核发育系间的相互亲缘关系,其中P和A系同属一个雌核发育系,F系起源于E系,A、D和E系可能分别独立地起源于不同的杂交事件,鉴定的微卫星分子标记为进行银鲫群体遗传学和进化遗传学研究,以及银鲫的分子标记育种和进行基因组作图提供了理想的工具。 相似文献
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5.
猕猴桃属种间体细胞杂种 总被引:10,自引:0,他引:10
利用PEG融合方法,分别进行了中华猕猴桃(Actinidia chinensis var.chinensis)(2n=2x=58)子叶愈伤组织来源的原生质体与美味猕猴桃(A.deliciosa var.deiciosa)(2n=6x=174)子叶愈伤组织原生质体、以及狗枣猕猴桃(A.kolomikta)(2n=2x=58)叶肉原生质体种间原生质体融合。结果表明:中华猕猴桃与美味猕猴桃融合的1个克隆和中华猕猴桃与狗枣猕猴桃融合的4个克隆的RAPD谱带分别具有双亲特异的DNA谱带;经流式细胞仪分析,前者细胞核倍性推测为8倍体,后者细胞核为3倍体、4倍体和5倍体。初步鉴定这5个克隆是猕猴桃属种间体细胞杂种。 相似文献
6.
R. J. Schnell C. M. Ronning R. J. Knight Jr. 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1995,90(2):269-274
Twenty-five accessions of mango were examined for random amplified polymorphic DNA (RAPD) genetic markers with 80 10-mer random primers. Of the 80 primers screened, 33 did not amplify, 19 were monomorphic, and 28 gave reproducible, polymorphic DNA amplification patterns. Eleven primers were selected from the 28 for the study. The number of bands generated was primer- and genotype-dependent, and ranged from 1 to 10. No primer gave unique banding patterns for each of the 25 accessions; however, ten different combinations of 2 primer banding patterns produced unique fingerprints for each accession. A maternal half-sib (MHS) family was included among the 25 accessions to see if genetic relationships could be detected. RAPD data were used to generate simple matching coefficients, which were analyzed phenetically and by means of principal coordinate analysis (PCA). The MHS clustered together in both the phenetic and the PCA while the randomly selected accessions were scattered with no apparent pattern. The uses of RAPD analysis for Mangifera germ plasm classification and clonal identification are discussed. 相似文献
7.
The role of random amplified polymorphic DNA (RAPD) markers in detecting intra-clonal genetic variability in vegetatively propagated UPASI-9 clone of tea (Camellia sinensis) was studied. Twenty five decamer primers were used, of which three did not amplify, three gave single bands and the rest of nineteen primers generated upto twelve bands (an average of 6.3 bands per primer). Twenty one primers exhibiting amplified products gave monomorphic banding patterns. Only one primer (OPE-17) gave a unique extra band of similar size in four plants. 相似文献
8.
The randomly amplified polymorphic DNA (RAPD) technique was employed in the development of a typing protocol for Listeria isolates, particularly Listeria monocytogenes strains. A single strain of L. monocytogenes was used and 200 random decamer primers were screened for their discriminatory abilities by visualizing the amplification products electrophoretically. Three candidate primers displaying potentially useful banding patterns were selected and tested against 52 L. monocytogenes strains, encompassing 11 serotypes, and 12 other strains representing five other Listeria spp. Thirty-four banding profiles were obtained with one particular primer. RAPD analysis allowed differentiation between Listeria spp. and was found to further subdivide strains of the same serotype. Where only one primer was used strains from different serotypes were occasionally found to produce identical banding profiles. RAPD analysis, which in our hands proved to be reproducible, shows much promise as a molecular alternative to traditional L. monocytogenes typing protocols. 相似文献
9.
《Aquatic Botany》2007,87(3):242-246
The information on diversity and spatial distribution of clones of an invasive clonal plant is crucial for the understanding of its clonal structure and invasive history. In this paper, random amplified polymorphic DNA (RAPD) markers were used to explore the clonal diversity and clonal structure of Eichhornia crassipes (Mart.) Solms in natural populations, and their possible effects on the plant success as an invader are also discussed. Five populations covering the entire distribution area in China were studied, sampling 43 individuals per population at an interval of 1 m in a sampling plot. Twelve RAPD primers produced 69 reproducible bands, with 22 being polymorphic. Only five RAPD phenotypes (clones) were detected in these five populations, but each population consisted of at least three clones, contrary to the traditional expectations that E. crassipes populations should be monoclonal. The diversity of clones within populations is thought to be mainly resulted from multiple introductions by humans. The evenness of distribution of clones varied slightly and most clones were widespread, suggesting clonal growth is the predominant mode of regeneration in all the populations. A single clone dominated each population and this clone might be the first one introduced into China or the genotype with a higher phenotypic plasticity, which could survive and reproduce via clonal growth in various habitats. The clones in each population were highly intermixed, especially in river populations, suggesting this species has a guerilla clonal structure which can be facilitated by water current. 相似文献
10.
Genetic Evidence for Gonochoristic Reproduction in Gynogenetic Silver Crucian Carp (Carassius auratus gibelio Bloch) as Revealed by RAPD Assays 总被引:14,自引:0,他引:14
Sex evolution has been a debating focus in evolutionary genetics. In lower vertebrates of reptiles, amphibians, and fish,
a species or a bioform reproduces either sexually or asexually but never both. A few species were found to consist of all
females in fish. These all-female species can propagate by asexual reproduction modes, such as gynogenesis and hybridogenesis.
However, the coexistence of sexuality and asexuality in a single species was recently noted only in a cyprinid fish silver
crucian carp, Carassius auratus gibelio. This fish had been demonstrated to be capable of gynogenesis stimulated by sperm from other related species. Surprisingly,
natural populations of this fish consist of a minor but significant portion (approx. 20%) of males. As different clones with
specific phenotypic and genetic characteristics have been found, and RAPD markers specific to each clone have recently been
identified, this fish offers many advantages for analyzing whether or not genetic recombination occurs between different clones.
In this study, artificial propagation was performed in clone F and clone D. Ovulated eggs from clone F were divided into two
parts and respectively inseminated with sperm from a clone D male and from a red common carp (Cyprinus carpio) male. The control clone D individuals were selected from gynogenetic offspring of clone D activated by sperm of red common
carp. The phenotype and sex ratio in the experimental groups were also observed. Using RAPD molecular markers, which allow
for reliable discrimination and genetic analysis of different clones, we have revealed direct molecular evidence for gonochoristic
reproduction in the gynogenetic silver crucian carp and confirmed a previous hypothesis that the silver crucian carp might
reproduce both gynogenetically and gonochoristically. Therefore, we conclude that the silver crucian carp possesses two reproductive
modes, i.e., gynogenetic and gonochoristic reproduction. The response mechanism of two reproductive development modes may
be the first discovery in vertebrates. Additionally, we discuss the evolutionary implication between gynogenetic and gonochoristic
reproduction modes and the contribution of the minor proportion of males to genetic flexibility in the gynogenetic silver
crucian carp.
Received: 5 January 2000 / Accepted: 3 August 2000 相似文献
11.
应用RAPD技术对吐鲁番地区火焰山及艾丁湖区域分离的15株土壤绿藻(chlorophyta)品系的遗传多样性及其亲缘关系进行探讨。结果表明:从20个随机引物中,筛选出多态性和重复性较好且谱带清晰的引物8个,这8个引物扩增出的DNA片段大多在300~2 000 bp之间,所形成的多态性位点数差距较大,显示该区域土壤绿藻具有较丰富的遗传多样性;15株土壤绿藻扩增共得到74条谱带,71条多态性带,其多态性比率为95.95%;聚类分析显示15株土壤绿藻明显地聚为2大类,与其来源相对应,即隶属于同一亚组或相近亚组的不同种基本归为一类,其种间关系与传统的形态学分类结果相吻合。 相似文献
12.
Polyploid gibel carp, Carassius auratus gibelio, is an excellent model system for evolutionary genetics owing to its specific genetic background and reproductive modes. Comparative karyotype studies were performed in three cultured clones, one artificially manipulated group, and one mated group between two clones. Both the clones A and P had 156 chromosomes in their karyotypes, with 36 metacentric, 54 submetacentric, 36 subtelocentric, 24 acrocentric, and six small chromosomes. The karyotype of clone D contained 162 chromosomes, with 42 metacentric, 54 submetacentric, 36 subtelocentric, 24 acrocentric, and six small chromosomes. All the three clones had six small chromosomes in common. Group G, being originated from the clone D by artificial manipulation, showed supernumerary microchromosomes or chromosomal fragments, in addition to the normal chromosome complement that was identical to the clone D. The offspring from mating between clones D and A had 159 chromosomes. Comparing with the clone A, the DA offspring showed three extra metacentric chromosomes. In addition, variable RAPD fingerprint patterns and unusual SCAR marker inheritance were, respectively, detected among individuals of artificial group G and in the mated DA offspring. Both the chromosome and molecular findings suggest that genome reshuffling might have occurred by manipulation or mating of the clones. 相似文献
13.
R. Bhatia K. P. Singh T. R. Sharma Tripta Jhang 《Plant Cell, Tissue and Organ Culture》2011,104(1):131-135
The genetic fidelity of in vitro-raised gerbera clones was assessed by using random amplified polymorphic DNA (RAPD) and inter-simple
sequence repeat (ISSR) markers. Out of 35 RAPD and 32 ISSR primers screened, only 12 RAPD and 10 ISSR primers produced clear,
reproducible and scorable bands. The 12 RAPD primers produced 54 distinct and scorable bands, with an average of 4.5 bands
per primer. The number of scorable bands for ISSR primers varied from 3 (ISSR-14) to 9 (ISSR-07), with an average of 5.5 bands
per primer. The number of bands generated per primer was greater in ISSR than RAPD. All banding profiles from micropropagated
plants were monomorphic and similar to those of the mother plant. A similarity matrix based on Jaccard’s coefficient revealed
that the pair-wise value between the mother and the in vitro-raised plantlets was 1, indicating 100% similarity. This confirmed
the true-to-type nature of the in vitro-raised clones. 相似文献
14.
Detection of Polymorphic DNA and Taxonomic Relationships Among 10 Wild Perennial Soybean Species Using Specific and Arbitrary Nucleotide Primers 总被引:1,自引:0,他引:1
Polymorphic DNA in complex genomes of agronomic crops can be detected using specific nucleotide and arbitrary primers and
the polymerase chain reaction (PCR). Nineteen accessions representing 10 species of the wild perennial soybean were evaluated
using 4 sets of specific primers and 3 sets of random amplified polymorphic DNAs (RAPD) primers. The potential of the RAPD
assays was further increased by combining two primers in a single PCR. The fragments generated by the two assays discriminated
10 wild species by banding profiles. The size of the amplified DNA fragments ranged from 100 to 2100 base pairs. The resolved
PCR products yielded highly characteristic and homogeneous DNA fingerprints. The fingerprints were useful not only for investigating
genetic variability but also for further characterizing the wild soybean species by detecting inter- and intra-specific polymorphisms,
constructing dendrograms defining the phylogenetic relationships among these species, and identifying molecular markers for
the construction of genetic linkage maps. Furthermore, unique markers distinguishing particular species were also identified.
Thus, it is expected that PCR will have great relevance for taxonomic studies.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
15.
Random amplified polymorphic DNAs (RAPD) analysis has been adapted to assess the degree of RAPD polymorphism within the genus
Hordeum to determine if this approach can distinguish wild and cultivated species. Nineteen wild and seven cultivated accessions
were evaluated using 4 random 10-mer primers. The potential of the RAPD assay was further increased by combining two primers
in a single polymerase chain reaction (PCR). RAPD fragments generated by two pairs of arbitrary 10-mer primers discriminated
six wild species and one cultivated species by banding profiles. The size of the amplified DNA fragments ranged from 150 to
2300 base pairs. 33 %percent of the fragments were common to both wild and cultivated species; 67% were specific to either
wild or cultivated species. The average difference in fragments was less within the species than among the species. By comparing
RAPD fingerprints of wild and cultivated barley, markers were identified among the set of amplified DNA fragments which could
be used to distinguish wild and cultivated Hordeum species.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
16.
R. Delourme A. Bouchereau N. Hubert M. Renard B. S. Landry 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,88(6-7):741-748
Bulked segregant analysis was employed to identify random amplified polymorphic DNA (RAPD) markers linked to the restorer gene (Rfo) used in theOgura radish cytoplasmic male sterility of rapeseed. A total of 138 arbitrary 10-mer oligonucleotide primers were screened on the DNA of three pairs of bulks, each bulk corresponding to homozygous restored and male sterile plants of three segregating populations. Six primers produced repeatable polymorphisms between paired bulks. DNA from individual plants of each bulk was then used as a template for amplification with these six primers. DNA polymorphisms generated by four of these primers were found to be completely linked to the restorer gene with the polymorphic DNA fragments being associated either with the fertility restorer allele or with the sterility maintainer allele. Pairwise cross-hybridization demonstrated that the four polymorphic DNA fragments did not share any homology. Southern hybridization of labelled RAPD fragments on digested genomic DNA from the same three pairs of bulks revealed fragments specific to either the male sterile bulks or to the restored bulks and a few fragments common to all bulks, indicating that the amplified sequences are low copy. The four RAPD fragments that were completely linked to the restorer locus have been cloned and sequenced to develop sequence characterized amplified regions (SCARs). This will facilitate the construction of restorer lines used in breeding programs and is the first step towards map-based cloning of the fertility restorer allele. 相似文献
17.
Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to investigate the genetic
structure of four subpopulations of Mystus nemurus in Thailand. The 7 RAPD and 7 ISSR primers were selected. Of 83 total RAPD fragments, 80 (96.39%) were polymorphic loci,
and of 81 total ISSR fragments, 75 (92.59%) were polymorphic loci. Genetic variation and genetic differentiation obtained
from RAPD fragments or ISSR fragments showed similar results. Percentage of polymorphic loci (%P), observed number of alleles,
effective number of alleles, Nei’s gene diversity (H) and Shannon’s information index revealed moderate to high level of genetic
variations within each M. nemurus subpopulation and overall population. High levels of genetic differentiations were received from pairwise unbiased genetic
distance (D) and coefficient of differentiation. Mantel test between D or gene flow and geographical distance showed a low
to moderate correlation. Analysis of molecular variance indicated that variations among subpopulations were higher than those
within subpopulations. The UPGMA dendrograms, based on RAPD and ISSR, showing the genetic relationship among subpopulations
are grouped into three clusters; Songkhla (SK) subpopulation was separated from the other subpopulations. The candidate species-specific
and subpopulation-specific RAPD fragments were sequenced and used to design sequence-characterized amplified region primers
which distinguished M. nemurus from other species and divided SK subpopulation from the other subpopulations. The markers used in this study should be useful
for breeding programs and future aquacultural development of this species in Thailand. 相似文献
18.
Chansiripornchai N Ramasoota P Bangtrakulnonth A Sasipreeyajan J Svenson SB 《FEMS immunology and medical microbiology》2000,29(3):221-225
Randomly amplified polymorphic DNA (RAPD) analysis was performed for the molecular genetic typing of 30 Salmonella enterica subsp. enterica strains isolated from chickens and ducks in Thailand. Six different primers were tested for their discriminatory ability. While some of the primers could only differentiate between the different serovars, the use of multiple primers showed that the RAPD method could also subdivide within a given serovar. The Ready-To-Go RAPD analysis beads used, resulted in reproducible and stable banding patterns. As the RAPD technique is simple, rapid and rather cheap, we suggest that it may be a valuable new tool for studying the molecular genetic epidemiology of S. enterica ssp. enterica, both inter- and intra-serovars. 相似文献
19.
陕西大豆资源遗传多样性及变异特点研究 总被引:2,自引:0,他引:2
利用42个PAPD引物对75份陕西大豆种质进行遗传多样性分析,共扩增出310个条带,平均每个引物扩增7.3个条带,多态性比率为96%;田间试验考察了13个农艺性状。陕西大豆的遗传多样性在秦岭南、北两个地区有所不同,秦岭北品种遗传多样性指数较高的性状数目和性状遗传多样性指数都大于秦岭南品种,RAPD分子标记遗传多样性指数也是秦岭北品种大于秦岭南品种,但秦岭南品种RAPD分子标记的遗传多样性指数较高的个数大于秦岭北品种。聚类分析将参试大豆材料分为三大类,基本上反映了材料的地理来源。主成分分析结果显示,前两个主成分反映了10.95%的遗传变异,基于前两个主成分值的二维散点图可以将两个地区的材料基本区分开来。AMOVA分析显示,陕西大豆品种个体间的遗传变异占总变异的92.06%,地区间的遗传变异占总变异的7.94%,二者都达到了极显著水平。研究结果表明,陕西大豆资源存在丰富的遗传多样性,秦岭北品种遗传多样性较高,但秦岭南品种有着广泛的微小变异。 相似文献
20.
Molecular characterization of RAPD and SCAR markers linked to the Tm-1 locus in tomato 总被引:9,自引:0,他引:9
T. Ohmori M. Murata F. Motoyoshi 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,92(2):151-156
We have cloned and sequenced six RAPD fragments tightly linked to the Tm-1 gene which confers tomato mosaic virus (ToMV) resistance in tomato. The terminal ten bases in each of these clones exactly matched the sequence of the primer for amplifying the corresponding RAPD marker, except for one in which the 5-endmost two nucleotides were different from those of the primer. These RAPD clones did not cross-hybridize with each other, suggesting that they were derived from different loci. From Southern-hybridization experiments, five out of the six RAPD clones were estimated to be derived from middle- or high-repetitive sequences, but not from any parts of the ribosomal RNA genes (rDNA), which are known to be tightly linked with the Tm-1 locus. The remaining clone appeared to be derived from a DNA family consisting of a few copies. These six RAPD fragments were converted to sequence characterized amplified region (SCAR) markers, each of which was detectable using a pair of primers having the same sequence as that at either end of the corresponding RAPD clone. All pairs of SCAR primers amplified distinct single bands whose sizes were the same as those of the RAPD clones. In four cases, the SCAR markers were present in the line with Tm-1 but absent in the line without it, as were the corresponding RAPD markers. In the two other cases, the products of the same size were amplified in both lines. When these SCAR products were digested with different restriction endonucleases which recognize 4-bp sequences, however, polymorphisms in fragment length were found between the two lines. These co-dominant markers are useful for differentiating heterozygotes from both types of homozygote. 相似文献