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1.
RAPD标记在紫菜遗传多样性检测和种质鉴定中的应用 总被引:42,自引:0,他引:42
用RAPD技术对4类紫菜(Porphyra yezoensis,P.haitanensis,P.katadni var.hemiphylla和P.oligospermatangia)的15个无性系丝状体进行了遗传多样履分析,从50个OPERON引物中经过初筛,其中6个引物可以扩增出稳定的可重复的图谱。这6个引物共扩增出了60条带,多态性比例达97.1%。根据RAPD结果将这15个无性了紫菜的DNA 相似文献
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Abdolreza Daneshvar Amoli Seyed Abolhasan Shahzadeh Fazeli Mehdi Aminafshar Naser Emam Jomeh Kashan Hamidreza Khaledi 《In vitro cellular & developmental biology. Animal》2018,54(4):265-271
Iranian Bactrian camel population is less than 100 animals. Iranian biological resource center produced more than 50 Bactrian camel fibroblast cell lines as a somatic cell bank for conservation animal genetic resources. We compared two type markers performance, including 14 random amplified polymorphic DNA (RAPDs) (dominant) and eight microsatellite (co-dominant) for cell line identification, individual identification and investigation genetic structure of these samples. Based on clarity, polymorphism, and repeatability, four RAPD primers were selected for future analysis. Four RAPD primers and eight microsatellite markers have generated a total of 21 fragments and 45 alleles, respectively. RAPD primers revealed fragment size between 150 to 2000 bp and gene diversity since 0.27 (IBRD) to 0.46 (GC10), with an average of 0.37. Microsatellite markers generated number of alleles per locus ranged from 3 to 11, with an average of 5.62 alleles. The observed heterozygosity ranged from 0.359 (IBRC02) to 0.978 (YWLL08), and expected heterozygosity ranged from 0.449 (IBRC02) to 0.879 (YWLL08). Bottleneck analysis and curve showed that Bactrian camel population did not experience a low diversity. RAPD profiles were especially suitable for investigation population genetics. All primers generated novel and polymorphic fragments. Briefly, our results show that a multiplex PCR based on these markers can still be valuable and suitable for authentication of cell lines, investigating gene diversity and conservation genetic resources in Bactrian camel, while new technologies are continuously developed. 相似文献
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RAPD polymorphisms in spring wheat cultivars and lines with different level of Fusarium resistance 总被引:4,自引:0,他引:4
Sun G Bond M Nass H Martin R Dong Z 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2003,106(6):1059-1067
Random amplified polymorphic DNA (RAPD) markers have been used to characterize the genetic diversity among 35 spring wheat cultivars and lines with different levels of Fusarium resistance. The objectives of this study were to determine RAPD-based genetic similarity between accessions and to derive associations between Fusarium head blight (FHB) and RAPD markers. Two bulked DNA from either highly resistant lines or susceptible lines were used to screen polymorphic primers. Out of 160 screened primers, 17 primers generated reproducible and polymorphic fragments. Genetic similarity calculated from the RAPD data ranged from 0.64 to 0.98. A dendrogram was prepared on the basis of a similarity matrix using the UPGMA algorithm, which corresponded well with the results of principal component analysis and separated the 35 genotypes into two groups. Association analysis between RAPD markers and the FHB index detected three RAPD markers, H19(1000), F2(500) and B1(2400), significantly associated with FHB-resistant genotypes. These results suggest that a collection of unrelated genotypes can be used to identify markers linked to an agronomically important quantitative trait like FHB. These markers will be useful for marker-assistant breeding and can be used as candidate markers for further gene mapping and cloning. 相似文献
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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
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M. Lorenz A. Weihe T. Borner 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,88(6-7):775-779
The technique of random amplified polymorphic DNA (RAPD) offers a broad range of applications in the investigation of plant genomes. A promising prospect is the use of RAPD products as genetic markers. We have investigated a possible organellar source of fragments in RAPD patterns of total DNA. Two nearly-isogenic lines of cytoplasmic male-sterile and male-fertile sugar beet (Beta vulgaris L.) were subjected to RAPD analysis with six different primers. Total, nuclear, mitochondrial (mt), and chloroplast (cp), DNA from each line were investigated. Reproducible DNA fingerprints could be obtained from both organellar DNAs. Differences in band patterns of mtDNA between cytoplasmic male-sterile and -fertile lines were observed with five out of six primers, whereas different cpDNA patterns were generated by one of the primers. Consequently, the RAPD technique can be used to discriminate between different cytoplasms. Clear evidence is provided for the organellar origin of fragments in genomic (total DNA) RAPD patterns. The consequences of these results for the interpretation of RAPD analyses are discussed. 相似文献
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RAPD markers in diversity detection and variety identification of Tibetan hulless barley 总被引:1,自引:0,他引:1
RAPD markers were generated from 6 groups of Tibetan hulless barley (23 varieties): Lasagoumang, QB, Zangqing, Guoluo, Dongqing,
and Hymalayia. Of the 48 fragments generated by 5 selected primers (among 68 primers), 44 appeared to be polymorphic (92%).
Cluster analysis was performed (RAPDistance 1.04). The 23 varieties were divided into 2 groups, and the molecular foundation
of genetic diversity was explored. In addition, to identify the varieties, one DNA fingerprint was constructed based on 17
bands amplified with S32 and 2 bands with S18. A specific RAPD fragment can be used to select for high and low β-glucan content
varieties.
Supported by the important program of Ministry of Science and Technology of P.R. China, “The Construction of Key Animal &
Plant Resource System of Southwest China”. 相似文献
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M. Yamagishi 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1995,91(6-7):830-835
Random amplified polymorphic DNA (RAPD) markers were utilized for the identification of Lilium species and inter-specific hybrids. The optimum annealing temperature of the polymerase chain reaction (PCR) for the RAPD assay in Lilium was 54 °C, which is relatively higher than the temperature used for other genera reported by previous researchers. Among 76 primers used to amplify genomic DNA by PCR, 18 primers (24%) generated polymorphic DNA fragments in Lilium species and hybrids. Cultivars were also identified by RAPD markers. Some amplified fragments were unique to species of each section and to hybrids derived from these species; that is, they were the section-specific DNA markers. Sections, Sinomartagon, Leucolirion b, Leucolirion a and Archelirion could be identified by 6 section-specific markers amplified with five primers. Seven inter-section hybrids showed the section-specific bands of both parental sections, indicating that these markers would be useful for identifying the parental sections of inter-section hybrids. 相似文献
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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
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U A Haemmerli U E Br?ndle O Petrini J M McDermott 《Molecular plant-microbe interactions : MPMI》1992,5(6):479-483
Genetic variation in 30 isolates of Discula umbrinella derived from beech, chestnut, and oak was assessed using randomly amplified polymorphic DNA (RAPD) and restriction fragment length polymorphic markers. Polymerase chain reaction amplifications with 17 primers produced 134 different DNA fragments. Three RAPD fragments were subsequently used for Southern hybridization. By these techniques up to four different individuals could be detected in the same leaf. The presence of several individuals within a single leaf indicates a finely tuned balance between the endophyte and its host. Cluster analysis of all arbitrary primed amplified DNA fragments showed that the isolates could be placed into four groups corresponding to their host origin. The high percentage of private RAPD variants within groups is consistent with low gene flow. 相似文献
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利用RAPD分子标记对三组三系杂交水稻及亲本的遗传分析和鉴定 总被引:19,自引:1,他引:18
利用RAPD技术,从248个随机寡聚核苷酸(10bp)中筛选出13个引物能在供试的三组三系杂交水稻及亲本间扩增出43条稳定性较好的多态性片段,其中6个引物能在供试材料间扩增出20个强的多态性标记。利用这些标记能有效地区分各组合中不育系、保持系、恢复系和F1,并能看出各组合中不育系与保持系、不育系与恢复系、F1与亲本间的遗传关系。
Abstract:A total of 248 arbitrary 10-mer oligonucleotide primers were screened using RAPD (random amplified polymorphic DNA) techniques with the genome DNA of three groups of three-line hybrid rice and their parents.Thirteen primers produced 43 polymorphism fragments.Six primers of them produced 20 obviously repeatable polymorphic markers among rice lines tested.Using this RAPD markers,the hybrid rice combinations (sterile-line,maintainer-line,restorer-line and F1)can be effectively identified,and the genetic relationship among them can be shown. 相似文献
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Isolates of Verticillium dahliae were sampled from different olive tree orchards in Morocco. These olive trees were located in different commercial culture locations in southern, central and northern Morocco. The isolates were characterized using genetic markers obtained after their DNA PCR amplification with random amplified polymorphic DNA (RAPD) primers. Among the 40 primers tested, 10 generated a total of 66 polymorphic fragments. Among the 38 isolates of V. dahliae tested, RAPD markers were successful in the characterization of groups based on their geographic origin. With the exception of one specific isolate, no correlation could be established among the isolates, based on the morphological appearance of the colony in culture. 相似文献
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Optimization of primer screening for evaluation of genetic relationship in 34 cultivars of rose through random amplified polymorphic
DNA (RAPD) markers was investigated. Four series of decamer primers were used for screening and optimization of RAPD analysis
between which A and N series performed good amplification of fragments as compared with other series. The primers OPN-07 and
OPN-15 produced maximum number of DNA fragments in Rosa hybrida cv. Anuraag. Some primer either did not produce amplification or produced very poor amplification. Further, ten selected
primers were used for genetic analysis of 34 rose cultivars. The primer OPN-15 amplified 21 fragments in all cultivars tested.
A total of 162 distinct DNA fragments (bands) ranging from 100 to 3400 base pairs were amplified by using 10 selected random
primers. The cluster analysis indicated that these rose cultivars formed nine clusters. 相似文献
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利用RAPD分析杜氏藻属(Dunaliella)嗜盐种间遗传多样性 总被引:8,自引:2,他引:6
本研究首次运用随机引物对杜氏藻属(Dunaliella)中6个嗜盐种的基因组DNA进行RAPD分析。筛选获得的6个有效引物共扩增出98个可重复的DNA片段,其中95条带具有多态性,多态性条带的频率为96.9%。根据系统进化树图将杜氏藻属6个种分别归于亲缘关系相对较远的2个类群中,且6个种与它们各自的生态分布联系不紧密。 相似文献
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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
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Development of DNA markers for identifying chrysanthemum cultivars generated by ion-beam irradiation
Tsukasa Shirao Kei-ichiro Ueno Tomoko Abe Tomoki Matsuyama 《Molecular breeding : new strategies in plant improvement》2013,31(3):729-735
Four chrysanthemum cultivars were generated through (carbon) ion-beam irradiation of the original ‘Jimba’ (Chrysanthemum morifolium Ramat.). The new cultivars had acquired a number of superior cultivation traits, while remaining identical to the commercially available ‘Jimba’ in appearance. In this study, polymerase chain reaction (PCR) assays were used to detect the mutated region of each strain, thereby allowing clear identification at the molecular level. PCR assays were performed with 446 primer sets, including random amplified polymorphic DNA (RAPD) primer sets (10-mer RAPD), arbitrarily primed (AP)-PCR primers based on retrotransposon-like sequences and modified RAPD primers (15-mer RAPD). 15-mer RAPD primers generated a 1.49-fold increased band number at high annealing temperatures compared with the original 10-mer RAPD primers and could thus be effective for detection of polymorphic patterns. Our results provide information on the mutated regions of these ion-beam-irradiated chrysanthemum cultivars. Thus, specific DNA markers could be used to improve identification of new cultivars of chrysanthemum as well as other clonal cultivars of horticultural and agricultural crops. 相似文献
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以陕西省杂交油菜研究中心选育的单显性核不育油菜分离群体为材料,利用集群分离法(BSA)对该油菜单显性核不育基因进行了RAPD分析。在随机选取的300个10碱基随机引物中,引物S243(5′CTATGCCGAC3′)在可育集团与不育集团间扩增出特异而可重复的1.5kb的多态性片段OPU-031500,而在细胞质雄性不育和其它核不育类型油菜中均未扩增出上述特异性片段,从而确证此RAPD标记OPU-031500。片段是与甘蓝型油菜单显性核不育基因连锁的。将该多态性片段克隆并测序,发现其序列与拟南芥的一段DNA序列高度同源。根据同源序列及测序结果设计两对特异引物(P1/P2和P3/P4),引物P3/P4在可育系中可扩增到约1.5kb的单一特异片断,而在不育系中无带,从而将RAPD标记转化为稳定可靠的SCAR标记。 相似文献
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Application of RAPD analysis for identification of Lactococcus lactis subsp. cremoris strains isolated from artisanal cultures 总被引:1,自引:0,他引:1
Randomly amplified polymorphic DNA (RAPD) was used for identification of Lactococcus lactis subsp. cremoris strains isolated 40 years ago from various dairy homemade products. Total genomic DNAs from six randomly chosen isolates and the reference strain Lactococcus lactis subsp. cremoris NIZO B64 were amplified using four different 10-mer primers. Although most RAPD fragments were common to all six isolates, a sufficient number of polymorphic fragments were also detected that allowed clear distinction of the isolates and the reference strain. The results indicate that RAPD analysis could be a useful and efficient method to distinguish Lactococcus lactis subsp. cremoris at the strain level and to detect genetic diversity. 相似文献
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Rout GR 《Zeitschrift für Naturforschung. C, Journal of biosciences》2006,61(1-2):118-122
Identified germplasm is an important component for efficient and effective management of plant genetic resources. Traditionally, plant identification has relied on morphological characters like growth habit, floral morphology like flower colour and other characteristics of the plant. Studies were undertaken for identification and genetic variation within 15 clones of Tinospora cordifolia through random amplified polymorphic DNA (RAPD) markers. Analysis was made using forty decamer primers. Out of them, 15 primers were selected and used for identification and genetic relationships within 15 clones. A total of 138 distinct DNA fragments ranging from 0.2 to 3.2 kb were amplified using 15 selected random primers. The genetic similarity was evaluated on the basis of presence or absence of bands. The genetic distance was very close within the clones. Thus, these RAPD markers have the potential for identification of species and characterization of genetic variation within the population. This study will be helpful to know the genetic background of the medicinal plants with high commercial value, and also provides a major input into conservation biology. 相似文献