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AFLP(扩增性片段长度多态性)是一种新的DNA分子标记。与RFLP、RAPD相比,AFLP具有在一次试验中可同时观察到大量的限制性片段的优点。本文阐述了AFLP的原理和方法,综述了AFLP目前在植物遗传育种研究中的应用进展,并对AFLP技术在植物遗传育种中的应用前景提出了初步设想。 相似文献
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Amplified Fragment Length Polymorphism (AFLP) Provides Molecular Markers for the Identification of Caladium bicolor Cultivars 总被引:4,自引:0,他引:4
LOH JIN PHANG; KIEW RUTH; KEE ANDREA; GAN LEONG HUAT; GAN YIK-YUEN 《Annals of botany》1999,84(2):155-161
Caladiums are popular ornamental plants that have not been wellstudied at the molecular level. Identification of species withinthe genus Caladium (Araceae) has been based primarily on morphology.However, the lack of comprehensive references makes identificationof Caladium cultivars extremely difficult. Amplified fragmentlength polymorphism (AFLP) analysis using 17 primer combinationswas carried out on two species of Caladium (C. bicolor and C.schomburgkii), including six cultivars of C. bicolor. Resultsshowed that AFLP can be used to distinguish these two speciesby their unique and different banding patterns. Unweighted PairGroup Method using Arithmetic Averages (UPGMA) permitted clusteranalysis of data from 17 selected primer combinations on sixcultivars of C. bicolor and one cultivar ofC. schomburgkii .It showed that closely related species can clearly be differentiatedand that genetic difference between cultivars can also be established.Unique AFLP molecular markers were detected for all the C. bicolorcultivars used. The use of AFLP has potential for preciselycharacterizing and identifying particular caladium cultivarsas well as for the registration of new cultivars. It will alsobe useful in future breeding programmes and systematics studies.Copyright 1999 Annals of Botany Company Araceae, Caladium species and cultivars, AFLP DNA fingerprinting, diversity, AFLP markers. 相似文献
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用扩增片段的长度多态性(amplified fragment length polymorphism,AFLP)标记分析研究了中国5个盾叶薯蓣居群30个个体的遗传多样性。筛选出9对AFLP引物,从中检测到14698条清晰可见的条带,其中多态性带12628条,多态性比率85.92%。Shannon信息指数(I)为0.3656±0.1721,物种水平的Nei基因多样性(H)为0.2322±0.2200。遗传变异分析表明,物种水平的遗传分化系数Gst为0.4827,说明其群体间存在一定的遗传分化,居群间的基因流Nm为0.5358,居群间遗传交换较小。聚类分析结果显示5个居群盾叶薯蓣有较为丰富的遗传变异,且与地理分布有相关性。 相似文献
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Hamid Badali Seyed Amir Yazdanparast Alexandro Bonifaz Bita Mousavi G. Sybren de Hoog Corné H. W. Klaassen Jacques F. Meis 《Mycopathologia》2013,175(5-6):505-513
Inter- and intraspecific genomic variability of 18 isolates of Veronaea botryosa originating from clinical and environmental sources was studied using amplified fragment length polymorphism (AFLP). The species was originally described from the environment, but several severe cases of disseminated infection in apparently healthy individuals have been reported worldwide. All tested strains of V. botryosa, identified on the basis of sequencing and phenotypic and physiological criteria prior to our study, were confirmed by AFLP analysis, yielding a clear separation of V. botryosa as a rather homogeneous group from related species. In vitro antifungal susceptibility testing resulted in MIC90s across all strains in increasing order posaconazole (0.25 μg/ml), itraconazole (1 μg/ml), voriconazole (4 μg/ml), terbinafine (4 μg/ml), caspofungin (8 μg/ml), anidulafungin (8 μg/ml), isavuconazole (16 μg/ml), amphotericin B (16 μg/ml), and fluconazole (32 μg/ml). Overall, the isolates showed a uniform pattern of low MICs of itraconazole and posaconazole, but high MICs for remaining agents. The echinocandins (caspofungin and anidulafungin) had no activity against V. botryosa. There was no statistically significant difference between susceptibilities of environmental (n = 11) and clinical (n = 7) isolates of V. botryosa (P > 0.05). 相似文献
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Amplified Fragment Length Polymorphism (AFLP) as a source of genetic markers for red algae 总被引:7,自引:0,他引:7
A recent PCR-based fingerprinting technique, amplified fragment length polymorphism (AFLP), was successfully applied to the
red alga Chondrus crispus Stackh. This is apparently the first account to describe the application of AFLP methodology to
an alga. Six isolates of C. crispus were analyzed by AFLP. A total of twenty-five primer pairs were screened and six primer
pairs were selected for further investigation. Both conservative and variable markers were identified within and between populations;
some markers were unique to individuals. As such, AFLP should prove useful as a source of genetic markers in algae for applications
as diverse as genome mapping to population genetic investigations.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
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Farida Bouftass Mustapha Labhilili Brahim Ezzahiri Aziz Ziouti 《Journal of Phytopathology》2010,158(2):111-116
The genetic variability and collection structure of the wheat leaf rust fungus Puccinia recondita collected from four agro‐ecological areas of Morocco, Abda‐doukala, Chaouia‐Tadla, Gharb and Tangérois were investigated by amplified fragment length polymorphism (AFLP) markers. A set of five AFLP primers combinations which generated 253 polymorphic loci were used. Hierarchical partitioning revealed that bread wheat collections of Puccinia recondita form a single collection. No significant variation was observed between durum wheat collections of Puccinia recondita; they maintained most of the genetic variability within rather among collections. The distribution pattern of genetic variation of Puccinia recondita collections seems to be the result of high gene flow and the mixed reproduction system. 相似文献
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A Study of Genetic Variation and Relationships within the Bamboo Subtribe Bambusinae using Amplified Fragment Length Polymorphism 总被引:10,自引:1,他引:9
Loh Jin Phang; Kiew Ruth; Set Ohn; Gan Leong Huat; Gan Yik-Yuen 《Annals of botany》2000,85(5):607-612
Taxonomic and systematic studies of the woody bamboos are traditionallybased on floral morphology, which can cause problems in identificationdue to the lack of, or infrequent, flowering. Limited studieshave been conducted using molecular techniques to overcome thisproblem. In this study, we used amplified fragment length polymorphisms(AFLPs) to conduct a study of four genera of bamboos (Bambusa,Dendrocalamus, Gigantochloa andThyrsostachys ) in the subtribeBambusinae. AFLP analysis using eight primer combinations wascarried out on 15 species of bamboo. Results showed that AFLPsdistinguish the different species by their unique banding patterns.Unique AFLPs were detected in 13 of the 15 species examined.The six Bambusa species examined separated into two clusters.The sixGigantochloa species studied formed a discrete clusterdiverging from one of the Bambusa clusters, whileThyrsostachyswas less similar to the Bambusa clusters. The similarity indexbetween B. lako and G. atroviolacea was the highest, suggestingthat B. lako is more appropriately included within the genusGigantochloarather than the genus Bambusa. The two Dendrocalamus speciesexamined were very different with D. brandisii clustering withinone of the Bambusa clusters and D. giganteus appearing as avery distant species. These results support the contention thatcritical study of the genus Dendrocalamus is required. The useof AFLPs for identification of particular bamboo species, aswell as for the study of relationships within the subtribe,will be useful for industrial purposes and for systematic studies.Copyright 2000 Annals of Botany Company Bamboo, Bambusinae, Bambusa, Gigantochloa, Dendrocalamus, Thyrsostachys, AFLP, diversity, AFLP markers 相似文献
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Although the use of molecular markers in population genetics of marine organisms is increasingly employed, methodologic limitations still hampered the research for some taxa, such as symbiotic cnidarians, including scleractinian corals. The development of molecular tools in scleractinian corals' studies is faced with a list of obstacles, such as high cost, labor, time consuming, contamination with foreign DNA, and markers with low resolution. The AFLP (amplified fragment length polymorphism) method, overcomes most of the obstacles listed above except of the difficulty of contamination by algal symbiont DNA. We compared the implication of two pre-DNA extraction treatments to obtain coral DNA free of algal contaminations, termed as CPEM, cell population enriched method, and TTEM, total tissue extraction method. The CPEM process result in pure coral DNA for all samples, but is time consuming, whereas in the TTEM process, approximately 25% to 18% of the samples are still contaminated by algal DNA. However, algal DNA contaminations in the PCR at 2.5 x 10(-1) ng level (approximately 100 algal cells) and below, did not amplify any new AFLP band or peak for neither radioactive nor florescence analyses. Therefore, even the TTEM process may be used because it is faster, easier to handle, and easily employed on a large amount of samples, with minimal contamination artifacts. When correctly employed, both methods are applicable to wide experimental manipulations. 相似文献
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We have utilized Amplified Fragment Length Polymorphisms (AFLP) in conjunction with chloroplast DNA (cpDNA) sequence data to study the genetic diversity in 53 accessions of Dioscorea dumetorum from six countries in West and Central Africa. Our results provide a comparison of the two marker systems with regards to their applicability to differentiate intraspecific genotypes and the grouping of the accessions based on localities of collection. A total of 1052 AFLP fragments (of which 94.1% were polymorphic) produced from twelve primer combinations indicate a relatively high level of polymorphism among the accessions. Three major genetic groups that do not strictly follow a geographic distribution pattern were identified using Neighbour-joining and the principal coordinate (PCo) analyses. Accessions from Togo showed higher numbers of private fragments and the highest percentage polymorphism (59.4%). The detection of highest genetic diversity in accessions from Nigeria and Togo and their relationship to other accessions suggest that these countries are the centre of origin and diversity of D. dumetorum. The moderately high genetic diversity (average of 61%) is suggesting great influence on the D. dumetorum germplasms through exchange and transfer of cultivars among local farmers in the sub-region. In contrast, DNA sequence data from the psbA-trnH and the rpoB-trnC chloroplast regions revealed no variation among accessions from the different localities and clearly differentiated by AFLP patterns. The results demonstrate the usefulness of the AFLP marker in generating high polymorphism in the D. dumetorum accessions from West and Central Africa and hence may be used for agronomic purposes. 相似文献
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Blanco-Conde Sara González-Cortés Carolina López-Medrano Ramiro Palacios-Gutiérrez Juan José Diez-Tascón Cristina Nebreda-Mayoral Teresa Sierra-García María Josefa Rivero-Lezcano Octavio Miguel 《Molecular biology reports》2020,47(5):3397-3405
Molecular Biology Reports - The increasing worldwide incidence of mycobacteriosis and the need to achieve improved clinical management makes nontuberculous mycobacteria (NTM) genotyping a useful... 相似文献
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Esteban A Leong SL Hocking AD Abarca ML Cabañes FJ Tran-Dinh N 《Current microbiology》2008,57(4):348-355
Microsatellite markers and the results of amplified fragment length polymorphism (AFLP) were compared in the characterization of 68 Aspergillus carbonarius and A. niger aggregate strains of differing ochratoxin-producing ability and from different geographic areas, isolated mainly from grapes and soil. AFLP was applied to both A. carbonarius and A. niger aggregate strains, and it clearly differentiated these species. Microsatellite markers were only applied to A. niger aggregate strains because of the species-specific nature of these markers. Both AFLP and microsatellite marker analyses were able to divide A. niger aggregate strains into the two recognized internal transcribed spacer (ITS)-5.8S rDNA RFLP types, N and T. Clustering of A. niger aggregate strains was similar in both AFLP and microsatellite analyses, yielding an additional separation of N type strains into two groups. Both microsatellite marker and AFLP analyses showed high levels of polymorphism in the A. niger aggregate (index of discriminatory power 0.991 and 1.0, respectively). Of the two techniques, microsatellite marker analysis was quicker and more straightforward to perform. In addition, microsatellite marker analysis is more reproducible, and the results can be expressed as quantitative data, making microsatellite markers a good candidate for use in large-scale studies of genetic diversity in A. niger aggregate species. 相似文献
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Sanjuana?Hernández-Delgado Netzahualcoyotl?Mayek-Pérez Gustavo?Emilio?Santos-Medrano Roberto?Rico-Martínez
We have used amplified fragment length polymorphisms (AFLP) to investigate the potential of this technique as a tool to measure genetic variability in eight species of freshwater rotifers: Brachionus calyciflorus, Lecane bulla, L. luna, L. quadridentata, Plationus patulus, Philodina acuticornis odiosa, Rotaria neptunia, and R. rotatoria. We used nine combinations of oligonucleotides. We observed a total of 806 amplified bands, 798 polymorphic and 8 monomorphic. The data were analyzed using cluster analysis with UPGMA, first within each set of oligonucleotide combination and finally using all nine combinations. Our best dendrogram clearly separated monogononts from digononts, and grouped the species of monogononts in the two genera. However, it grouped R. neptunia with P. acuticornis odiosa rather than with R. rotatoria. These results are discussed in view of recent works in the literature measuring genetic variability and discussing the phylogeny of the Rotifera. 相似文献
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Amplified fragment length polymorphisms (AFLP) were used to study the inheritance of shell color in Argopecten irradians.
Two scallops, one with orange and the other with white shells, were used as parents to produce four F1 families by selfing and outcrossing. Eighty-eight progeny, 37 orange and 51 white, were randomly selected from one of the
families for segregation and mapping analysis with AFLP and microsatellite markers. Twenty-five AFLP primer pairs were screened,
yielding 1138 fragments, among which 148 (13.0%) were polymorphic in two parents and segregated in progeny. Six AFLP markers
showed significant (P < 0.05) association with shell color. All six loci were mapped to one linkage group. One of the markers, F1f335, is completely
linked to the gene for orange shell, which we designated as Orange1, without any recombination in the progeny we sampled.
The marker was amplified in the orange parent and all orange progeny, but absent in the white parent and all the white progeny.
The close linkage between F1f335 and Orange1 was validated using bulk segregation analysis in two natural populations, and
all our data indicate that F1f335 is specific for the shell color gene, Orange1. The genomic mapping of a shell color gene
in bay scallop improves our understanding of shell color inheritance and may contribute to the breeding of molluscs with desired
shell colors. 相似文献
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Genomic Relatedness of Chlamydia Isolates Determined by Amplified Fragment Length Polymorphism Analysis 总被引:4,自引:0,他引:4 下载免费PDF全文
Adam Meijer Servaas A. Morr Adriaan J. C. Van Den Brule Paul H. M. Savelkoul Jacobus M. Ossewaarde 《Journal of bacteriology》1999,181(15):4469-4475
The genomic relatedness of 19 Chlamydia pneumoniae isolates (17 from respiratory origin and 2 from atherosclerotic origin), 21 Chlamydia trachomatis isolates (all serovars from the human biovar, an isolate from the mouse biovar, and a porcine isolate), 6 Chlamydia psittaci isolates (5 avian isolates and 1 feline isolate), and 1 Chlamydia pecorum isolate was studied by analyzing genomic amplified fragment length polymorphism (AFLP) fingerprints. The AFLP procedure was adapted from a previously developed method for characterization of clinical C. trachomatis isolates. The fingerprints of all C. pneumoniae isolates were nearly identical, clustering together at a Dice similarity of 92.6% (+/- 1.6% standard deviation). The fingerprints of the C. trachomatis isolates of human, mouse, and swine origin were clearly distinct from each other. The fingerprints of the isolates from the human biovar could be divided into at least 12 different types when the presence or absence of specific bands was taken into account. The C. psittaci fingerprints could be divided into a parakeet, a pigeon, and a feline type. The fingerprint of C. pecorum was clearly distinct from all others. Cluster analysis of selected isolates from all species revealed groups other than those based on sequence data from single genes (in particular, omp1 and rRNA genes) but was in agreement with available DNA-DNA hybridization data. In conclusion, cluster analysis of AFLP fingerprints of representatives of all species provided suggestions for a grouping of chlamydiae based on the analysis of the whole genome. Furthermore, genomic AFLP analysis showed that the genome of C. pneumoniae is highly conserved and that no differences exist between isolates of respiratory and atherosclerotic origins. 相似文献
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Restriction Fragment Length Polymorphism and Random Amplified Polymorphic DNA Analysis of Chickpea Accessions 总被引:1,自引:0,他引:1
Genetic diversity analysis was carried out in chickpea accessions using restriction fragment length polymorphism (RFLP) and
random amplified polymorphic DNA (RAPD) techniques. RFLP analysis using 26 Pst I sub-genomic clones on ten chickpea accessions
in 130 probe-enzyme combinations detected polymorphism with only two clones. Pst I clones, CG 141 detected polymorphism in
ICC 4918 and Pusa 209 while CG 500 detected polymorphism in Pusa 261, ILC 26 and in ILC 13326. These clones detected very
few polymorphic markers. Analysis using 10 Eco RI clones on twelve chickpea accessions have shown better hybridisation signal
and one clone detected polymorphism in Pusa 256. RFLP analysis of both cultivated and wild Cicer species using heterologous
DNA probe Cab3C revealed polymorphism only in wild Cicer species (Cicer reticulatum L., JM 2100). RAPD analysis of 13 chickpea
accessions which includes mutants of C 235 and E100Y showed greater degree of polymorphism with 1 - 5 unique DNA bands for
all the accessions. Phylogenetic analysis of the RAPD data helped to group the accessions. C 235 and its mutants were found
to be closely grouped while E100Y and its mutant E100Ym grouped apart. Desi and kabuli chickpea accessions however, could
not be separately grouped.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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Intraspecific Differentiation of Vibrio vulnificus Biotypes by Amplified Fragment Length Polymorphism and Ribotyping 总被引:3,自引:0,他引:3 下载免费PDF全文
The intraspecific genomic relatedness of 80 Vibrio vulnificus isolates, 44 of biotype 1 and 36 of biotype 2, from different geographic origins and sources was evaluated by ribotyping and AFLP (amplified fragment length polymorphism) fingerprinting. Ribopatterns of DNAs digested with KpnI and hybridized with an oligonucleotide complementary to a highly conserved sequence in the 23S rRNA gene revealed up to 19 ribotypes in the species, which were different for the two biotypes. Sixteen different ribotypes were found within biotype 1 strains from clinical and environmental sources, and only three, recovered mainly from diseased eels, were found within biotype 2. Within this biotype, 96% of the strains showed the same ribopattern. The closest similarity was shown by the strains coming from the same eel farm, irrespectively of biotype. AFLP fingerprints obtained by selective PCR amplification of HindIII-TaqI double-restricted DNA fragments exhibited a strain-specific pattern which allowed the finest differentiation of subgroups within the eel-pathogenic isolates sharing the same ribopattern. Both techniques revealed good genetic markers for intraspecific differentiation of V. vulnificus. Ribotyping clearly separated the eel-pathogenic strains from the clinical and environmental isolates, whereas AFLP enabled the monitoring of individual strains and therefore constitutes one of the most discriminative tools for epidemiological and ecological studies. 相似文献