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1.
An influence of some Random Amplified Polymorphic DNA (RAPD) reaction factors on resulting banding pattern and the ability
of RAPD technique to detect DNA polymorphism among six economically important pea cultivars was tested. Relatively high level
of DNA polymorphism among peas was observed, using polyacrylamide/urea gels and silver staining. Altogether 13 arbitrarily
designed primers produced 313 amplification products. In addition 59 polymorphisms were found. These polymorphisms can serve
as potential genetic markers. RAPD data were processed using cluster analysis and plotted as dendrogram. Each tested cultivar
was clearly distinguished from the others. Moreover,Pisum sativum andP. sativum subsp.arvense cultivars were separated into 2 different clusters, according to their systematic relationships. 相似文献
2.
Genetic variation detected with RAPD markers among upland and lowland rice cultivars (Oryza sativa L.) 总被引:1,自引:1,他引:0
L. -X. Yu H. T. Nguyen 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,87(6):668-672
Genetic variation of nine upland and four lowland rice cultivars (Oryza sativa L.) was investigated at the DNA level using the randomly amplified polymorphic DNA (RAPD) method via the polymerase chain reaction (PCR). Forty-two random primers were used to amplify DNA segments and 260 PCR products were obtained. The results of agarosegel electrophoretic analysis of these PCR products indicated that 208 (80%) were polymorphic. All 42 primers used in this experiment were amplified and typically generated one-to-four major bands. Only two primers showed no polymorphisms. In general, a higher level of polymorphism was found between japonica and indica subspecies while fewer polymorphisms were found between upland and lowland cultivars within the indica subspecies. A dendrogram that shows the genetic distances of 13 rice cultivars was constructed based on their DNA polymorphisms. Classification of rice cultivars based on the results from the RAPD analysis was identical to the previous classification based on isozyme analysis. This study demonstrated that RAPD analysis is a useful tool in determining the genetic relationships among rice cultivars. 相似文献
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5.
“Candidatus Liberobacter,” the uncultured bacterium associated with citrus Huanglongbing (HLB) disease, is an α-Proteobacteria, and two species, “Candidatus L. africanum” and “Candidatus L. asiaticum,” have been characterized by sequence analysis of the 16S rDNA and β operon (rplKAJL-rpoBC) genes. These genes were isolated by PCR and random cloning of DNA from infected plants. However, this strategy is laborious
and allowed selection of only three Liberobacter DNA fragments. In this paper, we described isolation of additional genes
using Random Amplified Polymorphic DNA (RAPD). In total, 102 random 10-mer primers were used in PCR reactions on healthy and
Liberobacter-infected plant DNA. Eight DNA bands amplified from infected plant DNA were cloned and analyzed. Six of them were
found to be part of the Liberobacter genome by sequence and hybridization experiments. On these DNA fragments, four genes
were identified: nusG, pgm, omp, and a hypothetical protein gene. These results indicate that RAPD can be used to clone DNA of uncultured organisms.
Received: 14 September 1998 / Accepted: 6 October 1998 相似文献
6.
Using randomly amplified polymorphic DNA for evaluating genetic relationships among papaya cultivars 总被引:2,自引:0,他引:2
J. I. Stiles C. Lemme S. Sondur M. B. Morshidi R. Manshardt 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1993,85(6-7):697-701
Summary We have applied the recently developed technique of random amplification of polymorphic DNA (RAPD) to the analysis of the relationships among ten cultivars of papaya (Carica papaya L.). Eleven ten-base synthetic oligonucleotides were chosen that gave multiple PCR amplification products using papaya DNA as template. These 11 primers amplified a total of 102 distinct fragments. Cultivars were scored for presence or absence of RAPD fragments and grouped by cluster analysis using simple matching coefficients of similarity. A dendrogram of the ten cultivars was constructed. Of the ten cultivars seven were of the Hawaiian type, and all of these grouped to one branch of the tree. Divisions within the Hawaiian, branch were mostly consistent with the known genetic background of these cultivars. Three non-Hawaiian, cultivars were also analyzed. The minimum similarity detected was 0.7 suggesting that the domesticated papaya germ plasm is quite narrow. Our results show that RAPD technology is a rapid, precise and sensitive technique for genomic analysis.Journal series No. 3692 of the Hawaii Institute of Tropical Agriculture and Human Resources 相似文献
7.
Elisabetta Schiliro Stefano Predieri Assunta Bertaccini 《Plant Molecular Biology Reporter》2001,19(3):271-272
Pyrus communis L. is the most important pear species for European production. Very few cultivars satisfy standards for fruit quality and
clonal fidelity; thus, accurate verification of cultivar identity for checking propagation material and patent protection
is important. We evaluated the randomly amplified polymorphic DNA (RAPD) technique for its ability to identify genetic differences
among standard pear (Pyrus communis L.) cultivars, William, Passa Crassana, and Conference, and three gamma-ray induced variants. To identify genotype-specific
markers, we used thirty 10-mer and two 11-mer sequences, annealing temperatures from 36–45°C, 2Taq polymerases (AmpliTaq and Stoffel fragment, both from former Perkin Elmer Cetus), and 2–4 replicate amplifications. Of the
32 primers (30 from Operon Technologies, Alameda, CA, USA), very few distinguished William from Passa Crassana, and only 1
could clearly differentiate all 3 cultivars. Two primers that did not reveal polymorphisms when used singly, generated polymorphic
patterns that distinguished standard from gamma-ray-treated material when used in combination. We show that RAPD analyses
can discriminate pear genotypes and suggest this technique as a reliable and inexpensive method for marker-facilitated screening
of propagation material and for patent protection. 相似文献
8.
Rapid estimation of genetic relatedness among heterogeneous populations of alfalfa by random amplification of bulked genomic DNA samples 总被引:10,自引:1,他引:9
Kangfu Yu K. P. Pauls 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1993,86(6):788-794
A procedure which involves the use of RAPD markers, obtained from bulked genomic DNA samples, to estimate genetic relatedness among heterogeneous populations is demonstrated in this study. Bulked samples of genomic DNA from several alfalfa plants per population were used as templates in polymerase chain reactions with different random primers to produce RAPD patterns. The results show that the RAPD patterns can be used to determine genetic distances among heterogeneous populations and cultivars which correspond to their known relatedness. The results also indicate that, by using ten primers with bulked DNA samples from ten individuals, 18–72 populations or cultivars can be distinguished from each other on the basis of at least one unique RAPD marker. We anticipate that DNA bulking and methods for comparing RAPD patterns will be very useful for identifying cultivars, for studying phylogenetic relationships among heterogeneous populations and for selecting parents to maximize heterosis in crosses. 相似文献
9.
M. B. Ratnaparkhe V. S. Gupta M. R. Ven Murthy P. K. Ranjekar 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1995,91(6-7):893-898
Randomly amplified polymorphic DNA (RAPD) markers were used for the identification of pigeonpea [Cajanus cajan (L.) Millsp.] cultivars and their related wild species. The use of single primers of arbitrary nucleotide sequence resulted in the selective amplification of DNA fragments that were unique to individual accessions. The level of polymorphism among the wild species was extremely high, while little polymorphism was detected within Cajanus cajan accessions. All of the cultivars and wild species under study could be easily distinguished with the help of different primers, thereby indicating the immense potential of RAPD in the genetic fingerprinting of pigeonpea. On the basis of our data the genetic relationship between pigeonpea cultivars and its wild species could be established.NCL Communication No. 6062 相似文献
10.
随机扩增多态DNA(RAPD)标记用于丁香品种遗传分析及品种分类 总被引:8,自引:0,他引:8
采用随机扩增多态DNA(RAPD)标记分析了15个丁香品种的DNA扩增产物。研究选用了16个随机引物,共扩增出96条带,其中55条带为可重复性条带,有价值条带大小多在517bp至1636bp之间。这些标记足以区分这些丁香品种。欧丁香(Syringavulgaris)与S.×hyacinthiflora间的相似系数为61.5%,欧丁香与S.×prestoniae间的相似系数为47.2%,S.×hyacinthiflora与S.×prestoniae间的相似系数为43.6%。结果表明,欧丁香与S.×hyacinthiflora亲缘关系最近。应用RAPD资料分析讨论了一些品种的起源。RAPD技术为丁香品种分类鉴定提供了可靠方法。 相似文献
11.
Random amplified polymorphic DNA (RAPD) markers were used to determine the genetic stability of long-term (more than 10 years)
micropropagated shoots of Japanese black pine (Pinus thunbergii Parl.). Thirty-six shoots consisting of three morphotypes (short, medium, and long needles) were randomly chosen from about
4,000 micropropagated shoots regenerated from the explants of a single nematode-resistant mother plant. Out of 126 primers
screened, 30 gave 134 clear reproducible bands. A total of 4,824 bands obtained from these studies exhibited no aberration
in RAPD banding patterns among the tested shoots. Our results show that regenerants from our plant micropropagation system
are genetically stable.
Received: 5 December 1997 / Revision received: 17 May 1998 / Accepted: 1 June 1998 相似文献
12.
J. I. Hormaza L. Dollo V. S. Polito 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,89(1):9-13
The Random Amplified Polymorphic DNA (RAPD) technique was used to amplify DNA segments, with the objective of finding markers linked to sex determination in the dioecious species, Pistacia vera. Progenies from two female parents pollinated by a common male parent were studied. Two bulks of DNA were made in each cross, one from males and one from females, by pooling an equal weight of fresh leaves from each individual contributing to the bulk prior to DNA extraction. DNA was extracted from each bulked sample and from each of the contributing individuals. DNA was also extracted from 14 cultivars of P. vera and from 94 open-pollinated, fewweeks-old P. vera seedlings of unknown sex. Seven hundred different decamer oligonucleotide primers were used to perform DNA amplification, with 1 of these (OPO08) producing a 945 bp amplification band that was present only in the bulked female samples and absent in the bulked male samples of the two crosses. The relationship between band presence and female sex expression was conserved in every individual obtained from the two crosses and in the 14 cultivars unrelated to the crosses. We propose that this band is tightly linked to the gene(s) that control sex determination in pistachio. The OPO08945 RAPD marker could be used in a breeding program to screen the gender of pistachio plants long before they reach reproductive maturity, resulting in considerable savings of time and economic resources. In order to verify that assumption we screened 94 additional seedlings with the OPO08 primer and obtained results consistent with a 11 male:female ratio. 相似文献
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14.
W. Cao G. Scoles P. Hucl R. N. Chibbar 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,98(3-4):602-607
Crop germplasm collections contain a considerable percentage of misclassified accessions which may affect the use of germplasm
for agricultural crop improvement. The objective of this study was to determine if random amplified polymorphic DNA (RAPD)
analysis could be used to reclassify misclassified Triticum accessions. Twelve accessions suspected to be misclassified, based on morphological characters, as either macha or vavilovii
wheat were studied using RAPD and cytological analyses. In the RAPD analysis, a dendrogram, based on Jaccard genetic similarity
coefficients, grouped 5 dicoccum-like, 1 timopheevii-like, and 6 monococcum-like accessions with Triticum dicoccum, T. timopheevii, and T. monococcum accessions, respectively. These results were confirmed by the cytological analysis. A RAPD marker specific to the D genome
was also detected. This study suggests that RAPD analysis can be used to classify germplasm and to distinguish some species
in Triticum.
Received: 12 June 1998 / Accepted: 18 August 1998 相似文献
15.
S. M. Udupa F. Weigand M. C. Saxena G. Kahl 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,97(1-2):299-307
The poor definition of variation in the ascochyta blight fungus (Ascochyta rabiei) has historically hindered breeding for resistance to the chickpea (Cicer arietinum L.) blight disease in West Asia and North Africa. We have employed 14 RAPD markers and an oligonucleotide probe complementary
to the microsatellite sequence (GATA)4 to construct a genotype-specific DNA fragment profile from periodically sampled Syrian field isolates of this fungus. By
using conventional pathogenicity tests and genome analysis with RAPD and microsatellite markers, we demonstrated that the
DNA markers distinguish variability within and among the major pathotypes of A. rabiei and resolved each pathotypes into several genotypes. The genetic diversity estimate based on DNA marker analysis within pathotypes
was highest for the least-aggressive pathotype (pathotype I), followed by the aggressive (pathotype II) and the most-aggressive
pathotype (pathotype III). The pair-wise genetic distance estimated for all the isolates varied from 0.00 to 0.39, indicating
a range from a clonal to a diverse relationship. On the basis of genome analysis, and information on the spatial and temporal
distribution of the pathogen, a general picture of A. rabiei evolution in Syria is proposed.
Received: 10 January 1998 / Accepted: 23 January 1998 相似文献
16.
An influence of some Random Amplified Polymorphic DNA (RAPD) reaction factors on resulting banding pattern and the ability of RAPD technique to detect DNA polymorphism among six economically important pea cultivars was tested. Relatively high level of DNA polymorphism among peas was observed, using polyacrylamide/urea gels and silver staining. Altogether 13 arbitrarily designed primers produced 313 amplification products. In addition 59 polymorphisms were found. These polymorphisms can serve as potential genetic markers. RAPD data were processed using cluster analysis and plotted as dendrogram. Each tested cultivar was clearly distinguished from the others. Moreover,Pisum sativum andP. sativum subsp.arvense cultivars were separated into 2 different clusters, according to their systematic relationships. 相似文献
17.
S. K. Chakrabarti D. Pattanayak D. Sarkar V. P. Chimote P. S. Naik 《Biologia Plantarum》2006,50(4):531-536
Variations in random amplified polymorphic DNA (RAPD) profiles from leaf, stem, root, and tuber tissues were observed in case
of two glasshouse grown potato cultivars using 40 decamer primers suggesting possible danger of cultivar misidentification.
Genomic DNA extracted from the above four tissues of four in vitro grown potato cultivars, however, produced more uniform RAPD fingerprints. A significant effect of random primers on fingerprint
uniformity was observed in case of both glasshouse and in vitro grown samples. A new concept of stability index for random primers based on homogeneity of RAPD profiles obtained from different
tissues of a single plant have been introduced. It is concluded that RAPD analysis of genomic DNA extracted from any tissue
of in vitro grown potato plants using 14 selected decamer primers could be used to develop RAPD fingerprints for identification of Indian
potato cultivars. 相似文献
18.
Random amplification of polymorphic DNA (RAPD) was used to analyze six species, three populations, and seven regional cultivars of barley. A unique pattern of amplified DNA products was obtained for each species of the genus Hordeum.High polymorphism of barley species was revealed. Specific fragments were found in most RAPD patterns; the fragments can be used as molecular markers of corresponding species and subspecies. Several other DNA fragments were shown to serve as molecular markers of the H genome. Specific RAPD patterns were obtained for each population and each cultivar of H. vulgaresensu lato. In total, variation between the populations and between the cultivars was substantially lower than between species. Cluster analysis (UPGMA) was used to estimate genetic distances between theHordeumspecies, between the H. spontaneumpopulations, and between regional H. vulgarecultivars and a dendrogram was constructed. 相似文献
19.
K. V. Chowdari A. P. Davierwala V. S. Gupta P. K. Ranjekar O. P. Govila 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,97(1-2):154-162
The potential of DNA markers such as microsatellites, minisatellites and RAPDs was investigated in pearl millet [Pennisetum glaucum (L.) R. Br] with respect to their abundance and variability. Southern analysis, using 22 different di-, tri-, tetra- and
penta-oligonucleotide probes and five minisatellite probes, identified (GATA)4 as the most useful probe for the detection of multiple polymorphic fragments among pearl millet cultivars and landraces from
India. The clustering patterns of pearl millet cultivars and landraces based on (GATA)4 and RAPD (randomly amplified polymorphic DNA) markers differed. The landraces, representing eight states in India, could
not be grouped based on their geographical distribution with the DNA markers. RAPD analysis revealed a high degree of genetic
diversity among the cultivars and landraces employed in this study. The probability of an identical match by chance for any
two genotypes using (GATA)4 and RAPDs was 3.02×10-20 for cultivars and 5.2×10-9 for landraces. The microsatellite (GATA)4 and RAPDs provide useful tools for genotype identification and for the assessment of genetic relationships in pearl millet.
Received: 19 October 1997 / Accepted: 9 December 1997 相似文献
20.
Tui Ray Indrajit Dutta Prasenjit Saha Sampa Das S.C. Roy 《Plant Cell, Tissue and Organ Culture》2006,85(1):11-21
An efficient micropropagation protocol produced large number of plants of the three elite banana (Musa spp.) cultivars Robusta (AAA), Giant Governor (AAA) and Martaman (AAB) from shoot tip meristem. The genetic relationships
and fidelity among the cultivars and micropropagated plants as assessed by random amplified polymorphic DNA (RAPD) and inter-simple
sequence repeat (ISSR) markers, revealed three somaclonal variants from Robusta and three from Giant Governor. A total of
5330 RAPD and 2741 ISSR fragments were generated with 21 RAPD and 12 ISSR primers in micropropagated plants. The percentage
of polymorphic loci by RAPD and ISSR were found to be 1.75, 5.08 in Robusta and 0.83, 5.0 in Giant Governor respectively.
Among the two marker systems used, ISSR fingerprinting detected more polymorphism than RAPD in Robusta and Giant Governor
with most of the primers showing similar fingerprinting profile, whereas Martaman revealed complete genetic stability. 相似文献