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1.
S. Y. Park H. N. Murthy D. Chakrabarthy K. Y. Paek 《In vitro cellular & developmental biology. Plant》2009,45(1):104-108
Somaclones exhibiting variations with flower characteristics were recovered from the tissue-culture-derived plants of Doritaenopsis. Two molecular techniques, random amplified polymorphic DNA (RAPD) and methylation-sensitive amplification polymorphism (MSAP)
analyses, were used to characterize the somaclones. RAPD analysis, using 100 randomly selected primers, failed to differentiate
variants and normal plants, even though some primers (six out of 100 primers) exhibited 6–10 distinct banding patterns. However,
MSAP analysis revealed the differences in the DNA methylation patterns in the normal and variant plants which were correlated
with phenotypic variation. In all, 311, 337, 366, and 343 fragments were obtained with normal and V1, V2, and V3 variant plants,
respectively; each representing recognition site cleaved by either or both of the isoshizomers were amplified using 12 combination
of primers. A total of 36 (11.6%), 77 (22.9%), 73 (19.9%), and 47 (13.7%) sites were found to be methylated at cytosine in
the genomes of normal and V1, V2, and V3 variant Doritaenopsis plants. This study demonstrates usefulness of MSAP to detect DNA methylation events in tissue cultured Doritaenopsis plants. 相似文献
2.
P. K. Mandal I. M. Santha S. L. Mehta 《Journal of plant biochemistry and biotechnology.》1996,5(2):83-86
RAPD patterns were studied in seven somaclones of Lathyrus sativus having contrasting characteristics alongwith the parent cultivar P-24. Out of 81 decamer random primers used, 5 did not amplify and 24 revealed DNA polymorphism while the rest generated monomorphic banding patterns. Eight unique bands were amplified with different primers in four different somaclones. With most of the informative primers differences were observed between somaclones and also between some of the somaclones and parent cultivar P-24. More than 90% similarity in the RAPD patterns was evident among the somaclones and the parent cultivar P-24. Though it was not possible to identify a particular somaclone with a single primer, a combination of two or more primers could be employed to identify a somaclone. 相似文献
3.
In the present study, two polymerase chain reaction (PCR)-based methods namely, randomly amplified polymophic DNA (RAPD) and
amplification fragment length polymorphism (AFLP) were employed to assess genetic variations, which may appeared, in tissue
culture-derived date palm (Phoenix dactylifera) offshoots. Analysis of RAPD banding patterns generated by PCR amplification using 37 random primers gave no evidences for
somaclonal variations and the percentage of polymorphic bands in a total of 259 scored bands was zero. Meanwhile, analysis
of AFLP banding patterns generated using 13 primer combinations pointed to minor genetic variations in the AFLP banding patterns.
The percentage of genetic variations (polymorphism) in tissue culture-derived date palm offshoots belonging to cultivars Sakkoty,
Gandila and Bertamoda was 2.6, 0.79 and 1 %, respectively, as revealed by AFLP analysis. The low percentage of genetic variations
confirms the genetic stability of tissue culture-derived dry date palm cultivars. 相似文献
4.
Molecular markers were used to detect the influence of high concentrations of 2,4 dichlorophenoxyacetic acid (2,4-D) in the callusing media on DNA variations in regenerated rice plants. Restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD) and polymerase chain reaction (PCR) based RFLP analysis were carried out on 12 somaclones of Oryza sativa L. cv. B-370. In vitro culture induced DNA variations were detected in the regenerated plants but the effect of high auxin concentration in the medium could not be revealed. In a second study, fingerprinting of 15 semi-dwarf, high yielding somaclones of B-370 was carried out using RAPD technique. Amplification using 20 random primers produced a total of 167 DNA bands out of which 97 bands were polymorphic. A total of 32 unique DNA bands were detected across all the somaclones and they could be grouped based on their similarity to B-370. RAPD analysis helped to reveal similarity or differences among the somaclones while fingerprinting using additional RAPD markers was not successful. 相似文献
5.
Araújo L.G. Prabhu A.S. Filippi M.C. Chaves L.J. 《Plant Cell, Tissue and Organ Culture》2001,67(2):165-172
Seventeen somaclones of upland rice cultivar IAC 47 showing different plant types, and either resistance or susceptibility to leaf blast, were utilized for random amplified polymorphic DNA (RAPD) analysis. Somaclones exhibited differences in reaction to isolates of Pyricularia grisea. Two somaclones (SC02 and SC04) were resistant to all three field isolates of somaclones, while the cultivar IAC 47 was susceptible. The inheritance study of two distinct plant types, one with erect bright green leaves and the other with droopy yellow green leaves, showed that a single possibly different, dominant gene governs each plant type. Of 32 random decamer primers utilized, OPA02 and OPD02 detected polymorphisms between somaclones showing erect bright green leaves and droopy yellow green leaves. Reliable grouping exhibiting 80% similarity was achieved with 17 primers. Leaf blast resistance to race IC-2 of P. grisea was associated with the plant type of erect bright green leaves. 相似文献
6.
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
7.
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
8.
Tapan Kumar Nondal Pradeep Kumar Chand 《In vitro cellular & developmental biology. Plant》2002,38(3):296-299
Summary Randomly amplified polymorphic DNA (RAPD) techniques were applied to assess genetic instability among micropropagated tea
[Camellia sinensis (L.) O. Kuntze] eultivar ‘T-78’. Out of 49 random 10-mer primers, 11 generated polymorphism in four out of 17 micropropagated
plants and one mother plant. A total of 221 bands, ranging from 525 bp to 2.5 kb, were produced by the 49 primers. Twenty-four
were polymorphic for those four plants. However, the remaining bands were monomorphic among all plants. Polymorphism among
those four plants showed an identifical banding pattern suggesting the occurrence of a single mutation. Our results demonstrated
that RAPD can be used successfully to determine the genetic instability among micropropagated plants which otherwise were
morphologically indistinguishable. 相似文献
9.
Variability for resistance to Fusarium solani culture filtrate and fusaric acid among somaclones in pea 总被引:1,自引:0,他引:1
Pea (Pisum sativum L.) somaclones of cultivars Adept, Komet and Bohatýr were obtained after selection in vitro with Fusarium solani filtrate and fusaric acid (FA). R2 regenerants were analysed by random amplification of polymorphic DNA (RAPD; OPAB4, P-14, UBC-556) and inter-retrotransposon amplification polymorphism (IRAP; Ogre) markers. Marker UBC-556 showed different banding patterns for each cultivar, but without specific bands for selected and control plants. Markers OPAB4, P14 and Ogre were useful for clear discrimination between selected and non-selected variants of all three cultivars. Flow cytometry analysis proved the same genome size of selected and non-selected pea lines. Therefore in vitro selection by pathogen derived agents could be the efficient method for obtaining of pea somaclones with increased resistance to F. solani. 相似文献
10.
Guillermo Pérez Andrew Mbogholi Fernando Sagarra Carlos Aragón Justo González Miriam Isidrón José Carlos Lorenzo 《In vitro cellular & developmental biology. Plant》2011,47(3):428-433
We previously reported a partial agricultural and amplified fragment length polymorphism (AFLP) characterization of two new
pineapple somaclones (P3R5 and Dwarf) derived from in vitro culture of the donor cv. Red Spanish Pinar. Both somaclonal variants showed different AFLP banding patterns compared to the
donor cultivar, although they were separated by less than 0.09 U of genetic distance. The present report shows data of various
indicators of morphology and physiology of P3R5 and Dwarf D leaves. The stoma diameter, number of stomata per square millimiter, diameter of leaf vascular tissue, thickness of the leaf
aquiferous parenchyma, and thickness of the leaf photosynthetic parenchyma were measured. The photosynthetic rate, the transpiration
rate, the water use efficiency, the internal leaf CO2 concentration, and the chlorophyll pigment contents were recorded as well. Between the somaclonal variant P3R5 and the donor
genotype, statistically significant differences were recorded in all indicators with the exception of the stoma diameter and
the photosynthetic rate. Comparing the somaclonal variant Dwarf and the cv. Red Spanish Pinar (donor), statistically significant
differences were also recorded in all parameters except in the stoma diameter and in the transpiration rate. This investigation
was performed to demonstrate that small changes in the pineapple DNA may result in relevant phenotypic modifications. 相似文献
11.
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
12.
“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
13.
ME, PGM and PGI electrophoretic banding patterns in 20 laboratory cultures representing 14 species ofTrichogramma were studied. Variations in PGM were found inT. exiguum, T. marylandense, andT. pretiosum. PGI also showed variation inT. exiguum, T. marylandense, T. minutum, andT. parkeri. However, ME variations were found only inT. pretiosum. Based on progeny analyses, we concluded that ME is a tetramer inTrichogramma with fast and slow alleles at a single locus, and that both PGM and PGI have a single locus and each has four alleles. PGM
is a monomer, but PGI is a dimer.
Résumé Les bandes électrophorétiques de l'enzyme malique, de la P.G.M. et de la P.G.I. ont été étudiées chez 20 souches de laboratoire représentant 14 espèces deTrichogramma. Des variations de la P.G.M. ont été trouvées chezT. exiguum, T. marylandense etT. pretiosum. La. P.G.I. montre aussi des variations chezT. exiguum, T. marylandense, T. minutum etT. parkeri. Par contre, des variations de l'enzyme malique ne sont trouvées que chezT. pretiosum. En nous basant, sur l'analyse de progénitures, nous avons conclu que l'enzyme malique est un tétramère chezTrichogramma comprenant un allèle “lent” et un alléle “rapide”, à un seul locus, et que la P.G.M. et la P.G.I. ont chacune un seul locus à quatre allèles. La P.G.M. est un monomère mais la P.G.I. est un dimère.相似文献
14.
Primers with higher G+C content produced better random amplified polymorphic DNA (RAPD) profiles in Nepenthes. The occurrence of clustered G's and C's in the center of the primer seemed also to influence the banding patterns. It was also observed that for certain polymerases, the use of different buffers other than that recommended by the manufacturer provided a better amplification profile for Nepenthes. 相似文献
15.
Molecular characterization and genetic mapping of random amplified microsatellite polymorphism in barley 总被引:10,自引:0,他引:10
J. A. Dávila Y. Loarce E. Ferrer 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,98(2):265-273
This study has analyzed the molecular basis and genetic behaviour of the polymorphism generated by the amplification of barley
genomic DNA with primers complementary to microsatellites. Primers anchored at the 5′ end, used alone or in combination with
arbitrary sequence primers, generated random amplified microsatellite polymorphisms (RAMPs). Unanchored primers were also
used as single primers in a microsatellite primed-PCR (MP-PCR). Twenty six randomly selected RAMP DNA fragments which showed
polymorphism between the cultivars Steptoe and Morex were cloned and sequenced. All sequences showed the expected repeated
motif at the end of the insert, with the number of repeats ranging from five to ten. Genomic sequences containing low numbers
of microsatellite motifs were preferentially amplified; therefore, only a fraction of the polymorphism could be attributed
to variation in the number of microsatellite motifs at the priming site. Some sequences contained either cryptic simple sequences
or members of families of repeated DNA. Polymorphism at the internal cryptic simple sequences was detected by RAMP bands inherited
as co-dominant markers. Four MP-PCR bands were cloned and sequenced. A number of repeats identical to the primer itself were
found at each end of the insert. Two allelic bands were polymorphic for an internal microsatellite. The potential use of cloned
bands as fingerprinting tools was investigated by employing them as hybridization probes in Southern blots containing digested
barley DNA from a sample of cultivars. RAMP probes produced complex hybridization band patterns. MP-PCR probes produced either
a highly variable single locus or low-copy number loci. Segregations for 31 RAMPs and three MP-PCR bands were studied in a
population of 70 doubled-haploids from the Steptoe/Morex cross. One third of all markers were co-dominantly inherited. Markers
were positioned on an RFLP map and found to be distributed in all barley chromosomes. The new markers enlarged the overall
length of the map to 1408 cM.
Received: 6 May 1998 / Accepted: 20 July 1998 相似文献
16.
Andrea Menéndez-Yuffá Dominique Barry-Etienne Benoît Bertrand Frédéric Georget Hervé Etienne 《Plant Cell, Tissue and Organ Culture》2010,102(3):297-307
Plants of Coffea arabica L. derived via somatic embryogenesis, namely, somaclones, were evaluated with C. arabica seedlings grown in the nursery. At the time of their transfer to the nursery, somaclones of C. arabica cvs. Caturra and Costa Rica 95 (Catimor) were smaller and less vigorous than seedlings of the same cultivars. Following an
initial slow growth for a period of 10 weeks, somaclones began to grow faster than seedlings until both groups of plants were
equal in size at 21 weeks (entire duration of growth in the nursery). Comparisons of aerial and root systems of 30-cm long
somaclones and seedlings of two cultivars revealed that plants of somaclones were more vigorous than seedlings, based on the
higher number of leaves (13–16 vs. 9), larger leaf area (1060–1280 vs. 730–890 cm2), and greater dry weight of aerial organs (8.5–12 vs. 7.0–7.5 g). For cv. Caturra, the root dry weight of somaclones was
significantly greater than that of seedlings (2.7 vs. 1.9 g) and was attributable to the large diameter roots (>0.5 mm).
Analysis of 176,000 F1 hybrid somaclones revealed that these exhibited more heterogeneous growth than did the seedlings derived from zygotic embryos;
moreover, there was a genotype effect. Almost 9–20% of somaclones required an additional 3–4 months of growth in the nursery,
and 8–12% were culled for other undesirable horticultural attributes. Only 0.10–0.23% of somaclones displayed variant phenotypes.
The observed somaclone vigor in the nursery was carried over to field performance as these plants were more precocious than
seedlings and yielded coffee beans 1 year earlier than seedlings. 相似文献
17.
A. Muthukumar A. Eswaran S. Mathiyazhagan Sanjeev Kumar 《Archives Of Phytopathology And Plant Protection》2013,46(15):1543-1550
Twenty-five Pythium isolates comprising five species viz., Pythium aphanidermatum, P. deliense, P. graminicola, P. heterothallicum and P. ultimum from different geographical locations of Tamil Nadu (Coimbatore, 4; Cuddalore, 6; Dindigul, 1; Dharmapuri, 1; Erode, 1; Madurai, 1; Namakkal, 7; Thanjavur, 1; Theni, 1; Thirunelveli, 1 and Vellore, 1) isolated from chilli crop were analysed with randomly amplified polymorphic DNA (RAPD) markers. Morphological and molecular characteristics of these different species were correlated with the RAPD. Polymerase chain reaction amplification of total genomic DNA with six random primers generated unique banding patterns depending on the primer and the isolate. The isolate I17 produced identical banding patterns, while other isolates produced dissimilar bands within the particular species, indicating the genetic diversity among the isolates within a species. Morphological characters were also different from each other even in isolate I17 which shared identical bands. Cluster analysis showed minimum and maximum per cent similarities among the tested Pythium species which ranged from 49 to 89%, respectively. RAPD markers were better suited for differentiating isolates within a species rather than species. 相似文献
18.
Relationships among <Emphasis Type="Italic">Leymus</Emphasis> species assessed by RAPD markers 总被引:1,自引:0,他引:1
The DNA genetic diversity of 40 accessions of genus Leymus was analyzed by random amplified polymorphic DNA (RAPD) markers. A total of 352 products were amplified by 34 10-mer arbitrary
primers, among which 337 products (95.74 %) were found to be polymorphic. 5–14 polymorphic bands were amplified by each polymorphic
primer, with an average of 9.91 bands. The data of 352 RAPD bands were used to generate Jaccard’s similarity coefficients
and to construct a dendrogram by means of UPGMA. Great genetic diversity in genus Leymus was observed, the genetic diversity among the different species more abundant than that of the different accessions, and
the different accessions in a species or the species from the same areas were clustered together. 相似文献
19.
Sheng-Ping L. Hwang Shing Kou Wang Su Fen Wei Li-Chuan Cheng Jeng Chang 《Marine biotechnology (New York, N.Y.)》1999,1(2):200-206
To investigate the potential of DNA polymerase α as a marker for DNA replication in phytoplankton, two gene fragments that
showed a high degree of similarity with eukaryotic DNA polymerase α were cloned from two strains of a diatom, Skeletonema costatum (Greville) Cleve. The gene fragments amplified with the polymerase chain reaction were 397 and 396 bp in length, respectively.
The deduced amino acid sequences showed 44% to 61% similarity to the corresponding regions of DNA polymerase α sequences of
eukaryotic organisms ranging from yeast to humans. The similarity was especially high in three evolutionarily conserved regions
within the amplified fragments. Further, hybridization patterns from Southern blotting confirmed that the amplified fragments
were an integral part on the genome of S. costatum. In batch cultures abundant messenger of DNA polymerase α appeared in the late exponential phase and the early stationary
phase. This pattern suggests that DNA polymerase α expression is associated with actively dividing cells.
Received October 27, 1998; accepted November 13, 1998. 相似文献
20.
AFLP-Based detection of DNA methylation 总被引:14,自引:0,他引:14
By using the isoschizomersHpa II andMsp I which display differential sensitivity to cytosine methylation, a modified amplified fragment length polymorphism (AFLP)
technique has been developed to investigate DNA methylation profiles in eukaryotic organims. Genomic DNA was digested with
a mixture ofEcoR I and one of the isoschizomers, and ligated to oligonucleotide adapters. After two rounds of selective PCR amplification,
followed by DNA separation on a Long Ranger gel electrophoresis, a subset of restriction fragments can be displayed on an
X-ray film. Comparison of AFLP banding patterns betweenHpa II andMsp I revealed the extent of DNA methylation. The technique has been successfully applied in this study to investigate DNA methylation
profiles of apple (Malus domestica cv. Gala) genomic DNA extracted from leaves of field-grown adult trees andin vitro-grown shoot cultures. The results showed that up to 25 percent of AFLP bands were derived from methylated sequences, and
among those, a few bands unique to either adult trees orin vitro shoots were observed. These results demonstrated that this protocol is effective in identifying methylated DNA profiles.
Both first authors have contributed equally to this work. 相似文献