共查询到20条相似文献,搜索用时 15 毫秒
1.
Sandip Das Jyothi Rajagopal Sabhyata Bhatia P. S. Srivastava Malathi Lakshmikumaran 《Journal of biosciences》1999,24(4):433-440
Genetic relationships were evaluated among nine cultivars ofBrassica campestris by employing random amplification of polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers. RAPDs
generated a total of 125 bands using 13 decamer primers (an average of 9.6 bands per assay) of which nearly 80% were polymorphic.
The per cent polymorphism ranged from 60–100%. AFLP, on the other hand generated a total of 319 markers, an average of 64
bands per assay. Of these, 213 were polymorphic in nature (66.8%). AFLP methodology detected polymorphism more efficiently
than RAPD approach due to a greater number of loci assayed per reaction. Cultivar-specific bands were identified, for some
cultivars using RAPD, and for most cultivars with AFLP. Genetic similarity matrix, based on Jaccard’s index detected coefficients
ranging from 0.42 to 0.73 for RAPD, and from 0.48 to 0.925 for AFLPs indicating a wide genetic base. Cluster analyses using
data generated by both RAPD and AFLP markers, clearly separated the yellow seeded, self-compatible cultivars from the brown
seeded, self-incompatible cultivars although AFLP markers were able to group the cultivars more accurately. The higher genetic
variation detected by AFLP in comparison to RAPD was also reflected in the topography of the phenetic dendrograms obtained.
These results have been discussed in light of other studies and the relative efficiency of the marker systems for germplasm
evaluation. 相似文献
2.
Aerides vandarum and Vanda stangeana are two rare and endangered vandaceous orchids with immense floricultural traits. The intergeneric hybrids were synthesized
by performing reciprocal crosses between them. In vitro germination response of the immature hybrid embryos was found to be
best on half-strength Murashige and Skoog medium supplemented with 20% (v/v) coconut water/liquid endosperm from tender coconut. Determination of hybridity was made as early as the immature seeds or
embryos germinated in vitro, using randomly amplified polymorphic DNA (RAPD) markers. Out of 15 arbitrarily chosen decamer
RAPD primers, two were found to be useful in amplification of polymorphic bands specific to the parental species and their
presence in the reciprocal crosses. However, a decisive profile that can identify the reciprocal crosses could not be provided
by RAPD. Amplification of the trnL-F non-coding regions of chloroplast DNA of the parent species and hybrids aided easy identification of the reciprocal crosses
from the fact that maternal inheritance of chloroplast DNA held true for these intergeneric hybrids. Subsequent restriction
digestion of the polymerase chain reaction (PCR) amplified trnL-F non-coding regions of chloroplast DNA also consolidated the finding. Such PCR-based molecular markers could be used for
early determination of hybridity and easy identification of the reciprocal crosses. 相似文献
3.
Anna Szczerbakowa Justyna Tarwacka Michał Oskiera Henryka Jakuczun Bernard Wielgat 《Acta Physiologiae Plantarum》2010,32(5):867-873
Interspecific somatic hybrids between a diploid potato clone DG 81-68 susceptible to Phytophthora infestans (Mont.) de Bary and a resistant diploid tuber-bearing species Solanum × michoacanum were generated and analyzed. About 30 regenerants displaying an intermediate morphology were obtained as a result of three
separate PEG-mediated fusion experiments. The RAPD analysis confirmed the hybridity of all the regenerants. About 50% of the
hybrid plants exhibited vigorous growth and were stable in culture, while the rest of them rooted poorly and grew slowly in
vitro. Most of the hybrid clones were at the tetraploid level (70%), while 30% of the clones examined were at the hexaploid
level. The S. × michoacanum (+) DG 81-68 hybrids with growth anomalies were aneuploid. The variation in late blight resistance of the hybrid clones was
found in detached leaflet tests, with enhanced resistance characteristic for three tetraploid hybrids. 相似文献
4.
Qualitative and quantitative characterization of RAPD variation among snap bean (Phaseolus vulgaris) genotypes 总被引:1,自引:0,他引:1
P. W. Skroch J. Nienhuis 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1995,91(6-7):1078-1085
Ten snap bean (Phaseolus vulgaris) genotypes were screened for polymorphism with 400 RAPD (random amplified polymorphic DNA) primers. Polymorphic RAPDs were scored and classified into three categories based on ethidium bromide staining intensity. An average of 5.19 RAPD bands were scored per primer for the 364 primers that gave scorable amplification products. An average of 2.15 polymorphic RAPDs were detected per primer. The results show that primer screening may reduce the number of RAPD reactions required for the analysis of genetic relationships among snap-bean genotypes by over 60%. Based on the analysis of the distribution of RAPD amplification, the same number of polymorphic RAPDs were amplified from different genotypes for all RAPD band intensity levels. A comparison of RAPD band amplification frequency among genotypes for the three categories of bands classified by amplification strength revealed a measurable difference in the frequencies of RAPDs classified as faint (weakly amplifying) compared to RAPD bands classified as bold (strongly amplifying) indicating a possible scoring error due to the underscoring of faint bands. Correlation analysis showed that RAPD bands amplified by the same primer are not more closely correlated then RAPD bands amplified by different primers but are more highly correlated then expected by chance. Pairwise comparisons of RAPD bands indicate that the distribution of RAPD amplification among genotypes will be a useful criterion for establishing RAPD band identity. For the average pairwise comparison of genotypes, 50% of primers tested and 15.8% of all scored RAPDs detected polymorphism. Based on RAPD data Nei's average gene diversity at a locus was 0.158 based on all scorable RAPD bands and 0.388 if only polymorphic RAPD loci were considered. RAPD-derived 1 relationships among genotypes are reported for the ten genotypes included in this study. The data presented here demonstrate that many informative, polymorphic RAPDs can be found among snap bean cultivars. These RAPDs may be useful for the unique identification of bean varieties, the organization of bean germplasm, and applications of molecular markers to bean breeding. 相似文献
5.
Two DNA fingerprinting techniques, random amplified polymorphic DNA (RAPD) and inter-retrotransposon amplified polymorphism
(IRAP), were used to characterize somaclonal variants of banana. IRAP primers were designed on the basis of repetitive and
genome-wide dispersed long terminal repeat (LTR) retrotransposon families for assessing the somaclonal variation in 2Musa clones resistant and susceptible toFusarium oxysporum f. sp.cubense race 4. RAPD markers successfully detected genetic variation within and between individuals of the clones. IRAP makers amplified
either by a single primer or a combination of primers based on LTR orientation successfully amplified different retrotransposons
dispersed in theMusa genome and detected new events of insertions. RAPD markers proved more polymorphic than IRAP markers. Somaclonal variation
seems to be the result of numerous indels occurring genome-wide accompanied by the activation of retroelements, as a result
of stress caused by micropropagation. It is concluded that characterization of the somaclonal variants requires more than
one DNA marker system to detect variation in diverse components of the genome. 相似文献
6.
RAPD markers for constructing intraspecific tomato genetic maps 总被引:8,自引:0,他引:8
The existing molecular genetic maps of the tomato, Lycopersicon spp, are constructed based on isozyme and RFLP polymorphisms between tomato species. These maps are useful for certain applications but have few markers that exhibit sufficient polymorphisms for intraspecific analysis and manipulations within the cultivated tomato. The purpose of this study was to investigate the relative potential of RAPD technology, as compared to isozymes and RFLPs, to generate polymorphic DNA markers within cultivated tomatoes. Sixteen isozymes and 25 RFLP clones that were known to detect polymorphism between L. esculentum and L. pennellii, and 313 random oligonucleotide primers were examined. None of the isozymes and only four of the RFLP clones (i.e., 16%) revealed polymorphism between the cultivated varieties whereas up to 63% of the RAPD primers detected one or more polymorphic DNA fragments between these varieties. All RAPD primers detected polymorphism between L. esculentum and L. pennellii genotypes. These results clearly indicate that RAPD technology can generate sufficient genetic markers exploiting sequence differences within cultivated tomatoes to facilitate construction of intraspecific genetic maps.Abbreviations RFLP
restriction fragments length polymorphism
- RAPD
random amplified polymorphic DNA
- PCR
polymerase chain reaction
- QTLs
quantitative trait loci 相似文献
7.
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
8.
Inheritance of RAPD markers in channel catfish (Ictalurus punctatus), blue catfish (I. furcatus), and their F1, F2 and backcross hybrids 总被引:3,自引:0,他引:3
The channel catfish ( Ictalurus punctatus ) has become the most important aquaculture species in the USA. A genetic linkage map in catfish is needed to improve efficiency of breeding by marker-assisted selection (MAS) and for identification of economically important genes such as disease resistance genes. To identify DNA-based genetic polymorphism, the present authors tested 42 randomly amplified polymorphic DNA (RAPD) primers for their utility in identifying genetic polymorphism in catfish. Out of these primers, 22 generated 171 highly reproducible RAPD markers, producing almost eight polymorphic bands per primer. The remaining 20 primers produced an additional 20 polymorphic bands. The RAPD markers were highly reproducible, transmitted to F1 hybrids, and segregated in F2 or backcross progeny in ratios that did not differ from Mendelian expectations. Because the interspecific hybrids of channel catfish and blue catfish are fertile, RAPD markers using the interspecific hybrid system will be useful for rapid construction of genetic linkage maps of catfish and for analysis of important quantitative trait loci. 相似文献
9.
K. K. Nkongolo 《Plant Systematics and Evolution》1999,215(1-4):229-239
Random amplified polymorphic DNA (RAPD) analysis was used to determine genetic relationships amongP. mariana (black spruce),P. rubens (red spruce), andP. glauca (white spruce) and to assess the degree of polymorphism within populations from different provenances and among spruce hybrids. Eleven arbitrary decamer primers were used to amplify genomic DNAs extracted from embryogenic cultures and seedlings. Species-specific RAPD markers were identified.Picea mariana andP. rubens showed similar RAPD profiles confirming their close genetic relationship. Species-specific RAPD markers were identified and were useful in distinguishing white spruce from black and red spruces. RAPD differentiation between populations within each species was small. The level of polymorphism was much higher in spruce hybrid populations than in the pure species. Cytological analysis ofP. mariana ×P. rubens hybrids showed normal mitotic behaviour at prophase, metaphase, anaphase, and telophase. All the hybrids analyzed from different cross combinations were euploids. 相似文献
10.
Interspecific somatic hybrids between a dihaploid potato clone H-8105 susceptible to Phytophthora infestans (Mont.) de Bary and a resistant diploid tuberizing species Solanum bulbocastanum were generated and analysed. Only ten regenerants displaying the intermediate morphology with dominating characteristics
of the wild parent (simple leaves, anthocyanin pigmentation) were produced in 15 weeks after a single PEG-mediated fusion
event. The RAPD patterns confirmed the hybridity of all of them. The hybrids rooted poorly and grew slowly in vitro. The cytological analysis revealed a high degree of aneuploidy in the hybrids with morphological and growth anomalies in vitro, while the morphologically normal hybrids were tetraploids. All the S. bulbocastanum (+) H-8105 hybrids were unstable in culture and three of them were consequently lost during three years of propagation in vitro. The possible reasons for instability of somatic hybrids between the distantly related species are discussed. 相似文献
11.
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
12.
High-yielding dwarf clones of Hevea brasiliensis are tolerant to wind damage and therefore useful for high-density planting. The identification of molecular markers for the dwarf character is very important for isolating true-to-type high-yielding dwarf hybrid lines in the early stage of plant breeding programs. We have identified a dwarf genome-specific random amplified polymorphic DNA (RAPD) marker in rubber tree. A total of 115 random oligonucleotide 10-mer primers were used to amplify genomic DNA by PCR, of which 19 primers produced clear and detectable bands. The primer OPB-12 generated a 1.4-kb DNA marker from both natural and controlled F1 hybrid progenies (dwarf stature) derived from a cross between a dwarf parent and a normal cultivated clone as well as from the dwarf parent; it was absent in other parent (RRII 118). To validate this DNA marker, we analyzed 22 F1 hybrids (13 with a dwarf stature and nine with a normal stature); the dwarf genome-specific 1.4-kb RAPD marker was present in all dwarf-stature hybrids and absent in all normal-stature hybrids. This DNA marker was cloned and characterized. DNA marker locus specificity was further confirmed by Southern blot hybridization. Our results indicate that Southern blot hybridization of RAPD using probes made from cloned DNA fragments allows a more accurate analysis of the RAPD pattern based on the presence/absence of specific DNA markers than dye-stained gels or Southern blot analysis of RAPD blots using probes made from purified PCR products. Detection of RAPD markers in the hybrid progenies indicates that RAPD is a powerful tool for identifying inherited genome segments following different hybridization methods in perennial tree crops. 相似文献
13.
J. B. dos Santos J. Nienhuis P. Skroch J. Tivang M. K. Slocum 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,87(8):909-915
Genetic similarity among 45 Brassica Oleracea genotypes was compared using two molecular markers, random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphisms (RFLPs). The genotypes included 37 broccolis (var. italica), five cauliflowers (var. botrytis) and three cabbages (var. capitata) which represented a wide range of commercially-available germplasm, and included open-pollinated cultivars, commercial hybrids, and inbred parents of hybrid cultivars. Fifty-six polymorphic RFLP bands and 181 polymorphic RAPD bands were generated using 15 random cDNA probes and 62 10-mer primers, respectively. The objectives were to compare RFLP and RAPD markers with regard to their (1) sampling variance, (2) rank correlations of genetic distance among sub-samples, and (3) inheritance. A bootstrap procedure was used to generate 200 random samples of size n (n=2,3,5,... 55) independently from the RAPD and RFLP data sets. The coefficient of variance (CV) was estimated for each sample. Pooled regressions of the coefficient of variance on bootstrap sample size indicated that the rate of decrease in CV with increasing sample size was the same for RFLPs and RAPDs. The rank correlation between the Nei-Li genetic similarity values for all pairs of genotypes (990) based on RFLP and RAPD data was 0.745. Differences were observed between the RFLP and RAPD dendrograms of the 45 genotypes. Overlap in the distributions of rank correlations between independent sub-samples from the RAPD data set, compared to correlations between RFLP and RAPD sub-samples, suggest that observed differences in estimation of genetic similarity between RAPDs and RFLPs is largely due to sampling error rather than due to DNA-based differences in how RAPDs and RFLPs reveal polymorphisms. A crossing algorithm was used to generate hypothetical banding patterns of hybrids based on the genotypes of the parents. The results of this study indicate that RAPDs provide a level of resolution equivalent to RFLPs for detemination of the genetic relationships among genotypes. 相似文献
14.
应用SRAP分子标记方法对冬枣×宁梨巨枣的子代进行了分子鉴定及遗传多样性分析。采用构建基因池的方法对SRAP分子标记引物进行筛选,从88对引物中筛选出15对多态性好、主带清晰的引物,并对子代进行了真实性鉴定及多态性分析。结果表明:(1)15对引物共产生95个多态性条带,平均每对引物产生6.3个多态性条带,显示了较高的多态性比率。(2)80个子代中44个具有父本特征带,鉴定为真杂种。子代遗传多样性及UPGMA聚类分析表明,子代个体与亲本间的遗传相似系数在0.55~0.98之间,个体差异明显。该研究结果为枣树杂交育种提供了重要的分子证据。 相似文献
15.
甘霖;于肖夏;鞠天华;于卓;蒙美莲;张自强;李长青 《植物研究》2013,33(2):243-247
为给马铃薯新品种选育提供可靠材料,采用ISSR分子标记对2个马铃薯杂交组合‘J07-4’ב陇薯6号’和‘J07-6’ב陇薯6号’杂交种F1无性繁殖株系的真实性进行了鉴定。结果从152个ISSR引物中筛选出适于‘J07-4’ב陇薯6号’杂种F1 5个无性株系鉴定的3个ISSR引物AF18550、AW75511和AW20617及‘J07 6’ב陇薯6号’杂种F1 7个无性株系鉴定的2个ISSR引物AW20607和AF18549;以子代中含有父本特征带为主要依据,鉴定出杂种F1共12个无性繁殖株系均为真实的杂交种,并建立了杂种F1不同无性株系间的ISSR指纹图。表明ISSR分子标记技术用于马铃薯杂种F1无性株系鉴定是可行的。 相似文献
16.
Randomly Amplified Polymorphic DNA markers (RAPD) were used to assess the hybrid identity of individuals sampled as Phlomis × termessi Davis. Out of 95 primers screened, 11 primers produced reproducible amplification patterns used for discrimination of P. × termessi and their parents. Eleven primers produced 81 bands. Forty two percent of the RAPD bands existed in parents. Of the 54 bands
found in P. lycia, 19 were found only in this species and 7 of these were monomorphic. Similarly, of 57 RAPD bands observed in P. bourgaei, 18 were found only in P. bourgaei and 6 of these were monomorphic. Among hybrid individuals, 35 of the 73 markers were monomorphic. Fifteen of these existed
in individual parents showing that parents were homozygous for these markers. Of the 35 monomorphic bands observed among hybrid
individuals, 5 were present in the samples of one of the parents and completely absent from the samples of the other; therefore,
additive inheritance is indicated. Of the 5 additive bands, 1 was inherited from P. bourgaei and 4 were inherited from P. lycia. Among 38 polymorhic markers observed in hybrid individuals, 9 were new and hybrid-specific. Pollen fertility was also investigated.
Mean pollen fertility for P. lycia and P. bourgaei was 93% and 97% respectively. However, mean pollen fertility for hybrids was 65% (±10.5). 相似文献
17.
S. M. Ahmed V. Verma P. H. Qazi M. M. Ganaie S. K. Bakshi G. N. Qazi 《Plant Systematics and Evolution》2005,256(1-4):75-87
The phylogenetic relationships among the three species of Tinospora found in India are poorly understood. Morphology does not fully help to resolve the phylogeny and therefore a fast approach
using molecular analysis was explored. Two molecular approaches viz Random Amplified Polymorphic DNA (RAPD) assay and restriction
digestion of ITS1-5.8S-ITS2 rDNA (PCR-RFLP) were used to evaluate the genetic similarities between 40 different accessions
belonging to three species. Of the 38 random primers used only six generated the polymorphism, while as three out of 11 restriction
enzymes used gave polymorphic restriction patterns. The average proportion of polymorphic markers across primers was 95%,
however restriction endonucleases showed 92% polymorphism. RAPD alone was found suitable for the species diversions. In contrast
PCR- RFLP showed bias in detecting exact species variation. The correlation between the two markers was performed by Jaccard's
coefficient of similarity. A significant (r= 0.574) but not very high correlation was obtained. Further to authenticate the
results obtained by two markers, sequence analysis of ITS region of ribosomal DNA (ITS1 and ITS2, including 5.8S rDNA) was
performed. Three independent clones of each species T. cordifolia, T. malabarica and T. crispa were sequenced. Phylogenetic relationship inferred from ITS sequences is in agreement with RAPD data. 相似文献
18.
陕油8号种子纯度的RAPD鉴定研究 总被引:5,自引:0,他引:5
从杂交油菜“陕油8号”及其亲本中提取基因组DNA,用100个RAPD随机引物进行扩增,从中筛选出3个可将亲本和子代区分的引物BA208、BA1090、BA497。BA208产生亲本互补的特征带BA208-1050bp、BA2081250bp;BA1090产生母本特征带BA1090-700bp,BA497产生父本特征带BA497-870bp,上述谱带均在子代中出现。以BA208产生的特征谱带作为分子标记对杂交油菜种子纯度鉴定得到了一致的结果,并与大田纯度检测结果一致。BA497可将“陕油8号”与当地4个主栽品种有效区分。此外,还对双引物共同鉴定杂交种子纯度问题进行了初步探讨。 相似文献
19.
Anna Szczerbakowa Danuta Bołtowicz Renata Lebecka Paweł Radomski Bernard Wielgat 《Acta Physiologiae Plantarum》2005,27(3):265-273
The morphological, cytological and molecular analyses of the plants regenerated after PEG-induced fusion between mesophyll
protoplasts from the dihaploid potato clone H-8105 and the wild tuberous disease-resistant species S. pinnatisectum, were performed. A single fusion experiment yielded 313 calli, although only two calli produced shoots. From the rooted shoots,
two stable clones (PT-01-1 and PT-01-2) exhibiting different vigor and habitat, were developed. The plants of PT-01-1 clone
grew slowly in vitro, produced tubers after transfer to soil but did not set flowers. In contrast, the plants of the vigorous clone PT-01-2 produced
both tubers and flowers after transfer to soil. The flower and tuber morphology of PT-01-1 and PT-01-2 regenerants was intermediate
in comparison to the parental species. Cytological analysis revealed that the PT-01-1 clone was pentaploid and the PT-01-2
clone was tetraploid. The molecular (RAPD) analysis confirmed hybridity of both clones. The preliminary tests on late blight
resistance of the hybrids showed no differences with a potato parent. 相似文献
20.
Comparison of RFLP and RAPD markers to estimating genetic relationships within and among cruciferous species 总被引:25,自引:0,他引:25
C. E. Thormann M. E. Ferreira L. E. A. Camargo J. G. Tivang T. C. Osborn 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,88(8):973-980
Restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) markers are being used widely for evaluating genetic relationships of crop germplasm. Differences in the properties of these two markers could result in different estimates of genetic relationships among some accessions. Nuclear RFLP markers detected by genomic DNA and cDNA clones and RAPD markers were compared for evaluating genetic relationships among 18 accessions from six cultivated Brassica species and one accession from Raphanus sativus. Based on comparisons of genetic-similarity matrices and cophenetic values, RAPD markers were very similar to RFLP markers for estimating intraspecific genetic relationships; however, the two marker types gave different results for interspecific genetic relationships. The presence of amplified mitochondrial and chloroplast DNA fragments in the RAPD data set did not appear to account for differences in RAPD- and RFLP-based dendrograms. However, hybridization tests of RAPD fragments with similar molecular weights demonstrated that some fragments, scored as identical, were not homologous. In all these cases, the differences occurred at the interspecific level. Our results suggest that RAPD data may be less reliable than RFLP data when estimating genetic relationships of accessions from more than one species. 相似文献