共查询到20条相似文献,搜索用时 31 毫秒
1.
Alejandro Bolívar-González Marta Valdez-Melara Andrés Gatica-Arias 《In vitro cellular & developmental biology. Plant》2018,54(6):576-589
Crop improvement of Coffea arabica L. (coffee) via mutagenesis could accelerate breeding programs; thus, the present study aimed to develop an in vitro protocol using the chemical mutagens sodium azide (NaN3) and ethyl methanesulfonate (EMS) on embryogenic cell suspensions of Arabica coffee variety Catuaí and, subsequently, to evaluate the responses of the resulting mutagenized tissues to salinity stress. Embryogenic suspension cultures were incubated with 0.0, 2.5, 5.0, or 10.0 mM NaN3 or 0.0, 185.2, 370.5, or 741.0 mM EMS. As the concentration of NaN3 or EMS increased, the survival of embryogenic suspension cultures decreased compared to controls. The median lethal dose (LD50) for NaN3 was 5 mM for 15 min and for EMS it was 185.2 mM for 120 min. Embryogenic suspension cultures treated with NaN3 or EMS were cultured on selective medium supplemented with 0, 50, 100, 150, 250, or 300 mM NaCl showed that 50 mM NaCl could be used as selection pressure. Plantlet growth and total amino acid content were affected by NaCl stress; some mutants had longer shoots and higher amino acid content than controls. Random amplified polymorphic DNA (RAPD) analysis was performed to determine whether the NaN3 or EMS treatments could induce genetic variability and resulted in identifiable polymorphic markers. A total of 18 10-mer primers were used to amplify genomic DNA of putative mutant and non-mutant arabica coffee embryogenic cultures and produced 50 scorable bands, of which 22% were polymorphic. 相似文献
2.
A set of 30 accessions of five Curcuma species-C. latifolia, C. malabarica, C. manga and C. raktakanta and 13 morphotypes (identified on the basis of morphological markers) of C. longa conserved in the In Vitro Genebank at National Bureau of Plant Genetic Resources, New Delhi, were subjected to RAPD analysis. Of the 200 RAPD primers
screened, 21 polymorphic primers were selected for further study. Mean genetic similarities based on Jaccard’s similarity
coefficient ranged from 0.18 to 0.86 in accessions of cultivated species, i.e., C. longa and from 0.25 to 0.86 in wild species. The dendrogram derived from the RAPD data corroborated the morphological classification
of the morphotypes. The efficiency of individual RAPD primers was also compared; primers OPC-20, OPO-06, OPC-01 and OPL-03
were adjudged highly informative in discriminating the germplasm of Curcuma. 相似文献
3.
Analysis of genetic stability of plants regenerated from suspension cultures and protoplasts of meadow fescue (Festuca pratensis Huds.) 总被引:4,自引:0,他引:4
M. P. Vallés Z. Y. Wang P. Montavon I. Potrykus G. Spangenberg 《Plant cell reports》1993,12(2):101-106
A cytological and molecular analysis was performed to assess the genetic uniformity and true-to-type character of plants regenerated from 20 week-old embryogenic suspension cultures of meadow fescue (Festuca pratensis Huds.), and compared to protoplastderived plants obtained from the same cell suspension. Cytological variation was not observed in a representative sample of plants regenerated directly from the embryogenic suspensions and from protoplasts isolated therefrom. Similarly, no restriction fragment length polymorphisms (RFLPs) were detected in the mitochondrial, plastid and nuclear genomes in the plants analyzed. Randomly amplified polymorphic DNA markers (RAPDs) have been used to characterise molecularly a set of mature meadow fescue plants regenerated from these in vitro cultures. RAPD markers using 18 different short oligonucleotide primers of arbitrary nucleotide sequence in combination with polymerase chain reaction (PCR) allowed the detection of pre-existing polymorphisms in the donor genotypes, but failed to reveal newly generated variation in the protoplast-derived plants compared to their equivalent suspensionculture regenerated materials.The genetic stability of meadow fescue plants regenerated from suspension cultures and protoplasts isolated therefrom and its implications on gene transfer technology for this species are discussed.Abbreviations PCR
polymerase chain reaction
- RAPD
random amplified polymorphic DNA
- RFLP
restriction fragment length polymorphism. 相似文献
4.
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
5.
Rakesh Singh C. Durga Prasad Vibha Singhal Gurinder Jit Randhawa 《Journal of plant biochemistry and biotechnology.》2002,11(2):109-112
PCR-based random amplified polymorphic DNA (RAPD) markers were employed to assess genetic diversity in 23 chickpea genotypes. Forty of the 100 random primers screened revealed polymorphism among the genotypes. Most of the primers revealed single polymorphic band, and only 14.1 2% of the products were polymorphic. Estimates of genetic similarity based on Jaccard’s coefficient ranged from 0.92 to 0.99, indicating narrow genetic variability among the genotypes based on RAPD markers.The 23 chickpea genotypes formed two major clusters in the dendrogram.The low RAPD polymorphism among chickpea genotypes suggests that more number of polymorphic primers need to be analysed to determine genetic relationships. It was observed that RAPD analysis employing 30 polymorphic primers could provide better estimates of genetic relationships in chickpea. 相似文献
6.
N. Isabel L. Tremblay M. Michaud F. M. Tremblay J. Bousquet 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1993,86(1):81-87
Summary The usefulness of random amplified polymorphic DNA (RAPD) in assessing the genetic stability of somatic embryogenesis-derived populations of black spruce [Picea mariana (Mill.) B.S.P.] was evaluated. Three arbitrary 11-mer primers were successfully used to amplify DNA from both in-vivo and in-vitro material. Twenty-five embryogenic cell lines, additional zygotic embryos and megagametophytes from three controlled crosses involving four selected genotypes of black spruce were used for the segregation analysis of RAPD variants. Ten markers were genetically characterized and used to evaluate the genetic stability of somatic embryos derived from three embryogenic cell lines (one cell line per cross, 30 somatic embryos per cell line). No variation was detected within clones. The utilization of RAPD markers both for the assessment of genetic stability of clonal materials and to certify genetic stability throughout the process of somatic embryogenesis is discussed. 相似文献
7.
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
8.
Sheila Jazmín Reyes-Zambrano Carlos Alberto Lecona-Guzmán Federico Antonio Gutiérrez-Miceli Víctor Manuel Ruiz-Valdiviezo Arely Anayansi Vargas-Díaz 《Phyton》2021,90(6):1727-1739
Agave americana L. callus were exposed to different concentrations of ethyl methanesulphonate (EMS) 0, 15, 30, 45 and 60 mM and to different times of exposure (2 and 4 h). The viability and capacity of shoot formation were shown to be affected when the callus were exposed to high concentrations (30–60 mM). Only the callus exposed to 15 mM EMS presented shoot formation; the exposure time of two hours produced the largest quantity of shoots regenerated per callus (21 shoots/callus). In order to generate somaclonal variants resistant to Fusarium oxysporum, a selection pressure was applied through of a culture filtrate (CF) of 100 ppm of the fungus. This was made in callus obtained in the treatment with 15 mM EMS during 2 h of exposure. The CF caused oxidation and necrosis in 71.25% of the callus; however, they were capable of generating shoots (3.5 shoots/callus). Molecular markers type RAPD, ISSR and DAMD were used to evaluate the genetic variation arising from the mutations caused by EMS on control plants and 16-month-old somaclonal variants. The polymorphic information content (PIC) for each one of the initiating groups was: 0.28 (DAMD), 0.09 (ISSR) and 0.14 (RAPD). DAMD revealed a greater percentage of polymorphism than RAPD and ISSR. Polymorphic bands were detected in the somaclonal variants. This indicated that the EMS caused genetic variation in the regenerated plants conferring resistance to them against Fusarium oxysporum. 相似文献
9.
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
10.
Tribulus terrestris is well known for its medicinal importance in curing urino-genital disorders. Amplified fragment length polymorphism (AFLP),
selective amplification of microsatellite polymorphic loci (SAMPL), inter-simple sequence repeat (ISSR) and randomly amplified
polymorphic DNA (RAPD) markers were used for the first time for the detection of genetic polymorphism in this medicinal herb
from samples collected from various geographical regions of India. Six assays each of AFLP and SAMPL markers and 21 each of
ISSR and RAPD markers were utilized. AFLP yielded 500 scorable amplified products, of which 82.9% were polymorphic. SAMPL
primers amplified 488 bands, 462 being polymorphic (94.7%). The range of amplified bands was 66 [(TC)8G + M-CAG] to 98 [(CA)6AG + M-CAC] and the percentage polymorphism, 89.9 [from (CT)4C (AC)4A + M-CTG] to 100 [from (GACA)4 + M-CTA]. The ISSR primers amplified 239 bands of 0.4–2.5 kb, 73.6% showed polymorphism. The amplified products ranged from
5 to 16 and the percentage polymorphism 40–100. RAPD assays produced 276 bands, of which 163 were polymorphic (59%). Mantel
test employed for detection of goodness of fit established cophenetic correlation values above 0.9 for all the four marker
systems. The dendrograms and PCA plots derived from the binary data matrices of the four marker systems are highly concordant.
High bootstrap values were obtained at major nodes of phenograms through WINBOOT software. The relative efficiency of the
four molecular marker systems calculated on the basis of multiplex ratio, marker index and average heterozygosity revealed
SAMPL to be the best. Distinct DNA fingerprinting profile, unique to every geographical region could be obtained with all
the four molecular marker systems. Clustering can be a good indicator for clear separation of genotypes from different regions
in well-defined groups that are supported by high bootstrap values. 相似文献
11.
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
12.
Identification of molecular markers in soybean comparing RFLP,RAPD and AFLP DNA mapping techniques 总被引:16,自引:0,他引:16
Jhy-Jhu Lin Jonathan Kuo Jin Ma James A. Saunders Hunter S. Beard Margaret H. MacDonald William Kenworthy George N. Ude Benjamin F. Matthews 《Plant Molecular Biology Reporter》1996,14(2):156-169
Three different DNA mapping techniques—RFLP, RAPD and AFLP—were used on identical soybean germplasm to compare their ability
to identify markers in the development of a genetic linkage map. Polymorphisms present in fourteen different soybean cultivars
were demonstrated using all three techniques. AFLP, a novel PCR-based technique, was able to identify multiple polymorphic
bands in a denaturing gel using 60 of 64 primer pairs tested. AFLP relies on primers designed in part on sequences for endonuclease
restriction sites and on three selective nucleotides. The 60 diagnostic primer pairs tested for AFLP analysis each distinguished
on average six polymorphic bands. Using specific primers designed for soybean fromEco RI andMse I restriction site sequences and three selective nucleotides, as many as 12 polymorphic bands per primer could be obtained
with AFLP techniques. Only 35% of the RAPD reactions identified a polymorphic band using the same soybean cultivars, and in
those positive reactions, typically only one or two polymorphic bands per gel were found. Identification of polymorphic bands
using RFLP techniques was the most cumbersome, because Southern blotting and probe hybridization were required. Over 50% of
the soybean RFLP probes examined failed to distinguish even a single polymorphic band, and the RFLP probes that did distinguish
polymorphic bands seldom identified more than one polymorphic band. We conclude that, among the three techniques tested, AFLP
is the most useful. 相似文献
13.
S. Jayasankar Richard E. Litz Raymond J. Schnell Andres-Cruz Hernandez 《In vitro cellular & developmental biology. Plant》1998,34(2):112-116
Summary Genomic DNA isolated from embryogenic cultures of two mango cultivars, ‘Hindi’ and ‘Carabao,’ that had been selected for resistance
to the culture filtrate ofColletotrichum gloeosporioides, was analyzed using Randomly Amplified Polymorphic DNA (RAPD).In vitro selection caused changes in RAPD markers in the selected embryogenic cultures with respect to the unchallenged control cultures
and the stock plants. The differences involved both the absence and the presence of additional RAPD markers in the resistant
lines, although the former was most commonly observed. The absence of differences between the unchallenged control of either
cultivar and DNA from the leaves of parent trees confirmed that the changes were not due to prolonged maintenance in liquid
cultures. 相似文献
14.
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
15.
Applicability of inter-simple sequence repeat polymorphisms in wheat for use as DNA markers in comparison to RFLP and RAPD markers 总被引:43,自引:0,他引:43
T. Nagaoka Y. Ogihara 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,94(5):597-602
Inter-simple sequence repeat polymorphic DNA (ISSR) was evaluated for its applicability as a genetic marker system in wheat.
PCR was carried out with primers that annealed to simple sequence repeats. The resultant products were subjected to agarose-gel
electrophoresis, and the banding patterns were compared among six wheat accessions containing diploid, tetraploid, and hexaploid
members. Out of 100 examined, 33 primers produced distinguishable as well as polymorphic bands in each of the six accessions.
Although most of the primers that gave distinct bands (30 primers out of 33) contained dinucleotide repeats, each of the primers
with tri-, tetra-, and penta-nucleotide motifs also yielded discrete bands. Primers based on (AC)n repeats gave the most polymorphic bands. In total, 224 polymorphic bands were found in the comparison between Einkorn wheats
whereas, on the average, 120 polymorphic bands were detected between common wheats. ISSR primers produced several times more
information than RAPD markers. The extent of band polymorphism was similar to that of RFLP markers, and greater than that
of RAPDs. The genetic relationships of wheat accessions estimated by the polymorphism of ISSR markers were identical with
those inferred by RFLP and RAPD markers, indicating the reliability of ISSR markers for estimation of genotypes. These polymorphic
bands are potential candidates as novel markers for use in linkage-map construction in wheat. The characteristic features
of ISSR markers, i.e. polymorphism, generation of information and ease of handling, suggest their applicability to the analysis
of genotypes as well as to the construction of PCR-based genome maps of wheats.
Received: 15 September 1996 / Accepted: 25 October 1996 相似文献
16.
Mohd Arif N. W. Zaidi Y. P. Singh Qazi Mohd Rizwanul Haq U. S. Singh 《Plant Molecular Biology Reporter》2009,27(4):488-495
Shisham (Dalbergia sissoo) is one of the most preferred timber tree species of South Asia. Two DNA-based molecular marker techniques, intersimple sequence
repeat (ISSR) and random amplified polymorphism DNA (RAPD), were compared to study the genetic diversity in this species.
A total of 30 polymorphic primers (15 ISSR and 15 random) were used. Amplification of genomic DNA of 22 genotypes, using ISSR
analysis, yielded 117 fragments, of which 64 were polymorphic. Number of amplified fragments with ISSR primers ranged from
five to ten and varied in size from 180 to 1,900 bp. Percentage polymorphism ranged from 0 to 87.5. The 15 RAPD primers produced
144 bands across 22 genotypes, of which 84 were polymorphic. The number of amplified bands varied from five to 13, with size
range from 180 to 2,400 bp. Percentage polymorphism ranged from 0 to 100, with an average of 58.3 across. RAPD markers were
relatively more efficient than the ISSR assay. The mental test between two Jaccard’s similarity matrices gave r ≥ 0.90, showing very good fit correlation in between ISSR- and RAPD-based similarities. Clustering of isolates remained more
or less the same in RAPD and combined data of RAPD and ISSR. The similarity coefficient ranged from 0.734 to 0.939, 0.563
to 0.946, and 0.648 to 0.920 with ISSR, RAPD, and combined dendrogram, respectively. 相似文献
17.
Turkey is very rich in local grape varieties. The solution to the problem of identifying local cultivars, which is considered an important deficiency for the region, will only be possible when they can be defined with molecular markers. Forty-nine local grapevine cultivars from ?anl?urfa (Turkey) were characterized with RAPD markers. Twenty-five decamer primers selected from 60 primers were used in this analysis. A total of 171 bands were obtained with the 25 primers, of which 112 were polymorphic; the level of DNA polymorphism was 65.49% in these local cultivars. Among the selected primers, OPA-18, OPO-07 and P-123 gave the maximum number of polymorphic bands (seven). Genetic relationships among these cultivars were determined with a similarity index and using a dendogram. Among the grape cultivars, the lowest similarity ratio (0.578) was observed among the Külahi-K?z?lbanki cultivars and the highest similarity ratio (0.908) was observed for the ?ilorut-D?külgen combination. The high similarity ratio among the grape cultivars of ?anl?urfa Province was also reflected in the dendogram. In general, no relationships were encountered between the genetic identification of the cultivars and their ampelographic properties. 相似文献
18.
P. W. J. Taylor J. R. Geijskes H.-L. Ko T. A. Fraser R. J. Henry R. G. Birch 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1995,90(7-8):1169-1173
Random amplified polymorphic DNA (RAPD) analysis using 10-mer oligonucleotide primers efficiently differentiated sugarcane cultivars and proved suitable for detecting gross genetic change such as that which can occur in sugarcane subjected to prolonged tissue culture, for example in protoplast-derived callus. However, RAPD analysis was not sufficiently sensitive to detect smaller genetic changes that occur during sugarcane genetic transformation. The length of DNA scored for polymorphism per primer averaged 13.2 kb, or 0.0001% of the typical sugarcane genome size of 1.2 × 107 kb (2C). RAPD analysis of sugarcane plants regenerated from embryogenic callus revealed very few polymorphisms, indicating that gross genetic change is infrequent during this tissue culture procedure, although epigenetic effects result in transient morphological changes in regenerated plants. More sensitive variations on the RAPD technique may increase the practicality of DNA-based screening of regenerated plant lines to reveal somaclonal variants. 相似文献
19.
The comparison of RFLP,RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis 总被引:71,自引:0,他引:71
Wayne Powell Michele Morgante Chaz Andre Michael Hanafey Julie Vogel Scott Tingey Antoni Rafalski 《Molecular breeding : new strategies in plant improvement》1996,2(3):225-238
The utility of RFLP (restriction fragment length polymorphism), RAPD (random-amplified polymorphic DNA), AFLP (amplified fragment length polymorphism) and SSR (simple sequence repeat, microsatellite) markers in soybean germplasm analysis was determined by evaluating information content (expected heterozygosity), number of loci simultaneously analyzed per experiment (multiplex ratio) and effectiveness in assessing relationships between accessions. SSR markers have the highest expected heterozygosity (0.60), while AFLP markers have the highest effective multiplex ratio (19). A single parameter, defined as the marker index, which is the product of expected heterozygosity and multiplex ratio, may be used to evaluate overall utility of a marker system. A comparison of genetic similarity matrices revealed that, if the comparison involved both cultivated (Glycine max) and wild soybean (Glycine soja) accessions, estimates based on RFLPs, AFLPs and SSRs are highly correlated, indicating congruence between these assays. However, correlations of RAPD marker data with those obtained using other marker systems were lower. This is because RAPDs produce higher estimates of interspecific similarities. If the comparisons involvedG. max only, then overall correlations between marker systems are significantly lower. WithinG. max, RAPD and AFLP similarity estimates are more closely correlated than those involving other marker systems.Abbreviations RFLP
restriction fragment length plymorphism
- RAPD
random-amplified polymorphic DNA
- AFLP
amplified fragment length polymorphism
- SSR
simple sequence repeat
- PCR
polymerase chain reaction
- TBE
Tris-borate-EDTA buffer
- MI
marker index
- SENA
sum of effective numbers of alleles 相似文献
20.
Polymerase chain reaction (PCR) has been used extensively in the construction of linkage maps for many cultivated crops including soybean, [Glycine max (L.) Merr]. In this study, four sets of oligonucleotide primer pairs of known genes (pearl millet Adh 1, nodule specific proline-rich protein, Drosophila homeobox, heat shock protein), several different combinations from kits A, D, E, and J of arbitrary primers and five primer pairs of soybean simple sequence repeats of varying length (Satt 9, Satt 20, Satt 42, Satt 64, and Satt 30) were utilized in PCR to identify molecular markers which were then used to construct a genetic linkage map. DNA for the PCR reactions was isolated from 65 recombinant inbred soybean lines resulting from crossing PI 290,136 and BARC-2 (Rj
4
), followed by self-pollination for seven generations without selection. Mapmaker 3.0, a computer package, was used for construction of the linkage map. A total of 43 polymorphic markers were identified; 30 markers were linked and distributed among 5 linkage groups while 13 markers were unlinked. Arbitrary primers revealed more polymorphisms than specific primers. A combination of arbitrary primers A5 and A18 revealed the maximum number of polymorphic bands. Five observed linkage groups can be expanded in future soybean research by using additional markers. 相似文献