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1.
Evidence of gene introgression in apple using RAPD markers 总被引：4，自引：0，他引：4
Summary A genomic remnant of Malus floribunda clone 821 introgressed into the cultivated apple M. x domestica Borkh. was identified using randomly amplified polymorphic DNA (RAPD) markers obtained by the polymerase chain reaction (PCR). Using a set of 59 oligonucleotide decamer primers, polymorphic DNA markers were identified among three pooled DNA samples. Based on the presence or absence of bands among bulked apple scab-resistant selections and cultivars, bulked scab-susceptible cultivars, and a M. floribunda clone 821 sample, one primer, A 15, identified amplified fragments in the scab-resistant bulked sample that was also unique to the M. floribunda clone 821. The unique band from M. floribunda clone 821 was amplified in four out of 17 scab-resistant selections/cultivars. This RAPD, designated OA15900, identifies an introgressed fragment that has as yet no known function. 相似文献
2.
Randomly amplified polymorphic DNA (RAPD) markers were applied in purity control of hybrid seed production of tomato (Lycopersicon esculentum Mill.). DNA from three commercial F1-hybrid cultivars and their parental lines was subjected to RAPD screening with 50 primers. Two of four primers which detected polymorphism between the parents tested, generated paternal-specific RAPDs, enabling a clear distinction to be made between hybrids and their maternal parents. In addition, combination of the polymorphic DNA products generated by these primers exhibited hybrid-specific patterns, enabling each cultivar to be identified. This result indicates the practical usefulness of RAPD markers in hybrid-tomato-seed purity-control tests and cultivar identification. The approach is advantageous in its rapidity and simplicity, particularly as an alternative for those cultivars for which lengthy and costly phenotypic tests are currently used. 相似文献
3.
Development of reliable PCR markers for the selection of the Vf gene conferring scab resistance in apple 总被引：6，自引：0，他引：6
Early selection of scab-resistant apple seedlings can be enhanced by the use of markers tightly linked to the Vf resistance gene. Two sequence characterized amplified regions (SCAR) markers have been obtained from previously described random amplified polymorphic DNA (RAPD) markers. AM19-SCAR is a codominant marker, while AM19-SCAR is dominant, as is the RAPD from which it was derived. A highly detailed map in the vicinity of the Vf gene was built through the cumulative analysis of about 600 seedlings from six different controlled crosses. The usefulness of these and other SCAR markers will be discussed in relation to combining the traditional phenotypic selection with MAS. The availability of two codominant, tightly linked markers flanking both sides of the resistance gene (AL07-SCAR and M18-CAPS) also makes it easy to identify the seedlings homozygous for the resistance gene. 相似文献
4.
Identification and characterization of RAPD and SCAR markers linked to anthracnose resistance gene in sorghum [Sorghum bicolor (L.) Moench] 总被引：1，自引：0，他引：1
Anthracnose, one of the destructive foliar diseases of sorghum growing in warm humid regions, is incited by the fungus Colletotrichum graminicola.The inheritance of anthracnose resistance was studied using the parental cultivars of Sorghum bicolor (L.) Moench, HC 136 (susceptible to anthracnose) and G 73 (anthracnose resistant). The F1 and F2 plants were inoculated with the local isolates of C. graminicola cultures. The F2 plants showed a segregation ratio of 3 (susceptible): 1(resistant) indicating that the locus for resistance to anthracnose in sorghum accession G 73 segregates as a recessive trait in a cross to susceptible cultivar HC 136. RAPD (random amplified polymorphic DNA) marker OPJ 011437 was identified as marker closely linked to anthracnose resistance gene in sorghum by bulked segregant analysis of HC 136 × G73 derived recombinant inbred lines (RILs) of sorghum. A total of 84 random decamer primers were used to screen polymorphism among the parental genotypes. Among these, only 24 primers were polymorphic. On bulked segregant analysis, primer OPJ 01 amplified a 1437 bp fragment only in resistant parent G 73 and resistant bulk. The marker OPJ 011437 was cloned and sequenced. The sequence of RAPD marker OPJ 011437 was used to generate specific markers called sequence characterized amplified regions (SCARs). A pair of SCAR markers SCJ 01-1 and SCJ 01-2 was developed using Mac Vector program. SCAR amplification of resistant and susceptible parents along with their respective bulks and RILs confirmed that SCAR marker SCJ 01 is at the same loci as that of RAPD marker OPJ 011437 and hence, is linked to anthracnose resistance gene. Resistant parent G 73 and resistant bulk amplified single specific band on PCR amplification using SCAR primer pairs. The RAPD marker OPJ 011437 was mapped at a distance of 3.26 cM apart from the locus governing anthracnose resistance on the sorghum genetic map by the segregation analysis of the RILs. Using BLAST program, it was found that the marker showed 100 per cent alignment with the contig{_}3966 located on the longer arm of chromosome 8 of sorghum genome. Therefore, these identified RAPD and SCAR markers can be used in the resistance-breeding program of sorghum anthracnose by marker-assisted selection.An erratum to this article can be found at 相似文献
5.
One‐hundred and twenty‐four amplified fragment length polymorphism (AFLP) and 49 random amplified polymorphic DNA (RAPD) markers have been used to distinguish between 20 and 23 commercial chicory cultivars, respectively. These were all Cichorium intybus var. foliosum F1 hybrids, currently used in hydroponic forcing. Five‐hundred and twenty RAPD primers (OPERON) were tested, of which 156 resulted in reproducible patterns and 26 yielded polymorphisms. Two‐hundred and fifty‐six AFLP primer‐combinations were tested and six combinations were selected for identification purposes. Similarity indices were measured and clustering has been done using pairwise comparison. Both types of marker provide similar conclusions. Two major clusters are formed, representing late and early cultivars. All cultivars were identified using 10 informative RAPD primers or three AFLP primer combinations. A low degree of polymorphism was detected between some early cultivars, suggesting a narrow genetic base in their breeding strategy. 相似文献
6.
The random amplified polymorphic DNA (RAPD) technique was adapted for segregation analysis in sugar beet. 83 IO/I individuals were scored with a set of 20 arbitrary decamer primers. 4 preliminary linkage groups could be established, enclosing 9 RAPD markers, 2 isozyme loci, a gene for the hypocotyl colour and a gene for resistance to the root knot nematode (Heterodera schachtii Schm.). 相似文献
7.
Summary DNA polymorphism among five Asparagus officinalis L. cultivars-Imperial, Snow, Steline, UC-157 and Larac, as detected by random amplified polymorphic DNA (RAPD), is reported. Thirty one decamer primers were tested. and twenty six of them yielded amplification products. Fourteen primers gave products with at least one polymorphic DNA fragment. Among a total of 119 amplified fragments 33 were polymorphic. These RAPD markers enabled the identification of asparagus cultivars. Unique markers for cultivars were: Snow-bands 475 bp, 772 bp, 412 bp, 935 bp and 820 bp amplified by primers D5, OPA-07, OPA-09, OPA-10 and OPA-18, respectively. Steline-bands 645 bp, 680 bp and 997 bp amplified by primers A32, OPA-03 and OPA-09, respectively. A band 903 bp, amplitied by primer OPA-12, is a marker for Imperial, and a band 420 bp, amplified by primer D52, is a marker for Larac. Cultivar UC-157 could be identified by a combination of shared polymorphic bands. The pairwise marker difference between cultivars ranged from 0.08 to 0.17. A phenogram of the genetic relationship based on RAPD fits with the known origin of the cultivars. 相似文献
8.
In pea, a single recessive gene (er) on linkage group 6 confers resistance to powdery mildew caused by Erysiphe pisi. The present study aims to identify molecular markers linked to the er gene. Screening of the powdery mildew‐resistant cultivar ‘DMR11’ and its susceptible nearisogenic line for polymorphism revealed linkage of two RAPD primers (OPO‐02 and OPU‐17) to the er gene and a sequence characterized polymorphic region (SCAR) primer, ScOPD‐10650 with er in a population of 83 F2 plants in the order: OPU‐17 ‐ er ‐ ScOPD‐10650 ‐ OPO‐02. The markers ScOPD‐10650 and OPU‐17 being coupled with the allele causing resistance would substantially increase the efficiency of marker‐assisted selection in peabreeding for powdery mildew. 相似文献
9.
A random amplified polymorphic DNA marker OPG17450 linked to the Ns gene that confers resistance of potato to potato virus S (PVS), was used to develop sequence‐characterized amplified region (SCAR) markers. After cloning and sequencing of OPG17450 new polymerase chain reaction (PCR) primers were designed to generate dominant (SCG17321) and codominant (SCG17448) markers. For SCG17448, polymorphism between susceptible and resistant genotypes was recovered after digestion of the marker with the restriction enzyme Muni. In addition to the band corresponding to ‘susceptible’ allele that does not contain the Muni cleavage site, two bands of approximately 251 bp and 197 bp were observed in the resistant genotypes. The usefulness of these SCAR markers was verified in diploid potatoes possessing the Ns locus from clone G‐LKS 678147/60, and in tetraploid potatoes derived from G‐LKS 678147/60 and from clone MPI 65118/3. 相似文献
10.
Development of SCAR markers linked to the Pm21 gene conferring resistance to powdery mildew in common wheat 总被引：39，自引：0，他引：39
Powdery mildew is an important disease in most of the wheat production areas of the world. The resistance gene Pm21 (6AL/6VS trans-location) derived from Haynaldia villosa confers resistance to all available isolates of Erysiphe (Blumeria) graminis f. sp. tritici in China and Europe. The objective of this study was to develop fast and reliable sequence characterized amplified region (SCAR) markers linked to the Pm21 gene. A random amplified polymorphic DNA (RAPD) marker for Pm21, OPH171400, was converted to SCAR markers after sequencing the two ends of the polymorphic DNA fragment. Two SCAR markers, SCAR1265 and SCAR1400, were developed to detect the Pm21 gene in different genetic backgrounds. The specific SCAR1265 marker enable large-scale accurate screening for the presence/absence of Pm21 allele. 相似文献
11.
A random amplified polymorphic DNA (RAPD) marker named OPC06-1900 was previously found linked to a fertility restorer gene (Rfw) for cytoplasmic male sterility (CMS) in radish (Raphanus sativus L.). The RAPD marker was converted to a dominant sequence characterized amplified region (SCAR) marker SCC06-1894 by molecular cloning and nucleotide sequencing. A BLAST search revealed that the SCAR marker SCC06-1894 showed significant homology to the corresponding regions of Arabidopsis and Brassica sulfate transporter genes. The presence of the intron and exon of the DNA fragment SCC06-1894 was demonstrated by comparing RT-PCR and PCR products. Thus, allele-specific oligonucleotide primers were designed to amplify the SCAR marker SCC06-415. PCR test with F2 plants and sequence analysis showed that SCC06-1894 and SCC06-415 were allelic, linked to Rfw/rfw gene at 8.0 cM. Nine oligonucleotide primers were designed based on a single radish nuclear restorer gene mRNA. A survey of these primer combinations by bulked segregant analysis (BSA) identified three polymorphisms. The three PCR-based markers were co-segregant in the coupling phase and distant from the Rfw gene by 1.4 cM. These specific markers distributed on both sides of the Rfw gene and will be helpful for breeding new rapseed (Brassica napus L.) restorer lines. 相似文献
12.
Marker assisted selection (MAS) may improve the efficiency of breeding downy mildew resistant cucumber cultivars. A study
was conducted to identify random amplified polymorphic DNA (RAPD) markers linked to the downy mildew resistance gene (dm) which would be suitable for MAS. A total of 145 F3 families from two populations (55 from the WI 1983G × Straight 8 population and 90 from the Zudm1 × Straight 8 population)
were evaluated over five locations in North America and Europe. Resistant and susceptible F3 families were identified and mean family resistance ratings were used to type individual F2 plants. No evidence for race differences in the pathogen (Psuedoperonospora cubensis (Berk. & Curt.) Rostow) between North America and Europe was found. Phenotypic correlations between locations ranged from
0.3 to 0.7. Of the 135 polymorphic RAPD markers identified from 960 primers, five were linked to dm - G14800, X151100, AS5800, BC5191100, and BC5261000. In the WI 1983G × Straight 8 population, G14800 was linked to dm at 16.5 cm, AS5800 at 32.8 cm, BC5191100 at 9.9 cm, and BC5261000 at 19.2 cm. In the Zudm1 ×Straight 8 population, G14800 was linked at 20.9 cm, X151100at 14.8 cm, AS5800 at 24.8 cm, and BC526_1000 at 32.9 cm. MarkersG14800 and BC5191100 were linked in repulsion to the dm allele, and X151100, AS5800, and BC5261000 were linked in coupling phase. These genetic markers may be exploited to develop an efficient MAS strategy for breeding resistant
cucumber cultivars.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
13.
AFLP fingerprinting in Medicago spp.: Its development and application in linkage mapping 总被引：3，自引：0，他引：3
G. Barcaccia E. Albertini S. Tavoletti M. Falcinelli F. Veronesi 《Plant Breeding》1999,118(4):335-340
Cultivated alfalfa (Medicago sativa L., 2n= 4x= 32) is one of the most important forage crops in temperate climates. The genus Medicago includes diploid species that are a valuable source of wild germplasm for studying the reproductive system of alfalfa and its abnormalities. A linkage map of an apomeiotic mutant of Medicago falcata (L.) Arcang. (2n= 2x= 16) that spanned 368.6 cM and included 29 amplified fragment length polymorphism (AFLP), 35 random amplified polymorphic DNA (RAPD) and three restriction fragment length polymorphism (RFLP) loci was constructed using a one-way pseudo-testcross mapping strategy. The success of such a strategy depends on the presence of sufficiently high levels of heterozygosity in the individual plant which is being mapped and on the informativeness of the marker system that is used. In general: (1) highly informative and reproducible RAPD and AFLP fingerprints were generated and several genome-specific primers selected; (2) of 67 marker loci mapped, 51 were arranged in 11 main linkage groups and eight additional couples of linked marker loci were detected; (3) mapping of an F1 population theoretically allowed a better estimation of linkage distances since it avoided segregation distortion (x2 analyses revealed segregation distortion in only 5.2% of marker loci); (4) the high frequency of unlinked marker loci obtained suggests that, in this alfalfa genotype, DNA markers are distributed throughout the genome. This type of genetic map should find application and prove useful in marker-assisted selection and map-based breeding programmes in meiotic mutants of alfalfa for which there is a lack of suitable genetic markers. 相似文献
14.
Squash silverleaf (SSL), caused by the silverleaf whitefly [Bemisia argentifolii (formerly known as Bemisia tabaci Gennadius, B strain)], is an important physiological disorder that affects squash (Cucurbita spp.) by reducing yield potential. Breeding squash with resistance to SSL disorder can be facilitated by using marker-assisted
selection (MAS). Resistance to SSL disorder, in Cucurbita pepo, is conferred by a single recessive gene (sl). The objective of this study was to identify molecular markers associated with resistance. A zucchini squash, SSL disorder
resistant breeding line, ‘Zuc76’ (sl/sl) and a SSL disorder susceptible zucchini cultivar ‘Black Beauty’ (Sl/Sl) were screened with 1,152 randomly amplified polymorphic DNA (RAPD) primers and 432 simple sequence repeat (SSR) markers
to identify polymorphisms. Using F2 and BC1 progeny segregating for SSL disorder resistance, three RAPD (OPC07, OPL07 and OPBC16) primers and one SSR (M121) marker were
found associated with sl. Fragments amplified by RAPD primer OPC07 was linked in coupling phase to sl, whereas RAPD primer OPL07 was linked in repulsion phase. RAPD primer OPBC16 and SSR marker M121 were co-dominant. The allelic
order of these loci was found to be M121–sl–OPC07–OPL07–OPBC16. The closest marker to sl is M121 with an estimated genetic distance of 3.3 cM. The markers identified in this study will be useful for breeding summer
squash (C. pepo) for SSL disorder resistance derived from zucchini squash breeding line ‘Zuc76’. 相似文献
15.
Jianhui Ji Bi Qin Haiyan Wang Aizhong Cao Suling Wang Peidu Chen Lifang Zhuang Yu Du Dajun Liu Xiue Wang 《Euphytica》2008,163(2):159-165
The powdery mildew resistance gene Pm6, transferred to common wheat from the tetraploid Triticum timopheevii, is effective in most epidemic areas for powdery mildew in China. RFLP probe BCD135 was previously associated with Pm6. In the present research, four STS primers (NAU/STSBCD135-1, NAU/STSBCD135-2, STS003 and STS004) were designed from the sequence data of BCD135. These primers were used for PCR amplification using the
genomic DNA of resistant near-isogenic lines with Pm6 and their recurrent parent, cv. Prins. No polymorphic product was observed using primers STS003 and STS004; however, primers
NAU/STSBCD135-1 and NAU/STSBCD135-2 amplified two and one bands, respectively, polymorphic between the resistant near-isogenic-lines and Prins. The two primers
were then used to amplify the F2 population from the cross IGV1-465 (FAO163b/7*Prins) × Prins. The amplification and the powdery mildew resistance identification
data were analyzed using the software Mapmaker 3.0. The results indicated that both NAU/STSBCD135-1 and NAU/STSBCD135-2 were closely linked to Pm6 with a genetic distance of 0.8 cM. A total of 175 commercial varieties without Pm6 from different ecological areas of China were tested using marker NAU/STSBCD135-2 and none of them amplified the 230 bp-specific band. This marker thus has high practicability and can be used in MAS of Pm6 in wheat breeding programs for powdery mildew resistance.
Jianhui Ji and Bi Qin contributed equally to this work. 相似文献
16.
The applicability of random amplified polymorphic DNA (RAPD) markers in the cultivated rubber tree, Hevea, was evaluated using 43 decamer oligonucleotide primers in a set of 24 clones selected in different South-East Asian countries. A total of 220 0.35–3.5 kb DNA fragments were amplified, of which 111 were polymorphic. Of these, 80 fragments (RAPD markers) which were repeatable and clearly scorable across all genotypes were used to estimate genetic distances among the clones tested. The estimated genetic distances ranged from 0.05 (RRII 308 and PB 5/51) to 0.75 (RRIC 100 and SCATC 88–13). A mean genetic distance of 0.5 indicates a rather high genetic variability among the tested clones. As expected, because of the breeding history of Hevea, UPGMA cluster analysis and Principal Coordinate Analysis (PCoA) indicated the absence of a distinct geographical grouping. The possible application of RAPD markers for clone identification and also for analysis of genetic relationships among Hevea clones is discussed. 相似文献
17.
L. De Benedetti G. Burchi A. Mercuri N. Pecchioni P. Faccioli T. Schiva 《Plant Breeding》2000,119(5):443-445
Random amplified polymorphic DNA (RAPD) markers were used to verify interspecific hybridization in Alstroemeria. Five putative interspecific hybrids and their parents were analysed by means of four preselected RAPD primers. The putative parentage was confirmed in four hybrids and was excluded in one that showed completely different RAPD patterns from its putative parents and a different phenotype. Our results demonstrated that this molecular technique is a powerful tool for verifying hybridity rapidly if the putative parents are given. This tool will allow screening of small immature seedlings for verification of hybridity and should improve the efficiency of breeding programmes. 相似文献
18.
Genetic variation among five elite winter barley cultivars (H. vulgare L.) currently grown in Bulgaria was assessed at the molecular level using restriction fragment length polymorphism (RFLP)
and randomly amplified polymorphic DNA (RAPD) markers. The present study sampled RFLPs in four well characterized multigene
families in barley: the seed storage protein loci; the 18S, 5.8S and 26S ribosomal DNA loci; the loci coding for 5S ribosomal
RNA and the loci coding subunit α of ATP-A complex in the mitochondrial genome. RFLPs were detected in three out of five investigated
chromosomal loci in the barley cultivars studied. RAPD assay using arbitrary 10-base primers was applied to generate amplified
length polymorphic markers in barley. Overall a total of 15 polymorphic phenotypes were found among the studied barley cultivars
by using 11 out of 25 tested primers. All RAPDs were considered as dominant genetic markers except for two, where PCR and
Southern blot
analysis indicated the presence of codominant amplification products. Five RAPD polymorphisms in F1 and F2 progenies of the cross between Alpha and Obzor were inherited in Mendelian fashion. The determined values for the genetic
variation proved a high genetic similarity among the tested cultivars. Genetic similarity (GS) calculated from RFLP and RAPD data ranged from 0.888 to 0.997 with a mean GS – 0.933.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
19.
Mahboob A. Chowdhury Chandra P. Andrahennadi Alfred E. Slinkard Albert Vandenberg 《Euphytica》2001,118(3):331-337
Resistance to ascochyta blight of lentil (Lens culinaris Medikus),caused by the fungus Ascochyta lentis, is determined by a single recessive gene, ral
2, in the lentil cultivar Indian head. Sixty F2 individuals from a cross between Eston (susceptible) and Indian head (resistant) lentil were analyzed for the presence of
random amplified polymorphic DNA (RAPD) markers linked to the ral
2gene, using bulked segregant analysis (BSA). Out of 800 decanucleotide primers screened, two produced polymorphic markers
that co-segregated with the resistance locus. These two RAPD markers, UBC2271290and OPD-10870, flanked and were linked in repulsion phase to the gene ral
2 at 12 cm and 16 cm, respectively. The RAPD fragments were converted to SCAR markers. The SCAR marker developed from UBC2271290 could not detect any polymorphism between the two parents or in the F2. The SCAR marker developed from OPD-10870 retained its polymorphism. The polymorphic RAPD marker UBC2271290 and the SCAR marker developed from OPD-10870 can be used together in a marker assisted selection program for ascochyta blight resistance in lentil.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
20.
This research was conducted to study the genetic diversity in safflower (Carthamus tinctorius L.) using agro-morphological traits and RAPD markers. Sixteen selected lines derived from landraces growing in various agro-climatic
regions of Iran along with four exotic genotypes were evaluated in a randomized complete block design with three replications
under field conditions. Days to emergence, days to initial flowering, days to flowering, days to maturity, plant height, branches
per plant, capitula per plant, seeds per capitulum, 1,000-seed weight, seed yield per plant, seed yield, and reaction to powdery
mildew (Leveillula taurica Arnaud) were evaluated in this study. Genetic diversity of the genotypes was assessed by RAPD markers. The results indicated
significant differences among genotypes for the agro-morphological traits and clustering based on these traits classified
the genotypes into five groups. Analysis of the RAPD markers revealed 15 polymorphic primers out of 50 used primers. Based
on RAPD data, the highest genetic similarity was observed between the cultivars of “AC Sunset,” “AC Sterling” from Canada
and the lowest relatedness observed between a local breeding line “E2428” and genotype “GE62923” from Germany. Cluster analysis based on RAPD markers and 54% coefficient of similarity divided the genotypes into five distinct
groups. Comparing the clusters based on agro-morphological traits with those from molecular markers showed slight similarities.
The finding of high genetic variation for agro-morphological traits and polymorphism at DNA level reveal that agronomic traits
can be improved by selection programs. 相似文献