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1.
Summary The first genetic linkage map of Japanese bunching onion (Allium fistulosum) based primarily on AFLP markers was constructed using reciprocally backcrossed progenies. They were 120 plants each of (P1)BC1 and (P2)BC1 populations derived from a cross between single plants of two inbred lines: D1s-15s-22 (P1) and J1s-14s-20 (P2). Based on the (P2)BC1 population, a linkage map of P1 was constructed. It comprises 164 markers – 149 amplified fragment length polymorphisms (AFLPs), 2 cleaved amplified polymorphic sequences (CAPSs), and 12 simple sequence repeats (SSRs) from Japanese bunching onion, and 1 SSR from bulb onion (A. cepa) – on 15 linkage groups covering 947 centiMorgans (cM). The linkage map of P2 was constructed with the (P1)BC1 population and composed of 120 loci – 105 AFLPs, 1 CAPS, and 13 SSRs developed from Japanese bunching onion and 1 SSR from bulb onion – on 14 linkage groups covering 775 cM. Both maps were not saturated but were considered to cover the majority of the genome. Nine linkage groups in P2 map were connected with their counterparts in P1 map using co-dominant anchor markers, 13 SSRs and 1 CAPS.  相似文献   

2.
In several autogamous and vegetatively propagated crops, DNA markers have been used for cultivar identification. However, allogamous crops such as bunching onion (Allium fistulosum L.) are recalcitrant to marker-aided cultivar identification, as well as hybrid seed purity tests, due to the high degree of genetic heterogeneity within each cultivar. To aid cultivar identification and ensure its accuracy in bunching onion, we proposed the “SSR-tagged breeding” scheme in our previous study. The feasibility of this scheme was investigated here using a landrace of bunching onion with two populations tagged with two or four selected SSR markers. Compared with a control population, no significant differences were detected in the agronomic traits of the SSR-tagged populations. The targeted SSR loci were genetically uniform within each population whereas other loci maintained high heterogeneity. These results demonstrate that the SSR-tagged breeding scheme, even with a very small number of markers, is efficient for the identification of newly bred cultivars, and consequently for F1 purity tests, in allogamous crops in which inbreeding depression is as severe as in bunching onion.  相似文献   

3.
Identification and characterization of microsatellites in eggplant   总被引:4,自引:0,他引:4  
T. Nunome    K. Suwabe    H. Iketani  M. Hirai  G. Wricke 《Plant Breeding》2003,122(3):256-262
The potential of microsatellite markers for use in genetic studies in eggplant, Solanum melongena, has been evaluated. A genomic library of eggplant was screened for GA and GT repeat motifs to isolate microsatellite clones. The frequency of each repeat motif in the eggplant genome was found to be every 3200 kb for GA repeats and every 820 kb for GT repeats. Sixty‐one per cent of GT repeats were found to directly flank AT repeats. A total of 37 polymerase chain reaction (PCR) primer pairs were designed, 23 of which amplified a single product or several products. The level of microsatellite polymorphism was evaluated by using S. melongena lines and related Solanum species. Two to six alleles per primer pair were displayed in the S. melongena lines and two to 13 alleles were displayed in the Solanum relatives. Seven microsatellites showed polymorphism between parental lines of the mapping population and segregated in a codominant Mendelian manner. These microsatellite loci were distributed throughout the linkage map.  相似文献   

4.
苎麻基因组微卫星的分离与鉴定   总被引:1,自引:0,他引:1  
以苎麻[Boehmeria nivea (L.) Gaud.]栽培品种芦竹青为材料,经MseⅠ酶切并与相应接头连接,然后与生物素标记的探针(CT)15杂交,再用链亲和素包被磁珠(Dynabeads M-280)捕捉杂交产物,以21碱基接头寡核苷酸为引物经PCR扩增获得了双链目的片段,回收300~1 000 bp DNA片段,然后克隆到pMD18-T载体上,转化至DH5α中,这样就构建了苎麻富含(GA)n微卫星的部分基因组文库。随机挑取36个克隆,以锚定简单重复序列为引物对其进行PCR筛选。测序分析了22个阳性克隆,每个克隆都含有微卫星DNA,阳性克隆率为61.1%,微卫星种类以与探针互补的(GA/CT)n为主,占83.3%,同时还存在(TG/AC)n和三碱基、六碱基为单元构成的苎麻微卫星,如(GAC)n、(ACG)n、(TCT)n、(TCG)n、(GAA)n、(CCGACG) n、(GAGAAA)n等。最后以18个微卫星位点设计了18对苎麻微卫星特异引物。GA/CT重复单元的长度从7到40范围内变化,平均为19.2,显示了它们将产生良好多态性,为一种强有力的遗传标记。苎麻微卫星DNA标记可广泛应用于苎麻基因型鉴定,基因和QTL分析,分子标记辅助育种,系谱分析等。  相似文献   

5.
Microsatellite or simple sequence repeat (SSR) markers are important tools for genetic analyses, especially those targeting diversity. The primary objective of this study was to develop robust oat‐based microsatellite markers from newly enriched genomic libraries to expand on a relatively small existing oat SSR toolbox. Microsatellite motifs characterized by (CA/GT), (AAT/TTA), (ATG/TAC) and (CATC/GTAG) repeats were targeted for enrichment. Preliminary screening showed that 90% of clones from the (CA/GT) and 79% of the clones from the (ATG/TAC) libraries contained repeats, while < 11% of the clones from (AAT/TTA) and (CATC/GTAG) libraries contained repeats. Subsequent sequencing of 1536 clones from the (CA/GT) and (ATG/TAC) libraries resulted in 539 and 578 SSRs for which primers were designed, respectively. A total of 246 SSRs were polymorphic across 11 oat lines. One hundred and twenty‐five of the markers produced highly reproducible assays that interrogated 369 alleles at 193 loci. Of these, 79 robust assays interrogated 146 codominant alleles. These markers will be useful for a wide range of genetic analyses in oat including assessment of diversity and marker‐assisted breeding.  相似文献   

6.
Summary Both random amplified polymorphic DNA and microsatellite repeat sequences were investigated as DNA markers for distinguishing hop cultivars. Microsatellite sequences converted to STS markers proved to be most successful. The relative abundance of microsatellite repeat sequences in the hop genome varied depending on the sequence class. Of the repeat types investigated the dinucleotide repeats (GA)n and (GT)n are the most highly represented in the hop genome. Microsatellite repeat sequences in hops have been shown to be highly polymorphic and are very informatives as STS molecular markers. A DNA typing system using sequence-tagged microsatellite site markers has been developed which will not only be useful for hop cultivar identification but also marker assisted breeding and quality control purposes.  相似文献   

7.
SSR markers targeting (GATA) n motifs are known to be highly polymorphic and useful in many organisms. (GATA) n motif specific SSR markers covering the whole rice genome are not available. The present study was carried out with an objective to identify class I rice microsatellites in the rice genome with (GATA)n motifs, in-silico, and validate their potential as molecular markers. A total of 243 such motifs were identified; 65 of these were present in the genic region, 59 were in the upstream region and the remaining motifs were found in the intergenic regions. Many of the (GATA) n motifs were found within and/or upstream of genes associated with biotic or abiotic stress tolerance. A total of 230 PCR-based markers targeting all the class I (GATA) n microsatellites were developed and 35 of these markers spread across the rice genome were validated in a set of 24 representative rice varieties belonging to five distinct cultivar groups. All the markers were polymorphic, with average polymorphism information content (PIC) value of 0.61, and the rice cultivars could be uniquely distinguished into different cultivar groups based on marker analysis. These informative markers targeting (GATA) n motifs representing a new set of markers in rice will be highly useful for genetic studies and marker-assisted selection. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Passoupathy Rajendrakumar, Akshaya Kumar Biswal and Kannabiran Sakthivel contributed equally.  相似文献   

8.
Microsatellite or simple sequence repeat (SSR) markers are valuable tools for many purposes, such as phylogenetic, fingerprinting and molecular breeding studies. However, such marker resources are unavailable in Assam tea (Camellia assamica ssp. assamica; Masters). With an objective to enrich the repertoire of microsatellite markers in traditional tea, 185 novel microsatellite (150 genomic and 35 genic) markers were identified from (GA)n‐enriched genomic libraries and public expressed sequence data in Assam tea. High‐quality 0.412‐Mb non‐redundant (NR) genomic data set derived from nucleotide sequencing of 1297 (GA)n‐enriched genomic positive clones and 2723 unigenes (1.33 Mb) predicted from 10 803 random public expressed sequence tags (ESTs) in C. assamica ssp. assamica were utilized for identification of genomic and genic microsatellite markers, respectively. The average number of alleles and polymorphic information content (PIC) recorded for the newly developed SSR markers were 6.17 and 0.398, respectively. The average observed (Ho) and expected (He) heterozygosity varied from 0.626 to 0.697, respectively. These markers were found to be highly transferable (74.5–100%) to cultivated (C. sinensis, C. assamica ssp. lasiocalyx) and five wild Camellia species. Genetic diversity coefficient detected a high level of divergence in 24 cultivated tea accessions (69.3%). Phylogenetic analysis revealed that major groupings were broadly in accordance with taxonomic classification of tea, and all the wild Camellia species remained as an out‐group. The high polymorphic content coupled with high rate of cross‐transferability demonstrates wider applicability of novel microsatellite markers in genotyping, genetic diversity, genome mapping and evolutionary studies in various Camellia species.  相似文献   

9.
L. R. Pinto    K. M. Oliveira    T. Marconi    A. A. F. Garcia    E. C. Ulian    A. P. de Souza   《Plant Breeding》2006,125(4):378-384
Microsatellites or simple sequence repeats (SSRs) are one of the most suitable markers for genome analysis as they have great potential to aid breeders to develop new improved sugarcane varieties. The development of SSR derived from expressed sequence tags (EST) opens new opportunities for genetic investigations at a functional level. In the present work, the polymorphism obtained with a subset of 51 EST–SSRs derived from sucest was compared with those generated by 50 genomic SSRs (gSSR) in terms of number of alleles, polymorphism information content, discrimination power and their ability to establish genetic relationships among 18 sugarcane clones including three Saccharum species (S. officinarum, S. barberi, S. sinense). The majority of EST–SSRs loci had four to six alleles in contrast to the seven to nine observed for the gSSRs loci. Approximately, 35% of the gSSRs had PIC values around 0.90 in contrast to 15% of the EST–SSRs. However, the mean discrimination power of the two types of SSR did not differ significantly as much as the average genetic similarity (GS) based on Dice coefficient. The correlation between GS of the two types of SSRs was high (r = 0.71/P = 0.99) and significant. Although differences were observed between dendrograms obtained with each SSR type, both were in good agreement with pedigree information. The S. officinarum clone IJ76‐314 was grouped apart from the other clones evaluated. The results here demonstrate that EST–SSRs can be successfully used for genetic relationship analysis, extending the knowledge of genetic diversity of sugarcane to a functional level.  相似文献   

10.
Lentil is the sixth most important pulse crop terms of production in the world, but the number of available and mapped SSR markers are limited. To develop SSR markers in lentil, four genomic libraries for (CA)n, (GA)n, (AAC)n and (ATG)n repeats were constructed. A total of 360 SSR primers were designed and validated using 15 Turkish lentil cultivars and genotypes. The most polymorphic repeat motifs were GA and CT, with a mean number of alleles per locus of 7.80 and 6.55, respectively. Seventy‐eight SSR primers amplified a total of 400 polymorphic alleles, whereas 71 SSR primers produced markers within the expected size range. For 78 polymorphic SSR primers, the average number of alleles per locus was 5.1 and PIC value ranged from 0.07 to 0.89, with an average of 0.58. A linkage map was constructed using 92 individual F2 plants derived from a cross between Karacada? × Silvan, with 47 SSR markers. The SSR markers developed in this study could be used for germplasm classification and identification and mapping of QTL in lentil.  相似文献   

11.
The induction of haploid plants from F1 hybrids between CMS shallot with Allium galanthum cytoplasm and common onion was examined. Starting with 535 unpollinated flowers cultured in B5 medium 25 seedlings from part henogenetic embryos were obtained of which 13 seedlings survived. Eleven seedlings were determined as haploid plants (2n = x = 8) and 2 seedlings were doubled haploid plants (2n = 2x = 16). All haploid and doubled haploid plants preserved chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) from A. galanthum. Segregation in different characters was observed among the haploid plants. The haploid and doubled haploid plants exhibited the different combinations of genes from shallot and common onion. Crossing of the doubled haploid plants with other shallot strains, common onion cultivars or related species may produce excellent F1 hybrids for bulb production. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

12.
An SSR-based molecular genetic map of cassava   总被引:7,自引:2,他引:7  
E. Okogbenin  J. Marin  M. Fregene 《Euphytica》2006,147(3):433-440
Summary Microsatellites or simple sequence repeats (SSR) are the markers of choice for molecular genetic mapping and marker-assisted selection in many crop species. A microsatellite-based linkage map of cassava was drawn using SSR markers and a F2 population consisting of 268 individuals. The F2 population was derived from selfing the genotype K150, an early yielding genotype from an F1 progeny from a cross between two non-inbred elite cassava varieties, TMS 30572 and CM 2177-2 from IITA and CIAT respectively. A set of 472 SSR markers, previously developed from cassava genomic and cDNA libraries, were screened for polymorphism in K150 and its parents TMS 30572 and CM 2177-2. One hundred and twenty two polymorphic SSR markers were identified and utilized for linkage analysis. The map has 100 markers spanning 1236.7 cM, distributed on 22 linkage groups with an average marker distance of 17.92 cM. Marker density across the genome was uniform. This is the first SSR based linkage map of cassava and represents an important step towards quantitative trait loci mapping and genetic analysis of complex traits in M. esculenta species in national research program and other institutes with minimal laboratory facilities. SSR markers reduce the time and cost of mapping quantitative trait loci (QTL) controlling traits of agronomic interest, and are of potential use for marker-assisted selection (MAS).  相似文献   

13.
Tagging of restorer genes for wild abortive (WA) CMS source by studying a 222 individual plants from a F2 population of a cross between IR58025A × IR42686R. The restorer line IR42686R that was used in this study had been previously derived through random mating composite population (RMCP) involving 12 parents facilitated by IR36 genetic male sterility. Four Rf genes were tagged to simple sequence repeats (SSR) markers on chromosomes 1, 7, 10, 12 by recessive class analysis. The recombination frequency between a positive marker and Rf locus was calculated using maximum likelihood estimator assuming that all the 46 extremely sterile individual plants were homozygous at the targeted Rf locus. The recombination frequency between the marker and the restorer trait were converted to genetic distances using Kosambi function. A new Rf locus designated as Rf7 on chromosome 12 was found to be linked to RM7003 at a genetic distance of 13.3 cM (LOD 6.12). We report here first, a new molecular marker (RM 6344) linked to Rf4 locus on chromosome 7 that was previously mapped by trisomic analysis. RM443 and RM315 were flanking the Rf3 gene at a genetic distance of 4.4 (LOD 10.29) and 20.7 cM (LOD 3.98) on chromosome 1, respectively. The Rf6 was flanked on both side with SSR markers RM258 and RM591 at a genetic distance of 4.4 (LOD 10.29) and 23.3 cM (LOD 3.39) located on chromosome 10. The random mating composite population is an excellent breeding approach to develop superior restorer lines and for pyramiding different Rf genes of different CMS systems. Rf genes tagged with closely linked SSR markers would be facilitating marker assisted selection (MAS) in hybrid rice breeding program by reducing time and workload for identifying potential restorers. L. Bazrkar and A. J. Ali equally contributed to this work.  相似文献   

14.
Bacterial artificial chromosome (BAC) libraries with large DNA fragment inserts have rapidly become the preferred choice for physical mapping. BAC-derived microsatellite or simple sequence repeats (SSRs) markers facilitate the integration of physical maps with genetic maps. The objective of this research was to identify chromosome locations of the BAC-derived SSR markers in tetraploid cotton. A total of 192 SSR primer pairs were derived from BAC clones of an Upland cotton genetic standard line TM-1 (Gossypium hirsutum L.). Metaphor agarose gel electrophoresis results revealed 76 and 59 polymorphic markers between TM-1 and 3–79 (G. barbadense) or G. tomentosum, respectively. Using deletion analysis method, we assigned 39 markers out of the 192 primer pairs to 17 different chromosomes or chromosome arms. Among them, 19 and 17 markers were localized to A-subgenomes (chromosome 1–13) and D-subgenomes (chromosome 14–26), respectively. The subgenome status for the remaining three markers remained unclear due to their two potential chromosome locations achieved by tertiary monosomic stocks deletion analysis. Chromosomal assignment of these BAC-derived SSR markers will help in integrating physical and cotton genetic linkage maps and thus facilitate positional candidate gene cloning, comparative genome analysis, and the coordination of chromosome-based genome sequencing project in cotton. Disclaimer: Mention of trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by USDA, ARS and does not imply its approval to the exclusion of other products or vendors that may also be suitable. The U.S. Government’s right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.  相似文献   

15.
Summary Generation of Simple Sequence Repeat (SSR) DNA markers was based on the construction of genomic DNA library of avocado (Persea americana M.). The library was screened with the four dinucleotide probes (AG), (AT), (GC) and (CA). Positive clones were sequenced to validate the presence of simple sequence repeats (SSR) and to generate polymerase chain reaction (PCR) primers based on the sequences flanking the simple sequence repeat. Twenty six different pairs of primers which yield a PCR product in the initial screening were synthesized. The SSR A1E11 was found to have eleven alleles while A3F8 has eight alleles. The SSRs in avocado were found to be inherited in a Mendelian fashion.Contribution from the Agricultural Research Organization, The Volcani Center, Bet-Dagan, Israel. No. 1335-E, 1994 series.  相似文献   

16.
17.
Summary Simple sequence repeat (SSR) markers linked to quantitative trait loci (QTL) associated with resistance to sorghum shoot fly, Atherigona soccata resistance were used to characterize the genetic and phenotypic diversity of 12 cytoplasmic male-sterile (CMS) and maintainers, 12 restorer lines, and 144 F1 hybrids. The genetic diversity was quite high among the shoot fly-susceptible parents and the hybrids based on them, as indicated by high polymorphic information content (PIC) values, while limited genetic diversity was observed among shoot fly-resistant lines. The phenotypic and genotypic dissimilarity analysis indicated that the shoot fly-resistant and -susceptible parents were 73.2 and 38.5% distinct from each other, and the morphological and genetic distances of certain resistant and susceptible cross combinations was more than their resistant or susceptible parents. Genetic variability among the groups was low (10.8%), but high within groups (89.2%). The genetic and morphological distances suggested that the F1 hybrids were closer to CMS (5 to 12% dissimilar) than the restorer (11 to 87% dissimilar), suggesting that CMS influences the expression of resistance to sorghum shoot fly. The SSR markers can be used to characterize the homologous traits in sorghum germplasm.  相似文献   

18.
Sequence databases were screened to identify simple sequence repeats (SSRs) in Brassica oleracea sequences. A total of 512 B. oleracea DNA sequences were screened and 43 potential SSRs were identified. Thirty-six primer pairs were designed to amplify target sequences. Of the 36 primer pairs, six failed to amplify fragments of expected sizes, and 17 primer pairs failed to generate polymorphisms. Thirteen SSRs were used to assess genetic similarity between 54 B. oleracea cultivars, belonging to 3 variteal groups (cabbage, cauliflower, and broccoli). Pairwise genetic similarities were calculated for cultivars, and a dendrogram of relationships was produced. All cabbage cultivars were distinguished from each other and clustered in two separate groups. Five cauliflower cultivars could not be distinguished with SSR markers used in the study. Three broccoli cultivars clustered with cauliflower cultivars, and two cauliflower cultivars grouped with broccoli cultivars. The varietal group with the narrowest genetic variation in the study was cauliflower (B. oleracea var. botrytis) followed by broccoli (B. oleracea var. italica) and cabbage (B. oleracea var. capitata) groups. Polymorphism information content (PIC) values and number of alleles produced per marker ranged between 0.25 to 0.86 and 1 to 8, respectively, for database derived SSR markers.  相似文献   

19.
In this study, we developed a total of 37 simple sequence repeat (SSR) markers from 11 bacterial artificial chromosome (BAC) clone sequences anchored on chromosome 12 of tomato available at Solanaceae Genomics Network. These SSR markers could group a set of 16 tomato genotypes comprising of Solanum lycopersicum, S. pimpinellifolium, S. habrochaites, and S. pennellii unambiguously according to their known species status. Clear subgroups of genotypes within S. lycopersicum were also observed. A subset of 16 SSR markers representing the 11 BAC clones was used for developing genetic linkage maps of three interspecific F2 populations produced from the crosses involving a common S. lycopersicum parent (CLN2498E) with S. pennellii (LA1940), S. habrochaites (LA407) and S. pimpinellifolium (LA1579). The length of the genetic linkage maps were 112.5 cM, 109.3 cM and 114.1 cM, respectively. Finally, an integrated genetic linkage map spanning a total length of 118.7 cM was developed. The reported SSR markers are uniformly distributed on chromosome 12 and would be useful for genetic diversity and mapping studies in tomato.  相似文献   

20.
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