Inheritance and mapping of 2n-egg production in diploid alfalfa

Genome ◽  
2000 ◽  
Vol 43 (3) ◽  
pp. 528-537 ◽  
Author(s):  
G Barcaccia ◽  
E Albertini ◽  
D Rosellini ◽  
S Tavoletti ◽  
F Veronesi
Keyword(s):  
Molecular Markers ◽  
Linkage Group ◽  
Egg Production ◽  
Seed Set ◽  
Bulked Segregant Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Aflp Markers ◽  
Putative Gene ◽  
2N Eggs

The production of eggs with the sporophytic chromosome number (2n eggs) in diploid alfalfa (Medicago spp.) is mainly associated with the absence of cytokinesis after restitutional meiosis. The formation of 2n eggs through diplosporic apomeiosis has also been documented in a diploid mutant of M. sativa subsp. falcata (L.) Arcang. (2n = 2x = 16), named PG-F9. Molecular tagging of 2n-egg formation appears to be an essential step towards marker-assisted breeding and map-based cloning strategies aimed at investigating and manipulating reproductive mutants of the M. sativa complex. We made controlled crosses between PG-F9 and three wild type plants of M. sativa subsp. coerulea (Less.) Schm. (2n = 2x = 16) and then hand-pollinated the F1 progenies with tetraploid plants of M. sativa subsp. sativa L. (2n = 4x = 32). As a triploid embryo block prevents the formation of 3x progenies in alfalfa because of endosperm imbalance, and owing to the negligible selfing rate, seed set in 2x-4x crosses was used to discriminate the genetic capacity for 2n-egg production. F1 plants that exhibited null or very low seed sets were classified as normal egg producers and plants with high seed sets as 2n-egg producers. A bulked segregant analysis (BSA) with RAPD (random amplified polymorphic DNA), ISSR (inter-simple sequence repeat), and AFLP (amplified fragment length polymorphism) markers was employed to identify a genetic linkage group related to the 2n-egg trait using one of the three F1 progenies. This approach enabled us to detect a paternal ISSR marker of 610 bp, generated by primer (CA)8-GC, located 9.8 cM from a putative gene (termed Tne1, two-n-eggs) that in its recessive form determines 2n eggs and a 30% recombination genomic window surrounding the target locus. Eight additional RAPD and AFLP markers, seven of maternal, and one of paternal origin, significantly co-segregated with the trait under investigation. The minimum number of quantitative trait loci (QTLs) controlling seed set in 2x-4x crosses was estimated by ANOVA and regression analysis. Four maternal and three paternal independent molecular markers significantly affected the trait. A paternal RAPD marker allele, mapped in the same linkage group of Tne1, explained 43% of the variation for seed set in 2x-4x crosses indicating the presence of a major QTL. A map of the PG-F9 chromosome regions carrying the minor genes that determine the expression level of 2n eggs was constructed using selected RAPD and AFLP markers. Two of these genes were linked to previously mapped RFLP loci belonging to groups 1 and 8. Molecular and genetic evidence support the involvement of at least five genes.Key words: Medicago spp., meiotic mutants, molecular markers.

scholarly journals Identification of Markers Linked to Seedlessness in Citrus kinokuni hort. ex Tanaka and Its Progeny Using Bulked Segregant Analysis

HortScience ◽  
2011 ◽  
Vol 46 (5) ◽  
pp. 693-697 ◽  
Author(s):  
Dario J. Chavez ◽  
José X. Chaparro
Keyword(s):  
Molecular Markers ◽  
Linkage Group ◽  
Bulked Segregant Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Dominant Gene ◽  
Segregation Ratio ◽  
Single Dominant Gene ◽  
Citrus Breeding

Citrus kinokuni ‘Mukaku kishu’ PI539530 and its progeny were studied to identify random amplified polymorphic DNA (RAPD) primers associated with seedlessness. Ninety-one F1 [(Robinson op) × C. kinokuni] individuals showed a 1:1 segregation ratio between seedless and seeded phenotypes with seedless as a single dominant gene. Bulked segregant analysis was used to identify markers associated with the seedless locus. Eighteen RAPD primers were mapped into a partial linkage group (≈55.8 cM length) with four RAPD primers flanking the seedless locus: OPAI11-0.8 at 8.7 cM, OPAJ19-1.0 at 8.4 cM, OPM06r-0.85 at 4.3 cM, and OPAJ04r-0.6 at 6.4 cM. The identification of molecular markers linked to C. kinokuni Fs seedless locus constitutes an important and major tool for citrus breeding and selection.

scholarly journals Bulked Segregant Analysis Identifies Molecular Markers Linked to Melampsora medusae Resistance in Populus deltoides

2000 ◽  
Vol 90 (9) ◽  
pp. 1039-1042 ◽  
Author(s):  
G. M. Tabor ◽  
T. L. Kubisiak ◽  
N. B. Klopfenstein ◽  
R. B. Hall ◽  
H. S. McNabb McNabb
Keyword(s):  
Molecular Markers ◽  
Leaf Rust ◽  
Populus Deltoides ◽  
Bulked Segregant Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Genetic Distances ◽  
Resistance Locus ◽  
The North ◽  
Melampsora Medusae ◽  
Central United States

In the north central United States, leaf rust caused by Melampsora medusae is a major disease problem on Populus deltoides. In this study we identified molecular markers linked to a M. medusae resistance locus (Lrd1) that was segregating 1:1 within an intraspecific P. deltoides family (C9425DD). Previous field results were confirmed in the controlled environment of a growth chamber through an excised whole-leaf inoculation method. Using bulked segregant analysis we identified two random amplified polymorphic DNA (RAPD) markers (OPG10340 and OPZ191800) that are linked to Lrd1. Based on segregation in a total of 116 progeny, the genetic distances between OPG10340 and OPZ191800 and the resistance locus were estimated as 2.6 and 7.4 Haldane centimorgans (cM), respectively. Multipoint linkage analyses strongly suggest the most likely order for these loci is Lrd1, OPG10340, and OPZ191800. These markers may prove to be instrumental in the eventual cloning of Lrd1, as well as for marker-assisted selection of leaf-rust resistant genotypes.

scholarly journals Characterization of genetic diversity of cactus species (Opuntia spp.) in Morocco by morphological traits and molecular markers

2017 ◽  
Vol 5 (2) ◽  
pp. 149-159 ◽  
Author(s):  
Y. El Kharrassi ◽  
M.A. Mazri ◽  
M.H. Sedra ◽  
A. Mabrouk ◽  
B . Nasser ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Molecular Markers ◽  
Morphological Traits ◽  
Polymorphic Information Content ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Group Method ◽  
Rapd Primer ◽  
Opuntia Spp ◽  
Issr Primers

The genetic diversity within and among 124 accessions of Opuntia spp. collected from different regions of Morocco was assessed using morphological descriptors and molecular markers. Based on 10 morphological traits, the accessions were separated into 3 main clusters; each cluster was containing accessions from different regions and species. Polymerase chain reaction (PCR) was then performed on 22 accessions from different regions and species, with 10 inter-simple sequence repeat (ISSR) primers and one random amplified polymorphic DNA (RAPD) primer. ISSR primers produced 66 bands overall, 64 (96.9 %) of which were polymorphic while 6 bands were generated by the RAPD marker, all polymorphic. The polymorphic information content (PIC) values ranged from 0.62 to 0.97, with an average of 0.82. The dendrogram of genetic differences generated using the unweighted pair-group method using arithmetic averages (UPGMA) method showed 7 different clusters at a similarity of 0.76, which was confirmed by the principal component analysis (PCA). The main conclusion of our work is the high genetic similarity between Opuntia ficus indica and Opuntia megacantha species in Morocco. Our results will be useful for plant breeding and genetic resource conservation programs.

Molecular Markers Linked to Phytophthora fragariae Resistance Genes in Strawberry

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 500a-500 ◽  
Author(s):  
K.M. Haymes ◽  
S.C. Hokanson ◽  
K. Salazar ◽  
J. Maas
Keyword(s):  
Molecular Markers ◽  
Resistance Genes ◽  
Bulked Segregant Analysis ◽  
Aflp Markers ◽  
Avirulence Genes ◽  
Phytophthora Fragariae ◽  
Red Stele ◽  
Strawberry Breeding ◽  
Direct Applicability ◽  
Selection Of

The commercial strawberry (Fragaria × ananassa Duch.) is susceptible to red stele root rot caused by the fungus Phytophthora fragariae Hickman var. fragariae. Characteristics of the disease are the reddened steles and “rat tail” appearance of the roots. Infected plants are dwarfed, exhibit wilting, and eventually die. Red stele resistance in strawberries and virulence of P. fragariae behave according to the gene-for-gene system. Resistance genes and their corresponding avirulence genes have been identified by screening plant roots for infection. The goal of this international research project with CPRO-DLO, The Netherlands, is to develop highly specific molecular markers for various Rpf resistance genes (Rpf1, Rpf2, Rpf3, and Rpf6) that confer resistance against P. fragariae. Bulked Segregant Analysis was used to identify RAPD and AFLP markers putatively linked to P. fragariae resistance genes. The bulked DNAs representing subsets of three F1 populations that segregated monogenically for either resistance or susceptibility to P. fragariae. The map of these Rpf loci was generated using JoinMap®. The RAPD markers linked to the resistance genes are being converted into SCAR markers, while the AFLP markers will be used as probes for the detection of P. fragariae. The application of the molecular markers linked to resistance genes will have direct applicability to strawberry breeding programs. Marker-facilitated selection of these resistance genes would allow an efficient means in the screening and selection of plant material containing these genes and help in the pyramiding for resistance to P. fragariae. Genetic improvement of the strawberry by the ability to pyramid resistance genes will contribute to the productivity of the strawberry industry worldwide.

scholarly journals Genome Relationship among Nine Species of Millettieae (Leguminosae: Papilionoideae) Based on Random Amplified Polymorphic DNA (RAPD)

2004 ◽  
Vol 59 (11-12) ◽  
pp. 868-873 ◽  
Author(s):  
Laxmikanta Acharya ◽  
Arup Kumar Mukherjee ◽  
Pratap Chandra Panda
Keyword(s):  
Molecular Markers ◽  
Genetic Relationship ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Similarity Index ◽  
Morphological Characteristics ◽  
Genome Relationship ◽  
Tephrosia Purpurea ◽  
Constituent Species

Random amplified polymorphic DNA (RAPD) marker was used to establish intergeneric classification and phylogeny of the tribe Millettieae sensu Geesink (1984) (Leguminosae: Papilionoideae) and to assess genetic relationship between 9 constituent species belonging to 5 traditionally recognized genera under the tribe. DNA from pooled leaf samples was isolated and RAPD analysis performed using 25 decamer primers. The genetic similarities were derived from the dendrogram constructed by the pooled RAPD data using a similarity index, which supported clear grouping of species under their respective genera, inter- and intra-generic classification and phylogeny and also merger of Pongamia with Millettia. Elevation of Tephrosia purpurea var. pumila to the rank of a species (T. pumila) based on morphological characteristics is also supported through this study of molecular markers.

Isolation of microsatellite and RAPD markers flanking the Yr15 gene of wheat using NILs and bulked segregant analysis

Genome ◽  
1999 ◽  
Vol 42 (6) ◽  
pp. 1050-1056 ◽  
Author(s):  
V Chagué ◽  
T Fahima ◽  
A Dahan ◽  
G L Sun ◽  
A B Korol ◽  
...  
Keyword(s):  
Resistance Gene ◽  
Stripe Rust ◽  
Rust Resistance ◽  
Rapd Markers ◽  
Bulked Segregant Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Rust Resistance Gene ◽  
Stripe Rust Resistance ◽  
Stripe Rust Resistance Gene

Microsatellite and random amplified polymorphic DNA (RAPD) primers were used to identify molecular markers linked to the Yr15 gene which confer resistance to stripe rust (Puccina striiformis Westend) in wheat. By using near isogenic lines (NILs) for the Yr15 gene and a F2 mapping population derived from crosses of these lines and phenotyped for resistance, we identified one microsatellite marker (GWM33) and one RAPD marker (OPA19800) linked to Yr15. Then, bulked segregant analysis was used in addition to the NILs to identify RAPD markers linked to the target gene. Using this approach, two RAPD markers linked to Yr15 were identified, one in coupling (UBC199700) and one in repulsion phase (UBC2121200). After Mapmaker linkage analysis on the F2 population, the two closest markers were shown to be linked to Yr15 within a distance of about 12 cM. The recombination rates were recalculated using the maximum likelihood technique to take into account putative escaped individuals from the stripe rust resistance test and obtain unbiased distance estimates. As a result of this study, the stripe rust resistance gene Yr15 is surrounded by two flanking PCR markers, UBC199700 and GWM33, at about 5 cM from each side.Key words: wheat, Triticum dicoccoides, Yr15 stripe rust resistance gene, genetic mapping, microsatellite markers, RAPD markers.

scholarly journals Associating Molecular Markers with Virus Resistance to Classify Sweetpotato Genotypes

2005 ◽  
Vol 130 (3) ◽  
pp. 355-359 ◽  
Author(s):  
M. Mcharo ◽  
D. LaBonte ◽  
R.O.M. Mwanga ◽  
A. Kriegner
Keyword(s):  
Molecular Markers ◽  
Qtl Analysis ◽  
Ipomoea Batatas ◽  
Rapd Markers ◽  
Dna Marker ◽  
Fragment Length Polymorphism ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Multivariate Techniques ◽  
Accuracy Rates

Molecular markers linked to resistance to sweetpotato chlorotic stunt closterovirus [SPCSV (genus Crinivirus, family Closteroviridae)] and sweetpotato feathery mottle virus [SPFMV (genus Potyvirus, family Potyviridae)] were selected using quantitative trait loci (QTL) analysis, discriminant analysis and logistic regression. Eighty-seven F1 sweetpotato [Ipomoea batatas (L.) Lam.] genotypes from a cross of `Tanzania' and `Wagabolige' landraces were used to generate DNA marker profiles for this study. Forty-five of the clones were resistant to SPCSV while 37 were resistant to SPFMV. A combination of 232 amplified fragment length polymorphism (AFLP) markers and 37 random amplified polymorphic DNA (RAPD) markers obtained were analyzed to determine the most informative markers. All three statistical procedures revealed that AFLP marker e41m33.a contributed the greatest variation in SPCSV resistance and RAPD marker S13.1130 accounted for most of the variation in SPFMV resistance. The power of discriminant and logistic analyses is that you do not need a parent-progeny population. An evaluation of these two models indicated a classification and prediction accuracy rates of 96% with as few as four markers in a model. Both multivariate techniques identified one important discriminatory marker (e44m41.j) for SPCSV and two markers (e41m37.a and e44m36.d) for SPFMV that were not identified by QTL analysis.

scholarly journals Identification of Random Amplified Polymorphic DNA (RAPD) Markers for Self-incompatibility Alleles in Hazelnut

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 591e-591
Author(s):  
Kirk W. Pomper ◽  
Anita N. Azarenko ◽  
Joel W. Davis ◽  
Shawn A. Mehlenbacher
Keyword(s):  
Rapd Markers ◽  
Uv Light ◽  
Bulked Segregant Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Self Incompatibility ◽  
Ethidium Bromide Staining ◽  
Young Leaves ◽  
Incompatibility Alleles ◽  
Selection Of

Random amplified polymorphic DNA (RAPD) markers were identified for self-incompatibility (SI) alleles that will allow marker-assisted selection of desired S-alleles and assist in cloning the locus responsible for the sporophytic SI displayed in hazelnut (Corylus avellana L.). DNA was extracted from young leaves collected from field-planted parents and 27 progeny of the cross OSU 23.017 (S1 S12) × VR6-28 (S2 S26). Screening of 10-base oligonucleotide RAPD primers was performed using bulked segregant analysis. DNA samples from six trees each were pooled into four “bulks,” one for each of the following: S1 S2, S1 S26, S2 S12, and S12 S26. “Super bulks” of twelve trees each for S1, S2, S12, and S26 then were created for each allele by combining the appropriate bulks. The DNA from these four super bulks and also the parents was used as a template in the PCR assays. Amplification products were electrophoresed on 2% agarose gels and photographed under UV light after ethidium bromide staining. 200 primers were screened and one RAPD marker each was identified for alleles S2 (OPI-07700) and S1 (OPJ-141700).

scholarly journals Identification and Mapping of Markers Linked to the Mi Gene for Root-knot Nematode Resistance in Peach

2005 ◽  
Vol 130 (1) ◽  
pp. 24-33 ◽  
Author(s):  
Anne M. Gillen ◽  
Fred A. Bliss
Keyword(s):  
Linkage Group ◽  
Prunus Persica ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Nematode Resistance ◽  
Linkage Maps ◽  
Linkage Groups ◽  
Root Knot Nematode ◽  
Repulsion Phase ◽  
Group 2

An F2 population from a single F1 plant from the cross of peach [Prunus persica (L.) Batsch] rootstock cultivars Harrow Blood (HB) × Okinawa (Oki) was used to locate the Mi locus, which conditions resistance to Meloidogyne incognita (race 1) (Kofoid and White) Chitwood. These data and comparison of common markers among published genetic linkage maps placed the Mi locus on Prunus L. linkage group 2. Two restriction fragment length polymorphisms (RFLPs) [linked at 4.8 and 6.8 centimorgan (cM), repulsion phase] and one random amplified polymorphic DNA (RAPD) marker (linked at 9.5 cM, coupling phase) were linked to Mi. The RAPD marker was cloned, sequenced, and converted to a polymerase chain reaction (PCR)-based cleaved amplified polymorphic sequence (CAPs) marker. Clones of resistance gene analogs (RGA) developed from Oki were highly polymorphic when used as RFLP probes. The RGA's mapped to four linkage groups but clustered on two of the four linkage groups, providing limited coverage of the genome. Even so, they may be useful as markers for disease resistance genes that occur in other populations. The linkage maps of the HB × Oki F2 population and a peach × almond (Prunus amygdalus Batsch) F2 population were colinear in certain regions, however, a significant number of markers mapped to different linkage groups among the two populations. The locus for the blood-flesh trait (red-violet mesocarp) mapped to the top of linkage group 4.

scholarly journals Marker Saturation of the Region Flanking the Gene NSV Conferring Resistance to the Melon Necrotic Spot Carmovirus (MNSV) in Melon

2002 ◽  
Vol 127 (4) ◽  
pp. 540-544 ◽  
Author(s):  
Mónica Morales ◽  
Marisol Luís-Arteaga ◽  
José María Álvarez ◽  
Ramon Dolcet-Sanjuan ◽  
Amparo Monfort ◽  
...  
Keyword(s):  
Linkage Group ◽  
Cucumis Melo ◽  
Bulked Segregant Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Necrotic Spot ◽  
Spot Virus ◽  
Susceptible Line ◽  
Length Polymorphisms ◽  
Amplified Fragment Length Polymorphisms ◽  
Melon Genome

The recessive allele (nsv) of the NSV gene confers resistance to the Carmovirus melon necrotic spot virus (MNSV) in melon (Cucumis melo L.). Using an F2 population obtained from the cross between the resistant Korean accession PI 161375 and a susceptible line of `Piel de Sapo', we have mapped the NSV locus to linkage group 11 (G11) of the melon genome. Additional markers closely linked to NSV were developed by bulked segregant analysis (BSA) using a doubled haploid progeny population derived from the same cross. A detailed map of the NSV region was constructed containing 10 markers spanning a distance of 17.7 cM. The nearest flanking markers to NSV were two amplified fragment length polymorphisms (AFLPs) (CTA/ACG-115 and CTA/ACG-120) and one random amplified polymorphic DNA (RAPD) (OPD08-0.80) separated by 5.9 cM. Two more markers, ACC/ACC-110 and OPX15-1.06, cosegregated with NSV.

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