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1.
Construction of a 550 kb BAC contig spanning the genomic region containing the apple scab resistance gene Vf 总被引:2,自引:0,他引:2
Patocchi A Vinatzer BA Gianfranceschi L Tartarini S Zhang HB Sansavini S Gessler C 《Molecular & general genetics : MGG》1999,262(4-5):884-891
A positional cloning project was started in apple with the aim of isolating the Vf resistance gene of Malus floribunda 821. Vf confers resistance against apple scab, the most important disease in apple orchards. A chromosome walk starting from two
molecular markers (M18-CAPS and AM19-SCAR) flanking Vf was performed, using a bacterial artificial chromosome (BAC) library containing inserts of the cultivar Florina, which is
heterozygous for Vf. Thirteen BAC clones spanning the region between the two markers were identified in nine chromosome walking steps. The size
of the resulting contig is approximately 550 kb. In order to map the Vf region in more detail, we analyzed over 2000 plants from different populations segregating for Vf with markers produced from BAC end sequences. In this way, we were able to restrict the possible location of the Vf gene to a minimum of five clones spanning an interval of approximately 350 kb.
Received: 4 July 1999 / Accepted: 16 September 1999 相似文献
2.
I. Kaloshian J. Yaghoobi T. Liharska J. Hontelez D. Hanson P. Hogan T. Jesse J. Wijbrandi G. Simons P. Vos P. Zabel V. M. Williamson 《Molecular & general genetics : MGG》1998,257(3):376-385
As part of a map-based cloning strategy designed to isolate the root-knot nematode resistance gene Mi, tomato F2 populations were analyzed in order to identify recombination points close to this economically important gene.
A total of 21 089 F2 progeny plants were screened using morphological markers. An additional 1887 F2 were screened using PCR-based
flanking markers. Fine-structure mapping of recombinants with newly developed AFLP markers, and RFLP markers derived from
physically mapped cosmid subclones, localized Mi to a genomic region of about 550 kb. The low frequency of recombinants indicated that recombination was generally suppressed
in these crosses and that crossovers were restricted to particular regions. To circumvent this problem, a population of Lycopersicon peruvianum, the species from which Mi was originally introgressed, that was segregating for resistance was developed. Screening of this population with PCR, RFLP
and AFLP markers identified several plants with crossovers near Mi. Recombination frequency was approximately eight-fold higher in the Mi region of the L. peruvianum cross. However, even within the wild species cross, recombination sites were not uniformly distributed in the region. By
combining data from the L. esculentum and L. peruvianum recombinant analyses, it was possible to localize Mi to a region of the genome spanning less than 65 kb.
Received: 15 July 1997 / Accepted: 1 October 1997 相似文献
3.
G. J. King F. H. Alston L. M. Brown E. Chevreau K. M. Evans F. Dunemann J. Janse F. Laurens J. R. Lynn C. Maliepaard A. G. Manganaris P. Roche H. Schmidt S. Tartarini J. Verhaegh R. Vrielink 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,96(5):699-708
Apple scab, caused by the fungus Venturia inaequalis (Cke.) Wint., is an important disease in commercial apple production. A mapping population of 155 individuals, derived from
a cross between the apple varieties ‘Prima’ (resistant)בFiesta’ (susceptible), was scored for response to the disease in
replicated field and glasshouse trials throughout Europe. Twenty data sets were selected and cluster analysis was used to
form a consensus score for the population fitting a 1 : 1 segregation ratio of resistance:susceptibility. The progeny were
scored with molecular markers. A detailed map covering 54 cM of the ‘Prima’ linkage group containing the Vf gene for scab resistance was constructed using 24 molecular markers linked to the resistance gene. One isoenzyme marker (Pgm-1), six RFLP markers and 17 RAPD markers formed a linkage group with the consensus measure of resistance to scab. Four marker
bridges were established with the corresponding ‘Fiesta’ linkage group with additional markers (one isozyme, one RFLP, three
RAPD and one AFLP). A low chi-square value indicated a good fit of the marker ordering, which was in close agreement with
previously reported linkage positions for some of the markers and Vf. Differences were observed in the ability of different scoring methods to resolve susceptible and resistant classes. The
results obtained for the consensus classification of resistance to scab for the population may suggest the presence of virulent
inocula at some sites, which could overcome the Vf gene for resistance. The consequences of relying on individual scoring occasions for studying Vf scab resistance are discussed in the context of linkage analysis, conventional breeding selection, and marker-assisted selection.
Received: 23 July 1997 / Accepted: 31 October 1997 相似文献
4.
G. Cnops B. den Boer A. Gerats M. Van Montagu M. Van Lijsebettens 《Molecular & general genetics : MGG》1996,253(1-2):32-41
The Arabidopsis tornado1 (trn1) mutation causes severe dwarfism combined with twisted growth of all organs. We present a chromosome landing strategy, using
amplified restriction fragment length polymorphism (AFLP) marker technology, for the isolation of the TRN1 gene. The recessive trn1 mutation was identified in a C24 transgenic line and is located 5 cM from a T-DNA insertion. We mapped the TRN1 locus to the bottom half of chromosome 5 relative to visible and restriction fragment length polymorphism (RFLP) markers.
Recombinant classes within a 3-cM region around TRN1 were used to build a high-resolution map in this region, using the AFLP technique. Approximately 300 primer combinations
have been used to test about 26 000 fragments for polymorphisms. Seventeen of these AFLP markers were identified in the 3-cM
region around TRN1. These markers were mapped within this region using individual recombinants. Four of these AFLP markers co-segregate with
TRN1 whereas one maps at one recombinant below TRN1. We isolated and cloned three of these AFLP markers. These markers identified two yeast artificial chromosome (YAC) clones,
containing the RFLP marker above and the AFLP marker below TRN1, demonstrating that these YACs span the TRN1 locus and that chromosome landing has been achieved, using an AFLP-based strategy.
Received: 25 April 1996 / Accepted: 26 June 1996 相似文献
5.
Resistance to submergence stress is an important breeding objective in areas where rice cultivars are subjected to complete
inundation for a week or more. The present study was conducted to develop a high-resolution map of the region surrounding
the submergence tolerance gene Sub1 in rice, which derives from the Indian cultivar FR13A. Submergence screening of 8-day-old plants of F3 families kept for 14 days submerged in 60 cm of water allowed an accurate classification of Sub1 phenotypes. Bulked segregant analysis was used to identify AFLP markers linked to Sub1. A population of 2950 F2 plants segregating for Sub1 was screened with two RFLP markers flanking the Sub1 locus, 2.4 and 4.9 cM away. Submergence tolerance was measured in the recombinant plants, and AFLP markers closely linked
to Sub1 were mapped. Two AFLP markers cosegregated with Sub1 in this large population, and other markers were localized within 0.2 cM of Sub1. The high-resolution map should serve as the basis for map-based cloning of this important locus, as it will permit the identification
of BAC clones spanning the region.
Received: 15 December 1999 / Accepted: 18 February 2000 相似文献
6.
Narrowing down the region of the<Emphasis Type="Italic"> Vf</Emphasis> locus for scab resistance in apple using AFLP-derived SCARs 总被引:4,自引:0,他引:4
Huaracha E Xu M Korban SS 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2004,108(2):274-279
A narrow-down strategy to restrict the Vf region, which controls resistance to the fungal disease apple scab in apple, to a genetic distance of 0.4 cM is presented. Using 11 AFLP-derived SCARs and three RAPD-derived SCARs, all linked to the Vf gene, we subjected 1,412 scab-resistant individuals from 16 mapping populations to genotype analysis. Eleven recombinant individuals were identified within a genetic distance of 0.9 cM around the Vf gene. Using these 11 recombinants, we achieved fine-resolution of several AFLP-derived SCAR markers surrounding the Vf gene, resulting in the following genetic linkage map: ACS-6 and ACS are located left of the Vf gene at genetic distances of 0.2 cM and 0.1 cM, respectively; ACS-7 and ACS-9 are inseparable from the Vf gene; ACS-8, ACS-10, and ACS-4 are located to the right of the Vf gene at genetic distances of 0.1 cM, 0.4 cM, and 0.5 cM, respectively; the remaining five SCARs—ACS-11, ACS-5, ACS-2, ACS-1, and AL07—are inseparable and are located right of the Vf gene at a genetic distance of 0.7 cM. By integrating this linkage data with our previous physical map, we generated a revised map of the narrowed-down region of Vf.Communicated by P. Langridge 相似文献
7.
Genetic mapping of the pear scab resistance gene Vnk of Japanese pear cultivar Kinchaku 总被引:1,自引:0,他引:1
Terakami S Shoda M Adachi Y Gonai T Kasumi M Sawamura Y Iketani H Kotobuki K Patocchi A Gessler C Hayashi T Yamamoto T 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2006,113(4):743-752
Pear scab (caused by Venturia nashicola) is one of the most harmful diseases of pears, especially Japanese and Chinese pear species. The molecular identification and early selection of resistant plants could greatly improve pear breeding. We have identified the position of the scab resistance gene, designated Vnk in an indigenous Japanese pear cultivar Kinchaku, within the pear genome by using simple sequence repeat (SSR) markers derived from pear and apple. The position of Vnk was identified in the central region of linkage group 1 of Kinchaku. Several amplified fragment length polymorphism (AFLP) markers linked to Vnk were obtained by bulked segregant analysis. Among them, the AFLP marker closest to Vnk was converted into a sequence tagged site (STS) marker. Four random amplified polymorphic DNA (RAPD) markers previously found to be loosely associated with Vnk (Iketani et al. 2001) were successfully converted into STS markers. Six markers (one SSR Hi02c07 and five STSs converted from AFLP and RAPD) showed tight linkages to Vnk, being mapped with distances ranging from 2.4 to 12.4 cM. The SSR CH-Vf2, which was isolated from a BAC clone of the contig containing the apple scab gene Vf, was mapped at the bottom of linkage group 1 in Kinchaku, suggesting that the Vnk and Vf loci are located in different genomic regions of the same homologous linkage group. 相似文献
8.
J.M. Soriano S.G. Joshi M. van Kaauwen Y. Noordijk R. Groenwold B. Henken W.E. van de Weg H.J. Schouten 《Tree Genetics & Genomes》2009,5(3):475-482
Apple scab, caused by the fungal pathogen Venturia inaequalis, is one of the most devastating diseases for the apple growing in temperate zones with humid springs and summers. Breeding
programs around the world have been able to identify several sources of resistance, the Vf from Malus floribunda 821 being the most frequently used. The appearance of two new races of V. inaequalis (races 6 and 7) in several European countries that are able to overcome the resistance of the Vf gene put in evidence the necessity of the combination of different resistance genes in the same genotype (pyramiding). Here,
we report the identification and mapping of a new apple scab resistance gene (Vd3) from the resistant selection “1980-015-25” of the apple breeding program at Plant Research International, The Netherlands.
This selection contains also the Vf gene and the novel V25 gene for apple scab resistance. We mapped Vd3 on linkage group 1, 1 cM to the south of Vf in repulsion phase to it. Based on pedigree analysis and resistance tests, it could be deduced that 1980-015-25 had inherited
Vd3 from the founder “D3.” This gene provides resistance to the highly virulent EU-NL-24 strain of race 7 of V. inaequalis capable of overcoming the resistance from Vf and Vg.
JMS and SGJ contributed equally to this work 相似文献
9.
High-resolution mapping of loci conferring resistance to sugarcane mosaic virus in maize using RFLP, SSR, and AFLP markers 总被引:14,自引:0,他引:14
M. L. Xu A. E. Melchinger X. C. Xia T. Lübberstedt 《Molecular & general genetics : MGG》1999,261(3):574-581
Sugarcane mosaic virus (SCMV) is one of the most important virus diseases of maize in Europe. Genetic analysis on backcross
five (BC5) progeny derived from the cross FAP1360A (resistant) × F7 (susceptible) confirmed that at least two dominant genes,
Scm1 and Scm2, are required for resistance to SCMV in the progeny of this cross. With the aid of RFLP and SSR marker analyses, Scm1 was mapped in the region of 8.7 cM – between the nucleolus organizer region (nor) and RFLP marker bnl6.29 on the short arm of chromosome 6, while Scm2 was mapped to an interval of 26.8 cM flanked by the RFLP markers umc92 and umc102 near the centromere region of chromosome 3. Both chromosome regions were further enriched for AFLP markers by successful
application of a bulked segregant analysis to this oligogenic trait. A total of 23 linked AFLP markers were identified, clustered
in chromosome regions adjacent to either Scm1 or Scm2. Seven AFLP markers linked to Scm1 resided within the nor-bnl6.29 interval, and one of them, E3M8-1, showed no recombination with Scm1. Three AFLP markers linked to Scm2 are located between umc92 and umc102.
Received: 13 October 1998 / Accepted: 18 January 1999 相似文献
10.
G. Schwarz W. Michalek V. Mohler G. Wenzel A. Jahoor 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,98(3-4):521-530
The complex Mla locus of barley determines resistance to the powdery mildew pathogen Erysiphe graminis f. sp. hordei. With a view towards gene isolation, a population consisting of 950 F2 individuals derived from a cross between the near-isogenic lines ‘P01’ (Mla1) and ‘P10’ (Mla12) was used to construct a high-resolution map of the Mla region. A fluorescence-based AFLP technique and bulked segregant analysis were applied to screen for polymorphic, tightly
linked AFLP markers. Three AFLP markers were selected as suitable for a chromosome-landing strategy. One of these AFLP markers
and a closely linked RFLP marker were converted into sequence-specific PCR markers. PCR-based screening of approximately 70 000
yeast artificial chromosome (YAC) clones revealed three identical YACs harbouring the Mla locus. Terminal insert sequences were obtained using inverse PCR. The derived STS marker from the right YAC end-clone was
mapped distal to the Mla locus.
Received: 17 July 1998 / Accepted: 9 August 1998 相似文献
11.
QTL Mapping for Frond Length and Width in Laminaria japonica Aresch (Laminarales, Phaeophyta) Using AFLP and SSR Markers 总被引:1,自引:0,他引:1
Fuli Liu Zhanru Shao Haining Zhang Jidong Liu Xiuliang Wang Delin Duan 《Marine biotechnology (New York, N.Y.)》2010,12(4):386-394
In Laminaria japonica Aresch breeding practice, two quantitative traits, frond length (FL) and frond width (FW), are the most important phenotypic
selection index. In order to increase the breeding efficiency by integrating phenotypic selection and marker-assisted selection,
the first set of QTL controlling the two traits were determined in F2 family using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. Two prominent L. japonicas inbred lines, one with “broad and thin blade” characteristics and another with “long and narrow blade” characteristics, were
applied in the hybridization to yield the F2 mapping population with 92 individuals. A total of 287 AFLP markers and 11 SSR markers were used to construct a L. japonica genetic map. The yielded map was consisted of 28 linkage groups (LG) named LG1 to LG28, spanning 1,811.1 cM with an average
interval of 6.7 cM and covering the 82.8% of the estimated genome 2,186.7 cM. While three genome-wide significant QTL were
detected on LG1 (two QTL) and LG4 for “FL,” explaining in total 42.36% of the phenotypic variance, two QTL were identified
on LG3 and LG5 for the trait “FW,” accounting for the total of 36.39% of the phenotypic variance. The gene action of these
QTL was additive and partially dominant. The yielded linkage map and the detected QTL can provide a tool for further genetic
analysis of two traits and be potential for maker-assisted selection in L. japonica breeding. 相似文献
12.
The genetic diversity and genetic relatedness of mei (Prunus mume; 2n = 16) were studied using amplified fragment length polymorphism (AFLP) markers. Eight EcoRI–PstI AFLP primer combinations were applied to 121 distinct genotypes of mei cultivars and related species. A total of 508 AFLP
product bands were produced, of which 382 were polymorphic. The unweighted pair group method with arithmetic averages analysis
was carried out based on these AFLP markers. From this analysis, “Qugeng Mei,” “Yan Mei,” “Chaodou Mei,” and mei cultivars
were seen to share the same P. mume genetic stem. The AFLP data were able to clearly discriminate P. mume from other species in the genus Prunus, with P. armeniaca aligning as its closest related species. Two major groups and nine subgroups of mei flower were identified, and there was
a strong coincidence of these AFLP-based groupings with the respective morphological characters of the accessions. The genetic
diversity of mei accessions was greatest in the Yunnan Province and decreased toward Eastern China and Japan, so supporting
the hypothesis that the southwest of China represents the genetic diversity center of the species. 相似文献
13.
Xiao L Zhao Z Du D Yao Y Xu L Tang G 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2012,124(5):903-909
The development of yellow-seeded cultivars in Brassica rapa (B. rapa) would improve the quality and quantity of available oil. The identification and mapping of the seed coat color gene may
aid in the development of yellow-seeded cultivars and facilitate introgression of the yellow-seeded gene into desirable Brassica napus (B. napus) lines through marker-assisted selection. In the current study, we investigated the inheritance of a yellow-seeded landrace
in B. rapa, “Dahuang”, originating from the Qinghai-Tibetan plateau. Genetic analysis revealed that the phenotype of the yellow-seeded
trait in Dahuang is controlled by one recessive gene, termed Brsc1. Mapping of the Brsc1 gene was subsequently conducted in a BC1 population comprised 456 individuals, derived from (Dahuang × 09A-126) × Dahuang. From a survey of 256 amplified fragment
length polymorphism (AFLP) primer combinations, 10 tightly linked AFLP markers were obtained. The closest AFLP markers flanking
Brsc1, Y10 and Y06, were 0.2 and 0.4 cM away, respectively. Subsequently, using simple sequence repeat (SSR) markers in the reference
map, the Brsc1 gene was mapped on A09 in B. rapa. Blast analysis revealed that seven AFLP markers showed sequence homology to A09 of B. rapa, wherein six AFLP markers in our map were in the same order as those in A09 of B. rapa. The two closest markers, Y10 and Y06, delimited the Brsc1 gene within a 2.8 Mb interval. Furthermore, Y05 and Y06, the two closest AFLP markers on one side linked to Brsc1, were located in scaffold000059 on A09 of B. rapa, whereas the closet AFLP marker on the opposite side of Brsc1, Y10, was located in scaffold000081 on A09 of B. rapa. Molecular markers developed from these studies may facilitate marker-assisted selection (MAS) of yellow-seeded lines in
B. rapa and B. napus and expedite the process of map-based cloning of Brsc1. 相似文献
14.
Aligning male and female linkage maps of apple (Malus pumila Mill.) using multi-allelic markers 总被引:12,自引:0,他引:12
C. Maliepaard F. H. Alston G. van Arkel L. M. Brown E. Chevreau F. Dunemann K. M. Evans S. Gardiner P. Guilford A. W. van Heusden J. Janse F. Laurens J. R. Lynn A. G. Manganaris A. P. M. den Nijs N. Periam E. Rikkerink P. Roche C. Ryder S. Sansavini H. Schmidt S. Tartarini J. J. Verhaegh M. Vrielink-van Ginkel G. J. King 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,97(1-2):60-73
Linkage maps for the apple cultivars ‘Prima’ and ‘Fiesta’ were constructed using RFLP, RAPD, isozyme, AFLP, SCAR and microsatellite
markers in a ‘Prima’בFiesta’ progeny of 152 individuals. Seventeen linkage groups, putatively corresponding to the seventeen
haploid apple chromosomes, were obtained for each parent. These maps were aligned using 67 multi-allelic markers that were
heterozygous in both parents. A large number of duplicate RFLP loci was observed and, in several instances, linked RFLP markers
in one linkage group showed corresponding linkage in another linkage group. Distorted segregation was observed mainly in two
regions of the genome, especially in the male parent alleles. Map positions were provided for resistance genes to scab and
rosy leaf curling aphid (Vf and Sd
1, respectively) for the fruit acidity gene Ma and for the self-incompatibility locus S. The high marker density and large number of mapped codominant RFLPs and some microsatellite markers make this map an ideal
reference map for use in other progenies also and a valuable tool for the mapping of quantitative trait loci.
Received: 17 November 1997 / Accepted: 9 December 1997 相似文献
15.
AFLP-derived SCARs facilitate construction of a 1.1 Mb sequence-ready map of a region that spans the Vf locus in the apple genome 总被引:2,自引:0,他引:2
The availability of high-density anchored markers is a prerequisite for reliable construction of a deep coverage BAC contig, which leads to creation of a sequence-ready map in the target chromosomal region. Unfortunately, such markers are not available for most plant species, including woody perennial plants. Here, we report on an efficient approach to build a megabase-size sequence-ready map in the apple genome for the Vf region containing apple scab resistance gene(s) by targeting AFLP-derived SCAR markers to this specific genomic region. A total of 11 AFLP-derived SCAR markers, previously tagged to the Vf locus, along with three other Vf-linked SCAR markers have been used to screen two apple genome BAC libraries. A single BAC contig which spans the Vf region at a physical distance of approximately 1,100 kb has been constructed by assembling the recovered BAC clones, followed by closure of inter-contig gaps. The contig is 4 ×deep, and provides a minimal tiling path of 16 contiguous and overlapping BAC clones, thus generating a sequence-ready map. Within the Vf region, duplication events have occurred frequently, and the Vf locus is restricted to the ca. 290 kb region covered by a minimum of three overlapping BAC clones. 相似文献
16.
Kikuchi S Taketa S Ichii M Kawasaki S 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2003,108(1):73-78
The hulled or naked caryopsis character of barley (Hordeum vulgare L.) is an important trait for edibility and to follow its domestication process. A single recessive gene, nud, controls the naked caryopsis character, and is located on the long arm of chromosome 7H. To develop a fine map around the nud locus efficiently, the HEGS (High Efficiency Genome Scanning) electrophoresis system was combined with amplified fragment length polymorphism (AFLP). From bulked segregant analysis of 1,894 primer combinations, 12 AFLP fragments were selected as linked markers. For mapping, an F2 population of 151 individuals derived from a cross between Kobinkatagi (naked type) and Triumph (hulled type) was used. Seven AFLP markers were localized near the nud region. A fine map was developed with one-order higher resolution than before, along with the seven anchor markers. Among the seven linked AFLP markers (KT1–7), KT1, KT2 and KT6 were co-dominant, and the former two were detected for their single-nucleotide polymorphisms (SNPs) in the same length of fragments after electrophoresis with the non-denaturing gels of HEGS. The nud locus has co-segregated with KT3 and KT7, and was flanked by KT2 and KT4, at the 0.3-cM proximal and the 1.2-cM distal side, respectively. Four of these AFLP markers were converted into sequence-characterized amplified region (SCAR) markers, one of which was a dominant marker co-segregating with the nud gene.Communicated by G. Wenzel 相似文献
17.
J. N. A. M. Rouppe van der Voort H. J. van Eck P. M. van Zandvoort H. Overmars J. Helder J. Bakker 《Molecular & general genetics : MGG》1999,261(6):1021-1031
A mapping strategy is described for the construction of a linkage map of a non-inbred species in which individual offspring
genotypes are not amenable to marker analysis. After one extra generation of random mating, the segregating progeny was propagated,
and bulked populations of offspring were analyzed. Although the resulting population structure is different from that of commonly
used mapping populations, we show that the maximum likelihood formula for a normal F2 is applicable for the estimation of
recombination. This “pseudo-F2” mapping strategy, in combination with the development of an AFLP assay for single cysts, facilitated
the construction of a linkage map for the potato cyst nematode Globodera rostochiensis. Using 12 pre-selected AFLP primer combinations, a total of 66 segregating markers were identified, 62 of which were mapped
to nine linkage groups. These 62 AFLP markers are randomly distributed and cover about 65% of the genome. An estimate of the
physical size of the Globodera genome was obtained from comparisons of the number of AFLP fragments obtained with the values for Caenorhabditis elegans. The methodology presented here resulted in the first genomic map for a cyst nematode. The low value of the kilobase/centimorgan
(kb/cM) ratio for the Globodera genome will facilitate map-based cloning of genes that mediate the interaction between the nematode and its host plant.
Received: 7 January 1999 / Accepted: 16 April 1999 相似文献
18.
A. Boudichevskaia H. Flachowsky A. Peil C. Fischer F. Dunemann 《Tree Genetics & Genomes》2006,2(4):186-195
A major scab resistance gene initially called Vr1 was identified in the apple cultivar “Regia” derived from the Malus scab resistance source R12740-7A (Russian seedling, RS). A codominant, multiallelic sequence characterized amplified region (SCAR) marker was developed from a random amplified polymorphic DNA marker identified by bulked-segregant analysis. Additional alleles of the AD13 marker locus proved to be informative for the analysis of genetic relationships within Malus including putative relatives of RS. Separate linkage maps were created for the two families derived from crosses with “Regia”. Using phenotypic data from the greenhouse scab tests, the recombination frequency between Vr1 and AD13-SCAR was between 6 and 17%. The Vr1 locus appeared to be closely linked to the Vx [Hemmat et al. J Am Soc Hortic Sci, 127:365–370, 2002], Vr2 [Patocchi et al. Theor Appl Genet, 109:1087–1092, 2004], and the Vh4 gene [Bus et al. Mol Breed, 15:103–116, 2005a]. Our linkage analysis of the molecular markers identified by Hemmat et al. [J Am Soc Hortic Sci, 127:365–370, 2002] for two scab resistance factors from RS (Vr and Vx) indicate that both genes are separated by a large distance on apple linkage group 2 [Boudichevskaia et al. Acta Hortic, 663:171–175, 2004]. This is in agreement with the results of Bus et al., [Mol Breed, 15:103–116, 2005a] who concluded that (1) the RS-derived gene Vh2 is identical to Vr, (2) the RS-derived gene Vh4 is identical to Vx and Vr1, (3) Vh2/Vr and Vh4/Vr1/Vx map on opposite sides of LG 2. One of our main goals was the verification of the Vr1-SCAR within a practical apple-breeding program. The utility of the AD13-SCAR was evident after 2 years under natural scab infection conditions in both families investigated. This is the first report about the confirmation of a molecular marker for a RS resistance factor in a 2-year field experiment. A multiplex polymerase chain reaction assay based on two codominant SCARs for Vf and Vr1 was tested in an apple progeny segregating for both genes. The result of the two-marker approach is discussed with respect to scab races, which are able to overcome the Vf resistance gene. 相似文献
19.
Construction and characterization of a bacterial artificial chromosome library of apple 总被引:8,自引:0,他引:8
B. A. Vinatzer H.-B. Zhang S. Sansavini 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,97(7):1183-1190
A bacterial artificial chromosome (BAC) library has been constructed from apple (Malus×domestica Borkh.) using the variety “Florina”, which is resistant to scab (Venturia inaequalis) by virtue of the Vf gene. Since apple leaves are rich in polyphenols, high-molecular-weight DNA was extracted from leaf nuclei with a protocol
adapted to apple. The nuclei were then embedded in agarose microbeads and partially digested by varying ratios of EcoRI to EcoRI methylase. The resulting DNA fragments were size-selected by pulsed-field gel electrophoresis, ligated to the BAC cloning
vector pECBAC1 and transformed into Escherichia coli cells by electroporation. A total of 36 864 recombinant clones (BACs) were obtained. The library has an average insert size
of 120 kb and represents approximately 5×apple haploid-genome equivalents. It was screened with six cDNA probes using the
chemiluminescent DIG system. An average of 4.4 clones was detected for each locus. The apple BAC library will be used to isolate
the Vf scab resistance gene through map-based cloning. In this connection the library was screened with a marker closely linked
to the Vf gene and six positive clones have been isolated. This library should thus be well suited for map-based gene cloning, in particular
for the isolation of the Vf gene and for the construction of a physical map of the apple genome.
Received: 19 February 1998 / Accepted: 30 April 1998 相似文献
20.
Paolo Galli Giovanni Antonio Lodovico Broggini Markus Kellerhals Cesare Gessler Andrea Patocchi 《Molecular breeding : new strategies in plant improvement》2010,26(4):561-572
The Rvi15 (Vr2) apple scab resistance locus found in the GMAL 2473 accession has been previously mapped to the top of the Linkage Group
2 (LG2) by analyzing 89 progeny plants of a cross between ‘Idared’ and GMAL 2473. A new population of 989 progeny plants,
derived from a cross between ‘Golden Delicious’ and GMAL 2473, has been analyzed with the two SSR markers CH02c02a and CH02f06,
previously found to be associated with Rvi15 (Vr2), and with two published markers derived from NBS sequences (ARGH17 and ARGH37) estimated to map close to the Rvi15 (Vr2) locus. ARGH17 and ARGH37, were found to be the closest markers to the resistance locus, bracketing it within an interval
of 1.5 cM. The SSRs mapped one on each side of Rvi15 (Vr2). CH02f06 mapped at 2.9 cM from ARGH37 while CH02a02a mapped at 1.7 from ARGH17. The position of Rvi15 (Vr2) respect to CH02a02a indicates that Rvi15 (Vr2) and Rvi4 (Vh4), a second apple scab gene mapped on the top of LG2, are two different resistance genes. In order to develop even more tightly
linked markers to Rvi15 (Vr2), ARGH17 was used as the starting point for chromosome walking through the Rvi15 (Vr2) homolog region of the cv. ‘Florina’. A single ‘Florina’ BAC clone, 36I17, was sufficient to span the homologous locus in
the new population’s recombinant progeny. Sequencing of the 36I17 BAC clone allowed identifying seven putative ORFs, including
two showing a TIR-NBS-LRR structure. Ten additional markers could be developed mapping within a 1.8 cM interval around the
Rvi15 (Vr2) resistance gene. ARGH17 and GmTNL1 markers, the latter also derived from NBS-LRR resistance gene homolog sequence, are the
closest markers to Rvi15 (Vr2) bracketing it within a 0.5 cM interval. The availability of 12 markers within the Rvi15 (Vr2) region, all within a small physical distance (kbp) in ‘Florina’, suggests that cloning of the Rvi15 (Vr2) apple scab resistance gene from GMAL 2473 will be possible. 相似文献