The genetic basis of resistance to apple scab [Venturia inaequalis (Cke.) Wint.] in the Russian apple seedling R12740-7A (Malus Mill. sp.) was investigated. Segregation ratios obtained in crosses with susceptible cultivars suggested that at least two genes were involved, and three foliar resistance reactions (chlorotic, stellate necrotic, and pit type) were observed after inoculation. DNA markers were identified for both the stellate necrotic (Vr) and pit type (no locus designation, Vx suggested) resistance phenotypes. Comparison of resistance phenotypes with marker segregation demonstrated that only two major dominant genes were present in R12740-7A, one producing the stellate necrotic lesion and the other the pit-type lesion. The chlorotic lesion could be attributed to either unclear expression of the resistance phenotype or to susceptible genotypes not contracting the disease. These markers along with a previously published marker for Vf were used to analyze inheritance of resistance in a Vr × Vf cross in advanced breeding material. The markers identified successfully all susceptible progeny, as well as apparent escapes and individuals possessing both Vf and Vr. Thus, the markers should be useful in future screening of segregating progeny and in the pyramiding of scab resistance genes in new cultivars.
Minou Hemmat, Susan K. Brown and Norman F. Weeden
Darlene M. Lawson, Minou Hemmat and Norman F. Weeden
Five morphological and developmental traits (branching habit, vegetative budbreak, reproductive budbreak, bloom time, and root suckering) were analyzed in a family obtained from the apple (Malus domestica Borkh) cross `Rome Beauty' × `White Angel'. The phenotypic variation in these traits was compared with a selected set of marker loci covering the known genome of each of the parents to locate genes with major effects on the traits. The contrasting branching habits of the two parents appeared to be controlled by at least two loci. One of these, Tb, governed the presence or absence of lateral branches, particularly on the lower half of shoots. The locus was heterozygous in `White Angel' and was mapped to a 5 CM interval on linkage group 6. At least one other locus conditioning spur-type branching appeared to be segregating, but the locus or loci could not be linked to segregating markers. The timing of initial vegetative growth was tightly associated with the chromosomal region in which the Tb gene is located and maybe a pleiotropic effect of this gene. Time of reproductive budbreak correlated with segregation at the isozyme marker, Prx-c, on linkage group 5. Variation in time of bloom and later stages in flower development appeared to be controlled by different genes not linked to Prx-c. The tendency to produce root suckers cosegregated with a marker on `White Angel' linkage group 1, suggesting control by a single locus, Rs. Data from a `Rome Beauty' x `Robusta 5' family provided additional information on the inheritance of these traits.
Minou Hemmat, Norman F. Weeden and Susan K. Brown
We mapped DNA polymorphisms generated by 41 sets of Simple Sequence Repeat (SSR) primers, developed independently in four laboratories. All primer sets gave polymorphisms that could be located on our `White Angel' x `Rome Beauty' map for apple [Malus sylvestris (L.) Mill. Var. domestica (Borkh.) Mansf.]. The SSR primers were used to identify homologous linkage groups in `Wijcik McIntosh', NY 75441-58, `Golden Delicious', and `Liberty' cultivars for which relatively complete linkage maps have been constructed from isozyme and Random Amplified Polymorphic DNA (RAPD) markers. In several instances, two or more SSRs were syntenic, and except for an apparent translocation involving linkage group (LG) 6, these linkages were conserved throughout the six maps. Twenty-four SSR primers were consistently polymorphic, and these are recommended as standard anchor markers for apple maps. Experiments on a pear (Pyrus communis L.) population indicated that many of the apple SSRs would be useful for mapping in pear. However some of the primers produced fragments in pear significantly different in size than those in apple.
Minou Hemmat, Norman F. Weeden and Susan K. Brown
Apple scab, Venturia inaequalis (Cke.) Wint., is one of the most damaging diseases of apples. Although fungicide sprays have been used to control the disease, genetic resistance in existing commercially important varieties would be desirable. Identification of molecular marker(s) would be helpful in devising biotechnological approaches to control the disease. We used bulk segregant analysis to identify RAPD markers that cosegregate or display a tight linkage with Vf gene in Prima × Spartan cross. Using this approach, we are saturating the region around the scab resistance gene for the purpose of bracketing the locus. We have identified several markers associated with the Vf locus. The closest markers have been isolated and sequenced to be used as SCARs. The relationship and distances of the markers with the Vf locus and other previously reported markers will be discussed.
Minou Hemmat, Norman F. Weeden, Frank S. Cheng and S.K. Brown
The positions of over 50 SSR loci and other sequence tagged sites (STSs) have been located on the linkage maps of five apple cultivars (Rome Beauty, White Angel, Golden Delicious, Liberty, McIntosh) and two New York accessions. In most cases, the primers used produced single amplification products, permitting identification of homologous loci in the different cultivars and the precise alignment of the linkage maps generated for each. Based on this information, we present a general linkage map for apple with STS markers on each linkage group. The map consists of 17 linkage groups (equal to the haploid chromosome number for the species) with over 500 markers. The positions of several resistant gene analogues have been located on this linkage map. None of these sequences map near genes conferring resistance to scab or powdery mildew. SSR loci exhibited a tendency to cluster in certain regions of the linkage map. This clustering slightly reduces their effectiveness as genome markers for comparative mapping or germplasm diversity. However, the SSR markers definitely displayed a high level of polymorphism, making them particularly useful for genetic studies.
Minou Hemmat, Norman F. Weeden, Herb S. Aldwinckle and Susan K. Brown
Bulked segregant analysis was used to identify RAPD markers that display tight linkage to the Vf gene in apple (Malus sp.) that confers resistance to five races of apple scab [Venturia inaequalis (Cke.) Wint.]. We identified several new RAPD markers linked to Vf. The most tightly linked marker in the test population, S52500, was cloned and sequenced. A linkage map of the Vf region was developed using these markers, RAPD markers previously described by other laboratories, and the isozyme locus Pgm-1. An assay was developed for Vf by multiplexing the two markers closely flanking the Vf locus. This assay has a theoretical `escape' value (discarding a resistant plant) of 3% and an error rate (selection of a susceptible plant) of 0.02%.
Minou Hemmat, Norman F. Weeden, Patrick J. Conner and Susan K. Brown
The columnar mutation `Wijcik McIntosh' has attracted much attention because of its compact growth habit, which is compatible with high-density plantings. Using bulked segregant analysis, we identified several randomly amplified polymorphic DNA (RAPD) markers that displayed a close linkage with the columnar locus (Co). The RAPD marker that displayed the closest linkage was end sequenced to develop a sequence tagged site for rapidly screening segregating populations. A simple sequence repeat (SSR) of (GA)17 was identified within the DNA fragment. Four allelic forms, including an apparent null allele, could be distinguished among the cultivars tested. The null allele displayed close linkage with Co in two progenies, and we used this marker to identify the location of the gene on the apple linkage map.