Identification of RAPD markers closely linked to the mlo-locus in barley

Developing resistance to powdery mildew, Erysiphe graminis f.sp. hordei, is a major goal of many barley breeding programmes. Several resistance genes have been tagged or mapped with molecular markers. The mlo gene confers durable resistance towards all known isolates of the pathogen. In this study, RAPD markers and bulked segregant analysis were used to determine PCR-based markers linked to the mlo-locus. Sixty doubled haploid lines from a cross between an isogenic line of ‘Ingrid’ carrying the mlo11 allele and a susceptible cv. ‘Pokko’ were used as plant material. Seven linked RAPD markers were found, the closest lying 1.6 cM away from the resistance gene. When eight barley varieties were assayed for the presence of this band, F4-980, it was found in the resistant varieties but not in the susceptible ones. The linked marker bands could be amplified from DNA-samples prepared by using three different methods, including a quick squash technique. PCR-based markers linked to the resistance gene can be used as tools for selection in breeding programmes.     

Linkage of Rust Resistance Genes from Wild Barley (Hordeum spontaneum) with Isozyme Markers

Eighty-three third backcross lines which comprise a set of near isogenic lines (NIL's) of the barley cultivar ‘Clipper’ but each carrying a different chromosomal segment from Hordeum spontaneum, marked with a distinct isozyme, were tested for resistance to three races of the barley leaf rust pathogen (Puccmia hordei). Fourteen lines showed resistance to at least one race and three showed resistance to all three races. The resistance in two of these lines was controlled by separate, single partially dominant genes. In one case the resistance gene named Rph1O was on chromosome 3 and linked (r = 0.15 ±0.05) with the isozyme locus Est2. In the second case, the gene (Rph11) was on barley chromosome 6 and linked (r = 0.07±0.02) with the isozyme locus Acp3 and (r = 0.11±0.02) with Dip2.     

Localization of the Laevigatum Resistance Gene MlLa against Powdery Mildew in the Barley Genome by the Use of RFLP Markers

RFLP markers which were previously assigned to chromosome 2 (2H) were found to detect polymorphism between the cv. ‘Pallas’ and a near isogenic line carrying the Laevigatum resistance gene MlLa. Linkage analysis carried out with two sets of DH lines derived from the crosses ‘RisøS’בSultan’ and ‘Alf’בVogelsanger Gold 2’ confirmed three DNA probes closely linked with the MlLa locus.     

Validation of a major QTL for scab resistance with SSR markers and use of marker-assisted selection in wheat

The objectives of this study were to validate the major quantitative trait locus (QTL) for scab resistance on the short arm of chromosome 3B in bread wheat and to isolate near‐isogenic lines for this QTL using marker‐assisted selection (MAS). Two resistant by susceptible populations, both using ‘Ning7840’ as the source of resistance, were developed to examine the effect of the 3BS QTL in different genetic backgrounds. Data for scab resistance and simple sequence repeat (SSR) markers linked to the resistance QTL were analyzed in the F_2:3 lines of one population and in the F_3:4 lines of the other. Markers linked to the major QTL on chromosome 3BS in the original mapping population (‘Ning7840’/‘Clark’) were closely associated with scab resistance in both validation populations. Marker‐assisted selection for the QTL with the SSR markers combined with phenotypic selection was more effective than selection based solely on phenotypic evaluation in early generations. Marker‐assisted selection of the major QTL during the seedling stage plus phenotypic selection after flowering effectively identified scab resistant lines in this experiment. Near‐isogenic lines for this 3BS QTL were isolated from the F₆ generation of the cross ‘Ning7840’/‘IL89‐7978’ based on two flanking SSR markers, Xgwm389 and Xbarc147. Based on these results, MAS for the major scab resistance QTL can improve selection efficiency and may facilitate stacking of scab resistance genes from different sources.     

Genetical Studies of Resistance of Powdery Mildew in Barley Lines Derived from Hordeum spontaneum Collected from Israel

Genetical studies on mildew resistance were carried out with Hordem spontaneum derived lines. A total of 28 lines (66 %) showed monofactorial segregation for mildew resistance, For 14 lines, a bifactorial mode of inheritance was found. In total fifty six mildew resistance genes take part in the inheritance of mildew resistance of the H. spontaneum derived lines, while the presence of known genes for mildew resistance (i.e, Ml-a.9 and Ml-p) was established only in two cases. Independent segregation from the Ml-a locus was found in 10 mbnofaetorial segregating lines, The genes conditioning mildew resistance in barley lines derived from the accessions 1B-54B, RS 170-47, RS 20-1. 1B-86B, RS 145-39 and 1B–152B of H. spontaneum were closely linked or alleles to the Ml-a locus, but shown to be different from 15 previously identified Ml-a alleles. It is suggested that these genes should be designated Ml-a16, Ml-18, Ml-19 Ml-20 and Ml-a21 respectively. No recombinants were found in test crosses when both parents carried genes/alleles of the Ml-a locus. In addition, polymorphism has been observed also for the Ml-a locus. In 4 lines mildew resistance was conditioned by two dominant complementary genes. For one of the 2 genes, conditioning mildew resistance of line RS 42-8 × OrioL a new locus was found located near the centromere of the long arm of chromosome 5, and should be designated Ml-i The potential use of H. spontaneum genes for mildew resistance in barley breeding is discussed.     

Identification of New Genes for Mildew Resistance of Barley at the Mia Locus in Lines Derived from Hordeum spontaneum

Genetical studies have been conducted with lines derived from H. spontaneum showing that these lines carry dominant mildew resistance genes located at or near the Mla, locus. The resistance spectra of the lines ‘RS170—10 × Piccolo A’, ‘Diamant × 1B-20’, ‘RS1—8 × Piccolo E’ and ‘Diamant × 1B-151’ obtained from 10 European and Israeli isolates differ from previously-identified Mia alleles. Therefore, it is suggested that these genes should be designated as Mla 25, Mla 26, Mla.27 and Mla 28, respectively. In addition, the RFLP-patterns of these lines and their crossing parents were studied by hybridization with probes MWG 1H036, MWG 1H060 and MWG 1H068, which are very closely linked to the Mia locus. Two double crossover events have been identified. The use of RFLP markers for the identification of mildew resistance genes is discussed.     

Identification of Resistance Genes Against Powdery Mildew in Four Accessions of Hordeum Vulgare SSP. Spontaneum

Summary Four newly detected accessions of wild barley (Hordeum vulgare ssp. spontaneum) resistant to powdery mildew caused by Blumeria graminis f. sp. hordei were studied with the aim of finding the number of genes/loci conferring the resistance of individual accessions, the type of inheritance of the genes and their relationships to the Mla locus. F₂ populations after crosses between the winter variety ‘Tiffany’ and four wild barley accessions and use of microsatellite DNA markers were focused on the identification of individual resistance genes/loci by means of their chromosomal locations. In PI466495, one locus conferring powdery mildew resistance was identified in highly significant linkage with the marker Bmac0213. This location is consistent with the known locus Mla on chromosome 1HS. In the other three accessions the resistance was determined by two independent loci. In PI466197, PI466297 and PI466461, one locus was identified on chromosome 1HS and three new loci were revealed on chromosomes 2HS (highly significant linkage with Bmac0134), 7HS (highly significant linkage with Bmag0021) and 7HL (significant linkage with EBmac0755). Our prospective aim is identification of further linked DNA markers and the exact location of the resistance genes on the barley chromosomes.     

Relationship between tolerance and resistance to pepper mottle virus in a cross between Capsicum annuum L. × Capsicum chinense Jacq.

Summary Progenies of a cross between pepper cultivar Delray Bell (Capsicum annuum L.) tolerant to pepper mottle virus (PeMoV) and P.I. 159236 (Capsicum chinense Jacq.), resistant to PeMoV were evaluated in greenhouse experiments. The F₁ generation was susceptible to PeMoV, whereas the backeross generations to both parents and the F₂ segregated in a ratio of 1:1. This indicates that PeMoV resistance is controlled at the same locus in both parents or at two closely linked loci. Each parent is homozygous for one locus and the homozygous condition of either allele results in failure to support virus; in heterozygous condition as observed in the F₁ generation, the effect of a single allele is insufficient even when one allele of each type is present, resulting in systemic infection.The abstract of this paper was presented at the IVth Eucarpia Capsicum meetings held in October at Wageningen, Netherlands.Florida Agricultural Experiment Stations Journal Series Paper No. 3175.     

Powdery-mildew-resistance genes Mla29 and Mla32 in H. spontaneum derived winter-barley lines

Allelism to the highly polymorphic Mia locus was demonstrated for the powdery-mildew resistance of two Hordeum spontaneum derived winter–barley lines, ‘110-4 × Sonja’ and ‘142–29 × Dura’, by testing the F2 progeny of crosses between these lines and the winter-barley cv. ‘Triton’ (Mlal3) with two appropriate isolates. The results were confirmed by RFLP analysis, using the probe MWG 1H036, which is very closely linked to the Mia locus. The designations Mla29 and Mla32 are proposed for the genes identified in the two lines.     

A novel barley scald resistance gene: genetic mapping of the Rrs15 scald resistance gene derived from wild barley, Hordeum vulgare ssp. spontaneum

Most genes for resistance to barley leaf scald map either to the Rrs1 locus on the long arm of chromosome 3H, or the Rrs2 locus on the short arm of chromosome 7H. Other loci containing scald resistance genes have previously been identified using lines derived from wild barley, Hordeum vulgare ssp. spontaneum. A single dominant gene conditioning resistance to scald was identified in a third backcross (BC3F3) line derived from an Israeli accession of wild barley. The resistance gene is linked to three microsatellite markers that map to the long arm of chromosome 7H; the closest of these loci, HVM49, maps 11.5 cM from the resistance gene. As no other scald resistance genes have been mapped to this chromosome arm, it is considered to be a novel scald resistance locus. As the Acp2 isozyme locus is linked to this scald resistance locus, at 17.7 cM, Acp2 is assigned to chromosome 7H. Molecular markers linked to the novel scald resistance gene, designated Rrs15, can be used in breeding for scald resistance.     

New molecular markers linked to qualitative and quantitative powdery mildew and scald resistance genes in barley for dry areas

A partial genetic linkage map was constructed on 71 doubled-haploid lines derived from a cross between the barley lines Tadmor and WI2291 with 181 molecular markers. The segregating population was used to detect markers linked to the gene Mlg conferring resistance to powdery mildew (Erysiphe graminis f. sp. hordei) and to genes for quantitative resistance to scald (Rhynchosporium secalis). The gene Mlg on chromosome 4H was flanked by two AFLP markers at a distance of 2.0 and 2.4 cM, respectively. QTLs for resistance to scald were detected on chromosomes 2H and 3H. This association of molecular markers with qualitative and quantitative disease resistance loci represents a valuable starting-point for marker-assisted selection. This revised version was published online in July 2006 with corrections to the Cover Date.     

Multigene Families of Powdery Mildew Resistance Genes in Locus Mla on Barley Chromosome 5

A review of data on powdery mildew resistance genes in the Mla locus of barley reveals that there are at least 12 clusters of genes present, each comprising one Mla gene and one or more closely linked, additional resistance genes. Tentative designations are listed for 16 additional resistance genes. Many sources of powdery mildew resistance in barley appear to harbour multigene families in the Mla region, not single ‘superior’genes.     

Allelic variation for the rust resistance gene <Emphasis Type="Italic">Lr34</Emphasis>/<Emphasis Type="Italic">Yr18</Emphasis> in Canadian wheat cultivars

The wheat (Triticum aestivum L.) gene Lr34/Yr18 conditions resistance to leaf rust, stripe rust, and stem rust, along with other diseases such as powdery mildew. This makes it one of the most important genes in wheat. In Canada, Lr34 has provided effective leaf rust resistance since it was first incorporated into the cultivar Glenlea, registered in 1972. Recently, molecular markers were discovered that are either closely linked to this locus, or contained within the gene. Canadian wheat cultivars released from 1900 to 2007, breeding lines and related parental lines, were tested for sequence based markers caSNP12, caIND11, caIND10, caSNP4, microsatellite markers wms1220, cam11, csLVMS1, swm10, csLV34, and insertion site based polymorphism marker caISBP1. Thirty different molecular marker haplotypes were found among the 375 lines tested; 5 haplotypes had the resistance allele for Lr34, and 25 haplotypes had a susceptibility allele at this locus. The numbers of lines in each haplotype group varied from 1 to 140. The largest group was represented by the leaf rust susceptible cultivar “Thatcher” and many lines derived from “Thatcher”. The 5 haplotypes that had the resistance allele for Lr34 were identical for the markers tested within the coding region of the gene but differed in the linked markers wms1220, caISBP1, cam11, and csLV34. The presence of the resistance or susceptibility allele at the Lr34 locus was tracked through the ancestries of the Canadian wheat classes, revealing that the resistance allele was present in many cultivars released since the 1970s, but not generally in the older cultivars.     

Linkage of dominant hypersensitive resistance to bean common mosaic virus to seed color in Phaseolus vulgaris L.

Summary Evaluation of Phaseolus vulgaris germplasm bank materials and progenies from a large number of crosses using red- or yellow-colored, BCMV-susceptible bean lines, crossed to purple- or grey/brown-colored, hypersensitive-resistant lines, suggested strong trait association between seed color and BCMV resistance. The cross of red-mottled I⁺I⁺ (susceptible) BAT 1255R to isogenic purple-mottled II (resistant) BAT 1255M was made to study the segregation of the two characters and to recover red-mottled resistant progenies. No recombinant genotypes were observed among 353 F₃ families inoculated with BCMV-NL3, suggesting that linkage of purple-mottled seed color and dominant BCMV resistance is very close.Contribution of the Centro Internacional de Agricultural Tropical.     

Inheritance of resistance to Pyrenochaeta lycopersici in tomato

Summary Tomato accessions (Lycopersicon sp.), along with commercial cultivars and breeding lines were grown in a field infested with the brown root rot (BRR) organism, Pyrenochaeta lycopersici and evaluated for resistance. Three L. esculentum Mill. accessions, P.I. 260397, P.I. 262906 and P.I. 203231, were resistant and were used as male parents in crosses designed to transfer resistance to tomatoes of fresh market type. Through analysis of parental generations and F₁ and F₂ progenies from three crosses the heritability of resistance in the broad sense was estimated to range from 25 to 43 percent. The minimum number of genes influencing resistance was estimated to be from 4 to 8.Florida Agricultural Experiment Stations Journal Series Paper no. 317.     

Identification of resistance gene analogs as markers of disease resistance loci in oats, using near-isogenic lines

Genomic sequences with features of the major class of disease resistance genes and which bear nucleotide‐binding leucine‐rich repeat sequences (resistance gene analogs; RGA) were tested as potential markers of crown rust resistance loci in hexaploid oats. Two collections of paired near‐isogenic lines carrying resistance to different isolates of crown rust, Puccinia coronata were screened. Two out of the four RGAs assayed showed restriction fragment length polymorphism (RFLP) between one line of each collection and its recurrent parent. The paired lines X466 and D494 were polymorphic for RGA III2.2 and the pair of lines X470 and D504 were polymorphic for RGA III2.18. The III2.18 polymorphism was located in the hexaploid map Avena byzantina cv. ‘Kanota’ × A. sativa cv. ‘Ogle’ in linkage group KO17 in a region previously associated with crown rust resistance. In addition, 220 random primers were used for random amplified polymorphic DNA (RAPD) analysis to screen the two sets of NILs. Only one polymorphic band was obtained that differentiated the paired lines X470 and D504 from their parents. The RAPD band was used as a probe and the relevant RFLP that differentiated the NILs X470 and D504 was found at 1.7 cM from the III2.18 marker in KO17. RFLP analysis using probes previously mapped in KO17 confirmed differences for X470 and D504 in the region around the III2.18 marker. These results suggest that the resistance locus shared by this pair of NILs is probably linked to the markers revealed by RGA III2.18. The use of RGAs as RFLP probes in the screening of NILs with differences in crown rust resistance has proved to be more effective than RAPDs for finding polymorphic markers possibly linked to resistance loci.     

AFLP-based STS markers closely linked to a fertility restoration locus (Rfm1) for cytoplasmic male sterility in barley

The Rfm1 gene restores the fertility of the msm1 and msm² male‐sterile cytoplasms in barley. Rfm1 is located on the short arm of chromosome 6H. To develop molecular markers tightly linked to Rfm1 for use in sophisticated marker‐assisted selection and map‐based cloning, an amplified fragment‐length polymorphism (AFLP) marker system with isogenic lines and a segregating BC₁F₁ population was used. Nine hundred primer combinations were screened and a linkage map was constructed around the Rfm1 locus by using 25 recombinant plants selected from 214 BC₁F₁ plants. Three AFLP markers were identified, e34m2, e46m19 and e48m17, linked to the locus. The most closely linked markers were e34m2, at 1.0 cM distally and e46m19, at 1.1 cM proximally. The two AFLP markers were converted to dominant STS markers. These markers should accelerate programmes for breeding restorer lines and will be useful for map‐based cloning.     

Genetic analysis and pyramiding of two gall midge resistance genes (Gm-2 and Gm-6t) in rice ( Oryza sativa L.)

Asian rice gall midge (Orseolia oryzae) is a major pest across much of south and southeast Asia. This pest is genetically diverse and many gall midge biotypes are known to exist in each country. During the last three decades, host plant resistance has proved to be the most effective mechanism of controlling the Asian rice gall midge. Seven genes conditioning resistance to gall midge larvae have been identified in rice (Oryza sativa) and are being used in cultivar improvement programs. However, some of these genes are rendered ineffective by new gall midge biotypes. Increased understanding of genetics, inheritance, allelic relationships and linkage is necessary to maximise the durability of major gene resistance by the pyramiding of these genes. The two genes, Gm-2 and Gm-6(t), are known to confer resistance against a number of biotypes in India and China, respectively. An F₃ population derived from a cross between Duokang #1 (donor of Gm-6(t)) and Phalguna (donor of Gm-2) was screened against Chinese gall midge biotype 4 at Guangdong, China, and Indian gall midge biotype 1 at Raipur, India. At each location, separately,a single gene governed resistance. The parallel segregation of 417 F₃progenies for both biotypes at two locations revealed that recombination had occurred between the two genes, establishing that the two genes are not allelic. However, the two genes Gm-2 and Gm-6(t), were found to be linked with a distance of ∼16.3 cM. A number of lines homozygous at one locus and segregating for the other locus were identified and selected. These lines were selfed to obtain lines homozygous for the favourable alleles at both loci (two locus pyramids). This is the first report on use of conventional host-pest interaction method for pyramiding two closely located Gm-resistance loci of dissimilar effects. The implications of deployment of these pyramids within and across country borders, with reference to the prevailing gall midge populations are discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Inheritance of resistance to Xanthomonas campestris pv. translucens in triticale

Summary Three triticale lines, Siskiyou, M2A-Beagle, and OK 77842 have been reported to possess resistance to bacterial leaf streak caused by Xanthomonas campestris, pv. translucens (Xct.). The three resistant lines were crossed to susceptible lines and crossed with each other. F₂, BC₁-F₁, BC₂-F₁ plants were inoculated with a mixture of two Xct strains. The segregation data indicate the presence of a single dominant gene in each of the three resistant lines to bacterial leaf streak. These three genes are either the same or closely linked herein designated as Xct₁.     

Genes for scald resistance from wild barley (Hordeum vulgare ssp spontaneum) and their linkage to isozyme markers

Summary Accessions of Hordeum vulgare ssp. spontaneum, the wild progenitor of barley, collected in Israel (70), Iran (15) and Turkey (6) were screened for seedling response to four isolates of Rhynchosporium secalis, the pathogen causing leaf scald in barley. Resistance was very common in the collection (77%) particularly among accessions from the more mesic sites (90%). The genetics of this resistance were investigated in fifteen backcross (BC₃) lines that contained an isozyme variant from H.v. ssp. spontaneum in a H.v. ssp. vulgare (cv. Clipper) background and were resistant to scald. Segregation in the BC₃F₂ families conformed with a single dominant resistance gene in 9 of the 15 lines. Scald resistance and the isozyme marker were closely linked in three of the BC₃-lines, loosely linked in four and unlinked in the remaining eight. Scald resistance genes were identified on barley chromosomes 1, 3, 4 and 6. Crosses between several of the scald resistant BC-lines together with the linkage data indicated that at least five genetically independent resistances are available for combining together for deployment in barley. The linkage of scald resistance in several BC₃-lines to the isozyme locus Acp2 is of special interest as this locus is highly polymorphic in wild barley.