Using molecular markers to pyramid genes for resistance to ascochyta blight and anthracnose in lentil (Lens culinaris Medik)

Ascochyta blight caused by the fungus Ascochyta lentis Vassilievsky and anthracnose caused by Colletotrichum truncatum [(Schwein.) Andrus & W.D. Moore] are the most destructive diseases of lentil in Canada. The diseases reduce both seed yield and seed quality. Previous studies demonstrated that two genes, ral1 and AbR1, confer resistance toA. lentis and a major gene controls the resistance to 95B36 isolate of C. truncatum. Molecular markers linked to each gene have been identified. The current study was conducted to pyramid the two genes for resistance to ascochyta blight and the gene for resistance to anthracnose into lentil breeding lines. A population (F_6:7) consisting of 156 recombinant inbred lines (RILs) was developed from across between ‘CDC Robin’ and a breeding line ‘964a-46’. The RILs were screened for reaction to two isolates (A1 and 3D2) ofA. lentis and one isolate (95B36) ofC. truncatum. χ² analysis of disease reactions demonstrated that the observed segregation ratios of resistant versus susceptible fit the two gene model for resistance to ascochyta blight and a single gene model for resistance to anthracnose. Using markers linked to ral1 (UBC 227₁₂₉₀), to AbR1(RB18₆₈₀) and to the major gene for resistance to anthracnose (OPO6₁₂₅₀),respectively, we confirmed that 11 RILs retained all the three resistance genes. More than 82% of the lines that had either or both RB18₆₈₀ and UBC227₁₂₉₀markers were resistant to 3D2 isolate and had a mean disease score lower than 2.5. By contrast, 80% of the lines that had none of the RAPD markers were susceptible and had a mean disease score of 5.8. For the case of A1 isolate of A. lentis, more than 74% of the lines that carriedUBC227₁₂₉₀ were resistant, whereas more than 79% of the lines that do not have the marker were susceptible. The analysis of the RILs usingOPO6₁₂₅₀ marker demonstrated that 11out of 72 resistant lines carried the marker, whereas 66 out of 84 susceptible lines had the marker present. Therefore, selecting materials with both markers for resistance to ascochyta blight and a marker for resistance to anthracnose can clearly make progress toward resistance in the population. This revised version was published online in July 2006 with corrections to the Cover Date.     

A new citrus linkage map based on SRAP,SSR, ISSR,POGP, RGA and RAPD markers

Sequence-related amplified polymorphism (SRAP), simple sequence repeats (SSR), inter-simple sequence repeat (ISSR), peroxidase gene polymorphism (POGP), resistant gene analog (RGA), randomly amplified polymorphic DNA (RAPD), and a morphological marker, Alternaria brown spot resistance gene of citrus named as Cabsr caused by (Alternaria alternata f. sp. Citri) were used to establish genetic linkage map of citrus using a population of 164 F₁ individuals derived between ‘Clementine’ mandarin (Citrus reticulata Blanco ‘Clementine) and ‘Orlando’ tangelo’ (C. paradisi Macf. ‘Duncan’ × C. reticulata Blanco ‘Dancy’). A total of 609 markers, including 385 SRAP, 97 RAPD, 95 SSR, 18 ISSR, 12 POGP, and 2 RGA markers were used in linkage analysis. The ‘Clementine’ linkage map has 215 markers, comprising 144 testcross and 71 intercross markers placed in nine linkage groups. The ‘Clementine’ linkage map covered 858 cM with and average map distance of 3.5 cM between adjacent markers. The ‘Orlando’ linkage map has 189 markers, comprising 126 testcross and 61 intercross markers placed in nine linkage groups. The ‘Orlando’ linkage map covered 886 cM with an average map distance of 3.9 cM between adjacent markers. Segregation ratios for Cabsr were not significantly different from 1:1, suggesting that this trait is controlled by a single locus. This locus was placed in ‘Orlando’ linkage group 1. The new map has an improved distribution of markers along the linkage groups with fewer gaps. Combining different marker systems in linkage mapping studies may give better genome coverage due to their chromosomal target site differences, therefore fewer gaps in linkage groups.     

Marker selection for <Emphasis Type="Italic">Fusarium</Emphasis> head blight resistance based on quantitative trait loci (QTL) from two European sources compared to phenotypic selection in winter wheat

Fusarium head blight (FHB) infects all cereals including maize and is considered a major wheat disease, causing yield losses and mycotoxin contamination. This study aimed to compare the realized selection gain from marker and phenotypic selection in European winter wheat. A double cross (DC) combined three FHB resistance donor-QTL alleles (Qfhs.lfl-6AL and Qfhs.lfl-7BS from ‘Dream’, and one QTL on chromosome 2BL from ‘G16-92’) with two high yielding, susceptible winter wheats, ‘Brando’ and ‘LP235.1’. The base population of 600 DC derived F₁ lines was on one hand selected for the respective QTLs by SSR markers (marker-selected cycle, CM), resulting in 35 progeny possessing different combinations of beneficial donor-QTL alleles. On the other hand it was selected phenotypically, only by FHB rating, and the best 20 lines were recombined and selfed (phenotypically selected cycle, CP). The variants CP, CM, and an unselected variant (C0) were tested at four locations by inoculation of Fusarium culmorum. Resistance was measured as the mean of multiple FHB ratings (0–100%). FHB severity was reduced through both phenotypic and marker selection by 6.2 vs. 5.0%, respectively. On a per-year basis, marker selection by 2.5% was slightly superior to phenotypic selection with 2.1%, because the first variant saved 1 year. Marker-selected lines were on average 8.6 cm taller than phenotypically selected lines. A high genetic variation within the marker-selected variant for FHB resistance and the high effect of a resistance-QTL allele on straw length indicate that additional phenotypic selection will further enhance selection gain.     

Inheritance of resistance to <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">melonis</Emphasis> races 0 and 2 in melon accession Tortuga

The inheritance of the resistance to Fusarium oxysporum f. sp. melonis (F.o.m.) races 0 and 2 in ‘Tortuga’, a Spanish cantalupensis accession, was studied from crosses of ‘Tortuga’ by the susceptible line ‘Piel de Sapo’ and the resistant one ‘Charentais-Fom1’ that carries the resistance gene Fom-1. The segregation patterns observed in the F₂ (‘Tortuga’ × ‘Piel de Sapo’) and the backcross (‘Piel de Sapo’ × (‘Tortuga’ × ‘Piel de Sapo’) populations, suggest that resistance of ‘Tortuga’ to races 0 and 2 of F.o.m. is conferred by two independent genes: one dominant and the other recessive. In the F2 derived from the cross between accessions ‘Tortuga’ and ‘Charentais-Fom1’, the lack of susceptible plants indicated that the two accessions are carrying the same resistance gene (Fom-1). The analysis of 158 F₂ plants (‘Tortuga’ × ‘Piel de Sapo’) with a Cleaved Amplified Polymorphic Sequence marker 618-CAPS, tightly linked to Fom-1 (0.9 cM), confirmed that ‘Tortuga’ also carries a recessive gene, that we propose to symbolize by fom-4.     

Residual macronutrient concentration and follower maize (<Emphasis Type="Italic">Zea mays</Emphasis>) crop performance in soilless growth medium previously cropped with six <Emphasis Type="Italic">Musa</Emphasis> genotypes

The nutrient mining abilities of six Musa genotypes: ‘Agbagba’, ‘PITA 22’, ‘Nsukka Local’, ‘FHIA 17’, ‘Fougamou’, and ‘BITA 7’, grown in organic medium formulated by decomposing rice husks with poultry manure in volume proportions of 3:2, were determined at the Teaching and Research Farm of the Department of Crop Science, University of Nigeria, Nsukka, Nigeria. Results indicated significant variation in nutrient mining with respect to Musa genotype and duration of growth. The medium previously cropped with ‘Agbagba’ recorded highest residual mean values for P and Ca, while the medium cropped with ‘Fougamou’ had the highest residual concentration of Mg and S. Residual amount of K was highest in the medium where ‘PITA 22’ and ‘Nsukka Local’ were previously grown. Peak residual concentration of P and K occurred in the medium in which Musa plants were grown only for 4 months, decreasing thereafter, in contrast to Ca, Mg, and S. Residual N tended to increase with longer growth duration of the Musa crops. Follower crop of maize grown in the medium previously cropped with Musa genotypes showed variable performance. While medium previously cropped with ‘PITA 22’ and ‘BITA 7’ favored growth attributes and leaf greenness of the follower maize plant, ‘Fougamou’ favored maize fresh weight attributes. Ectomycorrhiza association was observed in medium previously cropped to ‘BITA 7’ and ‘PITA 22’. Generally, ‘Agbagba’ and ‘Fougamou’ seemed to be the most reticent nutrient miners while uptake of P and K appeared to be low 4 months after planting (MAP) in contrast to Ca, Mg, and S uptake. However, substrate previously cropped with ‘PITA 22’, ‘BITA 7’ and ‘Fougamou’ gave rise to superior growth and fresh weight attributes, respectively, of the maize, the follower crop. A prospect of utilizing the ectomycorrhiza association observed with some genotypes to upgrade Musa yields and those of associated or follower crops exist.     

Sequence characterized amplified region markers tightly linked to the dwarf mosaic resistance gene <Emphasis Type="Italic">mdm1 (t)</Emphasis> in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.)

Maize dwarf mosaic is one of the devastating and wide spread viral diseases in the world. The present investigation was carried out to develop DNA markers closely linked to the resistance gene mdm1 (t). Linkage between the markers and phenotypes was confirmed by analyzing an F₂ population obtained from a cross between a resistant parent ‘Huangzaosi’ and a susceptible parent ‘Mo17(478)’. Four AFLP markers were found in the maize dwarf mosaic resistant plants. By using (BSA) bulked segregant analysis, two of the four AFLP markers were transformed into Sequence-characterized amplified regions markers (SCARs), nominated Rsun-1 and Rsun-2. The two amplified fragment length polymorphism (AFLP) markers, RHC-1and RHC-2, from the amplification products of primer combination E-AGC/M-CAA and E-AGC/M-GAA, showed linkage with the mdm1 (t) gene in a genetic distance 1.6 and 2.0 cM, respectively. The results indicate that the new SCAR markers will be valuable to distinguish resistant plants from susceptible plants in plantlets growing in seedbeds. The markers developed in this study are suitable for marker-assisted selection for maize dwarf mosaic resistance.     

A dominant gene for garnet brown seed coats at the <Emphasis Type="Italic">Rk</Emphasis> locus in ‘Dorado’ common bean and mapping <Emphasis Type="Italic">Rk</Emphasis> to linkage group 1

The color of the seed coats of ‘Dorado’ (Phaseolus vulgaris L.) is garnet brown (dark red kidney bean color) and differs from most other dry bean varieties in the Honduran red bean market class. A genetic investigation of the color of ‘Dorado’ (same as DOR364) and G19833 (Liborino market class) seed coats was conducted. Crosses with genetic tester stocks demonstrated that the gene for garnet brown (GB) in ‘Dorado’ was not allelic with the R gene for dominant red (oxblood) seed coat. An allelism test between the ‘Dorado’ gene for GB seed coat and the rk ^drv gene for recessive expression of GB demonstrated allelism. We propose the gene symbol for Rk ^r for the ‘Dorado’ GB seed coat color gene. Rk ^r expresses partial dominance over Rk, where Rk ^r/Rk expresses a paler and highly variable intermediate red color. The interactions of Rk ^r, rk ^drv, and c ^u are discussed. Segregation analysis in the mapping population made up of DOR364 (same as ‘Dorado’) × G19833 recombinant inbred lines showed that the Rk ^r gene mapped to linkage group 1. The new allele at Rk was located at a distance of 17 cM from the RFLP marker Bng130 with a LOD > 3.0.     

Identification of RAPD and SCAR markers linked to northern leaf blight resistance in waxy corn (Zea mays var. ceratina)

Exserohilum turcicum causes northern corn leaf blight (NCLB), an important disease occurring in maize producing areas throughout the world. Currently, the development of cultivars resistant to E. turcicum seems to be the most efficient method to control NCLB damage. Marker-assisted selection (MAS) enables breeders to improve selection efficiency. The objective of this work was to identify random amplified polymorphic DNA (RAPD) and sequence characterized amplified region (SCAR) markers associated with NCLB resistance. Bulked segregant analysis (BSA) was used to search for RAPD markers linked to NCLB resistance genes, using F₂ segregating population obtained by crossing a susceptible inbred ‘209W’ line with a resistant inbred ‘241W’ line. Two hundred and twenty-two decamer primers were screened to identify four RAPD markers: OPA07₅₂₁, OPA16₄₅₇, OPB09₅₂₀, and OPE20₅₃₆ linked to NCLB resistance phenotype. These markers were converted into dominant SCAR markers: SCA07₄₉₆, SCA16₄₂₀, SCB09₄₆₄, and SCE20₄₂₉, respectively. The RAPD and SCAR markers were developed successfully to identify NCLB resistant genotypes in segregating progenies carrying NCLB resistant traits. Thus, the markers identified in this study should be applicable for MAS for the NCLB resistance in waxy corn breeding programs.     

Identification of QTLs for resistance to anthracnose to two <Emphasis Type="Italic">Colletotrichum</Emphasis> species in pepper

Pepper (Capsicum spp.) anthracnose caused by Colletotrichum spp. is a serious disease damaging pepper production in Asian monsoon regions. For QTL mapping analyses of anthracnose resistance, an introgression BC₁F₂ population was made by interspecific crosses between Capsicum annuum ‘SP26’ (susceptible recurrent parent) and Capsicum baccatum ‘PBC81’ (resistant donor). Both green and red fruits were inoculated with C. acutatum ‘KSCa-1’ and C. capsici ‘ThSCc-1’ isolates and the disease reactions were evaluated by disease incidence, true lesion diameter, and overall lesion diameter. On the whole, distribution of anthracnose resistance was skewed toward the resistant parent. It might indicate that one or two major QTLs are present. The introgression map consisting of 13 linkage groups with a total of 218 markers (197 AFLP and 21 SSR), covering a total length of 325 cM was constructed. Composite interval mapping analysis revealed four QTLs for resistance to ‘KSCa-1’ and three QTLs for ‘ThSCc-1’ isolate, respectively. Interestingly, the major QTLs (CaR12.2 and CcR9) for resistance to C. acutatum and C. capsici, respectively, were differently positioned but there were close links between the minor QTL CcR12.2 for C. capsici and major QTL CaR12.2 as well as the minor QTL CaR9 for C. acutatum and major QTL CcR9. These results will be helpful for marker-assisted selection and pyramiding two different anthracnose-resistant genes in commercial pepper breeding.     

Development and characterization of common black bean lines resistant to anthracnose,rust and angular leaf spot in Brazil

Anthracnose, rust and angular leaf spot caused by Colletotrichum lindemuthianum, Uromyces appendiculatus and Pseudocercospora griseola, respectively, are economically important diseases affecting the common bean production in Brazil. The BIOAGRO/UFV bean breeding program developed Rudá-R, a dry bean line with ‘carioca’ seed type, containing the following disease resistance genes: Co-4, Co-6 and Co-10 (anthracnose); Ur-ON (rust) and Phg-1 (angular leaf spot). To transfer this combination of disease resistance genes present in Rudá-R to a black-seeded bean, a backcrossing program aided by molecular markers was conducted, involving Rudá-R (donor genitor) and Diamante Negro (recurrent genitor). Forty black-seeded BC₃F_3:6 lines were obtained with combinations of at least three markers linked to the indicated disease resistance genes. The lines were evaluated for resistance to the three mentioned pathogens. Eight of the lines were homozygous and resistant to all four evaluated races of C. lindemuthianum, but susceptible to race 2047. Four of the lines were homozygous and resistant to two races of U. appendiculatus. Twenty of the lines were homozygous and resistant to the two races of P. griseola tested. Grain yield of the BC₃F_3:6 lines was evaluated during the ‘winter’ season of 2006 and the ‘dry’ season of 2007. All lines had statistically equal or higher yields than Rudá-R and Diamante Negro. Lines were identified that not only were high yielding but also resistant to the three pathogens tested. These lines are potential genotypes for further testing and for release as new black common bean varieties.     

Characterization of segregation distortion on chromosome 3 induced in wide hybridization between indica and japonica type rice varieties

We previously surveyed chromosomal regions showing segregation distortion of RFLP markers in the F₂ population from the cross between a japonica type variety ‘Nipponbare’ and an indica type variety ‘Milyang23’, and showed that the most skewed segregation appeared on the short arm of chromosome 3. By comparison with the marker loci where distortion factors were previously identified, this region was assumed to be a gametophytic selection-2 (ga2) gene region. To evaluate this region, two near isogenic lines (NILs) were developed. One NIL had the ‘Nipponbare’ segment of this region on the genetic background of ‘Milyang23’ (NIL9-23), and the other NIL had the ‘Milyang23’ segment on the genetic background of ‘Nipponbare’ (NIL33-18). NIL9-23 and ‘Milyang23’, NIL33-18 and ‘Nipponbare’, and ‘Nipponbare’ and ‘Milyang23’ were respectively crossed to produce F₁ and F₂ populations. The F₁ plants of NIL9-23 × ‘Milyang23’ and NIL33-18 × ‘Nipponbare’ showed high seed fertility and the same pollen fertility as their parental cultivars, indicating that ga2 does not reduce seed and pollen fertility. Segregation ratio of a molecular marker on the ga2 region in the three F₂ populations was investigated to clarify whether segregation distortion occurred on the different genetic backgrounds. Segregation distortion of the ga2 region appeared in the both F₂ populations from the NIL9-23 and ‘Milyang23’ cross (background was ‘Milyang23’ homozygote) and the ‘Nipponbare’ and ‘Milyang23’ cross (background was heterozygote), but did notin the F₂ population from the NIL33-18 and ‘Nipponbare’ cross (background was ‘Nipponbare’ homozygote). This result indicates that ga2 interacts with a ‘Milyang23’ allele(s) on the different chromosomal region(s) to cause skewed segregation of the ga2 region. In addition, segregation ratio was the same between the F₂ populations from NIL9-23 × ‘Milyang23’ and ‘Nipponbare’ × ‘Milyang23’ crosses, suggesting that the both genotypes, ‘Milyang23’ homozygote and heterozygote, of gene(s) located on the different chromosomal region(s) have the same effect on the segregation distortion. This revised version was published online in July 2006 with corrections to the Cover Date.     

Development of molecular markers linked to the <Emphasis Type="Italic">Fom-1</Emphasis> locus for resistance to Fusarium race 2 in melon

Fusarium wilt, caused by Fusarium oxysporum f. sp. melonis (F.o.m), is a worldwide soil-borne disease of melon (Cucumis melo L.). The most effective control measure available is the use of resistant varieties. Resistance to races 0 and 2 of this fungal pathogen is conditioned by the dominant gene Fom-1. An F₂ population derived from the ‘Charentais-Fom1’ × ‘TRG-1551’ cross was used in combination with bulked segregant analysis utilizing the random amplified polymorphic DNA (RAPD) markers, in order to develop molecular markers linked to the locus Fom-1. Four hundred decamer primers were screened to identify three RAPD markers (B17₆₄₉, V01₅₇₈, and V06₁₀₉₂) linked to Fom-1 locus. Fragments amplified by primers B17₆₄₉ and V01₅₇₈ were linked in coupling phase to Fom1, at 3.5 and 4 cM respectively, whereas V06₁₀₉₂ marker was linked in repulsion to the same dominant resistant allele at 15.1 cM from the Fom-1 locus. These RAPDs were cloned and sequenced in order to design primers that would amplify only the target fragment. The derived sequence characterized amplified region (SCAR) markers SB17₆₄₅ and SV01₅₇₄ (645 and 574 bp, respectively) were present only in the resistant parent. The SV06₁₀₉₂ marker amplified a band of 1092 bp only in the susceptible parent. These markers are more universal than the CAPS markers developed by Brotman et al. (Theor Appl Genet 10:337–345, 2005). The analysis of 24 melon accessions, representing several melon types, with these markers revealed that different melon types behaved differently with the developed markers supporting the theory of multiple, independent origins of resistance to races 0 and 2 of F.o.m.     

Resistance to <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> race 1 in the lettuce cultivars Grand Rapids and Salinas-88

The objective of this work was to check the possible allelism between two sources of resistance to the root-knot nematode Meloidogyne incognita race 1 in lettuce (‘Grand Rapids’ and ‘Salinas-88’). The experiments were carried out in greenhouses, in expanded 128-cell polystyrene trays filled with commercial substrate. Lettuce cultivars ‘Salinas 88’ and ‘Grand Rapids’ were tested along with the populations F₁ (‘Grand Rapids’ × ‘Salinas-88’), F₂ (‘Grand Rapids’ × ‘Salinas-88’), F₃ (‘Grand Rapids’ × ‘Salinas-88’), and with F₄ families derived from the latter population. Seedlings were inoculated 15 days after sowing with a nematode egg suspension equivalent to 30 eggs ml⁻¹ of substrate. Plants were evaluated for apparent gall incidence, gall scores, egg mass scores and extracted egg numbers 45 days after the inoculation date. There was evidence that two different genes are involved in control of resistance to M. incognita race 1 in lettuce cultivars Grand Rapids and Salinas-88. Lines with higher levels of nematode resistance than either Grand Rapids or Salinas-88 could be selected in the F4 generation of the cross between these resistant parental lines.     

Molecular mapping of a gene conferring resistance to Aphanomyces root rot (black root) in sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.)

Caused by Aphanomyces cochlioides Drechsler, Aphanomyces root rot is a serious disease of sugar beet (Beta vulgaris L.), for which sources of resistance are scarce. To identify the segregation pattern of the rare resistance trait found in Japanese sugar beet line ‘NK-310mm-O’, F₁ and BC₁F₂ seedings, drawn from a cross between ‘NK-310mm-O’ and susceptible line ‘NK-184mm-O’, were inoculated with zoospores and their survival evaluated in the greenhouse. Resistance segregation followed was that of a single dominant gene, which was designated Acr1 (Aphanomyces cochlioides resistance 1). Molecular markers tightly linked to Acr1 were identified by bulked segregant analysis of two BC₁F₂ populations. Fourteen AFLP markers linked to Acr1 were identified, the closest located within ±3.3 cM. Three F₅ lines and two BC₂F₁ lines, selected on the basis of their Acr1-AFLP markers, were tested for their resistance to Aphanomyces root rot in a highly infested field. Results indicated that Acr1 conferred significant resistance to Aphanomyces root rot at the field level. Based on its linkage with CAPS marker tk, a representative marker for chromosome III, Acr1 was located on this chromosome. The clear linkage between tk and Rhizomania resistance trait Rz1, suggests the clustering of major disease resistance genes on chromosome III.     

Dent corn genetic background influences QTL detection for grain yield and yield components in high-oil maize

Despite the well-recognized importance of grain yield in high-oil maize (Zea mays L.) breeding and production, few studies have reported the application of QTL mapping of such traits. An inbred line of high-oil maize designated ‘GY220’ was crossed with two dent maize inbred lines to generate two connected F_2:3 populations with 284 and 265 F_2:3 families. Our main objective was to evaluate the influence of genetic background on QTL detection of grain yield traits through comparisons between the F_2:3 populations. The field experiments were conducted during the spring in Luoyang and summer in Xuchang, Henan, China. Two genetic linkage maps were constructed with a genetic distance of 2111.7 and 2298.5 cM using 185 and 173 polymorphic SSR markers, respectively. In total, 18 and 15 QTL were detected for six grain yield traits in the two populations. Only one common QTL marker was shared between the two populations. A QTL cluster associated with five traits was identified at bin 1.05–1.06, including the shared QTL for 100GW, which demonstrated the largest effect (16.7%). Among the detected QTL, 12 digenic interactions were identified. Our results reflect the substantial influence of dent maize genetic background on QTL detection of grain yield traits.     

RAPD markers linked to a clubroot-resistance locus in Brassica rapa L.

Linkage of random amplified polymorphic DNA (RAPD) markers with resistance genes to clubroot (Plasmodiophora brassicae Wor.) in Brassica rapa L. was studied in a doubled haploid (DH population obtained by microspore culture. Thirty-six DH lines were obtained from F₁ plants from a cross between susceptible ‘Homei P09’ and resistant ‘Siloga S2’ plants. ‘Homei P09’ was a DH line obtained by microspore culture of the Chinese cabbage variety ‘Homei’, which is highly responsive in microspore culture. The resistant line ‘Siloga S2’ was obtained by two rounds of selfing of the fodder turnip ‘Siloga’. Three RAPD markers, RA12-75A, WE22B and WE49B, were found to be linked to a clubroot-resistance locus. These three markers were linked in the DH lines and an F₂ population and should be useful for marker-assisted selection in breeding programs. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of molecular markers associated with resistance to squash silverleaf disorder in summer squash (<Emphasis Type="Italic">Cucurbita</Emphasis> <Emphasis Type="Italic">pepo</Emphasis>)

Squash silverleaf (SSL), caused by the silverleaf whitefly [Bemisia argentifolii (formerly known as Bemisia tabaci Gennadius, B strain)], is an important physiological disorder that affects squash (Cucurbita spp.) by reducing yield potential. Breeding squash with resistance to SSL disorder can be facilitated by using marker-assisted selection (MAS). Resistance to SSL disorder, in Cucurbita pepo, is conferred by a single recessive gene (sl). The objective of this study was to identify molecular markers associated with resistance. A zucchini squash, SSL disorder resistant breeding line, ‘Zuc76’ (sl/sl) and a SSL disorder susceptible zucchini cultivar ‘Black Beauty’ (Sl/Sl) were screened with 1,152 randomly amplified polymorphic DNA (RAPD) primers and 432 simple sequence repeat (SSR) markers to identify polymorphisms. Using F₂ and BC₁ progeny segregating for SSL disorder resistance, three RAPD (OPC07, OPL07 and OPBC16) primers and one SSR (M121) marker were found associated with sl. Fragments amplified by RAPD primer OPC07 was linked in coupling phase to sl, whereas RAPD primer OPL07 was linked in repulsion phase. RAPD primer OPBC16 and SSR marker M121 were co-dominant. The allelic order of these loci was found to be M121–sl–OPC07–OPL07–OPBC16. The closest marker to sl is M121 with an estimated genetic distance of 3.3 cM. The markers identified in this study will be useful for breeding summer squash (C. pepo) for SSL disorder resistance derived from zucchini squash breeding line ‘Zuc76’.     

The use of wheat/goatgrass introgression lines for the detection of gene(s) determining resistance to septoria tritici blotch (Mycosphaerella graminicola)

At the IPK Gatersleben a series of 85 bread wheat (T. aestivum)/goatgrass (Aegilops tauschii) introgression lines was developed recently. Based on the knowledge that chromosome 7D of this particular Ae. tauschii is a donor of resistance to septoria tritici blotch (Mycosphaerella graminicola), a sub-set of thirteen chromosome 7D introgression lines was investigated along with the susceptible recipient variety ‘Chinese Spring’ (CS) and the resistant donor line ‘CS (Syn 7D)’. The material was inoculated with two Argentinian isolates of the pathogen (IPO 92067 and IPO 93014) at both the seedlings (two leaf) and adult (tillering) stages at two locations over 2 years (2003, 2004). The resistance was effective against both isolates and at both developmental stages, and the resistance locus maps to the centromeric region of chromosome arm 7DS. On the basis of its relationship with the microsatellite marker Xgwm44, it is likely that the gene involved is Stb5. Stb5 is therefore apparently effective against M. graminicola isolates originating from both Europe and South America.     

Evaluation of leaf rust resistance genes Lr1 , Lr9 , Lr24 , Lr47 and their introgression into common wheat cultivars by marker-assisted selection

Epidemiological field controls in different Italian locations and seedling evaluations of the ‘Thatcher’ near-isogenic lines (NILs) carrying the leaf rust resistance genes Lr1, Lr9, Lr24 and Lr47 were conducted during 5 years of testing. These genes confirmed their effectiveness in both field and greenhouse conditions. Moreover a backcross program was carried out by using as recurrent parents the susceptible high-quality common wheat cvs ‘Bolero’, ‘Colfiorito’, ‘Serio’ and ‘Spada’ and the ‘Thatcher’ NILs carrying the above mentioned genes as donor parents. The progenies of different cross combinations were selected by both resistance tests and marker assisted selection using molecular markers (STS, SCAR, CAPS) closely linked to Lr genes: a complete cosegregation was observed between the resistance genes used and the corresponding molecular markers.     

Microsatellite mapping of complementary genes for purple grain colour in bread wheat (Triticum aestivum) L.

Complementary genes for purple grain colour Pp1, Pp2, Pp3 (now designated Pp1, Pp3b, Pp3a, respectively) were mapped using crosses between purple-grained hexaploid wheats ‘Purple Feed’ – Pp1Pp1/Pp2Pp2 (Pp1Pp1/Pp3bPp3b), ‘Purple’ – Pp1Pp1/Pp3Pp3 (Pp1Pp1/Pp3aPp3a) with non-purple-grained cultivars ‘Novosibirskaya 67’ (‘N67’) and ‘Saratovskaya 29’ (‘S29’). The genes Pp2 (Pp3b) and Pp3 (Pp3a) were inherited as monofactorial dominant when purple-grained wheats were crossed to ‘N67’. Both were mapped in the centromeric region of the chromosome 2A. Therefore, they were suggested being different alleles at the same locus and designated Pp3a and Pp3b. In the crosses between purple-grained wheats and ‘S29’ a segregation ratio of 9 (purple) to 7 (non purple) was obtained suggesting a complementary interaction of two dominant genes, Pp1 and Pp3. To map Pp1 as a single gene, the influence of the other Pp gene was taken into consideration by determining the Pp3 genotype of the F₂ plants. The gene was mapped on chromosome 7BL, about 24 cM distal to the centromere. The Pp1gene was shown to be non allelic to the Rc-1 (red coleoptile) and Pc (purple culm) genes, contrary to what was previously suggested. The colouration caused by the Pp genes has no effect on pre-harvest sprouting.