Molecular mapping of black rot resistance locus Xca1bo on chromosome 3 in Indian cauliflower (Brassica oleracea var. botrytis L.)

Black rot is the most devastating disease of cauliflower worldwide causing severe damage to crop. The identification of markers linked to loci that control resistance can facilitate selection of plants for breeding programmes. In the present investigation, F₂ population derived from a cross between ‘Pusa Himjyoti’, a susceptible genotype, and ‘BR‐161’, a resistant genotype, was phenotyped by artificial inoculation using Xcc race 1. Segregation analysis of F₂ progeny indicated that a single dominant locus governed resistance to Xcc race 1 in ‘BR‐161’. Bulk segregant analysis in resistant and susceptible bulks of F₂ progeny revealed seven differentiating polymorphic markers (three RAPD, two ISSR and two SSR) of 102 markers screened. Subsequently, these markers were used to genotype the entire F₂ population, and a genetic linkage map covering 74.7 cM distance was developed. The major locus Xca1bo was mapped in 1.6‐cM interval flanked by the markers RAPD 04₈₃₃ and ISSR 11₆₃₅. The Xca1bo locus was located on chromosome 3. The linked markers will be useful for marker‐assisted resistance breeding in cauliflower.     

RAPD and SCAR markers for resistance to acochyta blight in lentil

Resistance to ascochyta blight of lentil (Lens culinaris Medikus),caused by the fungus Ascochyta lentis, is determined by a single recessive gene, ral ₂, in the lentil cultivar Indian head. Sixty F₂ individuals from a cross between Eston (susceptible) and Indian head (resistant) lentil were analyzed for the presence of random amplified polymorphic DNA (RAPD) markers linked to the ral ₂gene, using bulked segregant analysis (BSA). Out of 800 decanucleotide primers screened, two produced polymorphic markers that co-segregated with the resistance locus. These two RAPD markers, UBC227₁₂₉₀and OPD-10₈₇₀, flanked and were linked in repulsion phase to the gene ral ₂ at 12 cm and 16 cm, respectively. The RAPD fragments were converted to SCAR markers. The SCAR marker developed from UBC227₁₂₉₀ could not detect any polymorphism between the two parents or in the F₂. The SCAR marker developed from OPD-10₈₇₀ retained its polymorphism. The polymorphic RAPD marker UBC227₁₂₉₀ and the SCAR marker developed from OPD-10₈₇₀ can be used together in a marker assisted selection program for ascochyta blight resistance in lentil. This revised version was published online in July 2006 with corrections to the Cover Date.     

Molecular markers linked to the Aegilops variabilis-derived root-knot nematode resistance gene Rkn-mn1 in wheat

Aegilops variabilis no. 1 is the only known source of resistance to the root‐knot nematode Meloidogyne naasi in wheat. Previous studies showed that a dominant gene, Rkn‐mn1, was transferred to a wheat translocation line from the donor Ae. variabilis. Random amplified polymorphic DNA (RAPD) analysis was performed on the wheat cultivar ‘Lutin’, on Ae. variabilis, on a resistant disomic addition line and on a resistant translocation line. For genetic and molecular studies, 114‐117 BC₃F₂ plants and F₃‐derived families were tested. Five DNA and one isozyme marker were linked to Rkn‐mn1. Three RAPD markers flanking the Rkn‐mn1 locus were mapped at 0 cM (OpY16_‐1065), 0.8 cM (OpB12_‐1320) and 1.7 cM (OpN20_‐1235), respectively. Since the Rkn‐mn1 gene remained effective, its introduction into different wheat cultivars by marker‐assisted selection is suggested.     

Evaluation of common wheat cultivars for tan spot resistance and chromosomal location of a resistance gene in the cultivar 'Salamouni'

A total of 50 wheat (Triticum aestivum L.) cultivars were evaluated for resistance to tan spot, using Pyrenophora tritici‐repentis race 1 and race 5 isolates. The cultivars ‘Salamouni’, ‘Red Chief’, ‘Dashen’, ‘Empire’ and ‘Armada’ were resistant to isolate ASC1a (race 1), whereas 76% of the cultivars were susceptible. Chi‐squared analysis of the F₂ segregation data of hybrids between 20 monosomic lines of the wheat cultivar ‘Chinese Spring’ and the resistant cultivar ‘Salamouni’ revealed that tan spot resistance in ‘Salamouni’ was controlled by a single recessive gene located on chromosome 3A. This gene is designated tsn4. The resistant cultivars identified in this study are recommended for use in breeding programmes to improve tan spot resistance in common wheat.     

A second gene for resistance to race 4 of Fusarium wilt in chickpea and linkage with a RAPD marker

Fusarium wilt caused by Fusarium oxysporum Schlechtend.: Fr f. sp. ciceris (Padwick) Matuo & Sato is a devastating disease of chickpea. The current study was conducted to determine the inheritance of the gene(s) for resistance to race 4 of fusarium wilt and to identify linked RAPD markers using an early wilting line, JG-62, as a susceptible parent. Genetic analysis was performed on the F₁s, F₂s and F₃ families from the cross of JG-62 × Surutato-77. The F₃ families were inoculated with a spore suspension of the race 4 wilt pathogen and the results were used to infer the genotypes of the parent F₂ plants. Results indicated that two independent genes controlled resistance to race 4. Linkage analysis of candidate RAPD marker, CS-27₇₀₀, and the inferred F₂ phenotypic data showed that this marker locus is linked to one of the resistance genes. Allelism indicated that the two resistance sources, Surutato-77 and WR-315, shared common alleles for resistance and the two susceptible genotypes, C-104 and JG-62, carried alleles for susceptibility. The PCR-based marker, CS-27₇₀₀, was previously reported to be linked to the gene for resistance to race 1 in a different population which suggested that the genes for resistance to races 1 and 4 are in close proximity in the Cicer genome. This revised version was published online in August 2006 with corrections to the Cover Date.     

Inheritance of angular leaf spot resistance in common bean line BAT 332 and identification of RAPD markers linked to the resistance gene

The existence of genetic variability for angular leaf spot (ALS) resistance in the common bean germplasm allows the development of breeding lines resistant to this disease. The BAT 332 line is an important resistance source to common bean ALS. In this work we determined the inheritance pattern and identified RAPD markers linked to a resistance gene present in BAT 332. Populations F₁, F₂,BCs and BCr derived from crosses between BAT 332 and cultivar Rudá were used. Rudá is a commercial cultivar with carioca type grains and susceptible to ALS. The resistance of BAT 332 to race 61.41 of the pathogen was confirmed. Segregation analysis of the plants indicated that a single dominant gene confers resistance. For identification of RAPD markers linked to the resistance gene, bulk segregant analysis (BSA) was used. Two RAPD markers,OPAA07₉₅₀ and OPAO12₉₅₀, linked in coupling phase at 5.10 and 5.83 cM of this gene, respectively, were identified. These molecular markers are important for common bean breeders and geneticists as source of genetic information and for marker assisted selection in breeding programs. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of a molecular marker linked to an Agropyron elongatum-derived gene Lr19 for leaf rust resistance in wheat

The leaf rust resistance gene Lr19, transferred from Agropyron elongatum into wheat (Triticum aestivum L.) imparts resistance to all pathotypes of leaf rust (Puccinia recondita f.sp. tritici) in South‐east Asia. A segregating F₂ population from a cross between the leaf rust resistant parent ‘HW 2046’ carrying Lr19 and a susceptible parent ‘Agra Local’ was screened in the phytotron against a virulent pathotype 77‐5 of leaf rust with the objective of identifying the molecular markers linked to Lr19. The gene was first tagged with a randomly amplified polymorphic DNA (RAPD) marker S73₇₂₈. The RAPD marker linked to the gene Lr19 which mapped at 6.4 ± 0.035 cM distance, was converted to a sequence characterized amplified region (SCAR) marker. The SCAR marker (SCS73₇₁₉) was specific to Lr19 and was not amplified in the near‐isogenic lines (NILs) carrying other equally effective alien genes Lr9, Lr28 and Lr32 enabling breeders to pyramid Lr19 with these genes.     

Inheritance and linkage of a gene for resistance to race 4 of fusarium wilt and RAPD markers in chickpea

Several races of Fusarium oxysporum Schlechtend.:Fr f. sp. ciceris (Padwick) Matuo and K. Sato cause economic losses from wilting disease of chickpea ( Cicer arietinum L.). While the genetics of resistance to race 1 have been reported, little is known of the genetics of resistance to race 4. We undertook a study to determine the inheritance of resistance and identified random amplified polymorphic DNA markers (RAPDs) linked to the gene for resistance. For the investigation, we used 100 F₅ derived F₇ recombinant inbred lines (RILs) that had been developed from the cross of breeding lines C-104 x WR-315. Results indicated that resistance is controlled by a single recessive gene. The RAPD markers previously shown to amplify fragments linked to race 1 resistance also amplified fragments associated with race 4 resistance. The RAPD loci, CS-27₇₀₀, UBC-170₅₅₀ and the gene for resistance to race 4 segregated in 1:1 ratios expected for single genes. Both RAPD markers were located 9 map units from the race 4 resistance locus and were on the same side of the resistance gene. Our results indicated that the genes for resistance to race 1 and 4 are 5 map units apart. The need to determine the genomic locations of race specific resistance genes and the possibility that these genes are clustered to the same genomic region should be investigated. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic mapping and inheritance of Russian wheat aphid resistance gene in accession IG 100695

The Russian wheat aphid (RWA), Diuraphis noxia (Kurdjumov), is an important pest of small‐grain cereals, particularly wheat, worldwide. The most efficient strategy against the RWA is to identify sources of resistance and to introduce them into susceptible wheat genotypes. This study was conducted to determine the mode of inheritance of the RWA resistance found in ICARDA accession IG 100695, to identify wheat microsatellite markers closely linked to the gene and to map the chromosomal location of the gene. Simple sequence repeat (SSR) marker scores were identified in a mapping population of 190 F₂ individuals and compared, while phenotypic screening for resistance was performed in F_2 : 3 families derived from a cross between ‘Basribey’ (susceptible) and IG 100695 (resistant). Phenotypic segregation of leaf chlorosis and rolling displayed the effect of a single dominant gene, temporarily denoted Dn100695, in IG 100695. Dn100695 was mapped on the short arm of chromosome 7D with four linked SSR markers, Xgwm44, Xcfd14, Xcfd46 and Xbarc126. Dn100695 and linked SSR markers may be useful for improving resistance for RWA in wheat breeding.     

User‐friendly markers linked to Fusarium wilt race 1 resistance Fw gene for marker‐assisted selection in pea

Fusarium wilt is one of the most widespread diseases of pea. Resistance to Fusarium wilt race 1 was reported as a single gene, Fw, located on linkage group III. The previously reported AFLP and RAPD markers linked to Fw have limited usage in marker‐assisted selection due to their map distance and linkage phase. Using 80 F₈ recombinant inbred lines (RILs) derived from the cross of Green Arrow × PI 179449, we amplified 72 polymorphic markers between resistant and susceptible lines with the target region amplified polymorphism (TRAP) technique. Marker–trait association analysis revealed a significant association. Five candidate markers were identified and three were converted into user‐friendly dominant SCAR markers. Forty‐eight pea cultivars with known resistant or susceptible phenotypes to Fusarium wilt race 1 verified the marker–trait association. These three markers, Fw_Trap_480, Fw_Trap_340 and Fw_Trap_220, are tightly linked to and only 1.2 cM away from the Fw locus and are therefore ideal for marker‐assisted selection. These newly identified markers are useful to assist in the isolation of the Fusarium wilt race 1 resistance gene in pea.     

RAPD and SCAR markers for powdery mildew resistance gene er in pea

In pea, a single recessive gene (er) on linkage group 6 confers resistance to powdery mildew caused by Erysiphe pisi. The present study aims to identify molecular markers linked to the er gene. Screening of the powdery mildew‐resistant cultivar ‘DMR11’ and its susceptible nearisogenic line for polymorphism revealed linkage of two RAPD primers (OPO‐02 and OPU‐17) to the er gene and a sequence characterized polymorphic region (SCAR) primer, ScOPD‐10₆₅₀ with er in a population of 83 F₂ plants in the order: OPU‐17 ‐ er ‐ ScOPD‐10₆₅₀ ‐ OPO‐02. The markers ScOPD‐10₆₅₀ and OPU‐17 being coupled with the allele causing resistance would substantially increase the efficiency of marker‐assisted selection in peabreeding for powdery mildew.     

Mapping of the Vrn-B1 gene in Triticum aestivum using microsatellite markers

An F₂ population segregating for the dominant gene Vrn‐B1 was developed from the cross of the substitution line ‘Diamant/'Miro‐novskaya 808 5A’ and the winter wheat cultivar ‘Bezostaya 1′. Microsatellite markers (Xgwm and Xbarc) with known map locations on chromosome 5B of common wheat were used for mapping the gene Vrn‐B1. Polymorphism between parental varieties was observed for 28 out of 34 microsatellite markers (82%). Applying the quantitative trait loci mapping approach, the target gene was mapped on the long arm of chromosome 5B, closely linked to Xgwm408. The map position of Vrn‐B1 suggests that the gene is homoeologous to other vernalization response genes located on the homoeologous group 5 chromosomes of wheat, rye and barley.     

Genetic analysis of karnal bunt resistance in 14 Mexican bread-wheat genotypes

Fourteen Mexican genotypes of bread wheat (Triticum aestivum L.) with good to moderate levels of resistance to Karnal bunt (Tilletia indica (Mitra)) were crossed with the highly susceptible cultivar WL711 to determine the genetic basis of resistance. The parents, F₁ F₂ and backcross populations of the 14 crosses were evaluated under artificial epiphytotic conditions during the 1993–94 season for Karnal bunt resistance. The F₁ data suggested that the resistance was dominant to partially dominant over susceptibility. The F₂ analysis of the segregation ratios in the F₂ and backcross generations indicated that the resistance in the wheat genotypes Luan, Attila, Vee #7/Bow, Star, Weaver, Milan, Sasia and Turacio/Chil is controlled by two genes. The resistance in genotypes Cettia, Irena, Turaco, Opata, Picus, and Yaco was found to be conditioned by a single dominant gene. The genotypes with two genes for resistance expressed a higher level of resistance than those with a single gene and, therefore, are better sources of resistance to Karnal bunt.     

Inheritance of insensitivity to culture filtrate of Pyrenophora tritici-repentis, race 2, in wheat (Triticum aestivum L.)

Tan spot of wheat is caused by the fungus Pyrenophora tritici‐repentis. On susceptible hosts, P. tritici‐repentis induces two phenotypically distinct symptoms, tan necrosis and chlorosis. This fungus produces several toxins that induce tan necrosis and chlorosis symptoms in susceptible cultivars. The objectives of this study were to determine the inheritance of insensitivity to necrosis‐inducing culture filtrate of P. tritici‐repentis, race 2, and to establish the relationship between the host reaction to culture filtrate and spore inoculation with respect to the necrosis component. The F₁, F₂, and BC₁F₁ plants and F_2:8 lines of five crosses involving resistant wheat genotypes ‘Erik’, ‘Red Chief’, and line 86ISMN 2137 with susceptible cultivars ‘Glenlea’ and ‘Kenyon’ were studied. Plants were spore‐inoculated at the two‐leaf stage. Four days later, the newly emerged uninoculated third leaf was infiltrated with a culture filtrate of isolate Ptr 92–164 (race 2). Reactions to the spore inoculation and the culture filtrate were recorded 8 days after spore inoculation. The segregation observed in the F₂ and BC₁F₁ generations and the F_2:8 lines of all crosses indicated that a single recessive gene controlled insensitivity to necrosis caused by culture filtrate. This gene also controlled resistance to necrosis induced by spore inoculation.     

Inheritance of resistance to Karnal bunt (Tilletia indica Mitra) in bread wheat (Triticum aestivum L.)

The mode of inheritance and allelic relationships among genes conferring resistance to Karnal bunt were studied in seven bread-wheat (six resistant and one susceptible) genotypes. The resistant genotypes originated in China (‘Shanghai#8’), Brazil (PF71131), the USA (‘Chris’), and Mexico (‘Amsel’, CMH77.308 and ‘Pigeon’). The susceptible line WL711 was from India. Evaluation of these wheat lines and all possible crosses among their F₁ and F₃ generations (about 100 progenies in each cross) revealed that two partially recessive genes conferred the resistance to Karnal bunt in ‘Pigeon’, whereas four partially dominant genes were present in the other genotypes. ‘Chris’, ‘Amsel’ and PF71131 carry one gene, whereas ‘Shanghai#8’ and CMH77.308 have two genes. ‘Chris’, ‘Amsel’, and PF71131 have different genes, whereas one gene was common to PF71131, CMH77.308 and ‘Shanghai#8’, and another to ‘Chris’ and CMH77.308. Gene symbols were formally designated to the resistant stocks. Resistance was incomplete and stable.     

Molecular mapping of Aegilops speltoides derived leaf rust resistance gene Lr28 in wheat

In a segregating homozygous F₂ population of bread wheat involving a leaf rust resistance gene Lr28 derived from Aegilops speltoides, six randomly amplified polymorphic DNA (RAPD) markers, three each in coupling and repulsion phase were identified as linked to Lr28, mapped to a region spanning 32 cM including the locus. The F₂ and F₃ populations were studied in the phytotron challenged with the most virulent pathotype 77-5 of leaf rust. A coupling phase linked RAPD marker S464₇₂₁ and a repulsion phase linked RAPD marker S326₅₅₀ flanked the gene Lr28 by a distance of 2.4± 0.016 cM on either side. The flanking markers genetically worked as co-dominant markers when analyzed together after separate amplification in the F₂ population by distinguishing the homozygotes from the heterozygotes and increased the efficiency of marker assisted selection by reducing the false positives and negatives. One of the three RAPD markers, S421₆₄₀ was converted to locus specific SCAR marker SCS421₆₄₀ which was further truncated by designing primers internal from both ends of the original RAPD amplicon to eliminate a non-specific amplification of nearly same size. The truncated polymorphic sequence characterized amplified region marker (TPSCAR) SCS421₅₇₀ was 70 bp smaller, but resulted in a single band polymorphism specific to Lr28 resistance. The TPSCAR marker was validated for its specificity to the gene Lr28 in nine different genetic backgrounds and on 43 of the 50 Lr genes of both native and alien origin, suggesting the utility of the SCAR markers in pyramiding leaf rust resistance genes in wheat.     

Molecular mapping determines that Hessian fly resistance gene H9 is located on chromosome 1A of wheat

Hessian fly [Mayetiola destructor (Say)] is one of the major insect pests of wheat (Triticum aestivum L.) worldwide. Hessian fly resistance gene H9 was previously reported to condition resistance to Hessian fly biotype L that is prevalent in many wheat‐growing areas of eastern USA and an RAPD marker, OPO05₁₀₀₀, linked to H9 in wheat was developed using wheat near‐isogenic lines (NILs). However, marker‐assisted selection (MAS) with RAPD markers is not always feasible. One of the objectives in this study was to convert an RAPD marker linked to the gene H9 into a sequence characterized amplified region (SCAR) marker to facilitate MAS and to map H9 in the wheat genome. The RAPD fragment from OPO05₁₀₀₀ was cloned, sequenced, and converted into a SCAR marker SOPO05₉₀₉, whose linkage relationship with H9 was subsequently confirmed in two F₂ populations segregating for H9. Linkage analysis identified one sequence tagged site (STS) marker, STS‐Pm3, and the eight microsatellite markers Xbarc263, Xcfa2153, Xpsp2999, Xgwm136, Xgdm33, Xcnl76, Xcnl117 and Xwmc24 near the H9 locus on the distal region of the short arm of chromosome 1A, contrary to the previously reported location of H9 on chromosome 5A. Locus Xbarc263 was 1.2 cM distal to H9, which itself was 1.7 cM proximal to loci Xcfa2153, Xpsp2999 and Xgwm136. The loci Xgwm136, Xcfa2153 and SOPO05₉₀₉ were shown to be specific to H9 and not diagnostic to several other Hessian fly resistance genes, and therefore should be useful for pyramiding H9 with other Hessian fly resistance genes in a single genotype.     

Chromosomal location of powdery mildew resistance gene Pm16 in wheat using SSR marker analysis

The use of resistant cultivars is a most economical way to control powdery mildew (Blumeria graminis f.sp. tritici) in wheat (Triticum aestivum L.). Identification of molecular markers closely linked to resistance genes can greatly increase the efficiency of pyramiding resistance genes in wheat cultivars. The objective of this study was to identify molecular markers closely linked lo the powdery mildew resistance gene Pm16. An F₂ population with 156 progeny was produced from the cross‘Chancellor’(susceptible) ב70281’ (resistant), A total of 45 SSR markers on chromosomes 4A and 5B of wheat and 15 SSRs on chromosome 3 of rice was used lo lest the parents, as well as the resistant and susceptible bulks: the resulting polymorphic markers were used to genotype the F₂ progeny. Results indicated that the SSR marker Xgwm159, located on the short arm of chromosome 5B, is closely linked to Pm16 (genetic distance: 5.3 CM). The cytogenetical data presented in an original report, in combination with this molecular analysis, suggests that Pm16 may he located on a translocated 4A.5BS chromosome.     

Novel male-specific molecular markers (MADC5, MADC6) in hemp

Decamer RAPD primers were tested on dioecious and monoecious hemp cultivars to identify sex-specific molecular markers. Two primers (OPD05 and UBC354) generated specific bands in male plants. These two DNA fragments were isolated, cloned and sequenced. Both markers proved to be unique, since no sequence with significant homology to OPD05₉₆₁ and UBC354₁₅₁ markers were found in databases. These markers were named MADC3 (OPD05₉₆₁) and MADC4 (UBC354₁₅₁) (Male-Associated DNA from Cannabis sativa). The markers were converted into sequence-characterized amplified region (SCAR) markers. The SCAR markers correlated with the sex of the segregating F₂ population and proved the tight linkage to the male phenotype. Results of F₂ plant population analysis suggest these markers are to be linked to the Y chromosome. This revised version was published online in August 2006 with corrections to the Cover Date.     

Location of genes for common bunt resistance in the European winter wheat cv. Trintella

The location of new genes for resistance to common bunt in wheat is valuable for gene pyramiding in breeding. For this purpose, the genetics of the relatively high level of resistance in the European winter wheat variety Trintella was investigated using a doubled haploid mapping population of a cross between Trintella and the susceptible variety Piko. The population was scored for bunt infection in the field for 2 years following inoculation with a mixture of teliospores of Tilletia tritici and T. laevis. A genetic map consisting of 29 linkage groups was constructed using polymorphic simple sequence repeat markers. This map was used for QTL analysis, and in both years, results indicated that resistance to common bunt could mostly be attributed to a gene on chromosome 1B, near to the centromere and closest to marker Xgwm273 on the short arm. Additionally, in 2008, smaller QTL effects were ascribed to chromosomes 7A and 7B, and another smaller QTL effect to chromosome 5B in 2009 only.