Molecular mapping of a new gene for resistance to rice blast (Pyricularia grisea Sacc.)

A single dominant blast resistance gene conferring resistance to a Korean rice blast isolate was identified in rice variety `Suweon 365'. We report the chromosomal localization and molecular mapping of this blast resistance gene designated as Pi-18, which confers resistance to Korean isolate `KI-313' of the blast pathogen. To know whether there is a relationship among genes conditioning resistance to location-specific isolates of the blast pathogen and thereby to identify linked markers to resistance gene for isolate KI-313 collected in Korea, RFLP markers previously reported to be linked to major blast resistance genes in different rice germplasm and other markers mapped to nearby regions were surveyed for polymorphism between a resistant (`Suweon 365') and a susceptible (`Chucheongbyeo') parent. Linkage associations of the RFLP markers with the resistance gene were verified using an F2 and F3 segregating population of known blast reaction. RFLP analysis showed that Pi-18 was located near the end of chromosome 11, linked to a single copy clone RZ536 at a distance of 5.4 centiMorgans (cM) and that this gene was different from Pi-1(t). An allelism test revealed that this gene was also different from Pi-k. Currently, a combination of RAPD and microsatellite primers is being employed to find additional markers in this region. Tightly linked DNA markers will facilitate selection for resistant genotypes in breeding programs and provide the basis for map based cloning of this new blast resistance gene. This revised version was published online in July 2006 with corrections to the Cover Date.     

Localisation and combination of resistance genes against soil-borne viruses of barley (BaMMV,BaYMV) using doubled haploids and molecular markers

Barley yellow mosaic virus disease caused by different strains of BaYMV and BaMMV is a major threat to winter barley cultivation in Europe. Different resistance genes against these viruses have been mapped and suitable PCR-based markers have been developed. In this respect doubled haploid (DH) populations proved to be advantageous as they facilitate a repeated test for resistance against all agents of the barley yellow mosaic virus complex and besides this, dominant marker systems are as informative as co-dominant ones in DHs due to the lack of heterozygous genotypes. Using DH populations resistance genes rym4, rym5, rym11, rym13, rym15 and the BaYMV/BaYMV-2 resistance of the barley cultivar ‘Chikurin Ibaraki 1’ have been mapped. DHs are also well suited to pyramiding resistance genes against BaMMV and BaYMV. Since homozygous recessive genotypes are more frequent in DHs than in segregating F₂ populations, DHs can be efficiently used to create broad-spectrum resistance and to extend the usability of partly overcome resistance genes. Results from employing two different strategies for pyramiding, based on one and two DH-steps, respectively, combining three recessive resistance genes, i.e. rym4/rym5, rym9 and rym11, are presented. The faster strategy based on one haploidy step resulted in the identification of all three and two-way combinations of the respective resistance genes.     

Identification of AFLP fragments linked to one recessive genic male sterility (RGMS) in rapeseed (Brassica napus L.) and conversion to SCAR markers for marker-aided selection

117AB is a recessive genic male sterility (RGMS) line in which the sterility is controlled by a duplicate recessive gene named ms, located at two separate loci. In the RGMS line, the genotype of the sterile plant (117A) is msmsmsms, and that of the fertile plant (117B) is Msmsmsms. The present study was aimed to identify DNA markers linked to the ms locus by amplified fragment length polymorphism (AFLP). From the survey of 512 AFLP primer combinations, 6 AFLP fragments (y1, k1, k2, k3, k4, k5) were identified as being tightly linked to the Ms locus. The genetic distances between the markers and the Ms locus were all less than 8 cM, among which two fragments, designated as k2 and k3, co-segregated with the target gene in the tested population. Fragment k2 was successfully converted into a sequence characterized amplified region (SCAR) marker. The markers detected could be valuable in marker-assisted breeding of RGMS in Brassica napus.     

Dwarfing and lethal genes in apple progenies

Summary Three dwarf types are described, early dwarf, determined by two recessive genes d ₁ and d ₃, in the double homozygous state, and an additional recessive gene d ₄ and crinkle dwarf and sturdy dwarf each also determined by a recessive gene.The gene l, for pale green lethal was found to be closely linked to the gene V _f , for scab resistance.There was evidence of a regrowth promoting gene, G, in some seedlings homozygous for d ₁ and d ₃.     

Construction of a genetic map of melon with molecular markers and horticultural traits, and localization of genes associated with ZYMV resistance

Abstract: A partial linkage map of melon was constructed from a cross between PI414723 and Dulce. Twenty-two SSR, 46RAPD, 2 ISSR markers and four horticultural markers [female flower form (a), Fusarium resistance, striped epicarp (st), and fruit flesh pH (pH)] were analyzed in an F₂/F₃ population to produce a map spanning 14 linkage groups. We report for the first time map positions for the st, a, and pH genes. One SSR marker was tightly linked to pH. Mapping the a gene for the female flower form to molecular linkage group 4 enabled the merging of the map of horticultural traits with the of molecular markers in this region. Using the 22 SSR markers of this map, two of the three postulated ZYMV resistance genes were located using a BC₁ population (PI414723 recurrent parent). One SSR marker was tightly linked to a ZYMV resistance gene, designated Zym-1. This revised version was published online in August 2006 with corrections to the Cover Date.     

The V f gene for scab resistance in apple is linked to sub-lethal genes

Summary V _f is the most widely used resistance gene in the breeding for scab resistant apple cultivars. Distorted segregation ratios for V _f -resistance have frequently been reported. Here we revealed that sub-lethal genes caused the distorted segregation. The inheritance of V _f was examined in six progenies by testing linked molecular markers. Three progenies showed distorted segregations that could be explained by three sub-lethal genes (sl1, sl2 and sl3), of which sl1, sl2 were closely linked to V _f . The s11 gene was located at about 14 cM from V _f and expressed itself only in the presence of another independently segregating sub-lethal gene sl3. Only the double homozygous recessive genotypes (sl1sl1 sl3sl3) were lethal, which occurred at first as dwarf and poor vigour plants during the first three months after germination. The sl2 gene was also linked to V _f and its lethality was expressed prior to seed germination and also required the homozygous recessive presence of sl3. The map position of sl3 has not yet been identified. The linkage of V _f to sub-lethal genes usually results in a shortage of V _f -resistant progenies. But in some exceptional crosses, it will lead to abundance of resistant seedling.An erratum to this article can be found at     

Molecular mapping of a stripe rust resistance gene in wheat cultivar Wuhan 2

Stripe rust (or yellow rust), caused by Puccinia striiformis f. sp. tritici, is one of the most destructive diseases of wheat worldwide. Growing resistant cultivars is the best approach to control the disease. To identify and map genes for stripe rust resistance in wheat cultivar ‘Wuhan 2', an F₂ population was developed from a cross between the cultivar and susceptible cultivar Mingxian 169. The parents, 179 F₂ plants and their derived F_2:3 lines were evaluated for responses to Chinese races CYR30 and CYR31 of the pathogen in a greenhouse. A recessive gene for resistance was identified. DNA bulked segregant analysis was applied and resistance gene analog polymorphism (RGAP) and simple sequence repeat (SSR) techniques were used to identify molecular markers linked to the resistance gene. A genetic map consisting of five RGAP and six SSR markers was constructed. The recessive gene, designated Yrwh2, was located on the short arm of chromosome 3B and flanked by SSR markers Xwmc540 and Xgwm566 at 5.9 and 10.0 cM, respectively. The chromosomal location of the resistance gene and its close marker suggest that the locus is different from previously reported stripe rust resistance genes Yr30, QYr.ucw-3BS, Yrns-B1, YrRub and QYrex.wgp-3BL previously mapped to chromosome 3B. Yrwh2 and its closely linked markers are potentially useful for developing stripe rust resistance wheat cultivars if used in combination with other genes.     

A new powdery mildew resistance gene: Introgression from wild emmer into common wheat and RFLP-based mapping

An Israeli accession (TTD140) of wild emmer, Triticum turgidum var. dicoccoides, was found resistant to several races of powdery mildew. Inoculation of the chromosome-arm substitution lines (CASLs) of TTD140, in the background of the Israeli common wheat cultivar ‘Bethlehem’ (BL), with five isolates of powdery mildew revealed that only the line carrying the short arm of chromosome 2B of wild emmer (CASL 2BS) exhibited complete resistance to four of the five isolates. To map and tag the powdery mildew resistance gene, 41 recombinant substitution lines, derived from a cross between BL and CASL 2BS, were used to construct a linkage map at the gene region. The map, which encompasses 69.5 cM of the distal region of chromosome arm 2BS, contains six RFLP markers, a morphological marker (glaucousness inhibitor, W1 ^I), and the powdery mildew resistance gene. Segregation ratios for resistance in F₂ of BL × CASL 2BS and in the recombinant lines, combined with the susceptability of F₁ progeny to all tested isolates, indicate that resistance is controlled by a single recessive allele. This alleleco-segregated with a polymorphic locus detected by the DNA marker Xwg516, 49.4 cM from the terminal marker Xcdo456. The new powdery mildew resistance gene was designated Pm26. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Genetic analysis of broad spectrum resistance to potyviruses using doubled haploid lines of pepper (Capsicum annuum L.)

Summary Genetic analysis of resistance to PVY in androgenetic doubled haploid lines, F1, F2 and backcross progenies of the Mexican pepper line, CM 334 (Capsicum annuum L.), was performed. Three reaction types were observed when seedlings were inoculated with several PVY strains of different pathotypes and with an American PeMV strain. Resistant genotypes never showed systemic symptoms although some individuals sporadically developed necrotic local lesions on inoculated cotyledons. Susceptible genotypes exhibited either a typical systemic mosaic or a systemic necrosis that caused the death of the inoculated seedlings. Segregation analyses indicated that resistance to pepper potyviruses in CM 334 is conferred by two genes. The first one, tentatively named Pr4, is dominant and confers the resistance to all now known pathotypes of PVY and to PeMV. The second one, tentatively named pr5, is recessive; it confers only the resistance to common strains of PVY. The systemic necrotic response is conferred by an independent dominant gene, tentatively named Pn1.     

Phenotypic and marker‐assisted pyramiding of genes for resistance to zucchini yellow mosaic virus in oilseed pumpkin (Cucurbita pepo)

We report on the identification of phenotypic and molecular markers for genes introgressed into oilseed pumpkin Cucurbita pepo from C. moschata germplasm originating in Nigeria, Portugal and Puerto Rico, which provide resistance against zucchini yellow mosaic virus (ZYMV) and on pyramiding these genes for improved and long‐lasting field protection of oilseed pumpkins. One SCAR and two SSR markers have been found for three dominant resistance genes, Zym‐0, Zym‐1 and Zym‐2. Characteristic reactions to ZYMV inoculation of plants carrying the recessive genes for resistance zym‐4* and zym‐6 have been defined. Described are procedures and results of pyramiding various combinations of these genes in oilseed pumpkin using the three markers and the specific phenotypic reactions to infection of some of these genes. The putative combination of all six resistance genes in one genotype resulted in a resistance that appeared to be at least as strong as or even stronger than that of the resistance source germplasm in C. moschata.     

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.     

The inheritance of stem rust and leaf rust resistance in some Ethiopian wheat collections

Summary Common and durum wheat populations obtained from Sweden and originally collected in Ethiopia were screened for resistance to steum rust and leaf rust. Resistant selections of common wheat were crossed and backcrossed with either stem rust susceptible RL6071, or leaf rust susceptible Thatcher. Genetic studies, based largely on tests of backcross F₂ families, showed that four of the selections had in common a recessive gene SrA. Plants with this gene were resistant (1⁺ infection type) to all stem rust races tested. This gene was neither Sr26 nor Sr29. The resistance of other selections, based on tests with an array of rust isolates, was due to various combinations of Sr6, 8a, 9a, 9d, 9c, 11, 13, 30, and 36. One of the selections had linked genes, Lr19/Sr25. Another selection had a dominant gene for resistance (;1 infection type) to all the races of leaf rust. With the possible exception of this gene for leaf rust resistance and SrA, no obviously new resistance was found.     

Identification of AFLP markers linked to the epistatic suppressor gene of a recessive genic male sterility in rapeseed and conversion to SCAR markers

A recessive epistatic genic male sterility (REGMS) two‐type line, 9012AB, has been used for rapeseed hybrid seed production in China. The male sterility of 9012AB is controlled by two recessive duplicate sterile genes (ms1 and ms2) interacting with one recessive epistatic suppressor gene (esp). Homozygosity at the esp locus (espesp) suppresses the expression of the recessive male sterility trait in homozygous ms1ms1ms2 ms2 plants. In this study, we used a combination of bulked segregant analyses and amplified fragment length polymorphism (AFLP) to identify markers linked to the suppressor gene in a BC₁ population. From the survey of 1024 AFLP primer combinations, eight markers tightly linked to the target gene were identified. The two closest markers flanking both sides of Esp, P9M5₃₇₀ and S16M14₇₈₀, had a genetic distance of 1.4 cM and 2.1 cM, respectively. The AFLP fragment from P4M8₁₉₀, which co‐segregated with the target gene was converted into a sequence characterized amplified region marker. The availability of linked molecular markers will facilitate the utilization of REGMS in hybrid breeding in Brassica napus.     

一个新的抗玉米矮花叶病基因的发现及初步定位

由SCMV引起的矮花叶病是我国的主要玉米病害之一, 鉴定和发掘新的抗病基因对于玉米抗病遗传育种具有重要意义。以抗病自交系海9-21和感病自交系掖478杂交的一个BC₂F₃群体为试验材料, 通过人工接种矮花叶病毒进行抗病性鉴定, 发现该分离群体中抗病植株与感病植株数符合1∶3的分离比例, 推测其抗病基因是由1对隐性基因控制。抗感池和SSR标记连锁分析表明, 存在一个新的玉米矮花叶病隐性抗病基因(或等位基因), 将该基因命名为scm3。scm3基因来源于抗病玉米自交系海9-21, 位于第3染色体短臂3.04~3.05区域, 在SSR标记umc1965和bnlg420之间, 遗传距离分别为45.7 cM和6.5 cM。连锁的标记还有umc1307、umc2265、bnlg2241和umc2166, 它们与scm3之间的遗传距离分别是8.3、13.3、15.5和19.7 cM, 这些SSR标记与scm3基因在染色体上的排列顺序为umc1965—scm3—bnlg420—umc1307—umc2265—bnlg2241—umc2166。     

Genetic characterization of anthracnose resistance genes Co-4 3 and Co-9 in common bean cultivar tlalnepantla 64 (PI 207262)

Tlalnepantla 64 (PI 207262) is an important source of genes for resistance to common bean anthracnose, caused by Colletotrichum lindemuthianum. However, these genes have not been fully characterized. Inheritance studies using crosses involving PI 207262 show that two independent genes confer resistance to anthracnose. Allelism tests showed that the genes are located at distinct loci from the previously identified resistance genes Co-1, Co-2, Co-3, Co-5, Co-6, and Co-10. Also, no segregation was observed in relation to Co-4, Co-4 ², Co-9, and to the gene present in cultivar Widusa, indicating that PI 207262 harbors alleles of these genes. We conclude that PI 207262 harbors two anthracnose resistance genes, Co-4 and Co-9. The Co-4 allele of PI 207262 would be different from Co-4 and Co-4 ² and it is proposed Co-4 ³ as the genetic symbol for this resistance allele. As PI 207262 is the parent of BAT 93, the Co-9 symbol represents the gene of both cultivars. Also, one allele of Co-9 gene was detected in cultivar Widusa.     

Identification of microsatellite markers linked to the blast resistance gene <Emphasis Type="Italic">Pi-1(t)</Emphasis> in rice

The present work was conducted to identify microsatellite markers linked to the rice blast resistance gene Pi-1(t) for a marker-assisted selection program. Twenty-four primer pairs corresponding to 19 microsatellite loci were selected from the Gramene database (www. gramene.org) considering their relative proximity to Pi-1(t) gene in the current rice genetic map. Progenitors and DNA bulks of resistant and susceptible families from F₃ segregating populations of a cross between the near-isogenic lines C101LAC (resistant) and C101A51 (susceptible) were used to identify polymorphic microsatellite markers associated to this gene through bulked segregant analysis. Putative molecular markers linked to the blast resistance gene Pi-1(t) were then used on the whole progeny for linkage analysis. Additionally, the diagnostic potential of the microsatellite markers associated to the resistance gene was also evaluated on 17 rice varieties planted in Latin America by amplification of the specific resistant alleles for the gene in each genotype. Comparing with greenhouse phenotypic evaluations for blast resistance, the usefulness of the highly linked microsatellite markers to identify resistant rice genotypes was evaluated. As expected, the phenotypic segregation in the F₃ generation agreed to the expected segregation ratio for a single gene model. Of the 24 microsatellite sequences tested, six resulted polymorphic and linked to the gene. Two markers (RM1233*I and RM224) mapped in the same position (0.0 cM) with the Pi-1(t) gene. Other three markers corresponding to the same genetic locus were located at 18.5 cM above the resistance gene, while another marker was positioned at 23.8 cM below the gene. Microsatellite analysis on elite rice varieties with different genetic background showed that all known sources of blast resistance included in this study carry the specific Pi-1(t) allele. Results are discussed considering the potential utility of the microsatellite markers found, for MAS in rice breeding programs aiming at developing rice varieties with durable blast resistance based on a combination of resistance genes. Centro Internactional de Agricultura Tropical (CIAT) institute where the research was carried out     

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.     

Inheritance of resistance to four cucurbit viruses in Cucurbita moschata

Cucurbita moschata cv. Nigerian Local has been used as a source of resistance to Zucchini yellow mosaic virus (ZYMV), Watermelon mosaic virus (WMV), Papaya ringspot virus W (PRSV-W) and Cucumber mosaic virus (CMV) in breeding both Cucurbita moschata and Cucurbita pepo. We used the F₁, F₂ and BC₁ generations derived from the cross C.-moschata cv. Waltham Butternut × Nigerian Local to study the inheritance of resistance to each of the viruses. We confirmed monogenic dominant resistance to ZYMV previously attributed to Zym, and we report monogenic dominant resistance to WMV and CMV which we propose to designate Wmv and Cmv, respectively. A single recessive gene, which we propose to designate prv, controls resistance to PRSV. DNA samples were extracted from a Waltham Butternut BC₁ F₁ population screened with ZYMV and analyzed using randomly amplified polymorphic DNA markers. No RAPD markers linked to ZYMV resistance were found. This revised version was published online in July 2006 with corrections to the Cover Date.     

Analysis of rice blast resistance gene <Emphasis Type="Italic">Pi</Emphasis>-<Emphasis Type="Italic">z</Emphasis> in rice germplasm using pathogenicity assays and DNA markers

The Pi-z gene in rice confers resistance to a wide range of races of the rice blast fungus, Magnaporthe oryzae. The objective of this study was to characterize Pi-z in 111 rice germplasm accessions using DNA markers and pathogenicity assays. The existence of Pi-z in rice germplasm was detected by using four simple sequence repeat (SSR) markers (RM527, AP4791, AP5659-1, AP5659-5) closely linked to Pi-z, and was verified using pathogenicity assays with an avirulent strain (IE1k) and two virulent races (IB33 and IB49). Among 111 germplasm accessions evaluated, 73 were found to contain the Pi-z gene using both SSR markers and pathogenicity assays. The remaining 38 germplasm accessions were found to be inconsistent in their responses to the blast races IB33, IEIk and IB49 with expected SSR marker alleles, suggesting the presence of unexpected SSR alleles and additional R gene(s). These characterized germplasm can be used for genetic studies and marker-assisted breeding for improving blast resistance in rice.