Genetic Analysis of Resistance to Leaf Rust and Yellow Rust in Spring Wheat Cultivar Kenya Kongoni.

The Kenyan wheat (Triticum aestivum L.) 'Kenya Kongoni' exhibits high levels of adult plant resistance (APR) to leaf rust (LR) and yellow rust (YR). We determined the genomic regions associated with LR and YR resistance in a population of 148 recombinant inbred lines generated from a cross between 'Avocet-YrA' and Kenya Kongoni. Field experiments to characterize APR to LR and YR were conducted in four and two Mexican or Uruguayan environments, respectively. A linkage map was constructed with 438 diversity arrays technology and 16 simple-sequence repeat markers by JoinMap 4.1 software. Genetic analyses showed that resistance to both rusts was determined by four to five APR genes, including Lr46/Yr29 and Sr2/Lr27/Yr30. Quantitative trait loci (QTL) analysis indicated that pleiotropic APR loci QYLr.cim-1BL corresponding to Lr46/Yr29 and QYLr.cim-7BL that is a putative novel QTL accounted for 5 to 57% and 12 to 35% of the phenotypic variation for resistance to LR and YR, respectively. These loci, in combination with another three LR QTL and two YR QTL, respectively, conferred high levels of resistance to both LR and YR in wheat under Mexican and Uruguayan environments. Among other detected QTL, QLr.cim-1DS, QLr.cim-2BL, and QYLr.icm-7BL may be new loci for APR to both rusts in common wheat.

Yr60, a Gene Conferring Moderate Resistance to Stripe Rust in Wheat.

Stripe rust, caused by Puccinia striiformis f. sp. tritici W., is a devastating disease of wheat worldwide. A new stripe rust resistance gene with moderate seedling and adult plant resistance was mapped using an F5 recombinant inbred line (RIL) population developed from the cross of the resistant parent 'Almop' with the susceptible parent 'Avocet'. The parents and RILs were phenotyped for seedling stripe rust response variation in a greenhouse and in field trials at Toluca, Mexico for 2 years. Almop showed moderate levels of resistance at both seedling and adult plant stages compared with the highly susceptible response of Avocet. The distribution of homozygous resistant, homozygous susceptible, and segregating RILs conformed to segregation at a single locus. Seedlings and adult plant responses were correlated, indicating that the same gene conferred resistance at both stages. A bulk segregant analysis approach with widely distributed simple sequence repeat (SSR) markers mapped the resistance gene to the distal region of the long arm of chromosome 4A. The SSR marker wmc776 cosegregated with this gene, whereas markers wmc219 and wmc313 were tightly linked and both located at 0.6 centimorgans. The resistance locus was designated Yr60.

Genetic Analysis of Resistance to Leaf Rust and Stripe Rust in Wheat Cultivar Francolin#1.

Leaf rust and stripe rust are important diseases of wheat and can be controlled by growing resistant varieties. We investigated the genetic basis of resistance to both rusts in 198 F5 recombinant inbred lines derived from a cross between 'Avocet' and 'Francolin#1'. The population was phenotyped in greenhouse and field, and genotyped with known gene-associated molecular markers. Seedling resistance of Francolin#1 to leaf and stripe rusts was attributed to the loosely linked genes Lr16 and YrF, respectively, with a recombination frequency of 0.36. Field segregation indicated that adult plant resistance (APR) to leaf and stripe rusts was conferred by three and five additive genes, respectively. Among them, Lr46/Yr29 was associated with resistance to both rusts in Francolin#1, Lr16 reduced field leaf rust severity by 8 to 9%, and YrF contributed to 10 to 25% reductions in stripe rust severity. The Lr16 region was also associated with a 5 to 16% reduction in stripe rust severity, which is likely due to its linkage with YrF or another unidentified stripe rust APR gene. Significant additive effects on stripe rust were detected between YrF and Yr29. We conclude that APR in Francolin#1 to leaf and stripe rusts involves a combination of seedling and APR genes.

Lr72 Confers Resistance to Leaf Rust in Durum Wheat Cultivar Atil C2000.

Leaf rust, caused by Puccinia triticina (Pt), has become a globally important disease for durum wheat (Triticum turgidum subsp. durum) since the detection of race group BBG/BN, which renders ineffective a widely deployed seedling resistance gene present in several popular cultivars including Mexican cultivars Altar C84 and Atil C2000. The resistance gene continues to play a key role in protecting durum wheat against bread wheat-predominant races since virulence among this race group has not been found. We developed F3 and F5 mapping populations from a cross between Atil C2000 and the susceptible line Atred #1. Resistance was characterized by greenhouse seedling tests using three Pt races. Segregation tests indicated the presence of a single gene, which was mapped to the distal end of 7BS by bulk segregant analysis. The closest marker, wmc606, was located 5.5 cM proximal to the gene. No known leaf rust resistance genes are reported in this region; this gene was therefore designated as Lr72. The presence of Lr72 was further investigated in greenhouse tests in a collection of durum wheat using 13 Pt races. It was concluded that at least one additional gene protects durum wheat from bread wheat-predominant Pt races.

Quantitative trait loci of stripe rust resistance in wheat.

Over thirty publications during the last 10 years have identified more than 140 QTLs for stripe rust resistance in wheat. It is likely that many of these QTLs are identical genes that have been spread through plant breeding into diverse backgrounds through phenotypic selection under stripe rust epidemics. Allelism testing can be used to differentiate genes in similar locations but in different genetic backgrounds; however, this is problematic for QTL studies where multiple loci segregate from any one parent. This review utilizes consensus maps to illustrate important genomic regions that have had effects against stripe rust in wheat, and although this methodology cannot distinguish alleles from closely linked genes, it does highlight the extent of genetic diversity for this trait and identifies the most valuable loci and the parents possessing them for utilization in breeding programs. With the advent of cheaper, high throughput genotyping technologies, it is envisioned that there will be many more publications in the near future describing ever more QTLs. This review sets the scene for the coming influx of data and will quickly enable researchers to identify new loci in their given populations.

QTL analysis of the spring wheat "Chapio" identifies stable stripe rust resistance despite inter-continental genotype × environment interactions.

Chapio is a spring wheat developed by CIMMYT in Mexico by a breeding program that focused on multigenic resistances to leaf rust and stripe rust. A population consisting of 277 recombinant inbred lines (RILs) was developed by crossing Chapio with Avocet. The RILs were genotyped with DArT markers (137 randomly selected RILs) and bulked segregant analysis conducted to supplement the map with informative SSR markers. The final map consisted of 264 markers. Phenotyping against stripe rust was conducted for three seasons in Toluca, Mexico and at three sites over two seasons (total of four environments) in Sichuan Province, China. Significant loci across the two inter-continental regions included Lr34/Yr18 on 7DS, Sr2/Yr30 on 3BS, and a QTL on 3D. There were significant genotype × environment interactions with resistance gene Yr31 on 2BS being effective in most of the Toluca environments; however, a late incursion of a virulent pathotype in 2009 rendered this gene ineffective. This locus also had no effect in China. Conversely, a 5BL locus was only effective in the Chinese environments. There were also complex additive interactions. In the Mexican environments, Yr31 suppressed the additive effect of Yr30 and the 3D locus, but not of Lr34/Yr18, while in China, the 3D and 5BL loci were generally not additive with each other, but were additive when combined with other loci. These results indicate the importance of maintaining diverse, multi-genic resistances as Chapio had stable inter-continental resistance despite the fact that there were QTLs that were not effective in either one or the other region.

Genetic Analysis of Adult Plant Resistance to Yellow Rust and Leaf Rust in Common Spring Wheat Quaiu 3.

Identifying and utilizing rust resistance genes in wheat has been hampered by the continuous and rapid emergence of new pathogen races. A major focus of many wheat breeding programs is achieving durable adult plant resistance (APR) to yellow (stripe) rust (YR) and leaf (brown) rust (LR), caused by Puccinia striiformis and P. triticina, respectively. This study aimed to determine the genetic basis of resistance to YR and LR in the common spring wheat 'Quaiu 3'. To that end, we evaluated 198 F5 recombinant inbred lines (RILs), derived from a cross of susceptible 'Avocet-YrA' with Quaiu 3, for APR to LR and YR in artificially inoculated field trials conducted in Mexico during the 2009 and 2010 growing seasons. High narrow-sense heritability (h2) estimates, ranging between 0.91 and 0.95, were obtained for both LR and YR disease severities for both years. The quantitative and qualitative approaches used to estimate gene numbers showed that, in addition to known resistance genes, there are at least two to three APR genes associated with LR and YR resistance in the RIL population. The moderately effective race-specific resistance gene Lr42 and the pleiotropic slow-rusting APR gene Lr46/Yr29 were found to interact with additional unidentified APR genes. The unidentified APR genes should be of particular interest for further characterization through molecular mapping, and for utilization by wheat breeding programs.

Analysis of leaf and stripe rust severities reveals pathotype changes and multiple minor QTLs associated with resistance in an Avocet × Pastor wheat population.

Leaf rust and stripe rust are important diseases of wheat world-wide and deployment of cultivars with genetic resistance is an effective and environmentally sound control method. The use of minor, additive genes conferring adult plant resistance (APR) has been shown to provide resistance that is durable. The wheat cultivar 'Pastor' originated from the CIMMYT breeding program that focuses on minor gene-based APR to both diseases by selecting and advancing generations alternately under leaf rust and stripe rust pressures. As a consequence, Pastor has good resistance to both rusts and was used as the resistant parent to develop a mapping population by crossing with the susceptible 'Avocet'. All 148 F(5) recombinant inbred lines were evaluated under artificially inoculated epidemic environments for leaf rust (3 environments) and stripe rust (4 environments, 2 of which represent two evaluation dates in final year due to the late build-up of a new race virulent to Yr31) in Mexico. Map construction and QTL analysis were completed with 223 polymorphic markers on 84 randomly selected lines in the population. Pastor contributed Yr31, a moderately effective race-specific gene for stripe rust resistance, which was overcome during this study, and this was clearly shown in the statistical analysis. Linked or pleiotropic chromosomal regions contributing to resistance against both pathogens included Lr46/Yr29 on 1BL, the Yr31 region on 2BS, and additional minor genes on 5A, 6B and 7BL. Other minor genes for leaf rust resistance were located on 1B, 2A and 2D and for stripe rust on 1AL, 1B, 3A, 3B, 4D, 6A, 7AS and 7AL. The 1AL, 1BS and 7AL QTLs are in regions that were not identified previously as having QTLs for stripe rust resistance. The development of uniform and severe epidemics facilitated excellent phenotyping, and when combined with multi-environment analysis, resulted in the relatively large number of QTLs identified in this study.

First Detection of Virulence in Puccinia triticina to Resistance Genes Lr27 + Lr31 Present in Durum Wheat in Mexico.

Leaf rust caused by Puccinia triticina Eriks. was observed during the 2007-2008 crop season in the state of Sonora, Mexico on previously resistant durum wheat (Triticum turgidum L.) cvs. Jupare C2001 (Lr27 + 31) and Banamichi C2004. Single uredinial isolates were made from disease samples collected in the field and tested mostly on 'Thatcher' differentials at seedling or adult-plant stages (1). The isolates were identified as a new race, BBG/BP, resembling race BBG/BN predominant on durum wheat (2), but with additional virulences to resistance genes Lr27 + Lr31 in Gatcher and adult-plant resistance gene Lr12. The new race was also identified in samples collected from durum wheat in the State of Nuevo Leon during the same season. The avirulence/virulence formula of race BBG/BP is Lr1, 2a, 2b, 2c, 3, 3ka, 3bg, 9, 13, 14a, 15, 16, 17, 18, 19, 21, 22a, 24, 25, 26, 28, 29, 30, 32, 35, 37/Lr10, 11, 12, 14b, 20, 23, 27 + 31, 33. Although virulences to Lr27 + Lr31 and Lr12 is known to occur in P. triticina races predominant on bread wheat (T. aestivum L.) (1), to our knowledge, this is the first report of such virulences in the pathogen population on durum wheat. Pure isolates of race BBG/BP are stored in the CIMMYT leaf rust collection. References: (1) R. P. Singh. Plant Dis. 75:790, 1991. (2) R. P. Singh et al. Plant Dis. 88:703, 2004.

Molecular Mapping of a Leaf Rust Resistance Gene on the Short Arm of Chromosome 6B of Durum Wheat.

Leaf rust, caused by Puccinia triticina, is an important disease of durum wheat (Triticum turgidum subsp. durum) worldwide, and the most effective way to control it is through the use of resistant cultivars. A partially dominant leaf rust resistance gene present in the International Maize and Wheat Improvement Center-derived Chilean cv. Guayacan INIA and its sister line Guayacan 2 was mapped to chromosome arm 6BS by identifying linked amplified fragment length polymorphisms (AFLPs) and mapping two of the molecular markers in common wheat (T. aestivum) linkage maps of the International Triticeae Mapping Initiative and Oligoculm × Fukuho-komugi populations. Comparison of infection type responses of the two resistant durums with common wheat testers carrying the previously mapped resistance genes Lr36 and Lr53 on 6BS, and their chromosomal positions, indicated that the resistance gene in durum wheat Guayacan INIA is a new leaf rust resistance gene, which was designated as Lr61. Gene Lr61 is effective against the P. triticina race BBG/BN predominant in northwestern Mexico and other races infecting durum wheat in various countries.

Identification and Molecular Characterization of Leaf Rust Resistance Gene Lr14a in Durum Wheat.

Leaf rust, caused by Puccinia triticina, is an important disease of durum wheat (Triticum turgidum subsp. durum) and only a few designated resistance genes are known to occur in this crop. A dominant leaf rust resistance gene in the Chilean durum cv. Llareta INIA was mapped to chromosome arm 7BL through bulked segregant analysis using the amplified fragment length polymorphism (AFLP) technique, and by mapping three polymorphic markers in the common wheat (T. aestivum) International Triticeae Mapping Initiative population. Several simple sequence repeat (SSR) markers, including Xgwm344-7B and Xgwm146-7B, were associated with the leaf rust resistance gene. Resistance response and chromosomal position indicated that this gene is likely to be Lr14a. The SSR markers Xgwm344-7B and Xgwm146-7B and one AFLP marker also differentiated common wheat cv. Thatcher from the near-isogenic line with Lr14a, as well as durum 'Altar C84' from durum wheat with Lr14a. This is the first report of the presence of Lr14a in durum wheat, although the gene originally was transferred from emmer wheat 'Yaroslav' to common wheat. Lr14a is also present in CIMMYT-derived durum 'Somateria' and effective against Mexican and other P. triticina races of durum origin. Lr14a should be deployed in combination with other effective leaf rust resistance genes to prolong its effectiveness in durum wheat.

First Report of Ramularia cercosporelloides on Carthamus tinctorius in Northwestern Mexico.

Severe yield losses caused by an unidentified fungal disease occurred on safflower in Sonora, Mexico from 2001 to 2006. Leaf spots were pale at the beginning and became sunken lesions, spots turned brown, and diseased tissue became necrotic. Under continuous presence of dew, new infections occurred and the number of lesions increased, coalesced, and eventually the entire leaf and plant turned brown and dried up. The fungus appears as a whitish mold covering the lesions. Observations with a dissecting microscope revealed fungal growth on both leaf surfaces. Spots had minute, gray-to-pink tufts emerging from the host tissues. Stromata were roughly spherical, 35 to 55 µm in diameter, developed under the epidermis near the edge of the spots from which conidiophores arose. Fascicles of conidiophores emerged through stomata from colorless or slightly yellowish stroma and were simple, colorless, or slightly yellowish, as much as 85 µm long and 2.5 to 3 µm wide, with a pigmented scar at the tip. Conidia, formed in chains at scar sites on conidiophores, were colorless, obclavate, 0 to 1 septate, approximately 24 to 32 µm long, and 4.5 to 7 µm wide at the broadest part with a pigmented scar either at the base or at both ends. On the basis of symptoms and fungal morphology, the pathogen was identified as Ramularia cercosporelloides U. Braun & Crous (=Cercosporella carthami) (1). Seven specimens were deposited at the Uppsala University Herbarium as Nos. UPS F-119998 to 120004. Conidia were transferred to water agar plates from symptomatic leaves after 8 days in a humid chamber. Ten single conidia were plated onto a Septoria tritici medium (4 g each of malt extract, yeast extract, and sucrose, and 18 g of agar per liter of water). Colonies of the fungus were white-to-light pink with irregular margins and very slow in growth. Inoculum was produced by transferring 2 ml of distilled water containing conidia onto petri plates filled with Septoria medium. Plates were incubated at 18°C for 12 days. Water suspension was amended with two drops of Tween 20 per liter of inoculum. Pathogenicity of five isolates was confirmed by spraying a suspension of 1.0 × 105 conidia/ml onto five pots of four direct-seeded adult plants of cv. S-518 per isolate. Plants were kept in a dew chamber (20°C) for 48 h under 16 h of darkness and 8 h of light and then in a greenhouse (20 to 24°C). After 2 weeks, leaves developed the characteristic spots, and R. cercosporelloides was isolated from symptomatic tissue. To our knowledge, this is the first report of R. cercosporelloides on safflower, not only in Mexico, but also on the American continent. Reference: (1) U. Braun. A monograph of Cercosporella, Ramularia and allied genera (Phytopathogenic Hyphomycetes). Vol 2. IHW-Verlag. Eching bei Munchen, 1998.

Effect of Leaf Rust on Grain Yield and Yield Traits of Durum Wheats with Race-Specific and Slow-Rusting Resistance to Leaf Rust.

Leaf rust, caused by Puccinia triticina, is an important disease of durum wheat (Triticum turgidum) in many countries. We compared the effectiveness of different types of resistance in International Maize and Wheat Improvement Center-derived durum wheat germ plasm for protecting grain yield and yield traits. In all, 10 durum wheat lines with race-specific resistance, 18 with slow-rusting resistance, and 2 susceptible were included in two yield loss trials sown on different planting dates in Mexico with and without fungicide protection under high disease pressure. Eight genotypes with race-specific resistance were immune to leaf rust. Durum wheat lines with slow-rusting resistance displayed a range of severity responses indicating phenotypic diversity. Mean yield losses for susceptible, race-specific, and slow-rusting genotypes were 51, 5, and 26%, respectively, in the normal sowing date trial and 71, 11, and 44% when sown late. Yield losses were associated mainly with a reduction in biomass, harvest index, and kernels per square meter. Slow-rusting durum wheat lines with low disease levels and low yield losses, as well as genotypes with low yield losses despite moderate disease levels, were identified. Such genotypes can be used for breeding durum wheat genotypes with higher levels of resistance and negligible yield losses by using strategies that previously have been shown to be successful in bread wheat.

New Genes for Leaf Rust Resistance in CIMMYT Durum Wheats.

Leaf rust, caused by Puccinia triticina, has become an important disease of durum wheat (Triticum turgidum) in Mexico since the detection in 2001 of BBG/BN, a new race virulent on all common cultivars and on more than 80% of CIMMYT's durum wheat collection. We investigated the genetic basis and diversity of resistance in nine durum genotypes that are highly resistant to the new race. These resistant durums were crossed with the susceptible cv. Atil C2000 and intercrossed in a half diallel arrangement. Five diverse sources of resistance were identified by evaluating parents, F1, F2, and F3 populations in greenhouse and/or field trials under artificial epidemics of race BBG/BN. The same pair of partially dominant complementary genes determined resistance in Jupare C2001, Hualita, and Pohowera. Somateria and Llareta INIA shared the same dominant resistance gene, whereas a partially dominant gene conferred resistance in two sister lines, Guayacan 2 and Guayacan INIA. A different partially dominant gene present in Storlom was linked in repulsion to another partially dominant gene in Camayo. These diverse resistance genes can be used effectively to control leaf rust, preferably by deploying them in combinations.