Pseudo-black Chaff Research Articles

Phenotypic and molecular characterization of pleiotropic resistance to rusts and powdery mildew in spring wheat

An attempt was undertaken to comprehend how phenotypic markers, leaf tip necrosis (LTN), and pseudo black chaff (PBC), are associated with pleiotropic resistance genes in wheat (Triticum spp.). Pleiotropic resistance to stripe rust (Puccinia striiformis f. sp. tritici), leaf rust (P. triticina), stem rust (P. graminis f. sp. tritici) and powdery mildew (Blumeria graminis f. sp. tritici) was evaluated in 57 Egyptian spring wheat cultivars at seedling stage in greenhouse and at adult plant stage in field during 2021/22 and 2022/23. At seedling stage, pleiotropic resistance rated moderately resistance (MR) was observed in five cultivars, Misr-3 (all three rusts), Misr-4 (stripe rust, leaf rust, powdery mildew), Giza-168 (leaf rust, stem rust, powdery mildew), Sakha-94 and Sids-13 (leaf rust, stem rust). At adult plant stage, Misr-4 outperformed all other evaluated cultivars, exhibiting a high level of pleiotropic adult plant resistance (PAPR) against all diseases investigated. High levels of PAPR were also noticed against the three rusts in nine cultivars, Misr-3, Sakha-93, Sakha-94, Sakha-95, Giza-156, Giza-168, Giza-171, Gemmeiza-10 and Sids-13, and moderate levels against powdery mildew in Misr-3, Sakha-95, Giza-156. Four PAPR genes were characterized in cultivars based on phenotypic and molecular markers. Phenotypic markers (LTN and PBC) were observed in 23 cultivars. Molecular marker csLV34 indicated the presence of Lr34/Yr18/Sr57/Pm38/Ltn1 in six cultivars, whilst marker Xwmc44 indicated that Lr46/Yr29/Sr58/Pm39/Ltn2 was present in five cultivars. Marker Xcfd71 indicated the presence of Lr67/Yr46/Sr55/Pm46/Ltn3 in 19 cultivars. Gene Sr2/Yr30/Lr27/Pbc was identified with marker csSr2 in three cultivars. A complete association between LTN and molecular markers was recorded, while a strong association (r = 0.73) was recorded between PBC and molecular markers. Findings demonstrated the reliability of phenotypic markers in predicting pleiotropic resistance in wheat, which would facilitate marker-assisted selection in breeding programs.

Enhanced stripe rust resistance obtained by combining Yr30 with a widely dispersed, consistent QTL on chromosome arm 4BL.

YrFDC12 and PbcFDC, co-segregated in chromosome 4BL, and significantly interacted with Yr30/Pbc1 to enhance stripe rust resistance and to promote pseudo-black chaff development. Cultivars with durable resistance are the most popular means to control wheat stripe rust. Durable resistance can be achieved by stacking multiple adult plant resistance (APR) genes that individually have relatively small effect. Chinese wheat cultivars Ruihua 520 (RH520) and Fengdecun 12 (FDC12) confer partial APR to stripe rust across environments. One hundred and seventy recombinant inbred lines from the cross RH520 × FDC12 were used to determine the genetic basis of resistance and identify genomic regions associated with stripe rust resistance. Genotyping was carried out using 55K SNP array, and eight quantitative trait loci (QTL) were detected on chromosome arms 2AL, 2DS, 3BS, 4BL, 5BL (2), and 7BL (2) by inclusive composite interval mapping. Only QYr.nwafu-3BS from RH520 and QYr.nwafu-4BL.2 (named YrFDC12 for convenience) from FDC12 were consistent across the four testing environments. QYr.nwafu-3BS is likely the pleiotropic resistance gene Sr2/Yr30. YrFDC12 was mapped in a 2.1-cM interval corresponding to 12Mb and flanked by SNP markers AX-111121224 and AX-89518393. Lines harboring both Yr30 and YrFDC12 displayed higher resistance than the parents and expressed pseudo-black chaff (PBC) controlled by loci Pbc1 and PbcFDC12, which co-segregated with Yr30 and YrFDC12, respectively. Both marker-based and pedigree-based kinship analyses revealed that YrFDC12 was inherited from founder parent Zhou 8425B. Fifty-four other wheat cultivars shared the YrFDC12 haplotype. These results suggest an effective pyramiding strategy to acquire highly effective, durable stripe rust resistance in breeding.

Adult Plant Slow Rusting Genes Confer High Levels of Resistance to Rusts in Bread Wheat Cultivars From Mexico.

Rust diseases continuously threaten global wheat production: stem rust, leaf rust, and yellow rust caused by Puccinia graminis f. sp. tritici, Puccinia triticina, and Puccinia striiformis f. sp. tritici, respectively. Recent studies indicated that the average losses from all these three rusts reached up to 15.04 million tons per year, which is equivalent to an annual average loss of around US $2.9 billion per year. The major focus of Mexican and worldwide breeding programs is the release of rust resistant cultivars, as this is considered the best option for controlling rust diseases. In Mexico, the emphasis has been placed on genes that confer partial resistance in the adult plant stage and against a broad spectrum of rust races since the 1970s. In this study, a set of the first-generation tall varieties developed and released in the 1940s and 1950s, the first semi-dwarfs, and other releases in Mexico, all of which showed different levels of rust resistance have been phenotyped for the three rust diseases and genotyped. Results of the molecular marker detection indicated that Lr34, Lr46, Lr67, and Lr68 alone or in different gene combinations were present among the wheat cultivars. Flag leaf tip necrosis was present in all cultivars and most were positive for brown necrosis or Pseudo Black Chaff associated with the Sr2 stem rust resistance complex. The phenotypic responses to the different rust infections indicate the presence of additional slow rusting and race-specific resistance genes. The study reveals the association of the slow rusting genes with durable resistance to the three rusts including Ug99 in cultivars bred before the green revolution such as Frontera, Supremo 211, Chapingo 48, Yaqui 50, Kentana 52, Bajio 52, Bajio 53, Yaqui 53, Chapingo 53, Yaktana Tardio 54, and Mayo 54 and their descendants after intercrossing and recombination. These slow rusting genes are the backbone of the resistance in the current Mexican germplasm.

Disease Resistance Evaluation of Elite CIMMYT Wheat Lines Containing the Coupled Fhb1 and Sr2 Genes.

Fusarium head blight (FHB) and stem rust are among the most devastating diseases of wheat worldwide. Fhb1 is the most widely utilized and the only isolated gene for FHB resistance, while Sr2 is a durable stem rust resistance gene used in rust-prone areas. The two loci are closely linked on the short arm of chromosome 3B and the two genes are in repulsion phase among cultivars. With climate change and the shift in Fusarium populations, it is imperative to develop wheat cultivars resistant to both diseases. The present study was dedicated to developing wheat germplasm combining Fhb1 and Sr2 resistance alleles in the International Maize and Wheat Improvement Center (CIMMYT)'s elite cultivars' backgrounds. Four recombinant inbred lines (RILs) in Hartog background that have the resistant Fhb1 and Sr2 alleles in coupled phase linkage were crossed with seven CIMMYT bread wheat lines, resulting in 208 lines. Molecular markers for both genes were employed in addition to the use of pseudo-black chaff (PBC) as a phenotypic marker for the selection of Sr2. At various stages of the selection process, progeny lines were assessed for FHB index, Fusarium damaged kernels (FDK), stem rust, and PBC expression as well as other diseases of interest (stripe rust and leaf spotting diseases). The 25 best lines were selected for CIMMYT's wheat breeding program. In addition to expressing resistance to FHB, most of these 25 lines have an acceptable level of resistance to other tested diseases. These lines will be useful for wheat breeding programs worldwide and potentially speed up the resistance breeding efforts against FHB and stem rust.

Validation and Marker-Assisted Selection of Stem Rust Resistance Gene Sr2 in Indian Wheat Using Gel-Based and Gel-Free Methods

Stem rust resistance gene Sr2 is an important slow-rusting gene which has provided resistance against stem rust for many years. The Sr2 gene has durable resistance against all stem rust pathogens including the Ug99 group of races. It shows recessive inheritance and is linked with two phenotypic markers, Pseudo Black Chaff (PBC) and High Temperature-Induced Seedling Chlorosis (HTISC). However, direct screening as well as phenotypic marker-based screening for selection of Sr2 is difficult due to the effect of environmental factors and genetic background. Microsatellite marker Xgwm533 linked to Sr2 gene is useful for rapid screening of large populations. Here we report validation and use of Xgwm533 for screening 21 wheat lines and 24 wheat genotypes for the presence of the Sr2 gene. Furthermore, we also report the presence of a new, smaller allele (97 bp) of Xgwm533 locus in a few genotypes that was also confirmed by sequencing. We also demonstrate a new SYBR green dye, melt-curve/profile-based assay for convenient screening of the Xgwm533 locus, where the presence of different alleles can be differentiated in a gel-free manner.

Phenotypes Conferred by Wheat Multiple Pathogen Resistance Locus, Sr2, Include Cell Death in Response to Biotic and Abiotic Stresses

The wheat Sr2 locus confers partial resistance to four biotrophic pathogens: wheat stem rust (Puccinia graminis f. sp. tritici), leaf rust (P. triticina), stripe rust (P. striiformis f. sp. tritici), and powdery mildew (Blumeria graminis f. sp. tritici). In addition, Sr2 is linked with a brown coloration of ears and stems, termed pseudo-black chaff (PBC). PBC, initially believed to be elicited by stem rust infection, was subsequently recognized to occur in the absence of pathogen infection. The current study demonstrates that the resistance response to stem rust is associated with the death of photosynthetic cells around rust infection sites in the inoculated leaf sheath. Similarly, Sr2-dependent resistance to powdery mildew was associated with the death of leaf mesophyll cells around mildew infection sites. We demonstrate that PBC occurring in the absence of pathogen inoculation also corresponds with death and the collapse of photosynthetic cells in the affected parts of stems and ears. In addition, Sr2-dependent necrosis was inducible in leaves by application of petroleum jelly or by heat treatments. Thus, Sr2 was found to be associated with cell death, which could be triggered by either biotic or abiotic stresses. Our results suggest a role for the Sr2 locus in controlling cell death in response to stress.

Characterization of stem, stripe and leaf rust resistance in Tajik bread wheat accessions

Stem rust [causal organism: Puccinia graminis f. sp. tritici (Pgt)], stripe rust [Puccinia striiformis f. sp. tritici (Pst)], and leaf rust [Puccinia triticina (Pt)] are important fungal diseases of wheat in Central Asia and worldwide. Therefore, identification of seedling and adult plant resistance (APR) genes is of major importance for the national wheat breeding program in many countries. The objectives of this study were to identify genes that confer seedling and APR resistances in widely grown wheat cultivars, landraces and advanced lines from Tajikistan. A total of 41 wheat accessions were inoculated with eleven races of Pgt, twelve races of Pst and nine races of Pt for postulation of Sr (stem rust), Yr (yellow or stripe rust), and Lr Lr (leaf rust) resistance genes at the seedling stage. In addition, all of the accessions were tested in field trials for the response to stem rust and stripe rust. Genes for seedling stem rust resistance (i.e. Sr5, Sr6, Sr11, Sr31, and Sr38), stripe rust resistance (Yr9, Yr17, and Y27), and leaf rust resistance (Lr16 and Lr26) were postulated in the Tajik wheat. The presence of the pleiotropic APR genes Sr2/Yr30/Lr27 (associated with pseudo-black chaff phenotype) and Lr34/Yr18/Sr57 (associated with leaf tip necrosis phenotype), and also Lr37 were assessed in the field and confirmed with linked molecular markers. In most of the wheat accessions, resistance genes could not be postulated because their infection types did not match the avirulence or virulence profile of the Pgt, Pst and Pt races tested. Six, seven, and nine accessions were identified that likely possess new genes for resistance to stem rust, stripe rust, and leaf rust, respectively, which have not been described previously. The research demonstrates the presence of effective seedling resistance and APR genes in widely grown wheat accessions that could facilitate further rust resistance breeding in the national wheat breeding program in Tajikistan.

Identification of recombinants carrying stripe rust resistance gene Yr57 and adult plant stem rust resistance gene Sr2 through marker‐assisted selection

AbstractMany stem rust resistance genes have been formally named in wheat. Adult plant stem rust resistance gene Sr2 was mapped in the short‐arm of chromosome 3B. Stripe rust resistance gene Yr57, identified in Aus91463, was mapped about 5 cM away from Sr2 based on its linkage with Sr2‐linked marker gwm533. The objective of this study was to combine Sr2 and Yr57 in a single genotype. A mapping population containing 107 recombinant inbred lines was developed from a cross between Aus91463‐Yr57 and Hartog‐Sr2. This population was tested at the seedling stage in the glasshouse for variation in stripe rust response, and high temperature induced Sr2‐linked seedling chlorosis. The RIL population was screened for Sr2‐linked pseudo black chaff phenotype at the adult plant stage in field. Five recombinants carrying Sr2 and Yr57 in coupling were detected using phenotypic and marker data. Four recombinants also carried leaf rust resistance gene Lr23 from Aus91463. These recombinants are being used as triple rust resistance source in the Australian Cereal Rust Control Program.

Identification of Resistance Sources to Wheat Stem Rust from Introduced Genotypes in Kenya

Stem rust Puccinia graminis Pers. f. sp. tritici of wheat is the most important disease in Kenya. Emergence of race Ug99 and other variants virulent to host resistance genes including Sr31 has rendered 95% of Kenyan cultivars susceptible. This study aimed to identify new sources of resistance to stem rust in a collection of exotic genotypes. Three hundred and sixteen wheat genotypes were screened at the Kenya Agricultural and Livestock Research Organization (KALRO) in Njoro for two seasons in 2015. The host reaction to disease was evaluated based on the modified Cobb scale. The relative Final Rust Severity (rFRS), Average Coefficient of Infection (ACI) and relative Area Under Disease Progress Curve (rAUDPC) were used to characterize the genotypes for stem rust resistance. Agronomic traits were also recorded. Six genotypes namely ALBW-100, ALBW- 204, EPCBW-261, EPCBW-295, PCHP-309 and PCHPBW-310 with significantly low ACI, rAUDPC and rFRS were identified. Thirty five genotypes showed Pseudo-Black Chaff (PBC) phenotype associated with resistant gene Sr2, a source of partial resistance in wheat. The genotypes also showed low disease severity (20-25%) and Moderately Susceptible (MS) – Susceptible (S) infection types in both seasons. Genotypes had significant differences (p ≤ 0.05) on plant height, 1000-kernel weight and number of tillers indicating genetic variation which could be exploited in breeding for resistance to stem rust. The negative relationship between agronomic variables involving plant height, spikelet length and 1000-kernel weight showed harmful effects of stem rust on plant characteristics including yield. The stem rust resistant genotypes with good agronomic traits could be introgressed into adapted Kenyan backgrounds while the genotypes showing presence of PBC could be utilized to develop durable stem rust resistant wheat. Inheritance studies to elucidate the exact genes conferring resistance to stem rust could be conducted for breeders to exploit their genetic variability.

Rapid Phenotyping Adult Plant Resistance to Stem Rust in Wheat Grown under Controlled Conditions.

Stem rust (SR) or black rust caused by Puccinia graminis f. sp. tritici is one of the most common diseases of wheat (Triticum aestivum L.) crops globally. Among the various control measures, the most efficient and sustainable approach is the deployment of genetically resistant cultivars. Traditionally, wheat breeding programs deployed genetic resistance in cultivars, but unknowingly this is often underpinned by a single seedling resistance gene, which is readily overcome by the pathogen. Nowadays, adult plant resistance (APR) is a widely adopted form of rust resistance because more durable mechanisms often underpin it. However, only a handful of SR APR genes are available, so breeders currently strive to combine seedling and APR genes. Phenotyping adult wheat plants for resistance to SR typically involves evaluation in the field. But establishing a rust nursery can be challenging, and screening is limited to once a year. This slows down research efforts to isolate new APR genes and breeding of genetically resistant cultivars.In this study, we report a protocol for rapid evaluation of adult wheat plants for resistance to stem rust. We demonstrate the technique by evaluating apanel of 16 wheat genotypes consisting of near isogenic lines (NILs) for known Sr genes (i.e., Sr2, Sr33, Sr45, Sr50, Sr55, Sr57, and Sr58) and three landraces carrying uncharacterized APR from the N. I. Vavilov Institute of Plant Genetic Resources (VIR). The method can be completed in just 10weeks and involves two inoculations: first conducted at seedling stage and a second at the adult stage (using the same plants). The technique can detect APR, such as that conferred by APR gene Sr2, along with pseudo-black chaff (the morphological marker). Phenotyping can be conducted throughout the year, and is fast and resource efficient. Further, the phenotyping method can be applied to screen breeding populations or germplasm accessions using local or exotic races of SR.

Genome-Wide Association Mapping for Leaf Tip Necrosis and Pseudo-black Chaff in Relation to Durable Rust Resistance in Wheat.

The partial rust resistance genes Lr34 and Sr2 have been used extensively in wheat (Triticum aestivum L.) improvement, as they confer exceptional durability. Interestingly, the resistance of Lr34 is associated with the expression of leaf tip necrosis (LTN) and Sr2 with pseudo-black chaff (PBC). Genome-wide association mapping using CIMMYT's stem rust resistance screening nursery (SRRSN) wheat lines was done to identify genotyping-by-sequencing (GBS) markers linked to LTN and PBC. Phenotyping for these traits was done in Ithaca, New York (fall 2011); Njoro, Kenya (main and off-seasons, 2012), and Wellington, India (winter, 2013). Using the mixed linear model (MLM), 18 GBS markers were significantly associated with LTN. While some markers were linked to loci where the durable leaf rust resistance genes Lr34 (7DS), Lr46 (1BL), and Lr68 (7BL) were mapped, significant associations were also detected with other loci on 2BL, 5B, 3BS, 4BS, and 7BS. Twelve GBS markers linked to the Sr2 locus (3BS) and loci on 2DS, 4AL, and 7DS were significantly associated with PBC. This study provides insight into the complex genetic control of LTN and PBC. Further efforts to validate and study these loci might aid in determining the nature of their association with durable resistance.

Genetic Gain from Phenotypic and Genomic Selection for Quantitative Resistance to Stem Rust of Wheat

Stem rust of wheat (Triticum aestivum L.) caused by Puccinia graminis f. sp. tritici Eriks. and E. Henn. is a globally important disease that can cause severe yield loss. Breeding for quantitative stem rust resistance (QSRR) is important for developing cultivars with durable resistance. Genomic selection (GS) could increase rates of genetic gain for quantitative traits, but few experiments comparing GS and phenotypic selection (PS) have been conducted. Our objectives were to (i) compare realized gain from GS based on markers only with that of PS for QSRR in spring wheat using equal selection intensities; (ii) determine if gains agree with theoretical expectations; and (iii) compare the impact of GS and PS on inbreeding, genetic variance, and correlated response for pseudo-black chaff (PBC), a correlated trait. Over 2 yr, two cycles of GS were performed in parallel with one cycle of PS, with each method replicated twice. For GS, markers were generated using genotyping-by-sequencing, the prediction model was initially trained using historical data, and the model was updated before the second GS cycle. Overall, GS and PS led to a 31 ± 11 and 42 ± 12% increase in QSRR and a 138 ± 22 and 180 ± 70% increase in PBC, respectively. Genetic gains were not significant but were in agreement with expectations. Per year, gains from GS and PS were equal, but GS led to significantly lower genetic variance. This shows that while GS and PS can lead to equal rates of short-term gains, GS can reduce genetic variance more rapidly. Further work to develop efficient GS implementation strategies in spring wheat is warranted.

Different QTLs are associated with leaf rust resistance in wheat between China and Mexico

The wheat line ‘Chapio’ is resistant to leaf rust, caused by Puccinia triticinia, and was derived from a breeding programme that focuses on multi-genic resistance to provide durability. This line was crossed with the susceptible ‘Avocet’ to develop an F6 recombinant inbred line population. The population was phenotyped for leaf rust severity in two environments each in Mexico and China. There were significant differences in the loci providing resistance between the two intercontinental regions. The Lr34 locus had large effects in both Mexico and China, highlighting its importance in providing a basis for broad-spectrum resistance. The Lr46 locus on chromosome 1BL and a 3D locus had effects in Mexico but not in China. Presence of Sr2 was determined by the phenotypic marker of pseudo-black chaff and was mapped to chromosome 3BS. This region was associated with a QTL that had strong effects in China but no significant effect in Mexico, as did a locus on chromosome 4B. Seedling tests on the parents indicated that the 3B locus was not the complimentary gene Lr27, but the 4B locus was in the same position as Lr31 (or Lr12). Further investigations showed that these loci worked independently and additively in adult plants. Chapio was bred for quantitative, non-race-specific resistance under strong phenotypic selection for leaf rust in Mexico. It is interesting that different QTLs contribute to this resistance in another country, and these results suggest that environmental effects, as well as race specificity, can play a role in expression of resistance.

Studies of the genetics of inheritance of stem rust resistance in bread wheat

Pgt race TTKSK (Ug99) has a wide virulence range with respect to currently grown wheat cultivars worldwide. Aspects of migration, mutation, recombination and selection in the pathogen have led to previously deployed stem rust resistance genes being ineffective. Race TTKSK has further evolved to acquire virulence for resistance genes such as Sr24 (race Ug99 + Sr24) and Sr36 (Ug99 + Sr36). Five resistant wheat lines (KSL-2, KSL-3, KSL-5, KSL-12 and KSL-19) which were resistant in tests during 2008, 2009 and 2010 were used as parents in crosses with stem rust susceptible line CACUKE to develop genetic populations for determining the inheritance of resistance to stem rust. F3 populations were evaluated at KARI Njoro in the 2012 off season and 2012 to 2013 main season. The adult plant stem rust responses were scored using the modified Cobb’s scale. The F2:3 lines of population(s) exhibiting qualitative variation were grouped as homozygous resistant (HR), segregating (Seg) and homozygous susceptible (HS). Heavy disease pressure was present during the cropping seasons with the check CACUKE displaying 90% susceptibility. Chi square analysis revealed that the segregation data in the parent KSL-2 did not deviate significantly from the single gene model (1:2:1) suggesting that the resistance to stem rust is conditioned by a single dominant gene. The Chi square test also revealed that the stem rust resistance in the parents KSL-3, KSL-5, KSL-12 and KSL-19 was conditioned by two genes. The families from the KSL-2 and KSL-3 crosses also segregated for the presence of the pseudo black chaff implying that the Sr2 gene could be present in the background of these wheat parents. The superior transgressive segregants identified in these crosses will be used in breeding. Keywords: Genetic, wheat, gene, resistance African Journal of Biotechnology Vol. 12(21), pp. 3153-3159

Identification and mapping in spring wheat of genetic factors controlling stem rust resistance and the study of their epistatic interactions across multiple environments

Stem rust (Puccinia graminis f. sp. tritici) is responsible for major production losses in hexaploid wheat (Triticum aestivum L.) around the world. The spread of stem rust race Ug99 and variants is a threat to worldwide wheat production and efforts are ongoing to identify and incorporate resistance. The objectives of this research were to identify quantitative trait loci (QTL) and to study their epistatic interactions for stem rust resistance in a population derived from the Canadian wheat cultivars AC Cadillac and Carberry. A doubled haploid (DH) population was developed and genotyped with DArT® and SSR markers. The parents and DH lines were phenotyped for stem rust severity and infection response to Ug99 and variant races in 2009, 2010 and 2011 in field rust nurseries near Njoro, Kenya, and to North American races in 2011 and 2012 near Swift Current, SK, Canada. Seedling infection type to race TTKSK was assessed in a bio-containment facility in 2009 and 2012 near Morden, MB. Eight QTL for stem rust resistance and three QTL for pseudo-black chaff on nine wheat chromosomes were identified. The phenotypic variance (PV) explained by the stem rust resistance QTL ranged from 2.4 to 48.8 %. AC Cadillac contributed stem rust resistance QTL on chromosomes 2B, 3B, 5B, 6D, 7B and 7D. Carberry contributed resistance QTL on 4B and 5A. Epistatic interactions were observed between loci on 4B and 5B, 4B and 7B, 6D and 3B, 6D and 5B, and 6D and 7B. The stem rust resistance locus on 6D interacted synergistically with 5B to improve the disease resistance through both crossover and non-crossover interactions depending on the environment. Results from this study will assist in planning breeding for stem rust resistance by maximizing QTL main effects and epistatic interactions.

English

Wheat is an important staple food crop in sub-Saharan Africa. However, the current wheat consumption of 900 000 tons in Kenya outweighs the wheat production of 350 000 tons given the high population growth and inflation. The stem rust currently poses the greatest threat to wheat production due to the emergence of the virulent race of thePuccinia graminis f. sp tritici, Ug99 (TTKS) and its variants Ug99 + Sr24 (TTKSK) andUg99 + Sr36 (TTKST) leading to about 70 to 100% yield losses. This study aimed at evaluating twenty-five wheat genotypes for both field and seedling resistance to stem rust. The genotypes were grown in an alpha lattice design and in two replicates both in the greenhouse and in the field at Kenya Agricultural Research Institute (KARI), Njoro.  The seedling stage infection types were scored based on Stakman et al. (1962) scale. At the adult plant stages, the stem rust disease severity was based on modified Cobb’s scale. The genotypes showed diverse seedling and adult plant resistance responses. The most resistant entries, KSL-2, KSL-3 and KSL-20 also exhibited the pseudo black chaff (PBC) trait implying they contain the Sr2 gene in their background; the basis of breeding for durable resistance to stem rust in wheat. These lines with high stem rust resistance could be backcrossed to the adapted and high yielding but susceptible Kenyan wheat varieties to avert further wheat yield declines.   Key words: Stem rust, wheat, Ug99, resistance, disease severity.

Identification and Evaluation of Sources of Resistance to Stem Rust Race Ug99 in Wheat.

The race Ug99 of Puccinia graminis f. sp. tritici causing stem rust disease of wheat was initially identified in Uganda in 1998. It was designated as TTKSK based on the North American nomenclature and has caused periodic losses to wheat crops in East Africa. Ug99 has recently moved out of Africa to Yemen and West Asia. The most effective approach to prevent losses from stem rust is through the deployment of resistant cultivars. More effective sources of resistance need to be identified and incorporated in the existing commercial cultivars. The first Stem Rust Resistance Screening Nursery (1stSRRSN) assembled by the International Maize and Wheat Improvement Center (CIMMYT) consisted of elite and advanced CIMMYT bread wheat lines and was evaluated for resistance to Ug99 in Njoro, Kenya for four consecutive seasons (2005 to 2007). Seedling reactions were determined in the greenhouse at the Cereal Disease Laboratory, St. Paul, MN. Two race-specific genes, Sr24 and Sr25, were found to confer resistance to Ug99, although Sr24 became ineffective to a mutant race of Ug99, TTKST, in 2007. Three previously uncharacterized genes, one each from synthetic wheat, Chinese germplasm, and other genetic backgrounds, were detected. Although 30% of the screened lines were susceptible in the seedling stage, these lines displayed various levels of adult plant resistance (APR) in the field tests. Presence of the APR gene Sr2, identified based on the pseudo-black chaff phenotype on glumes and darkened internode, was common in wheat lines with APR. The information on the resistance identified in the 1stSRRSN constitutes an important source for breeding wheat for durable resistance.

BAC-derived markers for assaying the stem rust resistance gene, Sr2, in wheat breeding programs

Durable broad-spectrum, adult-plant stem rust resistance in wheat conferred by the Sr2 gene has remained effective against Pucciniagraminis f. sp tritici worldwide for more than 50 years. The Sr2 gene has been positioned on the physical map of wheat to the distal 25% portion of the short arm of chromosome 3B. Selection for this gene in wheat breeding programs within Australia has been performed so far through the use of the linked pseudo black chaff (PBC) phenotype and of the microsatellite markers Xgwm389 and Xgwm533 that flank the gene. The molecular markers flank a genetic interval of approximately 4 cM equating to a physical distance of over 10 Mbp. Recently, a 3B-specific BAC library was developed and a physical map established for this region. Analysis of the sequence of minimal tiling path-BAC clones within the region containing the Sr2 gene enabled the development of three new markers that were mapped within the Xgwm389–Xgwm533 genetic interval and tightly linked to the Sr2 gene. Screening a wide range of germplasm containing the Sr2 gene with these markers demonstrated their usefulness for marker-assisted selection in Australian wheat breeding programs.

Evaluation of seedling and adult plant resistance to stem rust in European wheat cultivars

A total of 105 European wheat cultivars were assessed for seedling and adult plant resistance (APR) to stem rust using an array of Australian isolates of Puccinia graminis f. sp. tritici. Twenty-seven cultivars were susceptible at both seedling and adult plant growth stages. Twelve catalogued seedling stem rust resistance genes (Sr7b, Sr8a, Sr8b, Sr9b, Sr9g, Sr11, Sr15, Sr17, Sr29, Sr31, Sr36 and Sr38) were detected in the remaining cultivars, and 13 cultivars carried additional seedling resistance genes that could not be postulated with the isolates used. Low levels of APR to stem rust were found in the cultivars Artaban, Forno, Mec, Mercia, Pandas and Vlada. Although the genetic identity of this APR was not determined, it was clear that the only designated stem rust APR gene Sr2 was not present in any of the cultivars tested based on the absence of the linked traits seedling chlorosis and pseudo black chaff. One of these cultivars, Forno, is believed to carry the leaf rust APR gene Lr34, previously reported to be associated with improved resistance to stem rust. A detailed genetic characterisation of the APRs in these cultivars will be needed to understand their modes of inheritance and relationships with catalogued stem rust resistance genes. Such knowledge may help in developing cultivars with effective gene combinations that confer higher levels of protection.

Fine genetic mapping fails to dissociate durable stem rust resistance gene Sr2 from pseudo-black chaff in common wheat (Triticum aestivum L.)

The broad-spectrum stem rust resistance gene Sr2 has provided protection in wheat against Puccinia graminis Pers. f. sp. tritici for over 80 years. The Sr2 gene and an associated dark pigmentation trait, pseudo-black chaff (PBC), have previously been localized to the short arm of chromosome 3B. In a first step towards the positional-based cloning of Sr2, we constructed a high-resolution map of this region. The wheat EST (wEST) deletion bin mapping project provided tightly linked cDNA markers. The rice genome sequence was used to infer the putative gene order for orthologous wheat genes and provide additional markers once the syntenic interval in rice was identified. We used this approach to map six wESTs that were collinear with the physical order of the corresponding genes on rice chromosome 1 suggesting there are no major re-arrangements between wheat and rice in this region. We were unable to separate by recombination the tightly linked morphological trait, PBC from the stem rust resistance gene suggesting that either a single gene or two tightly linked genes control both traits.