Stripe rust is a worldwide constraint to wheat production. The rust pathogens are assumed to have originated in the Caucasus, from which they disseminated into Western Europe and Asia (Zhukovsky 1965, Euphytica, 14; Stubbs 1985, The Cereal Rusts II). Rust surveys are a useful means to provide information on distribution. More than 400 wheat fields were monitored for rust incidence and severity and for collection of samples at 20 locations in Georgia during 2010-2013. The majority of wheat fields were occupied by Russian cultivars and Bezostaya-1 was the most common, followed by Copper and American cv. Jagger. Yellow rust was the most widely distributed rust, with>65% of fields showing its presence. In 2009 yellow rust incidence was moderate to high. Abundant overwintering inoculum, susceptible cultivars and favorable conditions resulted in severe epidemics in late May and early June 2010. Mean field incidence and severities were 74.8 and 84.6% in the Kakheti zone, and 70 and 68.2% in Kvemo Kartli. Incidence was lower in the following years due to drought and high late spring temperatures. Bezostaya 1 and Jagger showed moderately susceptible reactions to all three rust, but Copper was moderately resistant. Accessions of Georgian endemic species T. carthlicum, T. timopheevi, T. macha, T. georgicum and T. monococcum were resistant to all three rusts.
Primary Author: Natsarishvili, Institute of Phytopathology and Biodiversity, Batumi Shota Rustaveli State University,Georgia
Keywords: Georgia, surveillance, stripe rust
Stripe rust (Puccinia striiformis f. sp. tritici) is a devastating disease of wheat production world-wide. Yr5 is a race-specific resistance gene effective to all races which have been identified in the U.S. Therefore, it has been increasingly used for cultivar development in the U.S. The goal of this study was to identify “breeder friendly” SNP markers associated with Yr5 through linkage mapping in a spring wheat recombinant inbred line (RIL) population and validate these markers with an additional RIL population, Pacific Northwest (PNW) wheat elite breeding materials, and a world-wide collection of spring wheat. RIL populations were developed from a cross between moderately susceptible experimental line WA8149 and two elite Yr5 donors, S0900317 and S0900163. Seventy PNW spring and winter wheat lines either carrying Yr5 or not, and 3,040 hexaploid spring wheat lines from a world-wide collection were used to further validate flanking markers for Yr5. Seedling resistance response to isolates Pstv-37 and Pstv-51 was evaluated for these RIL populations. These RILs were genotyped with 46 KASP markers located on chromosome 2B in addition to previously reported Yr5 linked markers, wmc175 (Murphy et al. 2009) and TaAffix65234.1.S1_at (McGrann et al. 2014). Linkage mapping was conducted by MapDisto v1.7.5. A total 10 markers localized Yr5 within 0.7 cM chromosome region in the WA8149/S0900317 population of 300 RILs. The same set of markers also localized Yr5 within 3 cM chromosome region in the WA8149/S0900163 population of 274 RILs. Newly identified KASP markers were closer to Yr5 than wmc175 and TaAffix65234.1.S1_at in these two populations. These markers were further validated with PNW winter and spring wheat and a world-wide collection of spring wheat. These KASP markers flanking Yr5 reliably selected for germplasm carrying Yr5 and will assist in pyramiding different resistances into breeding lines to develop more durable stripe rust resistant cultivars.
Primary Author: Nauoka, Department of Crop and Soil Sciences, Washington State University, USA
Keywords: stripe rust, Yr5, SNP
The Sr2 gene has been used extensively in bread wheat improvement for durable stem rust resistance. Interestingly, the resistance of Sr2, associated with the pleotrophic gene Pbc expressed as pseudo-black chaff (PBC), is tightly linked with Yr30/Lr27/Pm genes conferring multiple disease resistance. The linkage map of chromosome 3BS revealed that Sr2 is 0.43cM away from lesion mimic (lm) locus. The RIL population (Yangmai#6 ? Sonalika) of 88 lines including parents where Sonalika carries Sr2 and lm while Yangmai#6 is deficient to both was evaluated for three years (2013-2016). The objective was to determine if this fragment is inherited as one unit and provides resistance to multiple diseases. Twenty four SSR markers distributed between 0.00 to 7.09cM on 3BS covering both Sr2 (5.57cM) and lm locus (6.0cM) were studied in the RIL population. Phenotyping was done for Sr2 associated PBC and lesion mimic along with disease severity for leaf rust, and spot blotch. Positive and significant correlations were observed between leaf stem rust resistance with Sr2 carrying PBC and lm. However, lines with lm either alone or with Sr2 (showing PBC) exhibited spot blotch susceptibility. The reverse situation does not hold not true where genotypes carrying Sr2 alone showed no correlation with spot blotch resistance. This indicates that the Sr2 complex is inherited as a single unit. Use of 24 SSR also suggest that Sr2 and lm loci are tightly linked and inherited together. The co-inheritance of Sr2 and lm ensures the stability and durability of rust resistance. However, the discouraging observation of spot blotch susceptibility due to lm gene suggests a limitation in achieving multiple disease resistance in environments where spot blotch is important. We identified two transgressive segregates in the population showing least expression of lm despite the presence of Sr2 and lm together.
Primary Author: Nawathe, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi 221005, India
In 2016 the bread wheat (BW) and durum wheat (DW) landrace accessions were evaluated against PstS2 and in 2017 against a mixture of PstS2 and warrior race in field inoculations at Izmir precision stripe rust phenotyping platform. Inoculation was carried out three times during seedling, tillering and booting stages using mixture of fresh spore and talcum powder. Adult-plant responses of tested accessions were recorded according to 0-9 scale once the flag leaf of the susceptible cultivar became fully susceptible. During 2016, out of 3319 BW accessions, 1135 (36%), 871 (28%) and 1133 (36%) were found resistant (1-3 scale), moderately resistant (4-6), and susceptible (7-9) to PstS2, respectively. Amongst the resistant accessions in 2016, 1043 (33%) remained resistant while 786 (25%) showed moderate resistant and 1310 (42%) became susceptible. In 2017, 43% of moderately resistant accessions showed susceptibility to warrior race and 57% remained resistant to moderately resistant. Within the susceptible accessions to PstS2 race in 2016, 22% showed resistance to the warrior race and the remaining were susceptible. In case of DW in 2016, 76% (553) of the accessions were resistant to PstS2, 23% (163) were moderately resistant and only 1% (7) were found susceptible. In 2017, 329 (46%) of the resistant accessions were found resistant, whereas 289 (40%) and 105 (15%) showed moderately resistance and susceptible reaction to Warrior race, respectively. The present data indicated that BW landraces were generally more susceptible to stripe rust than DWs. Susceptibility of both BW and DW accessions to Warrior race indicated that most likely some of the uncharacterized resistance genes which conferred resistance to PstS2 were ineffective against the warrior race. Sources of resistance to both races were identified in both BW and DW. Genetic architecture of identified sources of resistance in present study requires further investigations.
Primary Author: Nazari, Turkey-ICARDA Regional Cereal Rust Research Center (RCRRC), ICARDA, Menemen, Izmir, Turkey
Keywords: stripe rust
The Himalayan region of Pakistan and China has been shown to be the centre of diversity of Puccinia striiformis, however, little is known about the Eastern part of the Himalayas. We studied the genetic structure of P. striiformis from Nepal and Bhutan in comparison with Pakistan through microsatellite genotyping of 66 isolates from Nepal (35 isolates) and Bhutan (31 isolates) collected during 2015 and 2016. Genetic analyses revealed a recombinant and highly diverse population structure in Bhutan and Nepal. A high level of genotypic diversity was observed for both Bhutan (0.92) and Nepal (0.67) with the detection of 53 distinct multilocus genotypes (MLGs) in the overall population; 28 for Bhutan and 27 for Nepal. Mean number of alleles per locus was higher in Bhutan (3.33) than Nepal (3.11), while the gene diversity was higher in Nepal (0.4279) than Bhutan (0.3552). A non-significant difference between the observed and the expected heterozygosity in both populations further confirmed the recombinant structure. Analyses of population subdivision revealed a low divergence between Nepal and Bhutan (FST=0.1009), along with the detection of certain common MLGs in both populations. The overall population was clearly divided into six genetic groups, with no geographical structure, confirmed by the distribution of multilocus genotypes over two countries, suggesting a potential role of migration. Comparison with the Pakistani P. striiformis population suggested a high genotypic diversity in Nepal (0.933) and Bhutan (0.959), though lower than the previously reported from Himalayan region of Pakistan (Mansehra; 0.997). The overall high diversity and recombination signature suggested the potential role of recombination in the eastern Himalayan region (Nepal and Bhutan), which needs to be considered during host resistance deployment and in the context of aerial dispersal of the pathogen.
Primary Author: Nazir, The University of Agriculture, Peshawar, Pakistan
Keywords: stripe rust
Rust diseases are among the most important affecting wheat because they are responsible for a significant yield reduction globally. Different types of conventional breeding approaches are currently underway to protect wheat from these diseases. The involvement of molecular genetics and biotechnology tools in conventional plant breeding sets new directions to develop crop varieties with desired traits more efficiently and accurately. An array of molecular markers linked to rust resistant genes and dense molecular genetic maps are now available for use. Marker assisted selection (MAS) is now a routine activity in various crops especially for agronomic traits that are otherwise difficult to tag like resistance to pathogens, insects, nematodes etc. Gene pyramiding involves the stacking of many genes leading to real-time expression of all genes in single variety to develop durable resistance. This method is gaining significant popularity as it would enhance the efficiency of conventional breeding methods and precise development of broad spectrum resistant capabilities. Keeping in view the significance of MAS, rust resistant wheat parental lines were selected and molecular information was tagged using gene linked markers through PCR. Conventional breeding plane was designed on the basis of molecular data and maximum crosses were made between high yielding susceptible and resistant wheat genotypes. Molecular screening and other yield parameters were keenly noted on each stage of segregating population. Three rust resistant genes i.e. Lr-34/Yr-18, Lr-46/Yr-29 and Lr-19 were successfully combined in three cross combinations. Twenty crosses were found positive for two resistant genes i.e. Lr-46/Yr-29 and Lr-19, Moreover, one cross was positive for Lr-34/Yr-18 and Lr-46/Yr-29, and one was positive for Lr-34/Yr-18 and Lr-19. Introduction of more genes is also continued to develop superior resistance against a wide range of rust pathogen in wheat.
Primary Author: Nazir, Agricultural Biotechnology Research Institute, Ayub Agricultural Research Institute (AARI), Faisalabad-Pakistan
Keywords: molecular markers, rusts
Puccinia triticina, the causal agent of wheat leaf rust, is a devastating disease that can cause up to 40% yield loss. During fungal infection the host plant recognizes pathogen effectors, which trigger a host defense response. Changes in the pathogen effectors due to host selection pressure are responsible for the rapid development of new rust races and make durable resistance hard to obtain. The objectives of this study are to identify and characterize wheat genes that are utilized by races differently throughout infection and to understand functions of these genes using gene silencing. Six races of leaf rust were inoculated on a susceptible wheat variety and tissue was collected at six days post inoculation. RNA was sequenced and 63 wheat genes were identified that showed varying expression in response to the six races. 54 of these genes were evaluated in a time course study from zero days to six days post inoculation with the same six races. Real-time PCR was then used to analyze the timing of expression during early infection. The characterized genes have proposed functions involved in plant defense and stress, energy and metabolism, protein transport, replication, and RNA binding. Majority of the candidate genes showed three main expression patterns. However, race specific expression was found in three wheat genes that are affected by race shifts on Lr2A, Lr2C, and Lr17A. Sixteen potential susceptibility genes were also identified. Host susceptibility genes could be altered to provide durable resistance. RNAi was used to silence seven wheat genes to further understand their roles in leaf rust infection. T0 and T1 plants have been obtained and confirmed for the gene of interest. T2 plants were inoculated and observed for changes in susceptibility.
Primary Author: Neugebauer, Department of Plant Pathology, Kansas State University, USA
Keywords: leaf rust, gene expression
Stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is a threat to wheat production worldwide. To manage this important disease, new sources of genetic resistance are needed and common wheat landraces are a potential source of such resistance. Landrace accessions from the USDA-ARS National Small Grains Collection were evaluated for seedling resistance to the Ug99 race group. To identify accessions most likely to carry novel resistance genes, a bulked segregant analysis (BSA) approach was used. Seven resistant accessions were crossed to a susceptible parent line and F3 families were tested against Pgt race TTKSK. The resistant plants were identified and grouped into two bulks per population. The bulks, along with the parents and F1 progeny, were genotyped with the 90K wheat iSelect SNP genotyping platform. Four of the populations appeared to segregate in a 1:1 phenotypic resistant/susceptible ratio, one in a 1:2 ratio, and two in 1:3 ratios. However, chi squared tests indicated the ratios were statistically the best fit for only two of the 1:1 segregating populations and one of the 1:3 segregating populations. Initial BSA results indicate the markers associated with reduced stem rust infection are located on wheat chromosomes 1DL and 2B. These mapping populations are being advanced for further evaluation to ascertain if novel resistance to the Ug99 stem rust race group is present.
Primary Author: Neugebauer, USDA-ARS
Keywords: stem rust, resistance genes
Effective control of stripe rust (YR) requires deployment of resistant cultivars. Adult plant resistance (APR) is preferred over all-stage resistance because of its putatively durable nature. Discovery of new sources of resistance is a priority to combat rapidly evolving Pst races. Genebanks contain untapped genetic diversity that likely harbor novel resistance genes. We examined a diverse panel of 300 lines sourced from the Vavilov Institute, including landraces, cultivars and breeding lines from 28 countries. The most virulent Pst pathotype in Australia (134 E16 A+,Yr17+,Yr27) was used for all experiments, where YR reactions were determined on seedlings in a greenhouse and on adult plants in a field disease nursery. A total of 54% of accessions displayed all-stage resistance and 33% displayed moderate to high levels of APR. Accessions were genotyped using the DArTseq genotyping platform and using an association mapping approach we identified genomic regions associated with YR resistance. These were aligned with previously reported QTL and cataloged resistance genes on a consensus map. This enabled identification of novel genomic regions. Accessions carrying high levels of APR were screened using markers linked to well-known APR genes (i.e. Yr18, Yr29 and Yr46). Twenty two accessions carrying potentially novel sources of APR to YR were identified. Our current efforts are aimed at further characterizing and validating these genetic resources against a wide array of pathotypes and environments around Australia.
Primary Author: Ng, The University of Queensland, Queensland Alliance for Agriculture and Food Innovation, Australia
Keywords: stripe rust, genebank, APR
Stem rust resistance gene Sr43, derived from tall wheatgrass (Thinopyrum ponticum), is effective against Ug99 lineage Pgt races. Previous studies indicated that Sr43 was located on large Th. ponticum 7el2 chromosome segments in 7D/7el2 translocation stocks KS10-2 and KS24-1. In the present work, we applied a recently-established chromosome engineering procedure to reduce the size of the alien chromosome carrying Sr43. KS10-2 was crossed and backcrossed to the Chinese Spring (CS) ph1b mutant. BC1F1 plants were screened for stem rust response and Ph1- associated molecular markers. Resistant BC1F1 plants homozygous ph1bph1b were further backcrossed to CS. The resulting population of 706 BC2F1 plants was screened for stem rust response and with six co-dominant SSR markers. Wheat lines RWG33 and RWG34 carry Sr43 on shortened alien segments that are about 15% of that in KS10-2. Two molecular markers closely linked to Sr43 were identified; one was an SSR marker and the other a STS marker based on sequences of deletion bin-mapped expressed sequenced tags in wheat. The two new wheat lines with Sr43 and closely-linked markers may provide new resources for combating the threat of race Ug99 and derivatives.
Primary Author: Niu, USDA-ARS, Northern Crop Science Laboratory