Institute of Molecular Biology and Biotechnology, Azerbaijan National Academy of Sciences
Shahriyar Sadigov, Alamdar Mammadov, Irada Huseynova
Rust of cereals are considered to be an important disease in many countries, including Azerbaijan. One of these is stem rust caused by Puccinia graminis f. sp. tritici (Pgt). Extensive research on the identification of wheat stem rust resistance genes and effectiveness of these genes in various geographical regions have been conducted. Genetic resistance is one of the most effective ways for controlling stem rust. Sixty-nine stem rust resistance genes (including 45 identified Sr genes and 24 genes with temporary designations) are registered in the Komugi Wheat Genetics Resource Database. It is important to use proper combinations of resistance genes in developing lasting resistance wheat. The main purpose of the study was to identify lines caring Sr11, Sr26 and Sr31 genes, which are effective to the predominant Pgt races in Azerbaijan. Durum and bread wheat genotypes differing in their disease resistance, productivity and other physiological traits were chosen from the wheat gene bank of the Research Institute of Crop Husbandry (Baku, Azerbaijan) for analysis. DNA extraction was carried out according to standard CTAB protocol. RT-PCR performed using KASP markers (KASP_6BL_BS0074288_51 and KASp_6BL_Tdurum contig55744_822) identified nine durum genotypes (out of 34 genotypes) and seven wheat genotypes (out of ten genotypes), caring Sr11. Using the dominant STS marker (Sr26#43) a diagnostic 207 bp amplicon for Sr26 gene, was observed in 11 of the 42 wheat genotypes tested. In eight of the 42 wheat genotypes tested, the diagnostic 1,110 bp amplicon was observed using the Lr26-Sr31-Yr9 locus specific marker iag95, characteristic of Sr31 gene located at 1BL.1RS translocation. For the first time, wheat germplasm in Azerbaijan was analyzed using KASP genotyping technology and genetic resources, and resulted in the identification of wheat lines with effective resistant to Puccinia graminis f. sp. tritici race TKTTF.
Wheat Dis. Res. Dept., Plant Pathol. Res. Inst., A.R.C., Sakha, Egypt
Les John Szabo
Stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is one of the most serious disease of wheat worldwide. The discovery of new Pgt races in Africa, Ug99 and its variants, brings a new threat to global wheat production. In this study, 50 single pustule stem rust samples, were collected during 2015-2016 from the International Stem rust Trap Nursery (ISRTN) and commercial wheat fields in Sakha, the most important wheat growing region in Egypt. SNP-genotyping was carried out at USDA-ARS Cereal Disease Laboratory. Infection and genotype data confirmed that none of these samples belonged to the Pgt Ug99 race group. Forty-five samples were successfully genotyped consisting of 12 multi-locus genotypes (MLGs). The majority (86.7%) of the samples belonged to three clades: 10 samples, clade III-B (MLG.04, race TTRTF) collected from Misr 3, Sakha 95 and Sids 14 wheat lines; 12 samples, clade IV-A.2 (MLG.06, race TKTTF) collected from Sr 5, Sr6, Sr7a, Sr7b, Sr8b, Sr9a, Sr9e, Sr10, Sr11, Sr15, Sr16 and Sr17 wheat lines; 17 samples, clade IV-E.2 (MLG.11, race TKKTF) from Sr13, Sr14, Sr19, SrMcN, Sr24, Misr 1, Misr 2, Sakha95 and Sids 12 wheat lines. Pgt samples belonging to clades IV-A.2 and IV-E.2 have been observed from Europe to the Middle East, and samples from clade III-B from the southern Caucasus Mountains, Middle East to northeast Africa. The remaining six samples collected from Sr12, Sr18, Sr20, Sr21, Sr22 and Sr25 wheat lines represent two new genotypes (MLG.14 and MLG.17) that have not been assigned to clades. MLG.14 was also observed in samples from Azerbaijan, Iraq and Eritrea. In contrast, this represents the first detection of MLG.17. These results suggest continued variability of the Pgt population in Egypt therefore, emphasizing the importance regularly monitoring to timely identify new races, and utilize this information in screening and identification of effective sources of resistance.
The University of Agriculture, Peshawar, Pakistan
Muhammad Khan, Aamir Iqbal, Sher Nawab, Sohail Ahmed, Muhammad Imtiaz, Sajid Ali
Resistance breeding for wheat leaf rust requires testing of breeding materials under field conditions, which must be complemented with diagnostic molecular makers. A set of 28 exotic wheat lines from advanced CIMMYT material along with three check varieties (Siran, Atta-Habib, Ghanimat-e-IBGE) were tested at three contrasting locations (Peshawar, Mansehra and Lakki-Marwat) and were genotyped with markers linked to three Lr genes (LrPr, Lr37, and Lr34). The overall leaf rust pressure was low during the wheat season of 2015-16, with the maximum disease observed at Lakki-Marwat (up to 70%), followed by Peshawar (up to 50%) and the minimum disease at Mansehra (up to 30%). Despite the overall low leaf rust pressure, the germplasm behaved variably in terms of leaf rust resistance as revealed through average co-efficient of infection (ACI). According to ACI value, 16 out of 28 genotypes were completely resistant, while few genotypes showed partial resistance. The maximum CI value was recorded for wheat line W-SA-87, which was 55 at Lakki Marwat, 33 at Peshawar and 15 at Mansehra, while 18 lines had CI value of zero across the three locations. Variability existed in yield parameters with W-SA-84, W-SA-78 and W-SA-79 producing the better grain yield. Genotyping with Lr linked markers viz., STS-7 (LrPr), SC-Y15 (linked with Lr37) and csLV34 (linked with Lr34) revealed that among the tested lines LrPr was the most frequent (83.8%), present in 26 lines; followed by Lr37 (77.4%), present in 24 lines, while Lr34 was present in 16 lines (71.1%). All three genes were detected in 45% of the germplasm. Cluster analysis grouped the germplasm in four clusters based on both phenotypic and molecular markers data. The information generated in the present study would be valuable in resistance breeding for a better control of leaf rust disease in Pakistan.
Mohamed 5 university/ICARDA
Amadou tidiane sall, meryem zaim, Ayed Al-abdallat, Gregor Gorjanc, Jesse Poland, Miloudi Nachit, Abdel karim Filali Maltouf, Bouchra Belkadi, Rodomiro Ortiz, Filippo Bassi
Durum wheat production is globally important, but grain yield has been stagnating in recent decades. In order to ensure that its production maintains the pace with increasing demand, breeding for high grain yield must be supported by molecular-based methods. Genomic estimated breeding values for selection and genome scan were assessed as molecular tools holding maximum potential for durum wheat breeding. Four recombinant inbred line populations bred by inter-mating elite were sown in yield trials at five sites. All progenies were characterized using "genotyping by sequencing" method. A consensus map was developed, and missing genotypes were imputed using a Hidden Markov model to reach a total of 1987 polymorphic markers. Models accounting for genotype environment interactions were used to estimate the genetic component of each measured trait. Hence, Bayesian ridge regression was used to determine the predicted values and their relative accuracy in several combinations, testing full-sibs and half-sibs as training population for grain yield and 1,000 kernel weight. The high level of accuracy achieved suggests that GEBV for selection holds great potential for durum wheat breeding, as long as full-sibs are used as training populations, in combination with statistical models that account for G?E. In order to test the exploitability of genome scan to guide breeding crosses, a separate genome-wide association study was conducted. 288 elite were sown in the south of Morocco and at two sites along the Senegal River for two years. These sites show a temperature differential of 10?C. Implementing a GE model facilitated identifying the most heat tolerant among the tested entries. 8,173 polymorphic SNPs were inquired, and several associations could be identified between markers and the ability to withstand the heat gradient. Hence, GWAS holds great potential to increase genetic gain in breeding via increased accuracy in determining the crosses to be made.
Department of BioEngineering, Birla Institute of Technology, Mesra, Ranchi, India
Manish Kumar, Kunal Mukhopadhyay
Bread wheat (Triticum aestivum L.) being the world's most popular edible cereal, plays a major role in global economy. Rust in wheat leaves, caused by Puccinia triticina, affects grain quality and severely retards its production worldwide. Micro(mi)RNAs are considered major components of gene silencing and so have a great role to play during stress. The present study focuses on identification of miRNAs, produced by host to suppress pathogen as well as delivered by pathogens to encounter host defence mechanism. Therefore, these miRNAs may be called as leaf rust responsive microRNAs. Small RNA and degradome libraries were prepared from a pair of near isogenic lines of wheat (HD2329, HD2329+Lr24), one set was mock inoculated while the other set was inoculated with urediniospores of leaf rust pathogen. Using these libraries as input a vast number of miRNAs rather a population of miRNAs were identified derived from wheat that were targeting genes mostly involved in functions like defense response, signal transduction, development, metabolism, and transcriptional regulation.
When reads specifically produced under pathogen inoculation were taken as input with Puccinia triticina genome sequences as reference, only three putative miRNA precursor loci were detected and the molecules produced were called miRNA-like molecules as their precursors lacked one or two criteria essential for a true miRNA precursor. The identified miRNAs were targeting genes like F-box protein, MAP kinase, calmodulin and susceptible antioxidant protein. We further identified the presence of argonaute and dicer like domains in Puccinia proteome available at FungiEnsembl which strengthens presence of RNAi-like activities in Puccinia.
In addition, differential expression of wheat as well as Puccinia small RNAs using stem loop RT-PCR under varying time points of disease progression (0-168 hpi) revealed their direct connection with stress responses.
All Russian Research Institute of Biological Plant Protection
Irina Petrovna Matveeva
Yellow rust caused by Puccinia striiformis West. is a harmful and dangerous disease in the south of Russia. Yield losses under optimum conditions on highly susceptible varieties can vary from 10 to 100%. During the growing season of 2017, cool weather with constant precipitation from the third decade of April to the first decade of June contributed to the intensive development of the pathogen. Surveys of the main winter wheat production areas in five agroclimatic zones of the region revealed that yellow rust was prevalent in all areas. The maximum development of P.striiformis was observed in southern submontane and western Priazovsky agroclimatic zones. Some varieties such as Grom, Yuka, Tanya, Anka had losses to yellow rust of up to 30-40 %. In the central and northern agroclimatic zones, the losses averaged 5%, whereas in the dry eastern steppe zone losses were only up to 1%. The build up of yellow rust inoculum in the region raises concerns that in 2018, under favorable weather conditions in spring, winter wheat crops could be infected with the disease, especially in the wetter agroclimatic zones.
The University of Jordan
Ayed Al Abdallat, Monther Tahat
Studies on whet stem rust (WSR) in Jordan are considered to be old. There was only one study conducted in the late 1980's by Abu-Blan and Duwayri (1989) to evaluate the infection of wheat cultivars with black stem rust disease (Puccinia graminis f. sp. tritici). Recently, reports of stem rust were published in Israel and Lebanon in 2010 and first report of Ug99 was reported in Egypt in 2014. The objectives of our research are to: (i) survey wheat growing areas for WSR in Jordan during the years 2017-2020, (ii) identification of WSR races isolated from Jordan morphologically and molecularly, (iii) analyze rust populations in terms of their response to known differential sets, pathotype distribution and diversity, (iv) screening the response of Jordanian wheat germplasm to the identified WSR strains, and (vi) study the population diversity of WSR races using RT-PCR and SNP genotyping. In 2017 a total of 270 fields of wheat and barley in the wheat and barley growing areas in Jordan were surveyed from March-May. The survey covered northern, middle, and southern parts of Jordan (arid and semi-arid regions). Altitude, longitude, and latitude data was recorded. Only few WSR pustules (n=4) were collected because the environmental conditions were not suitable for the disease to develop. On the other hand, wheat stripe rust was very common in the wheat growing areas mainly at the southern parts of the country. Other fungal plant pathogens were also reported including smuts, spots, blotches, powdery mildew, crown rot, fusarium head blight, and flag smut.
Institute of Crop Science and Resource Conservation, Rheinische Friedrich-Wilhelms-University of Bonn
Mohammad Wali,Salari, Kobra, Yusefi, Mohammad, Yusefi, Gul Mohammad, Ajir, Wakil Ahmad, Sarhadi, Jens, L?on, , , , , , , , , , , , , , , , , ,
Bread wheat is a staple food in Afghanistan. Breeding for improving yield and its components in Afghan bread wheat without using new molecular methods such as marker-assisted selection (MAS) and quantitative trait loci (QTL) mapping approaches is difficult. Therefore study of genetic analysis by focus on yield and its components as first steps is necessary. Genetic analyses were performed on a winter wheat core collection of 20 accessions and commercial varieties sampled from different regions of Afghanistan and twenty agronomic traits were evaluated over three years under fully irrigated, rain-fed and drought treatments. Grain yield was the most important trait to water deficit and was highly correlated with other agronomic traits. The germplasm was structured into two sub-populations. Field plots of the genotypes were treated to one of three treatments including full irrigation, rain supplied and rain-sheltered. A randomized complete block design with three replicate was used every year of the trial. For every agronomic trait, variance components, heritability (h2) and genetic correlations was calculated. Results of the study showed that these genotypes may be good source for national breeding programs. The multiple statistical in this study showed that results of genetics correlation and regression analysis are same. Further analysis of these traits with additional experimental data to attain persuasive conclusion is suggested.
State Key Laboratory of Crop Stress Biology for Arid Areas, Northwest A&F University, Yangling, Shaanxi, P. R. China
Qilin,Wang, Jianhui, Wu, Qingdong, Zeng, Lili, Huang, Dejun, Han, Zhensheng, Kang, , , , , , , , , , , , , , , , , ,
The combination of several additive, partial resistance genes has been proposed as a preferred strategy to breed wheat cultivars with acceptable levels of durable resistance. The German winter wheat cv. Centrum has displayed high levels of adult plant stripe rust resistance (APR) in field environments for many years. One hundred and fifty one F2:7 RILs were developed from a cross between susceptible landrace Mingxian169 and Centrum to determine the inheritance of the APR resistance. The RILs and parents were evaluated for maximum disease severity (MDS) in the field during the 2015-2016 and 2016-2017 cropping seasons. Affymetrix 35K single nucleotide polymorphism (SNP) arrays were used to genotype the RILs and parents. In addition, the 660K SNP array was used to genotype bulked extreme pools and parents for saturation mapping. Four stable QTL were detected in all tested environments using inclusive composite interval mapping (ICIM); namely QYrCEN.nwafu-4AL, QYrCEN.nwafu-4BS, QYrCEN.nwafu-7BL, and QYrCEN.nwafu-7DS. QYrCEN.nwafu-4BS and QYrCEN.nwafu-7DS were contributed by MX169, QYrCEN.nwafu-4AL and QYr-CEN.nwafu-7BL were contributed by Centrum. QYrCEN.nwafu-7BL and QYrCEN.nwafu-4AL appear to represent new APR loci based on map comparisons. QYrCEN.nwafu-4BS contributed by MX169 also seems to represent a new locus. QYrCEN.nwafu-7DS is likely Yr18. Although MX169 was fully susceptible in our nurseries it is positive for the widely used marker csLV34. Reference lines carrying Yr18 are moderately resistant. Our hypothesis is that MX169 probably carries the inhibitor reported in Chinese landrace varieties by Wu et al. (2015, Plant Breeding 134: 634-640). SNP markers within these QTL were converted to KASP markers and validated in a subset of 120 diverse lines. These KASP markers should be useful for marker-assisted selection to improve stripe rust resistance in breeding programs.
Ayub Agricultural Research Institute, Faisalabad, Pakistan
Muhammad,Idrees, Faqir, Muhammad, Arshad, Mehmood, Majid, Nadeem, Saleem-ur, Rehman, Makhdoom, Hussain, Javed, Ahmad, , , , , , , , , , , , , , , ,
Under changing climatic conditions, the emergence of new diseases or new races of existing diseases is a serious threat to global wheat production. Particularly, the presence of wheat blast in Bangladesh and stem rust race Ug99 in Iran, created a fearsome and intractable situation for Pakistan. A study was planned for monitoring and surveillance of the wheat blast and rust pathogens in wheat growing districts of Punjab, Pakistan during the cropping season 2016-17 as vigilance program. During the survey, one hundred and seventy one wheat fields of upper and central Punjab region were monitored and two types of Rusts (Leaf Rust & Yellow Rust) were recorded in varying intensity on different varieties of wheat. Out of 171 locations 86 spots were free from both types of rusts i.e. Leaf Rust & Yellow Rust, while the remaining locations were found to be infected with both leaf and yellow rust. However, all the surveyed fields were free from the stem rust infestation. Among the infected fields, 23 were infected by Leaf Rust while 63 fields were infected by Yellow Rust.The susceptible type of rust attack was noticed on old/ banned/ unapproved wheat varieties. Moderately resistant to resistant reaction was observed on newly approved varieties. The rust infected samples having S or MS type infection were collected for race analysis. Similarly, blast suspected samples were analyzed in laboratory and none of the tested samples showed the presence of wheat blast pathogen, which indicates no need to panic but vigilant in future.