Instituto Nacional de Investigaci?n Agropecuaria (INIA) La Estanzuela
Paula Silva, Clara Pritsch, Miguel Raffo, Silvia Pereyra, Silvia German
Wheat stem rust (SR), caused by Puccinia graminis f. sp. tritici, (Pgt) is considered one of the most destructive diseases of the wheat crop. As Sr24 and Sr31 are the most widely used resistance genes in the Southern Cone of America, wheat crops in this region is under threat of SR outbreaks posed by the potential migration of virulent Pgt Ug99-lineage races (Ug99+). Efforts have to be made to develop adapted lines resistant to Ug99+. Genes Sr26, Sr32 and Sr39 are effective to both Ug99+ and local races of the pathogen. This work is aimed to pyramid two and three of the resistance genes in two locally adapted wheat cultivars (G?nesis 2375 and G?nesis 6.87). Donor lines of Sr26, Sr32 and Sr39 (developed by I. Dundas, University of Adelaide, Australia) and molecular markers Sr26#43, csSr32#1 and Sr39#22r (developed by R. Mago et al., University of Adelaide) were used. Lines with two-gene combinations were developed in two steps. First, tree-way crosses were made by crossing heterozygous F1 plants (derived from crossings donor lines) to either one of the two adapted wheat cultivars. Subsequently, tree-way F1 plants were genotyped and only those with two-gene combinations were backcrossed (BC) twice to the adapted cultivars. Among three-way F1 plants, two-genes combinations were confirmed for Sr26+Sr32 (8 out of 31), Sr26+Sr39 (2 of 115) and Sr32+Sr39 (26 out of 103). In the BC1F1 generation, Sr26+Sr32, Sr26+Sr39 and Sr32+Sr39 combinations corresponded with 9, 9 and 45 out of 99, 27 and 241 plants, respectively. In 2017, 1345 BC2F1 plants are being grown to obtain BC2F2. We plan to intercross plants with two-gene combinations to obtain lines with the three genes which will be used as sources of resistance to develop cultivars with presumably longer lasting resistance to wheat SR.
University of Minnesota
Ayele Badebo, Worku Bulbula, Matthew Rouse, Yue Jin
Our research objective is to identify new resistance genes in durum wheat that are effective against TTKSK and other significant stem rust pathogen races that could be utilized in durum breeding. We characterized 8,000 accessions for stem rust response in the field (Debre Zeit, Ethiopia, and St. Paul, MN). Accessions with resistant to moderately resistant responses in multiple field evaluations were evaluated at the seedling stage for resistance to races TTKSK, TRTTF, TTTTF, JRCQC, TKTTF, and six representative U.S. races. We identified 438 durum accessions resistant to moderately resistant in all field evaluations. Among the field-resistant accessions, 273 were resistant to all races used in seedling evaluations. Accessions susceptible at the seedling stage are being evaluated for the presence of adult plant resistance genes. The highest frequencies of resistant lines include landraces from East and North Africa (Ethiopia and Egypt) and advanced breeding lines and cultivars from North America (Mexico and USA). DNA markers will be performed to identify the presence of durum stem rust resistance genes, including Sr13, Sr8155B1, Sr11, and Sr8a. Nineteen resistant accessions were selected to investigate the genetics of TTKSK and TRTTF resistance. Results from evaluating F2 and F2:3 generations from biparental crosses revealed that resistance to race TTKSK was conferred mostly by one or two genes with dominant and recessive actions. Additional resistance genes were identified when populations were evaluated against race TRTTF. A bulk segregant analysis approach is being used to map the resistance in selected lines using the 90K SNP platform.
ICAR- Indian Institute of Wheat & Barley Research, Karnal-132001, India
Vinod Tiwari, DP Singh, RP Gangwar, GP Singh
The changing climatic conditions are affecting wheat production in major agro-ecological zones in India, namely, north western plains(NWPZ), north eastern plains(NEPZ), central (CZ) and peninsular zone(PZ) where the reproductive phase has to endure higher temperatures. Also, the prevalence and virulence of rust pathotypes and other diseases are affected. To address such challenges, development of wheat for climate resilience was initiated following shuttle breeding approach for incorporating heat stress tolerance as well as resistance to wheat rusts. During 2010-16, a total of 583 elite lines were evaluated against prevalent pathotypes of stripe rust 78S84, 110S119, 110S84 and 46S119; leaf rust 12-2(1R5), 12-5(29R45), 77-2(109R31-1), 77-5(121R63-1), 77-9(121R60-1) and 104-2 (21R55) and stem rust 11(79G31), 40A(62G29), 42(19G35), 122(7G11) and 117-6(37G19) of which 108 promising entries were identified. These lines were evaluated for disease response in multilocational Initial Plant Pathological Screening Nursery (IPPSN) against prevalent races of all three rusts. Based on average coefficient of infection (15.0 ACI), 42 (39%), 104 (96%) and 90(83%) entries were found resistant to different races of stripe, leaf and stem rusts, respectively. Based on performance in multiplication yield trials, 29 entries were contributed in national coordinated evaluation system on Wheat & Barley which resulted in release of four wheat cultivars DBW71(Yr9+27+,Lr26+,Sr2+5+31+), DBW107(Yr9+,Lr26+3+,Sr31+), DBW110(Yr2+, Lr13+10+,Sr13+11+) and DBW93(Yr9+, Lr26+23+, Sr31+) for commercial cultivation in NWPZ, NEPZ, CZ and PZ, respectively. These cultivars are becoming popular among farmers due to their yield advantage, resistance to diseases, tolerance to high temperature and better quality traits. Also, DBW 129 was screened in multiple disease screening nursery (MDSN) and observed resistant to all rusts, leaf blight, powdery mildew, flag smut and shoot fly. The adoption of the newly developed cultivars for deployment of differential genes for resistance would lead to reduction in disease pressure and bring higher profitability to farmers in different agro-ecological zones in India.
The Federal Research Center Institute of Cytology and Genetics, The Siberian Branch of the Russian Academy of Sciences
Vyacheslav Piskarev, Irina Leonova, Ekaterina Bukatich, Elena Salina
Stem and leaf rusts affect the winter and spring wheat in the Novosibirsk region. During 2008-2017 leaf rust incidence was generally moderate, from 20 to 40%. A leaf rust outbreak occurred in 2015 when incidence increased up to 80%. Leaf rust severity on the 'Thatcher' NILs ranged from immune or resistant to highly susceptible host response with maximum severity of 90S. Lines carrying genes Lr17, Lr18, Lr24, Lr29, Lr35, Lr37, Lr44, and LrW remained almost free of infection for the whole time of inspection. Genes Lr12, Lr13, Lr28, Lr34, and Lr38 exhibited moderate resistance but they did not provide sufficient level of resistance in favorable conditions. Since race-specific genes Lr24 and Lr29 are still effective in the neighboring Novosibirsk and Omsk regions, they might be recommended for breeding purposes in Western Siberia.
In 2016 stem rust was more prevalent and widespread in the region than ever before. Disease incidence ranged between 4.5 - 60% with high severity up to 80S in six fields from seven observed locations. The 4th ISRTN and varieties carrying Sr31 of West Siberian germplasm were assessed in field trials to monitor the virulence of the local population. There was no virulence to Sr9b, Sr9e, Sr20, Sr28, Sr29, Sr33, Sr39, Sr40, SrWld, Sr2 complex. Possible virulence to Sr6, Sr11, Sr12, Sr13, Sr17, Sr24, Sr25, Sr30, Sr31, Sr35, Sr38, Sr44, Sr57 was observed with low frequency. Entries genotyped for gene Sr31 were scored as MS and S. However, follow up race analysis work is needed to determine the actual stem rust races present and confirm the suspected possible observed virulence on Sr31.
University of Delhi South Campus, New Delhi
Towards understanding the molecular mechanisms of heat stress tolerance, we have analyzed heat stressed substractive cDNA libraries and undertaken genome-wide transcriptome exploration for genes associated with spike photosynthetic efficiency during thermal stress. The photosynthetic efficiencies of Aegilops tauschii and Ae. speltoides were also compared. While the former displayed nearly complete recovery of PSII, the adverse effect was more pronounced in the latter. Functional characterization of heat stress-associated transcription factors and thermal stress-associated proteins was also undertaken e.g. TaHSF, TabZIP, TaZnF and TaMIPS, and TaLTPs in the Indian wheat germplasm. Functional characterization of the three heat stress transcription factors was upregulated under high temperatures and other abiotic stresses. They also showed early flowering and better performance with respect to their growth and yield after heat stress. Additionally, we have identified various interacting components associated with thermal stress-mediated plant signaling partners during thermal stress.
Agriculture & Agri-Food Canada
Wheat cultivar McNair 701 carries resistance gene SrMcN and is used as a differential line to identify Pgt races using the international letter code nomenclature. The inheritance and location of the resistance gene has not been characterized. We developed a doubled haploid (DH) population from cross LMPG/McNair 701 to study the genetics and chromosomal location of SrMcN. A DH population inoculated with race QCCJB segregated 100 resistant : 94 susceptible, a 1:1 ratio (?2=0.186, P=0.666, NS) indicative of segregation at a single locus. This gene was mapped to chromosome 2DL using the Infinium 90k platform. The map position of SrMcN was similar to that of Sr54, one of two genes previously found in Norin 40. Comparison of stem rust seedling reactions using 12 diverse Pgt races indicated that McNair 701 and an Sr54 line derived from Norin 40 had an identical pattern of responses and similar low infection types (IT=12-) to races LCBNB and QCCJB. Based on the chromosomal location on 2DL and identical seedling responses to Sr54, it is likely that the resistance gene in McNair 701 formerly known as SrMcN is Sr54. This finding will be confirmed by a test of allelism.
Laboratory of Molecular Plant Physiology, Biotechnology Center of Borj Cedria (CBBC)
Mahmoud Gargouri, Hesham A.Y Gibriel, Richard B. Todd, Michael F. Seidl, Gerrit H.J. Kema
Septoria tritici blotch disease, caused by the fungus Zymoseptoria tritici, is a major threat to global wheat production. With the recent advances in high-throughput DNA-based technologies, Z. tritici has become a powerful model system for the discovery of candidate determinants that underlie virulence and host specialization. Although a few important virulence/regulatory genes have been identified, a global understanding of the larger regulatory network has not been developed. Therefore, to uncover the transcriptional regulatory networks of the infection cycle and most particularly the regulatory hubs that control the switch between the biotrophic and necrotrophic phases, we applied an integrated approach combining transcriptomics, proteomics, and metabolomics analyses based on the identification of plant and fungal transcription factors and regulators, which we characterized from the newly annotated genome sequence of the reference isolate IPO323 (Grandaubert et al., 2015) and using datasets from Rudd et al. (2015). Bread wheat transcription factors and regulators were identified by querying the proteome and subsequent categorization from the Plant Transcription Factor database (PTFDB). Similarly, Z. tritici transcription factors and regulators were identified and categorized using the PFAM TF family databases, and following fungal transcription factor rules as outlined by Todd et al. (2014) and rules we developed for fungal transcription regulators. Insights into transcription factors and regulators will enable synthetic biology approaches to alter the Z. tritici-wheat interaction and lead to rewiring of the regulatory networks thereby turning off the fungal infection process. Beyond providing insights into the regulatory systems-levels involved in Z. tritici-wheat interaction, we believe that our dataset and approach sets the stage for an emerging series of studies that will decipher the dynamic regulatory networks in other plant-pathogen interactions.
Bangladesh Agricultural Research Institute
Paritosh Malaker, Mostofa Reza, Abdul Hakim, Krishna Roy, Rabiul Islam, Thakur Prashad Tiwari, Pawan Kumar Singh, Arun Kumar Joshi
The major diseases of wheat in Bangladesh are leaf blight and leaf rust. Yellow rust occurs occasionally with sporadic infection in the northern parts whereas stem rust was observed only in 2014. So far the country is free of Pgt race Ug99. Wheat blast, a devastating head disease, was first reported in 2016. Currently, about 65% of the wheat area in Bangladesh is covered by leaf rust resistant varieties and about 30% of the area is covered by Ug99 resistant varieties. Surveillance and monitoring of diseases is conducted regularly. In 2017, 102 sites were surveyed of which 52% had leaf rust infection. The data were uploaded to the Wheat Rust Tool Box. A separate surveillance and monitoring of wheat blast was conducted on 421 farmers? fields in 24 districts. Different levels of blast incidence were recorded in 77 fields. The Wheat Research Centre in Bangladesh works with CIMMYT and BGRI to develop high yielding rust resistant varieties. This includes screening for response to Ug99 at KALRO, Kenya. However, the current major concern of wheat is wheat blast. The popular variety BARI Gom 26 is highly susceptible to this disease and no current cultivar in Bangladesh carries an acceptable level of resistance to blast. During 2016-17, 20 varieties and advanced lines from Bangladesh and 80 from CIMMYT Mexico, were evaluated. One elite breeding line, BAW 1260, showed resistance (<10% severity) in multiple environment tests and is also resistant to leaf blight and stem rust. This line carries the 2NS translocation from Aegilops ventricosa and will be released soon for commercial cultivation. Pre-release seed multiplication is already underway for rapid dissemination. Among recently released wheat varieties BARI Gom 30 and BARI Gom 32 are moderately tolerant to blast and are being promoted for wider adoption by farmers.
Panjab University Chandigarh
Sterol 14?-Demethylase Cytochrome P450 (CYP51) protein involved in ergosterol biosynthesis pathways is a crucial target for efficient fungicidal compounds. However, the recognition mechanism and dynamic behavior of CYP51 in wheat leaf rust pathogen, Puccinia triticina is still obscure. Previously, a mutation at codon 134 (Y134F) was reported in five European isolates of P. triticina, the structural basis of this mutation remain unclear. To address this problem, CYP51 wild type protein and its variant proteins were successfully modeled using I-TASSER, an ab initio based structure prediction pipeline. To gain valuable insights into structure-function behavior for the binding wild-type and mutant-type proteins, individually generated protein models was subjected to 50ns molecular dynamics (MD) simulations run. Observably, this comparative protein-ligand interaction analysis and binding free energy results revealed that impact of mutation on the thermodynamics and conformational stability of the CYP51 protein is negligible. In present study, we carried out structure-based molecular docking and identified potent novel fungicidal compounds from four different databases and libraries. Consequently through MD simulation and thermodynamic integration, four novel compounds such as CoCoCo54211 (CoCoCo database),ZINC04089470(ZINC database), Allyl pyrocatechol 3,4 diacetate (Natural compound library) and 9-octadecenoic acid (Traditional Chinese Medicine database) has been predicted as potent fungicidal compound against CYP51 with XPGlidedocking score of -11.41, -12.52, -7.40 and -7.55 kcal/mol, respectively. These compounds were found to directly bond to heme group of CYP51, subsequently disturbing the stability and survival of fungus and can be used to control leaf rust in wheat.
The Ohio State University
Nelly Arguello-Blanco, Mao Huang
Genomic selection facilitates rapid cycling through a breeding cycle by eliminating the need to phenotype prior to selecting superior parents and crossing among them. In winter wheat we can now complete a cycle of GS in about 12 months and two greenhouse seasons. Season consists of planting F1s from the previous cycle and selfing to obtain F2 seed. The second season involves planting and genotyping the F2s, predicting their value with GS, selecting and crossing the best, and harvesting the F1 seed. Our breeding program has completed five cycles of GS in one population primarily for grain yield, over the past five years. We have completed three cycles of GS for resistance to Fusarium Head Blight in a second population. Genotyping was done using genotyping-by-sequencing. This provides an opportunity to assess the changes in the population that have occurred as a result of this rapid cycling. These include 1) changes in genomic estimated breeding values for grain yield and FHB resistance, 2) effect of selection and drift on allele frequencies including fixation, 3) effect of selection on diversity and genetic relationships, and 4) changes in linkage disequilibrium. We are conducting these analyses and will present the results.