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.
Department of Botany, Faculty of Biology, Government College Murree, Pakistan
Barkat Ali, Muhammad Fayyaz, Atiq ur Reman Rattu, Abdul Samad Mumtaz, Muhammad Imtiaz
The new arrival of wheat rust pathotypes through migration during wheat cropping season requires regular monitoring to secure wheat production. In the present study, we collected leaf rust (Puccinia triticina Eriks.) infected wheat leaves from three major wheat growing provinces of Pakistan in the year 2014 to assess the haplotype diversity of P. triticina (Pt) isolates. The rDNA ITS sequence data of collected isolates was used in NCBI BLAST analysis. The blast hits showed best matches with Pt accessions EU014050 (Iran), JN120331 (Iran), JX533577 (Iran), AY956549 (Iran), DQ417412 (Czech Republic), DQ417418 (Israel), DQ417413 (Slovakia) and AF511083 (Louisiana). However, in cluster analysis, the Pakistani isolates showed strong bootstrap support with only Iranian and Indian (races 77-5 & 104-4) accessions that indicated eastward migratory mode of Pt pathotypes in Pakistan through westerly wind patterns. The predominant genotype DQ417412 (similar in alignment with AY956549 from Iran) overcome the resistance of top Pakistan mega varieties Seher06, Inqilab91, Kiran95, SKD1, TJ83 and NIFA-Batoor. Hence, the ITS based information remains a rapid molecular tool for pathogen surveillance across countries and continents.
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.
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.
National Agronomic Research Institute of Algeria
Among the many biotic constraints to wheat production in Algeria, rusts and in particular yellow rust (Puccinia striiformis), are among the most prevalent diseases that occur mostly all over the northern part of the country. Yellow rust has become now sporadic due to the exploitation of effective resistant genes in different forms and combinations (from CIMMYT and ICARDA). Earlier, durable resistance was probably due to many genes, such Yr18, Yr9, Yr27 and Yr1.
Yellow rust appeared as epidemic in 2004, over 600 000 ha of bread wheat ; severity exceeded 70%. Yields from affected fields of Hidhab a susceptible variety did not exceed 5.2 q/ha, while resistant cultivars yielded up to 48 q/ha. Monitoring of the pathogen virulence factors and their changes provides basic information for the development of an early warning system. This experiment was carried out in 5 Eastern Algeria locations. 30 lines of a standard set of yellow rust and 14 near-isogenic lines from ICARDA were sown in 2-m rows in 2014 and 2015. According to the results, virulence on Heines Kolben (Yr2), Kalyansona (Yr2), Lee (Yr7), Avocet R (YrA), Federation*4/Kavkaz (Yr9), Yr6/6*Avocet ?S?, Yr7/6*Avocet ?S?, Yr9/6*Avocet "S", Yr17/6*Avocet "S", TP1295 (Yr25) and YrSU was common during those two seasons. The frequency of virulence on plants with Yr2, Yr6, Yr7, Yr9 or YrA and Yr27 was up to 80%. No virulence was observed on plants with Yr1, Yr3, Yr4, Yr5, Yr8, Yr10, Yr15 and Yr18 genes. This material was extensively used in our breeding programs and several new cultivars are in the on farm trials where a participatory selection approach is used. All resistant and performing new varieties are being spread for replacement of most old susceptible ones.
National Institute of Agricultural Research
Nsarellah Nasserlhaq, Wuletaw Tadesse, Ahmed Birouk
In the context of climate change, drought is one of the most important and complex abiotic stresses affecting crop production worldwide. The adoption of an appropriate technological package, principally drought tolerant varieties, may overcome these challenges to meet global food security needs for the rapidly growing human population, particularly in developing countries. Therefore, this research was carried out to identify efficient phenotypic and genetic selection criteria to identify drought tolerant wheat varieties. In this perspective, 200 diverse elite bread wheat lines from ICARDA and CIMMYT were evaluated under four Moroccan environments during the 2015 and 2016 seasons for yield and 15 agro-physiological traits. The same set of genotypes was genotyped using 15k SNPs. Significant environment and genotype environment interaction effects were observed for yield. Average yield reached 3.18t/ha and ranged from 2.45 to 4.27t/ha. The secondary traits were mostly dominated by the environment effect (p<0.001). Based on correlation and regression analysis between grain yield and phenotypic data, the biomass, grain number per m<sup>2</sup> and to a lesser extent fertile spikes number and thousand kernel weights (depending of drought scenarios) can be more reliable traits than yield for the identification of drought tolerant genotypes. Moreover, the ground cover and canopy temperature depression can be used as supplementary criteria for more accurate selection. Slow selection on the basis of phenotypic traits may be accelerated and improved by using molecular markers. The genetic analysis highlighted significant SNPs and identified new QTLs linked to yield and the most efficient phenotypic traits under drought conditions. These findings could be useful for breeding drought-resistant wheat cultivars using marker-assisted selection to accumulate these favorable alleles of SNPs associated with yield-related traits to increase grain yield.
University of the Free State and Seed-Co
Vicky Coetzee, Cornelia M. Bender, Renée Prins, Zacharias A. Pretorius
Notwithstanding the re-emergence and importance of wheat stem rust caused by Puccinia graminis f. sp. tritici (Pgt), the degree of protection provided by different types of resistance has not been carefully investigated in contemporary studies. Seven wheat entries were exposed to stem rust infection and fungicide response in a split-plot field experiment over two seasons. Severe epidemics of Pgt race PTKST, generated by frequent inoculation of spreader rows within and around the trial, developed in both years. By comparing grain yield in rusted and fungicide sprayed plots, varieties SC Nduna (Sr31) and SC Stallion (Sr2+Sr31) sustained mean yield losses of 28.8% and 20.7%, respectively. From entries with adult plant resistance (APR), Kingbird recorded a loss of 10.1% as compared to W1406 (19.5%) and W6979 (15.4%). Grain yield of SC Sky which exhibits all stage resistance (ASR) was reduced by 6.4% over the two seasons. The highest yield loss (47.9%) was measured for Line 37, the susceptible control. A significant linear relationship occurred between percentage yield loss and AUDPC in both seasons (R2=0.99 and 0.83). This study showed that not all sources of APR to stem rust provided the same level of protection under severe disease pressure. In the absence of virulence for SC Sky, ASR conferred the most protection.
Sathguru Management Consultants
Kanan Vijayaraghavan, Venugopal Chintada, Rituparna Majumder, Richa Kapur, K. Aishwariya Varadan
South Asia has the highest "wheat dependent" low income community in the world. Stem rust and blast are recognized as the most damaging disease of wheat in the region producing 19% of the world's wheat. In order to combat the potential threat the national research centers were geared up to track the real time movement of wheat diseases, generate disease incidence data and create an enabling environment to boost wheat research in the region through streamlined efforts and enhanced SAARC tool box deployed six years ago.
Recent data (2016-17) from the tool box has shown a significant increase in the data records captured in this region compared to previous years. This has been possible because of heightened awareness amongst the scientists and with the continuous capacity building through pre-season and in-season surveillance trainings organized by Sathguru in collaboration with National Wheat Research Institutes at various levels.
The model is helping partner institutes to be self-sufficient for generating, maintaining wheat disease surveillance data in national and global databases and exchanging real time information with stakeholders. The application have been widely deployed and competently being used by 95% of rust surveillance teams in the wheat fields of SAARC region.
The study will focus on how national research center's judicious decision of carrying out diligent surveillance during the season contributed to safeguarding wheat crops in their respective nations through increased vigilance on emergence of new races and targeted introduction of regionally resistant varieties. Further using this data scientist's can aim to strategize their wheat research for identification of resistant varieties and eventually resulting in increased productivity addressing food security of the region.
Institute of Evolution and the Department of Evolutionary and Environmental Biology, University of Haifa, Israel
Dina Raats, Lin Huang, Valeria Bocharova, Jorge Dubcovsky, Abraham Korol, Tzion Fahima
Wild emmer wheat (Triticum dicoccoides, (DIC)) is an important source of resistance to stripe rust due to presence of Puccinia striiformis in its natural habitats with high humidity and relatively low temperatures that are favorable for stripe rust development. Previously, we showed that DIC accessions from northern Israel were highly resistant to stripe rust. According to the rust responses of three DIC accessions (G25, H52, G303) and mapping of the resistance to relatively close, but different, genetic positions on chromosome 1BS, three different resistance genes were assumed to be present. However, the development of additional critical recombinants and new higher resolution genetic maps for these three genes in subsequent work led us to place YrH52 and YrG303 in the same genetic interval as Yr15, suggesting that the three putative genes are allelic or identical. The recent cloning of Yr15 allowed us to test this hypothesis using an EMS mutagenesis approach. We sequenced the Yr15 locus in five yrH52 and three yrG303 susceptible mutants and identified missense point mutations associated with the susceptible phenotype in each one. Thus, YrH52 and YrG303 may not be new genes. Further work is under way to determine if these genes are allelic or identical.