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 Plant Molecular Biology, University of Delhi South Campus
L-myo-inositol phosphate synthase (MIPS; EC 188.8.131.52) have been involved in abiotic stress tolerance and its disruption leads to spontaneous cell death and enhanced tolerance to pathogen. However, its molecular mechanism underlying role of MIPS in growth, immunity and abiotic stress tolerance remains unknown. To delve deeper into the conserved molecular mechanism of MIPS action during growth and stress condition, we characterized the overexpression transgenic of TaMIPS and mutant lines of AtMIPS1. Subsequent, transcriptome analysis revealed the activation of ET/JA dependent immune response in transgenic and SA defense response in mutant. Pull-down analysis revealed the interaction of TaMIPS2 with ethylene synthesis (ACO) and signaling protein (CTR1) component. Due to the established role of ethylene during the skotomorphogenesis, we investigated the effect of myo-inositol phosphate synthase role in ethylene response during hook formation. Our results thus suggest the requirement of MIPS for ethylene response and regulating the growth and immunity.
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.
Wheat Research Institute, Faisalabad, Pakistan
Javed Ahmad, Abid Mahmood
Wheat is an important cereal crop and staple food in Pakistan. Most of the wheat is cultivated late after cotton, rice and sugarcane. Introduction of long duration Bt cotton varieties further pushed its sowing to late December or even early January. Late sowing of wheat crop results in yield loss in the Punjab province. A study has been conducted in experimental fields of Wheat Research Institute, AARI, Faisalabad, Pakistan to find out the possible reasons of low grain yields in late sown crop. Twelve experimental wheat lines were planted on seven sowing dates starting from 1st November to 30th December with ten days interval. The experimental design was a factorial combination of seven sowing dates as main plots and twelve varieties/ genotypes as subplots in a split-plot design with three replications. Effect of temperature on several crop growth stages was studied. Mean minimum temperature during the month of December, 2016 and January, 2017 remained below 5?C and mean maximum was more than 30?C during March 2017. Weather conditions experienced by the crop at each developmental stage were compared with the optimum conditions required on that specific stage in each sowing date. It was revealed that in late sown crop, different phonological/growth stages of the crop and yield components and grain yield were affected negatively. It was concluded from the study that the late sown crop suffered from two types of temperature stresses. The late sown crop faced low temperature stress at starting phase which result in delayed germination and low tillering. At caryopsis formation and grain filling the same crop face high temperature stress causing reduced grain formation and shriveled grains due to enforced maturity. Sowing of wheat at proper time i.e., by the end of November was recommended to fetch maximum yields.
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.
College of Plant Protection, Northwest A&F University, China
Yuanyuan Zhao, Shuxia Zuo, Dan Zheng, Lili Huang, Zhengshen Kang
Wheat stripe rust, caused by basidiomycete fungus Puccinia striiformis f. sp. tritici (Pst), is a damaging disease worldwide. The recent discovery demonstrated the fungus depends on living wheat and aecial hosts, mainly barberry (Berberis) species, to complete its life cycle. In China, we determined that, under natural conditions, the sexual cycle of Pst occurs based on collections of Pst isolates from the diseased barberry in the past three years. However, no direct evidence to support whether barberry plays a role in spreading inoculums to wheat field to cause stripe rust was detected. In the present study, we recovered 103 Pst samples from natural-infected B. shensiana in the western Shaanxi in spring 2016, and also collected 107 Pst isolates from neighboring wheat fields. Phenotype and genotype of the two Pst populations were tested using a set of Chinese differential hosts for Pst and SSR markers, respectively. The phenotype tests showed that 57 race types produced from the barberry-derived Pst populations, consisting of 58 known races, such as CYR 34, CYR32, G22-14, and Su11-14-3, and 45 new races. Many of the two Pst populations shared the same race types. The genotype tests indicated the barberry-derived Pst population produced a rich genotype, obviously higher than the wheat-derived Pst populations. The seven same genotypes were found on 40 isolates of the former and 26 of the latter. Our results provide evidence to support that sexual cycle of Pst occurs regularly in nature in China and that barberry provides inoculums to neighboring wheat fields, triggering stripe rust infections in the spring. This could be a reason why the Chinese Pst populations represent extreme genetic diversity.
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.
QAAFI, The University of Queensland
Robert McIntosh, Peng Zhang, Sami Hoxha, Adnan Riaz, Burkhard Steuernagel, Brande Wulff, Evans Lagudah, Lee Hickey, Sambasivam Periyannan
Wheat is one of the most important staple food and agricultural crop cultivated worldwide. To meet the demands of the raising human population, global wheat production has to be increased which is however declined due to appearance of highly virulent strains of Puccinia striiformis f. sp. tritici (Pst) fungus causing stripe rust disease. Globally, the incidence of stripe rust is effectively managed through the deployment of host plant mediated genetic resistance. But as the resistance present in the current wheat cultivars are ineffective, new sources of resistance particularly from pathogen unexposed genetic resources are of urgent need to prevent stripe rust epidemics. Landrace collections with rich genetic diversity and being less exposed to prevalent pathogen are of valuable source for resistance to new pathogens. In this study, a total of 295 landrace accessions collected by the famous Russian botanist Vavilov was screened for stripe rust resistance using the two predominant lineage Pst strains of Australia. Six accessions with good resistance against the two aggressive Pst strains were selected for genetic characterization and for utilization in global wheat breeding. Characterisation of these novel resistance were undertaken using combination of conventional and advanced genetic tools. While the conventional approach involves the traditional map based gene cloning, the other tool is the recently identified rapid method based on mutagenesis, targeted gene capture and next generation sequencing called "MutRenSeq". Subsequently, the identified novel resistant traits were transferred into elite wheat cultivars through the combination of linked molecular markers and speed breeding techniques. Thus along with the identification of novel resistance, elite wheat cultivars with broad spectrum stripe rust resistance were also generated through the use state of art techniques to sustain global wheat production from the rapidly evolving stripe pathogens.
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.