Location: Plant Science Research2018 Annual Report
1a. Objectives (from AD-416):
Crop improvement is a balancing act requiring simultaneous selection for multiple diverse traits, including resistance to a range of diseases, to develop superior new cultivars. One of the diseases that is a subject of investigation here (Fusarium head blight) continues to cause significant economic losses to the U.S. wheat crop, while another (stem rust) has the potential to do so. Similarly, crown rust continues to be a significant disease of oat. The overall goal of this project is to use genetic engineering technologies to develop novel molecular variants of specific genes and validate that they, as well as a previously identified spontaneous mutation, improve resistance to these particular diseases in wheat and oat. The approaches for improving disease resistance will generate novel resources and knowledge for protecting wheat against both FHB and stem rust, and oat against crown rust, in a manner that complements current breeding efforts for both diseases. These research activities will be coupled with the coordination of a service activity that provides a conduit for Midwestern hard red spring wheat breeders to evaluate jointly their advanced germplasm for agronomic quality and disease resistance at multiple locations. Combining basic and applied research in this manner will ensure that new wheat and oat cultivars retain high yield and quality while also being protected from current and potential disease threats. To achieve project goals, three objectives will be pursued: Objective 1: Evaluate a novel wheat genome deletion that improves Fusarium head blight resistance in adapted hard red spring wheat under field conditions. Sub-Objective 1.A. Evaluate the effect of genetic background on Fusarium head blight resistance conferred by a novel genome deletion. Sub-Objective 1.B. Evaluate the effect of pyramiding the deletion and the partial FHB resistance gene Fhb1 on suppression of FHB. Sub-Objective 1.C. Evaluate the effect of the deletion on agronomic performance in contemporary hard red spring wheat. Objective 2: Establish efficient transformation systems in parallel for wheat and oats, and improve disease resistance by endogenous gene disruption and foreign gene addition. Sub-Objective 2.A. Validate candidate rust susceptibility genes in the model grass Brachypodium. Sub-Objective 2.B. Disrupt stem rust susceptibility genes in wheat. Sub-Objective 2.C. Disrupt crown rust susceptibility genes in oat. Objective 3: Coordinate the Hard Red Spring Wheat Uniform Regional Performance Nursery Program.
1b. Approach (from AD-416):
Objective 1 seeks to enhance Fusarium head blight resistance in hard red spring wheat by introducing a unique genome deletion that improves resistance to this disease. We will determine if the deletion will improve resistance in other susceptible wheat genotypes. Near-isogenic lines of two susceptible hard red spring wheat cultivars that either possess or do not possess the deletion have been developed. These lines will be evaluated in Fusarium head blight nurseries at several locations to determine if lines with the deletion exhibit improved Fusarium head blight resistance when compared to the lines that do not possess it. We will test whether the deletion, when paired with a Fusarium head blight resistance gene, enhances Fusarium head blight resistance synergistically. Near-isogenic lines of two Fusarium head blight-susceptible hard red spring wheat cultivars that possess either the resistance gene alone or the gene together with the deletion will be evaluated in Fusarium head blight nurseries to determine if lines with both the deletion and the resistance gene exhibit superior resistance compared to the lines with resistance agene alone. We will examine how the deletion affects agronomic performance. The deletion has been introduced into diverse hard red spring wheat breeding lines. These near-isogenic lines and the original parents will be grown in field plots at several locations. Agronomic traits will be measured in the near-isogenic lines and compared to their parents to determine if the deletion has a detrimental effect on them. Objective 2 seeks to improve resistance to wheat stem rust and oat crown rust. We will employ the model grass Brachypodium as a testbed to test if mutating certain genes enhances resistance to these diseases. Genome editing using the CRISPR/Cas9 system will be used to perturb the genes, which are known or thought to enhance resistance to pathogens in other plant species when disrupted, in Brachypodium. Plants with confirmed mutations in the genes will be inoculated with the pathogens that cause wheat stem rust and oat crown rust, to confirm that their disruption improves resistance to these diseases. We will build on these results by creating, in wheat, mutations in the genes that enhance stem rust resistance in Brachypodium, and determining whether they also improve stem rust resistance in wheat. We will also create mutations in these same genes but in oats, to determine whether enhanced crown rust resistance can be obtained. Objective 3 will provide hard red spring wheat breeding programs in the upper Midwest an annual opportunity to have their advanced wheat germplasm evaluated for performance at more than a dozen field sites in fives states and Canada. The advanced lines are planted in replicated plots at these locations, and agronomic trait data on the germplasm are obtained by colleagues at each location.
3. Progress Report:
This is a new research project that was initiated on March 19, 2018, replacing project 5062-21000-030-00D. Research associated with project objectives has been initiated. This includes harvesting greenhouse seed increases of near-isogenic wheat lines produced in University of Minnesota, North Dakota State University, and South Dakota State University genetic backgrounds, and making reselections for superior plants. These near-isogenic lines harbor a novel genome deletion that improves Fusarium head blight resistance. They have been created to determine whether the deletion confers disease resistance in different genetic backgrounds, to test whether the deletion, when pyramided with the Fusarium head blight resistance gene Fhb1, measurably increases disease resistance, and to evaluate the effect of the deletion on agronomic performance. Plasmids for gene editing in the model grass Brachypodium distachyon have been developed, and transformation experiments have been initiated to use gene editing to disrupt disease susceptibility genes as a strategy for improving resistance to stem rust. Further, the implementation of a highly efficient transformation protocol in wheat has been initiated. Methodical testing of different parameters and variables has been conducted to develop the method for use in transgene alteration of disease resistance in wheat. As well, attempts to develop an efficient transformation method for oats have been conducted, and high rates of transformation based on transient expression of a reporter gene have been obtained. Lastly, the first stages of the coordination of the 2018 Hard Red Spring Wheat Uniform Regional Performance Nursery, including developing the list of germplasm entries for this year’s evaluations, and organizing seed distribution to the program’s location cooperators, have been completed.