Use of Pyrosequencing Technology to Genotype Imidazolinone-Tolerant Wheat

Cultivars of several cereal crops have been developed with acetohydroxyacid synthase (AHAS) insensitivity to imidazolinone herbicides and are now an important tool for weed management. Options for screening for imidazolinone resistant lines include direct herbicide application, biochemical assays for AHAS activity and DNA-based methods. Herbicide and biochemical assays for AHAS activity provide limited information as to mutation copy number and provide no information as to the genome on which the mutation is located without extensive test crossing. Wheat (Triticum aestivum L.) can have between one and six copies of the resistant acetohydroxyacid synthase on any of three genomes. A novel DNA-based screening protocol is described here in which pyrosequencing is used to screen for the S653N imidazolinone tolerant mutation in wheat. One assay is shown to successfully detect zero to four copies of the S653N mutation, while additional assays can detect the presence of S653N in individual wheat genomes. All of these assays are based on a single 298-bp PCR fragment and can be easily scaled up or down depending on the number lines that need to be screened. Potential applications include detection of mutant copy number in segregating populations, and the selection of parental lines with genome specific mutant composition

Mapping Stripe Rust Resistance in a BrundageXCoda Winter Wheat Recombinant Inbred Line Population

A recombinant inbred line (RIL) mapping population developed from a cross between winter wheat (Triticum aestivum L.) cultivars Coda and Brundage was evaluated for reaction to stripe rust (caused by Puccinia striiformis f. sp. tritici). Two hundred and sixty eight RIL from the population were evaluated in replicated field trials in a total of nine site-year locations in the U.S. Pacific Northwest. Seedling reaction to stripe rust races PST-100, PST-114 and PST-127 was also examined. A linkage map consisting of 2,391 polymorphic DNA markers was developed covering all chromosomes of wheat with the exception of 1D. Two QTL on chromosome 1B were associated with adult plant and seedling reaction and were the most significant QTL detected. Together these QTL reduced adult plant infection type from a score of seven to a score of two reduced disease severity by an average of 25% and provided protection against race PST-100, PST-114 and PST-127 in the seedling stage. The location of these QTL and the race specificity provided by them suggest that observed effects at this locus are due to a complementation of the previously known but defeated resistances of the cultivar Tres combining with that of Madsen (the two parent cultivars of Coda). Two additional QTL on chromosome 3B and one on 5B were associated with adult plant reaction only, and a single QTL on chromosome 5D was associated with seedling reaction to PST-114. Coda has been resistant to stripe rust since its release in 2000, indicating that combining multiple resistance genes for stripe rust provides durable resistance, especially when all-stage resistance genes are combined in a fashion to maximize the number of races they protect against. Identified molecular markers will allow for an efficient transfer of these genes into other cultivars, thereby continuing to provide excellent resistance to stripe rust

Jointed goatgrass genetics

Jointed goatgrass (Aegilops cylindrica) is a weed that causes serious problems in winter wheat fields in the western United States. Jointed goatgrass and wheat have a close genetic relationship and share many traits. To date, no herbicide has been developed that can selectively remove jointed goatgrass from conventional wheat. However, current herbicide-resistant wheat technology will provide producers with a tool to control jointed goatgrass in the crop. Concerns remain, though, that the resistance genes could move from wheat into jointed goatgrass populations. Understanding the genetic relationship between wheat and jointed goatgrass will help with the development of management strategies that minimize the potential for gene flow between the two species

Identification of a candidate gene for the wheat endopeptidase \u3ci\u3eEp-D1\u3c/i\u3e locus and two other STS markers linked to the eyespot resistance gene \u3ci\u3ePch1\u3c/i\u3e

Wheat is prone to strawbreaker foot rot (eyespot), a fungal disease caused by Oculimacula yallundae and O. acuformis. The most effective source of genetic resistance is Pch1, a gene derived from Aegilops ventricosa. The endopeptidase isozyme marker allele Ep-D1b, linked to Pch1, has been shown to be more effective for tracking resistance than DNA-based markers developed to date. Therefore, we sought to identify a candidate gene for Ep-D1 as a basis for a DNA-based marker. Comparative mapping suggested that the endopeptidase loci Ep-D1 (wheat), enp1 (maize), and Enp (rice) were orthologous. Since the product of the maize endopeptidase locus enp1 has been shown to exhibit biochemical properties similar to oligopeptidase B purified from E. coli, we reasoned that Ep-D1 may also encode an oligopeptidase B. Consistent with this hypothesis, a sequence-tagged-site (STS) marker, Xorw1, derived from an oligopeptidase B-encoding wheat expressed-sequence-tag (EST) showed complete linkage with Ep-D1 and Pch1 in a population of 254 recombinant inbred lines (RILs) derived from a cross between wheat cultivars Coda and Brundage. Two other STS markers, Xorw5 and Xorw6, and three microsatellite markers (Xwmc14, Xbarc97, and Xcfd175) were also completely linked to Pch1. On the other hand, Xwmc14, Xbarc97, and Xcfd175 showed recombination in the W7984 × Opata85 RIL population suggesting that recombination near Pch1 is reduced in the Coda/Brundage population. In a panel of 44 wheat varieties with known eyespot reactions, Xorw1, Xorw5, and Xorw6 were 100% accurate in predicting the presence or absence of Pch1 whereas Xwmc14, Xbarc97, and Xcfd175 were less effective. Thus, linkage mapping and a germplasm survey suggest that the STS markers identified here should be useful for indirect selection of Pch1

Genetic Dissection of QTL Associated with Grain Yield in Diverse Environments

Wheat (Triticum aestivum L.) breeding programs strive to increase grain yield; however, the progress is hampered due to its quantitative inheritance, low heritability, and confounding environmental effects. In the present study, a winter wheat population of 159 recombinant inbred lines (RILs) was evaluated in six trials under rainfed, terminal drought, and fully-irrigated conditions, over four years. Quantitative trait locus/loci (QTL) mapping was conducted for grain yield main effect (GY) and the genotype × environment interaction (GEI) effect. A total of 17 QTL were associated with GY and 13 QTL associated with GEI, and nine QTL were mapped in the flanking chromosomal regions for both GY and GEI. One major QTL Q.Gy.ui-1B.2, explaining up to 22% of grain yield, was identified in all six trials. Besides the additive effect of QTL associated with GY, interactions among QTL (QTL × QTL interaction), QTL × environment, and QTL × QTL × environment were also observed. When combining the interaction effects, QTL Q.Gy.ui-1B.2 along with other QTL explained up to 52% of the variation in grain yield over the six trials. This study suggests that QTL mapping of complex traits such as grain yield should include interaction effects of QTL and environments in marker-assisted selection

Mapping stripe rust resistance in a BrundageXCoda winter wheat recombinant inbred line population

A recombinant inbred line (RIL) mapping population developed from a cross between winter wheat (Triticum aestivum L.) cultivars Coda and Brundage was evaluated for reaction to stripe rust (caused by Puccinia striiformis f. sp. tritici). Two hundred and sixty eight RIL from the population were evaluated in replicated field trials in a total of nine site-year locations in the U.S. Pacific Northwest. Seedling reaction to stripe rust races PST-100, PST-114 and PST-127 was also examined. A linkage map consisting of 2,391 polymorphic DNA markers was developed covering all chromosomes of wheat with the exception of 1D. Two QTL on chromosome 1B were associated with adult plant and seedling reaction and were the most significant QTL detected. Together these QTL reduced adult plant infection type from a score of seven to a score of two reduced disease severity by an average of 25% and provided protection against race PST-100, PST-114 and PST-127 in the seedling stage. The location of these QTL and the race specificity provided by them suggest that observed effects at this locus are due to a complementation of the previously known but defeated resistances of the cultivar Tres combining with that of Madsen (the two parent cultivars of Coda). Two additional QTL on chromosome 3B and one on 5B were associated with adult plant reaction only, and a single QTL on chromosome 5D was associated with seedling reaction to PST-114. Coda has been resistant to stripe rust since its release in 2000, indicating that combining multiple resistance genes for stripe rust provides durable resistance, especially when all-stage resistance genes are combined in a fashion to maximize the number of races they protect against. Identified molecular markers will allow for an efficient transfer of these genes into other cultivars, thereby continuing to provide excellent resistance to stripe rust

Perceived Consequences of Herbicide-Tolerant and Insect-Resistant Crops on Integrated Pest Management Strategies in the Western United States : Results of an Online Survey

We conducted an online survey to assess the potential effects of herbicide-tolerant (HT) and insect-resistant (IR) crops on integrated pest management (IPM) practices in the Western United States For HT crops, participants perceived a decrease in several IPM practices, including crop and herbicide rotations and the combined use of multiple weed control strategies. The most serious potential consequences were considered to be a shift in weed species composition and development of herbicide-resistant weeds. For IR crops, respondents perceived a beneficial reduction in application of both broad-spectrum and selective insecticides. The most significant issues for IR crops were believed to be potential development of target pest resistance and difficulties with management of insect refuges. The survey results support the need for continued emphasis on comprehensive strategies in IPM education programs to prolong the usefulness of HT and IR crops.This paper received grant funding from USDA's Western Integrated Pest Management Center to support a work group on this topic

ZemetraRobertCropSoilScienceMappingStripeRust.pdf

A recombinant inbred line (RIL) mapping population developed from a cross between winter wheat (Triticum aestivum L.) cultivars Coda and Brundage was evaluated for reaction to stripe rust (caused by Puccinia striiformis f. sp. tritici). Two hundred and sixty eight RIL from the population were evaluated in replicated field trials in a total of nine site-year locations in the U.S. Pacific Northwest. Seedling reaction to stripe rust races PST-100, PST-114 and PST-127 was also examined. A linkage map consisting of 2,391 polymorphic DNA markers was developed covering all chromosomes of wheat with the exception of 1D. Two QTL on chromosome 1B were associated with adult plant and seedling reaction and were the most significant QTL detected. Together these QTL reduced adult plant infection type from a score of seven to a score of two reduced disease severity by an average of 25% and provided protection against race PST-100, PST-114 and PST-127 in the seedling stage. The location of these QTL and the race specificity provided by them suggest that observed effects at this locus are due to a complementation of the previously known but defeated resistances of the cultivar Tres combining with that of Madsen (the two parent cultivars of Coda). Two additional QTL on chromosome 3B and one on 5B were associated with adult plant reaction only, and a single QTL on chromosome 5D was associated with seedling reaction to PST-114. Coda has been resistant to stripe rust since its release in 2000, indicating that combining multiple resistance genes for stripe rust provides durable resistance, especially when all-stage resistance genes are combined in a fashion to maximize the number of races they protect against. Identified molecular markers will allow for an efficient transfer of these genes into other cultivars, thereby continuing to provide excellent resistance to stripe rust