Effect of EPSPS gene copy number and glyphosate selection on fitness of glyphosate-resistant Bassia scoparia in the field.

The widespread evolution of glyphosate-resistant (GR) Bassia scoparia in the U.S. Great Plains poses a serious threat to the long-term sustainability of GR sugar beet. Glyphosate resistance in B. scoparia is due to an increase in the EPSPS (5-enolpyruvyl-shikimate-3-phosphate) gene copy number. The variation in EPSPS gene copies among individuals from within a single GR B. scoparia population indicated a differential response to glyphosate selection. With the continued use of glyphosate in GR sugar beet, the effect of increasing glyphosate rates (applied as single or sequential applications) on the fitness of GR B. scoparia individuals with variable EPSPS gene copies was tested under field conditions. The variation in EPSPS gene copy number and total glyphosate rate (single or sequential applications) did not influence any of the reproductive traits of GR B. scoparia, except seed production. Sequential applications of glyphosate with a total rate of 2214 g ae ha-1 or higher prevented seed production in B. scoparia plants with 2-4 (low levels of resistance) and 5-6 (moderate levels of resistance) EPSPS gene copies. Timely sequential applications of glyphosate (full recommended rates) can potentially slow down the evolution of GR B. scoparia with low to moderate levels of resistance (2-6 EPSPS gene copies), but any survivors (highly-resistant individuals with ≥ 8 EPSPS gene copies) need to be mechanically removed before flowering from GR sugar beet fields. This research warrants the need to adopt ecologically based, multi-tactic strategies to reduce exposure of B. scoparia to glyphosate in GR sugar beet.

Pathotypes Detected Among Populations of Pratylenchus neglectus Collected From Montana.

The root lesion nematode, Pratylenchus neglectus, is one of the most damaging nematodes to affect wheat worldwide. The nematode is widely distributed in Montana, primarily affecting winter wheat within the state. Managing the nematode primarily involves rotation to resistant and moderately resistant crops (peas, lentils, and barley). A nematode survey was conducted across the state nearly 10 years after an initial survey, to reassess the nematode threat and assess the impact of changing trends in crop rotations. To assess the broad applicability of rotation crops to control P. neglectus across Montana, greenhouse trials were conducted to challenge rotational crops using eight populations of P. neglectus collected from geographically diverse locations across the state. In the trials, conducted with four Montana crops, a significant interaction was detected between crop and nematode population (analysis of variance P < 0.001). Populations from Hill, Dawson, and Chouteau counties were found to be pathogenic on barley. Male nematodes were detected in seven of the eight pot culture populations, and these were confirmed to be P. neglectus by morphological and molecular methods. These results suggest a re-evaluation of barley and lentils as a management option for P. neglectus in Montana, as pathotypes for each exist within the state.

Competition Between Fusarium pseudograminearum and Cochliobolus sativus Observed in Field and Greenhouse Studies.

Among root pathogens, one of the most documented antagonisms is the suppression of Cochliobolus sativus by Fusarium (roseum) species. Unfortunately, previous studies involved single isolates of each pathogen and thus, provided no indication of the spectrum of responses that occur across the respective species. To investigate the variability in interactions between Cochliobolus sativus and Fusarium pseudograminearum, field and greenhouse trials were conducted that included monitoring of spring wheat plant health and monitoring of pathogen populations via quantitative real-time polymerase chain reaction. The interactions between two isolates of C. sativus and four isolates of F. pseudograminearum were explored in three geographically distinct wheat fields. To complement field trials and to limit potentially confounding environmental variables that are often associated with field studies, greenhouse trials were performed that investigated the interactions among and between three isolates of C. sativus and four isolates of F. pseudograminearum. Across field locations, C. sativus isolate Cs2344 consistently and significantly reduced Fusarium populations by an average of 20.1%. Similarly, F. pseudograminearum isolate Fp2228 consistently and significantly reduced C. sativus field populations by an average of 30.9%. No interaction was detected in the field between pathogen species with regards to disease or crop losses. Greenhouse results confirmed a powerful (>99%), broadly effective suppression of Fusarium populations by isolate Cs2344. Among greenhouse trials, additional isolate-isolate interactions were observed affecting Fusarium populations. Due to lower C. sativus population sizes in greenhouse trials, significant Fusarium suppression of C. sativus was only detected in one isolate-isolate interaction. This study is the first to demonstrate suppression of Fusarium spp. by C. sativus in field and greenhouse settings. These findings also reveal a complex competitive interaction between these two pathogen species that was previously unknown.

Assessment and Management of Root Lesion Nematodes in Montana Wheat Production.

Root lesion nematodes (Pratylenchus spp.) hinder dryland wheat production worldwide. Montana, a leading U.S. wheat production region, has climatic conditions favorable for Pratylenchus spp. A 2006-2007 statewide soil survey revealed damaging populations of Pratylenchus neglectus, primarily in winter wheat production areas of Montana, whereas P. thornei was not found. Analyses of wheat yields in infested fields revealed negative correlations between yields and spring nematode populations (all P < 0.05 and all R2 > 0.2). Statewide yield losses due to root lesion nematodes were an estimated 12 and 15% for winter wheat in 2006 and 2007, respectively. A subsequent study conducted in 2008 to 2009 revealed significant differences in reproductive success of P. neglectus among seven rotation treatments (P < 0.001). Nematode populations persisted from spring to fall under fallow, barley, pea, and camelina; increased under winter wheat and canola; and decreased under lentil. Populations were sustained through winter following winter wheat and barley but declined following canola, camelina, pea, lentil, and fallow. A screening of 19 barley lines for resistance to P. neglectus revealed significant variation in resistance among entries (P < 0.001), with 'Harrington' barley displaying promising levels of resistance. Development of resistant wheat cultivars remains a principal goal in managing this nematode.

Distribution and Prevalence of Fusarium Crown Rot and Common Root Rot Pathogens of Wheat in Montana.

Distribution of Fusarium crown rot (FCR) and common root rot (CRR) pathogens associated with wheat (Triticum aestivum) in 91 fields in Montana were determined during the 2008 and 2009 crop seasons using real-time quantitative polymerase chain reaction (qPCR) and conventional isolation methods. Correlations (P < 0.001) were found between detection methods for both diseases. FCR was detected in 57% of the fields and CRR was detected in 93% of the fields surveyed. Percent incidence based on isolation from individual tillers was Bipolaris sorokiniana (15%), F. culmorum (13%), and F. pseudograminearum (8%). FCR populations were highly variable across the regions and were not detected in any fields from the Gb5 soil types of Judith Basin and Fergus counties. The spatial distributions of FCR and CRR were affected by elevation, soil type, and temperature. High FCR populations were associated with spring wheat crops rather than winter wheat based on qPCR (P < 0.001). FCR and CRR could produce yield losses in a range of 3 to 35%. This study is the first time that qPCR was used to survey these two pathogen groups, and the merits and weakness of qPCR relative to traditional isolation methods are discussed.

Population Dynamics Between Fusarium pseudograminearum and Bipolaris sorokiniana in Wheat Stems Using Real-Time qPCR.

Fusarium pseudograminearum and Bipolaris sorokiniana are causal agents of Fusarium crown rot and common root rot, respectively, of wheat and cause significant losses worldwide. Understanding the population dynamics between these two pathogens at late stages of wheat development is needed. The effect of F. pseudograminearum and B. sorokiniana inocula applied singly or in mixtures at seeding to spring wheat 'Hank' was measured using seedling stand, grain yield, and pathogen populations in the first internode at heading, milk, and harvest stage of wheat development using real-time quantitative polymerase chain reaction. High and low rates of F. pseudograminearum inoculum reduced B. sorokiniana populations in field trials but B. sorokiniana inoculations did not affect F. pseudograminearum populations. Populations of both pathogens increased from heading until harvest, with F. pseudograminearum colonizing lower internodes earlier than B. sorokiniana. Neither pathogen prevented infection by the other in the first internode of wheat stems. Inoculations increased incidence of infection and co-infection relative to natural settings observed for both pathogens. At the seedling stage, both fungi, individually or combined, reduced the seedling stands when compared with a noninoculated control for the three location-years. Grain yield and F. pseudograminearum populations were inversely correlated, while B. sorokiniana populations were not correlated with yield.

Influence of Beet necrotic yellow vein virus on Sugar Beet Storability.

Rhizomania caused by Beet necrotic yellow vein virus (BNYVV) and storage losses are serious sugar beet production problems. To investigate the influence of BNYVV on storability, six sugar beet cultivars varying for resistance to BNYVV were grown in 2005 and 2006 in southern Idaho fields with and without BNYVV-infested soil. At harvest, samples from each cultivar were placed in an outdoor ventilated pile in Twin Falls, ID and were removed at 40-day intervals starting at the end of October. After 144 and 142 days in storage, sugar reduction across cultivars averaged 20 and 13% without and 68 and 21% with BNYVV for the 2005 and 2006 roots, respectively. In the December samplings, frozen root area was 1 and 2% without and 25 and 41% with BNYVV for the 2005 and 2006 roots, respectively. Root rot was always worse with stored roots from BYNVV-infested soil in December, January, and February samplings. Root weight loss was variable in 2005; however, in 2006, an increase in weight reduction always was associated with BNYVV-infested roots. In order to prevent losses in rhizomania-infested areas, cultivars should be selected for storability as well as rhizomania resistance.

Viability and maturation of Aphanomyces cochlioides oospores.

Plasmolysis, tetrazolium bromide staining and microscopic appearance were tested for their usefulness in determining viability of oospores of Aphanomyces cochlioides. For comparison, three lethal treatments were employed to contrast the reaction of dead oospores and untreated, presumably viable oospores. Few oospores stained with tetrazolium bromide, even though plasmolysis and microscopic appearance indicated that 85% were viable. Cytoplasm of viable oospores was densely organized and uniformly granular (DOUG), whereas cytoplasm of oospores exposed to lethal treatments was loosely organized and non-uniformly granular (LONG). Dose-response bioassay experiments were conducted with untreated oospores of varying inoculum densities or with mixtures of untreated DOUG and heat-treated LONG oospores in varying proportions. The number of DOUG oospores was correlated (R(2) = 0.62, P < 0.001) with severity of damping-off of sugar beet seedlings caused by A. cochlioides. Thus, the granular appearance of cytoplasm offered a fast, easy and reliable indicator of viability of A. cochlioides oospores. Tests with newly formed oospores/oogonia showed that >80% harvested at 3-4 d after inoculation of hypocotyls stained with tetrazolium, but by 8-9 d <10% stained, apparently because of declining permeability of the spore wall to tetrazolium as oospores matured.