Properties of the tan spot necrosis toxin.

Cultivar-specific toxic metabolites of Pyrenophora tritici-repentis are involved in the appearance of necrotic and chlorotic foliar lesions characteristic of wheat tan spot. A necrosis-inducing proteinaceous toxin was purified from isolate 86-124 of P. tritici-repentis, sequenced by gas phase amino-acid microsequencing, and characterized by circular dichroism (CD) spectroscopy and isoelectric focusing. The purified protein had a similar amino acid composition and molecular weight as previously reported. A sequence of 101 continuous amino acids was obtained from overlapping proteolytic fragments, but the amino terminus was blocked and 9 to 16 amino acids could not be sequenced. A secondary structure prediction based on the amino acid sequence indicates a beta sheet protein with little alpha helix, which is in agreement with the structure determined by CD spectroscopy. Sequence analysis indicated the presence of a possible membrane adhesion site and several possible phosphorylation sites that may be involved in phytotoxicity. Additional pathogen isolates are presently being assessed for symptomology on wheat differentials to find candidates for biochemical analyses. (Meinhardt and Zhang)

An electrolyte leakage assay for tan spot necrosis.

Transmission electron microscopy demonstrated that leaf tissue from toxin-sensitive wheat quickly suffered membrane degradation after exposure to necrosis toxin. In an effort to develop a more rapid and quantitative bioassay for necrosis toxin activity, we found that the conductivity of leaching solutions increased for at least 8 h after toxin exposures of 4 h or more in toxin-sensitive wheat, but not in water controls or insensitive genotypes. Electrolyte leakage from sensitive wheat increased with toxin concentration between 1 and 20 µg/ml, the highest concentration tested. This bioassay, which can be completed in 1 day, should facilitate studies of necrosis toxin mode of action. (Rasmussen, Kwon, and Freeman)

Epidemiology.

The epidemiology of wheat diseases, with emphasis on tan spot and Fusarium head blight, is being studied with several approaches. Greenhouse-grown plants set out in a wheat field for 24 h show discrete foliar infection periods, so temporally continuous environmental variables can be analyzed. Similarly, Fusarium inoculum can be enumerated by washing wheat heads after a 24 h exposure. A common protocol is being followed in nine locations in North America to help ensure measurement of inoculum in a head blight epidemic. In an effort to improve disease warning systems, an artificial neural network was developed to distinguish tan spot infection periods. The model used environmental data as input and was 85 % accurate in a validation test. In comparison, a stepwise logistic regression model was 69 % correct, and a multivariate discriminant model was 50 % correct. A second neural-network model was developed to predict wheat leaf wetness status under various conditions of input information and crop density. Predictions of leaf wetness in a validation data set were 82 to 91 % accurate and duration estimates were correct within an average of 0.6 to 1.9 h. The accuracy of this model was equivalent to that of other models that predict dew period, but the model also can account for leaf wetness caused by rainfall. (Francl, Panigrahi, Jordahl, and De Wolf)

Conidiogenesis of Pyrenophora tritici-repentis.

Conidiogenesis of P. tritici-repentis occurs in culture under alternating light and dark cycles with conidiophores formed in light and conidia in dark. Leaf area of the tan spot-susceptible spring wheat line `ND495' was measured nondestructively, and plants were inoculated with isolate Pti2. After a 24-h wet period, plants were held in a growth chamber for 10 days. Plants then were subjected to various light and wetness regimes lasting from 12 to 96 h. Percent disease severity was estimated at the end of each treatment, and conidia/sq mm of diseased area were estimated from leaves blended in water (Riaz et al. Phytopath 81:1298-1302). Conidia failed to form prior to treatment or after 24 to 96 h in a continuous light, continuous wet environment. However, unlike the diurnal requirement for conidiogenesis in vitro, conidia formed after 12 h or longer in the dark. After 96 h of alternating 8 h dark, wet and 16 h light, dry cycles, 1.7 conidia/mm were produced. Thus, conidiophores can be initiated on wheat leaves in an unsaturated atmosphere in the light and a wet period during darkness is sufficient for conidiogenesis. (Francl, Jordahl, and De Wolf)

Publications.

Francl LJ and Jordahl JG. 1994. Refined procedures for inoculating wheat seedlings and rating their reaction to Pyrenophora tritici-repentis. Plant Dis 78:745-748.

Francl LJ. 1995. Challenge of bioassay plants in a monitored outdoor environment. Can J Plant Path 17:138-143.

Francl LJ, DeWolf E, and Jordahl JG. 1995. Conidiation of Pyrenophora tritici-repentis. Phytopath 85:1044.

Francl LJ, Panigrahi S, and Padhi T. 1995. Neural network models that predict leaf wetness. Phytopath 85:1128.

Freeman T, Rasmussen J, Francl L, and Meinhardt S. 1995. Wheat necrosis induced by Pyrenophora tritici-repentis toxin. In: Proc Microscopy Soc of America. Jones and Begell Publ, NY. Pp. 990-991

Meinhardt SW, Zhang H-F, Jordahl JG, and Francl LJ. 1995. Characterization of the necrosis toxin of Pyrenophora tritici-repentis strain 86-124. Phytopath 85:1199.

Rasmussen JB, Francl LJ, Meinhardt S, Kwon C, and Freeman TP. 1995. Effects of the necrosis toxin produced by Pyrenophora tritici-repentis on the membranes of wheat. Phytopath 85:1161.


OHIO

THE OHIO STATE UNIVERSITY

Ohio Agricultural Research and Development Center, Wooster, OH 44691, USA.

Kim G. Campbell, Robert W. Gooding, David M. Jordan, Larry D. Herald, Richard Minyo, Dimuth Siritunga, and Emily Stowe (Department of Horticulture and Crop Science); Patrick Lipps and A.L Johnston (Department of Plant Pathology); and P. Finney, C. Gaines, D. Gualberto, C. Bergman, J. Donelson, G. Bains, L. Andrews, B. Renner, A. Bugaj, J. Kinney, S. Croskey, T. Andrews, L. Fleege, and M. Raeker (USDA-ARS Soft Wheat Quality Laboratory).

Growing conditions and production in Ohio, 1994-95.

Compiled by R.W. Gooding.

Growing Conditions. The winter of 1994-95 began with mild weather and ample precipitation. Moderate temperatures delayed the onset of dormancy until later than normal. Precipitation amounts increased through January and February, affording the crop adequate snowfall for insulation. The season's coldest weather was in February with temperatures as low as -7_F reported at some locations in Ohio. With the onset of spring, stands were found to be generally healthy throughout the state.

Cool, wet conditions during the spring contributed to high levels of wheat yellow mosaic virus in susceptible wheat lines. Wet conditions during flowering resulted in severe head scab throughout the wheat growing regions of Ohio in 1995. Wheat planted in corn stubble was found to be particularly at risk.

Wheat harvest began during the first week of July. Because of high temperatures, humidity, and generally favorable working conditions, the harvest was essentially completed in 4 weeks.

Production. Wheat production in Ohio increased by nearly 8 % compared to 1994 levels. Harvested acres of 1.2 million combined with an average yield of 61 bu/acre resulted in an overall production of 73.8 million bushels of wheat in 1995. In early June, the estimated statewide wheat yield was 70 bu/acre. Following harvest, the average yield was recorded to be 61 bu/acre. On a statewide basis, the estimated yield loss was 11 million bushels. Added to this loss were the dockage most farmers received at the elevators for low test weights (45-57 lb/bu) and shriveled grain. Most reports indicated a $0.30/bu dockage was common. Additionally, certified seed producers cleaned out over 30 % of the seed to achieve acceptable germination standards (30 % loss on 1.5 million bushels of seed wheat). Thus, $43 million was lost in production (at $3.90/bu price), dockage amounted to $22 million, and loss of seed wheat production was $3.8 million . The cost to the millers and bakers from mycotoxin contamination is still unknown.

USDA ARS Soft Wheat Quality Lab.

Research. We completed study of milling-, biochemical-, Mixograph-, gliadin and glutenin sub unit-, and sugar snap cookie-traits for a second year and straight dough bread traits to associate with RFLP markers for 78 lines of a `hard x soft' wheat cross. We continued density studies of wheat to compare with other flour milling and end-use properties. We also continued research using the NIRS-6500 to determine its potential to predict important flour milling and end-use properties. We have determined that grain shrivelling could be determined by instrument. It was found that the Single Kernel Hardness Tester did not accurately reflect relative kernel `hardness' when measuring kernels at various moisture levels, or for various degrees of kernel shrivelling. Also, we demonstrated that SKHT raw data can relate highly to kernel softness by milling. We developed formula substitution for chlorination of flour in cake and other related products. Modification studies of a flour hydration procedure to make it more sensitive to low levels of field sprouting were continued.

Evaluation. Evaluation of 7,950 micro-, 1600 advanced-, and about 300 drill-strip nursery samples from 18 public and 12 private breeding programs was completed. We installed movable-tier shelving for the second of three rooms. We completed a fourth year of evaluating uniform nurseries from sub-location composites. Statistical treatment will now be completed cooperatively with geneticists.

Ohio Soft Red Winter Wheat Performance Test.

Ohio is a leading state in the production of soft red winter wheat. The selection of superior cultivars has the potential to add millions of dollars in revenue to farmers' income. The Ohio State University annually tests public and private wheat varieties under field conditions to provide information on the comparative performance of cultivar traits. Field studies are conducted across several Ohio environments to compare agronomic performance on yield, test weight, heading date, plant height, lodging, and diseases present. In cooperation with the USDA-ARS Soft Wheat Quality Laboratory, grain quality evaluations provide information on milling and baking properties.

Table 1. Reaction of soft red winter wheat varieties to disease in Ohio, revised 1995.

__________________________________________________________________________________________

Septoria nodoru m Wheat

Powdery Leaf leaf glume yellow

Brand Variety mildew rust blotch blotch mosaic

__________________________________________________________________________________________

AGRA GR863 MR MR S S MR

GR876 MR R MR R MR

GR915 R MR R R R

GR933 S MS MS MS R

GR942 MR S S S S

Agripro Twain MR R S S ó

Sawyer MS MR MS MR S

Pontiac S S S S S

Clemens S R MS MR S

Certified Becker VS S MS S MR

Caldwell S MS S VS S

Cardinal S S MR MR MR

Clark MS S S S MR

Dynasty MS S S S R

Excel S S S S MR

Freedom MR R MR MR MR

Glory MR S MS MS R

Grant S MR S S MR

Hopewell MR S MS MS R

Jackson MR S MR MR S

Madison MR MS MS MS R

Wakefield MR S MR MS MS

Countrymark 544 R MR MS S MS

558 R MS MS MS S

568 MR S S MS MR

Greenland GL9240 S ó ó ó S

GL9400 MR R MS S MS

Hytest Succession R MR S MS S

Nosco Classic RW151 MR MR S MS MS

Table 1 (continued). Reaction of soft red winter wheat varieties to disease in Ohio, revised 1995.

__________________________________________________________________________________________

Septoria nodoru m Wheat

Powdery Leaf leaf glume yellow

Brand Variety mildew rust blotch blotch mosaic

__________________________________________________________________________________________

Pioneer 2510 MS R MR MR R

2545 MR S S S R

2548 MR MS S MS S

2550 S R MS MS MR

2552 R S MR MS R

2555 S MS MR MR ó

2571 MR R S S S

Rupp RS927 S S S S S

Shur Grow SG1550 MS MS MS MS S

Steyer Hoppes S S MS MS MR

Kline R MR MR MR R

McLane R MS MR MR S

Neuleib S MS MS ó ó

Podach MS MS S MS S

Rowland R R S MR R

Stine 501 R MR R R MR

Terra SR204 MR MR MS MS MS

SR205 S MS MS MS R

Thompson TS4020 R MR S MS S

TS5020 R S S MS S

Voris V8040 S S MS MS R

Wellman W9140 MS MR MR ó ó

W9350 MS MS S MS S

W9420 MR S S MS MR

W9540 R S MS MS R

__________________________________________________________________________________________

R = resistant; MR = moderately resistant; MS = moderately susceptible; S = susceptible; and VS = very susceptible; ó = no data available. Data obtained from disease ratings during 1993, 1994, and/or 1995. Changes in resistance or susceptibility from one year to the next are expected because of the presence of, absence of, or change in races of pathogens.