Comparing Genetic Variation within Red Winter Wheat Populations with and without Image-Based Optical Sorter Selection

Head scab is historically a devastating disease affecting not just all classes of wheat but also barley and other small grains around the world. Fusarium head blight (FHB), or head scab, is caused most often by Fusarium graminearum (Schwabe), (sexual stage – Gibberella zeae) although several Fusarium spp. can cause the disease. This study was conducted to determine the effect of mass selection for FHB resistance using an image-based optical sorter. lines were derived from the C0 and C2 of two populations to compare genetic variation within populations with and without sorter selection. Our overall hypothesis is that sorting grain results in improved Fusarium head blight resistance. Both of the used wheat derived line populations have genetic variation, and population 1 has more than population 17. They are significantly different from each other for fusarium damged kernel (FDK), deoxynivalenol (DON), and other FHB traits. Although both populations are suitable to be grown for bulks, population 1 seems better since it has more genetic variation as well as lower FDK and DON, and earlier heading date. Lines within each population were significantly different and some lines in each population had significantly lower FDK and DON after selection using an optical sorter. Some lines had significant reduction in both FDK and DON, and some others had either FDK or DON reduction. Lines of population 1 that had significant reduction, were more numerous than in population 17, and FDK and DON reduction were greater.

Specific Detection of the Wheat Blast Pathogen (Magnaporthe oryzae Triticum) by Loop-Mediated Isothermal Amplification

Wheat blast, caused by the Magnaporthe oryzae Triticum pathotype, is an economically important fungal disease of wheat. Wheat blast symptoms are similar to Fusarium head scab and can cause confusion in the field. Currently, no in-field diagnostic exists for M. oryzae Triticum. Loop-mediated isothermal amplification (LAMP) primers were designed to target the PoT2 and MoT3 loci, previously shown to be specific for M. oryzae and M. oryzae Triticum, respectively. Specificity was determined using 158 M. oryzae strains collected from infected wheat and other grasses and representing geographic and temporal variation. Negative controls included 50 Fusarium spp. isolates. Sensitivity was assessed using 10-fold serial dilutions of M. oryzae Triticum gDNA. PoT2- and MoT3-based assays showed high specificity for M. oryzae and M. oryzae Triticum, respectively, and sensitivity to approximately 5 pg of DNA per reaction. PoT2 and MoT3 assays were tested on M. oryzae Triticum-infected wheat seed and spikes and identified M. oryzae and M. oryzae Triticum, respectively, using a field DNA extraction kit and the portable Genie II system. The mitochondrial NADH-dehydrogenase (nad5) gene, an internal control for plant DNA, was multiplexed with PoT2 and MoT3 and showed results comparable with individual assays. These results show applicability for M. oryzae Triticum field surveillance, as well as identifying nonwheat species that may serve as a reservoir or source of inoculum for nearby wheat fields.

Effect of temperature and pH on the growth of Fusarium spp. causing Fusarium head blight (FHB) in wheat

Fusarium head blight (FHB) or head scab is emerging as a destructive disease affecting the quantity and quality of wheat worldwide. Several Fusarium spe-cies have been associated with the disease but their composition varies among geographical regions and years. Climatic factors like temperature, pH and humidity influence the growth, survival and infestation of Fusarium species. In the present study, response of thirty six isolates of three Fusarium spp. viz F. graminearum, F. oxysporum and F. pallidoroseum (F. semitectum) to different temperature and pH was assessed by analysing their in vitro growth rate (mm/day) on potato dextrose agar (PDA) medium. We found that all the isolates responded differentially but interestingly isolates of F. graminearum showed better tolerance at broader range of temperature and pH. This attributes make F. graminearum a widely distributed and potent pathogen of wheat.

FgIlv5 is required for branched-chain amino acid biosynthesis and full virulence in Fusarium graminearum

In this study, we characterized FgIlv5, a homologue of the Saccharomyces cerevisiae keto-acid reductoisomerase (KARI) from the important wheat head scab fungus Fusarium graminearum. KARI is a key enzyme in the branched-chain amino acid (BCAA, including leucine, isoleucine and valine) biosynthetic pathway that exists in a variety of organisms from bacteria to fungi and higher plants, but not in mammals. The FgILV5 deletion mutant ΔFgIlv5-4 failed to grow when the culture medium was nutritionally limited for BCAAs. When grown on potato-dextrose agar plates, ΔFgIlv5-4 exhibited a significant decrease in aerial hyphae formation and red pigmentation. Conidia formation was also blocked in ΔFgIlv5-4. Exogenous addition of 1 mM isoleucine and valine was able to rescue the defects of mycelial growth and conidial morphogenesis. Cellular stress assays showed that ΔFgIlv5-4 was more sensitive to osmotic and oxidative stresses than the wild-type strain. In addition, virulence of ΔFgIlv5-4 was dramatically reduced on wheat heads, and a low level of deoxynivalenol production was detected in ΔFgIlv5-4 in wheat kernels. The results of this study indicate that FgIlv5 is involved in valine and isoleucine biosynthesis and is required for full virulence in F. graminearum.

The mycotoxins dash 4-deoxynivalenol, zearalenone and aflatoxin dash in weather-damaged wheat harvested 1983-1985 in south-eastern Queensland

A survey for various mycotoxins was carried out on samples of all wheat delivered to nine storage and marketing depots in south-eastern Queensland, selected as most likely to receive mycotoxin-contaminated grain. All wheat was surveyed during 1983, when the degree of weather damage was high. Samples of the poorest grade of wheat from these depots were also surveyed in 1984 and 1985. The surveys included all regions where head scab of wheat caused by Fusarium graminearum Schwabe Group 2 had been reported to occur at significant levels. 4-Deoxynivalenol was detected in nearly all pooled samples representing bulk wheat at concentrations ranging from traces of <0.01 up to 1.7 mg kg-l. The highest concentration of zearlenone detected in a pooled wheat sample was 0.04 mg kg-l. In a few samples representing individual wheat deliveries and with up to 2.8% by weight of pink grains, 4-deoxynivalenol concentrations ranged up to 11.7 mg kg-1 and zearalenone up to 0.43 mg kg-l. Aflatoxins B1, B2, G1 and G2 were detected in only one pooled sample of wheat, at a total aflatoxin concentration of 0.003 mg kg-1. Ochratoxin A, sterigmatocystin and T-2 toxin were not detected. Higher concentrations of mycotoxins were found in the poorer grades of wheat.