Carbon Amendments Influence Composition and Functional Capacities of Indigenous Soil Microbiomes

Soil nutrient amendments are recognized for their potential to improve microbial activity and biomass in the soil. However, the specific selective impacts of carbon amendments on indigenous microbiomes and their metabolic functions in agricultural soils remain poorly understood. We investigated the changes in soil chemical characteristics and phenotypes of Streptomyces communities following carbon amendments to soil. Mesocosms were established with soil from two field sites varying in soil organic matter content (low organic matter, LOM; high organic matter, HOM), that were amended at intervals over nine months with low or high dose solutions of glucose, fructose, malic acid, a mixture of these compounds, or water only (non-amended control). Significant shifts in soil chemical characteristics and antibiotic inhibitory capacities of indigenous Streptomyces were observed in response to carbon additions. All high dose carbon amendments consistently increased soil total carbon, while amendments with malic acid decreased soil pH. In LOM soils, higher frequencies of Streptomyces inhibitory phenotypes of the two plant pathogens, Streptomyces scabies and Fusarium oxysporum, were observed in response to soil carbon additions. Additionally, to determine if shifts in Streptomyces functional characteristics correlated with microbiome composition, we investigated whether shifts in functional characteristics of soil Streptomyces correlated with composition of soil bacterial communities, analyzed using 16S rRNA gene sequencing. Regardless of dose, community composition differed significantly among carbon-amended and non-amended soils from both sites. Carbon type and dose had significant effects on bacterial community composition in both LOM and HOM soils. Relationships among microbial community richness (observed species number), diversity, and soil characteristics varied among soils from different sites. These results suggest that manipulation of soil resource availability has the potential to selectively modify the functional capacities of soil microbiomes, and specifically to enhance pathogen inhibitory populations of high value to agricultural systems

Carbon Amendments Induce Shifts in Nutrient Use, Inhibitory, and Resistance Phenotypes Among Soilborne Streptomyces

Carbon amendments are used in agriculture for increasing microbial activity and biomass in the soil. Changes in microbial community composition and function in response to carbon additions to soil have been associated with biological suppression of soilborne diseases. However, the specific selective impacts of carbon amendments on microbial antagonistic populations are not well understood. We investigated the effects of soil carbon amendments on nutrient use profiles, and antibiotic inhibitory and resistance phenotypes of Streptomyces populations from agricultural soils. Soil mesocosms were amended at intervals over 9 months with low or high dose solutions of glucose, fructose, a complex amendment, or water only (non-amendment control). Over 130 Streptomyces isolates were collected from amended and non-amended mesocosm soils, and nutrient utilization profiles on 95 different carbon substrates were determined. A subset of isolates (n = 40) was characterized for their ability to inhibit or resist one another. Carbon amendments resulted in Streptomyces populations with greater niche widths, and increased growth efficiencies as compared with Streptomyces in non-amended soils. Shifts in microbial nutrient use and growth capacities coincided with positive selection for Streptomyces antibiotic inhibitory phenotypes in carbon-amended soils, resulting in populations dominated by phenotypes that combine both antagonistic capacities and a generalist lifestyle. Carbon inputs resulted in populations that on average were more resistant to one another than populations in non-amended soils. Shifts in metabolic capacities and antagonistic activity indicate that carbon additions to soil may selectively enrich Streptomyces antagonistic phenotypes, that are rare under non-nutrient selection, but can inhibit more intensively nutrient competitors, and resist phenotypes with similar functional traits. These results shed light on the potential for using carbon amendments to strategically mediate soil microbial community assembly, and contribute to the establishment of pathogen-suppressive soils in agricultural systems

Identification of seedling resistance in wild oat relatives against oat crown rust

Faculty advisor: Melania FigueroaThis research was supported by the Undergraduate Research Opportunities Program (UROP)

Transgenic wheat expressing a barley class II chitinase gene has enhanced resistance against Fusarium graminearum

Fusarium head blight (FHB; scab), primarily caused by Fusarium graminearum, is a devastating disease of wheat worldwide. FHB causes yield reductions and contamination of grains with trichothecene mycotoxins such as deoxynivalenol (DON). The genetic variation in existing wheat germplasm pools for FHB resistance is low and may not provide sufficient resistance to develop cultivars through traditional breeding approaches. Thus, genetic engineering provides an additional approach to enhance FHB resistance. The objectives of this study were to develop transgenic wheat expressing a barley class II chitinase and to test the transgenic lines against F. graminearum infection under greenhouse and field conditions. A barley class II chitinase gene was introduced into the spring wheat cultivar, Bobwhite, by biolistic bombardment. Seven transgenic lines were identified that expressed the chitinase transgene and exhibited enhanced Type II resistance in the greenhouse evaluations. These seven transgenic lines were tested under field conditions for percentage FHB severity, percentage visually scabby kernels (VSK), and DON accumulation. Two lines (C8 and C17) that exhibited high chitinase protein levels also showed reduced FHB severity and VSK compared to Bobwhite. One of the lines (C8) also exhibited reduced DON concentration compared with Bobwhite. These results showed that transgenic wheat expressing a barley class II chitinase exhibited enhanced resistance against F. graminearum in greenhouse and field conditions

Overexpression of defense response genes in transgenic wheat enhances resistance to Fusarium head blight

Fusarium head blight (FHB) of wheat, caused by Fusarium graminearum and other Fusarium species, is a major disease problem for wheat production worldwide. To combat this problem, large-scale breeding efforts have been established. Although progress has been made through standard breeding approaches, the level of resistance attained is insufficient to withstand epidemic conditions. Genetic engineering provides an alternative approach to enhance the level of resistance. Many defense response genes are induced in wheat during F. graminearum infection and may play a role in reducing FHB. The objectives of this study were (1) to develop transgenic wheat overexpressing the defense response genes α-1-purothionin, thaumatin-like protein 1 (tlp-1), and β-1,3-glucanase; and (2) to test the resultant transgenic wheat lines against F. graminearum infection under greenhouse and field conditions. Using the wheat cultivar Bobwhite, we developed one, two, and four lines carrying the α-1-purothionin, tlp-1, and β-1,3-glucanase transgenes, respectively, that had statistically significant reductions in FHB severity in greenhouse evaluations. We tested these seven transgenic lines under field conditions for percent FHB disease severity, deoxynivalenol (DON) mycotoxin accumulation, and percent visually scabby kernels (VSK). Six of the seven lines differed from the nontransgenic parental Bobwhite line for at least one of the disease traits. A β-1,3-glucanase transgenic line had enhanced resistance, showing lower FHB severity, DON concentration, and percent VSK compared to Bobwhite. Taken together, the results showed that overexpression of defense response genes in wheat could enhance the FHB resistance in both greenhouse and field conditions

Cultural control practices for Fusarium head blight: Problems and solutions

Reduced tillage practices have been adopted worldwide in agriculture. The implementation of conservation tillage, leaving crop residues at the soil surface following the harvest of crops is essential to protect soils vulnerable to erosion, however these practices have contributed directly to the upsurge of Fusarium head blight of wheat and barley. While researchers have made significant progress in the identification and incorporation of genetic resistance, and in the identification and delivery of effective fungicides to cereal crops, Fusarium head blight remains a recalcitrant problem. It would seem that we can not reduce the threat of future Fusarium head blight epidemics without addressing the underlying origin of the problem, Fusarium -infested crop residues. Given the limitations of current agricultural practices, we are challenged to find ways to reduce the inoculum potential of Fusarium -infested residues without removing them from the soil surface

Disease Management and Estimated Effects on DON (Deoxynivalenol) Contamination in \u3ci\u3eFusarium\u3c/i\u3e Infested Barley

Fusarium head blight (FHB or scab) economically devastates barley production. FHB is predominantly caused by Fusarium graminearum and has resulted in major reductions in the quality of barley in the United States. The most common source of economic loss is through development of potent mycotoxins in the grain, the most prominent of which, in the United States, is deoxynivalenol (DON). DON levels can be managed through a variety of techniques. This study presents the estimate of the statistical relationship among DON contamination in barley, FHB incidence and severity, and a variety of disease management techniques. Data from 22 field studies and a survey of barley producers are used to estimate the relationship. Fungicide applications reduce DON in barley in general and via complementary interactions with the barley cultivar. Genetic FHB resistance in barley varieties is an important determinant of DON levels, as well as previous crop and factors related to time and location. Taking care to avoid rotations with FHB host crops immediately prior to barley is also important to reduce DON levels in barley. These become key inputs into barley producer decisions for evaluating the economic value of adopting FHB management techniques

Genetic and phenotypic characterization of Xanthomonas species pathogenic of wheat in Uruguay.

Bacterial diseases affecting wheat production worldwide have been an issue of growing concern in the last decade. Nevertheless, the main bacterial diseases of wheat in Uruguay remain largely uninvestigated. To identify bacterial pathogens associated with diseased wheat fields, 61 fields were surveyed from 2017 to 2019 in western Uruguay, yielding a collection of strains identified as Xanthomonas spp. by 16S rDNA sequencing. These strains were further characterized via multilocus sequence analysis (MLSA) and typing (MLST), as well as in planta pathogenicity assays. MLSA grouped 44 strains with reference strains for Xanthomonas translucens pv. undulosa, the pathovar predominantly associated with bacterial leaf streak of wheat (BLS). To evaluate the genetic diversity among strains, MLST was applied, revealing a low diversity among Uruguayan strains identified as X. translucens pv. undulosa. In addition, 17 strains in the collection were assigned to a separate clade distant from the X. translucens species, grouping together with previously unreported Xanthomonas strains isolated from wheat in Minnesota, USA. In planta pathogenicity assays were performed on BLS susceptible wheat seedlings. The 44 X. translucens pv. undulosa strains caused greasy, dark brown necrosis symptoms typical of BLS, while the 17 non-translucens Xanthomonas sp. strains caused distinctly different dry tan necrosis symptoms. These results reveal that the main bacterial pathogen affecting wheat crops in Uruguay is X. translucens pv. undulosa, the causal agent of BLS; and that other pathogenic Xanthomonas species are associated with wheat crops in South and North America.Agencia Nacional de Iinvestigación e InnovaciónComisión Sectorial de Investigación Científic