H. Arnold Bruns

Biocontrol of aflatoxin in corn by inoculation with non-aflatoxigenic Aspergillus flavus isolates

Abstract The ability of two non-aflatoxigenic Aspergillus flavus Link isolates (CT3 and K49) to reduce aflatoxin contamination of corn was assessed in a 4-year field study (2001–2004). Soil was treated with six wheat inoculant treatments: aflatoxigenic isolate F3W4; two non-aflatoxigenic isolates (CT3 and K49); two mixtures of CT3 or K49 with F3W4; and an autoclaved wheat control, applied at 20 kg ha−1. In 2001, inoculation with the aflatoxigenic isolate increased corn grain aflatoxin levels by 188% compared to the non-inoculated control, while CT3 and K49 inoculation reduced aflatoxin levels in corn grain by 86 and 60%, respectively. In 2002, the non-toxigenic CT3 and K49 reduced aflatoxin levels by 61 and 76% compared to non-inoculated controls, respectively. In 2001, mixtures of aflatoxigenic and non-aflatoxigenic isolates had little effect on aflatoxin levels, but in 2002, inoculation with mixtures of K49 and CT3 reduced aflatoxin levels 68 and 37% compared to non-inoculated controls, respectively. In 2003 and 2004, a low level of natural aflatoxin contamination was observed (8 ng g−1). However, inoculation with mixtures of K49 + F3W4 and CT3 + F3W4, reduced levels of aflatoxin 65–94% compared to the aflatoxigenic strain alone. Compared to the non-sclerotia producing CT3, strain K49 produces large sclerotia, has more rapid in vitro radial growth, and a greater ability to colonize corn when artificially inoculated, perhaps indicating greater ecological competence. Results indicate that non-aflatoxigenic, indigenous A. flavus isolates, such as strain K49, have potential use for biocontrol of aflatoxin contamination in southern US corn.

Dynamics of Mycotoxin and Aspergillus flavus Levels in Aging Bt and Non-Bt Corn Residues under Mississippi No-Till Conditions

Mycotoxin and Aspergillus flavus levels in soil-surface corn debris left by no-till agriculture methods (stover, cobs, and cobs with grain) were determined during the December-March fallow period for near-isogenic Bt and non-Bt hybrid corn. By December, average mycotoxin levels in non-Bt corn were many times higher in cobs with grain than in grain harvested in September (total aflatoxins, 774 vs 211 ng/g; total fumonisins, 216 vs 3.5 microg/g; cyclopiazonic acid, 4102 vs 72.2 microg/g; zearalenone, 0.2 vs < 0.1 microg/g). No trichothecenes were detected. Levels of mycotoxins and A. flavus propagules were approximately 10- to 50-fold lower in cobs without grain and stover, respectively, for all mycotoxins except zearalenone. Mycotoxin levels in corn debris fractions decreased during winter but began to rise in March. Levels of all mycotoxins and A. flavus propagules were lower in harvested grain and debris from Bt than non-Bt corn, but differences were significant (p < 0.05) only for aflatoxins.

Agricultural practices altered soybean seed protein, oil, fatty acids, sugars, and minerals in the Midsouth USA

Information on the effects of management practices on soybean seed composition is scarce. Therefore, the objective of this research was to investigate the effects of planting date (PD) and seeding rate (SR) on seed composition (protein, oil, fatty acids, and sugars) and seed minerals (B, P, and Fe) in soybean grown in two row-types (RTs) on the Mississippi Delta region of the Midsouth USA. Two field experiments were conducted in 2009 and 2010 on Sharkey clay and Beulah fine sandy loam soil at Stoneville, MS, USA, under irrigated conditions. Soybean were grown in 102 cm single-rows and 25 cm twin-rows in 102 cm centers at SRs of 20, 30, 40, and 50 seeds m-2. The results showed that in May and June planting, protein, glucose, P, and B concentrations increased with increased SR, but at the highest SRs (40 and 50 seeds m-2), the concentrations remained constant or declined. Palmitic, stearic, and linoleic acid concentrations were the least responsive to SR increases. Early planting resulted in higher oil, oleic acid, sucrose, B, and P on both single and twin-rows. Late planting resulted in higher protein and linolenic acid, but lower oleic acid and oil concentrations. The changes in seed constituents could be due to changes in environmental factors (drought and temperature), and nutrient accumulation in seeds and leaves. The increase of stachyose sugar in 2010 may be due to a drier year and high temperature in 2010 compared to 2009; suggesting the possible role of stachyose as an environmental stress compound. Our research demonstrated that PD, SR, and RT altered some seed constituents, but the level of alteration in each year dependent on environmental factors such as drought and temperature. This information benefits growers and breeders for considering agronomic practices to select for soybean seed nutritional qualities under drought and high heat conditions.

Implications of Bt Traits on Mycotoxin Contamination in Maize: Overview and Recent Experimental Results in Southern United States

Mycotoxin contamination levels in maize kernels are controlled by a complex set of factors including insect pressure, fungal inoculum potential, and environmental conditions that are difficult to predict. Methods are becoming available to control mycotoxin-producing fungi in preharvest crops, including Bt expression, biocontrol, and host plant resistance. Initial reports in the United States and other countries have associated Bt expression with reduced fumonisin, deoxynivalenol, and zearalenone contamination and, to a lesser extent, reduced aflatoxin contamination in harvested maize kernels. However, subsequent field results have been inconsistent, confirming that fumonisin contamination can be reduced by Bt expression, but the effect on aflatoxin is, at present, inconclusive. New maize hybrids have been introduced with increased spectra of insect control and higher levels of Bt expression that may provide important tools for mycotoxin reduction and increased yield due to reduced insect feeding, particularly if used together with biocontrol and host plant resistance.

Physiological responses of glyphosate-resistant and glyphosate-sensitive soybean to aminomethylphosphonic acid, a metabolite of glyphosate.

Aminomethylphosphonic acid (AMPA) is formed in glyphosate-treated glyphosate-resistant (GR) and glyphosate-sensitive (GS) soybean [Glycine max (L.) Merr.] plants and is known to cause yellowing in soybean. Although, AMPA is less phytotoxic than glyphosate, its mode of action is different from that of glyphosate and is still unknown. Greenhouse studies were conducted at Stoneville, MS to determine the effects of AMPA on plant growth, chlorophyll content, photosynthesis, nodulation, nitrogenase activity, nitrate reductase activity, and shoot nitrogen content in GR and GS soybeans. AMPA was applied to one- to two-trifoliolate leaf stage soybeans at 0.1 and 1.0 kg ha(-1), representing a scenario of 10% and 100% degradation of glyphosate (1.0 kg ae ha(-1) use rate) to AMPA, respectively. Overall, AMPA effects were more pronounced at 1.0 kg ha(-1) than at 0.1 kg ha(-1) rate. Visual plant injury (18-27%) was observed on young leaves within 3d after treatment (DAT) with AMPA at the higher rate regardless of soybean type. AMPA injury peaked to 46-49% at 14 DAT and decreased to 17-18% by 28 DAT, in both soybean types. AMPA reduced the chlorophyll content by 37%, 48%, 66%, and 23% in GR soybean, and 17%, 48%, 57%, and 22% in GS soybean at 3, 7, 14, and 28 DAT, respectively. AMPA reduced the photosynthesis rate by 65%, 85%, and 77% in GR soybean and 59%, 88%, and 69% in GS soybean at 3, 7, and 14 DAT, respectively, compared to non-treated plants. Similarly, AMPA reduced stomatal conductance to water vapor and transpiration rates at 3, 7, and 14 DAT compared to non-treated plants in both soybean types. Photosynthesis rate, stomatal conductance, and transpiration rate recovered to the levels of non-treated plants by 28 DAT. Plant height and shoot dry weight at 28 DAT; nodulation, nitrogenase activity at 10 DAT, and nitrate reductase activity at 3 and 14 DAT were unaffected by AMPA. AMPA reduced root respiration and shoot nitrogen content at 10 DAT. These results suggest that a foliar application of AMPA could indirectly reduce photosynthesis through decreased chlorophyll content in GR and GS soybean up to 14 DAT, but affected plants can recover to normal growth by 28 DAT.

Effects of Furrow Irrigation on Corn in the Humid Sub-tropical Mississippi Delta

Corn (Zea mays L.) production in the Mississippi Delta has nearly doubled since 1990 but is more susceptible to aflatoxin and fumonisin contamination when grown under drought. Four corn hybrids -- two Bt and two non-Bt -- were grown at Stoneville, MS under irrigated and non-irrigated treatments in 1999, 2000, and 2001. Furrow irrigation was applied at a rate of 1 inch per application during growth stages R1, R3, and R5 in 1999, R1 in 2000, and R1 and R3 in 2001. Irrigation increased grain yields in 2 out of 3 years. More irrigation treatments in 2000 and 2001 would have likely benefited yields. Yields in 2000 were lower than comparable treatments in 1999 and 2001 due to less weight per kernel indicating drought stress occurred during later reproductive growth (R4 to black-layer). Kernel weights were generally higher in both irrigated and non-irrigated treatments in 2001 than they were in 1999 or 2000. This compensated for having fewer kernels per ear, which likely resulted from drought stress at growth stage V12 in 2001. Differences in test weights were observed among years but not irrigation treatments. The Bt hybrids did not differ from the non-Bt hybrids in yield or mycotoxin levels. Aflatoxin and fumonisin contamination did not differ among years or irrigation treatments. Levels were below maximum acceptable concentrations for both mycotoxins. The hybrid N79-L3 had significantly lower fumonisin levels than the other hybrids.

Aflatoxin and Fumonisin in Corn (Zea mays) Infected by Common Smut Ustilago maydis

Corn infected with Ustilago maydis (common smut) produces galls that are valued as a delicacy in some cultures. During a 4-year period, aflatoxin levels in asymptomatic kernels of smutted ears were, on average, 45-fold higher than in kernels harvested from smut-free control ears and 99-fold higher than in smut galls. Aflatoxin levels in smut galls were lower than in kernels of smut-free control corn in all years combined. Fumonisin levels in asymptomatic kernels harvested from smutted ears were 5.2-fold higher than in kernels from smut-free control ears and 4.0-fold higher than in smut galls. Fumonisin levels in smut galls were not significantly different than in kernels of smut-free control corn. These studies indicate that, although corn smut was relatively free of the mycotoxins studied, the asymptomatic kernels of those ears contained mycotoxins at levels much higher than usually considered safe for direct human consumption.

Corn Yields Benefit in Rotations with Cotton

Continuous cotton (Gossypium hirsutum L.) was the primary crop for the Mississippi Delta until recently. Corn (Zea mays L.) is now grown on about 1 million acres in Arkansas, Mississippi, and Louisiana, usually in rotation with cotton. This research evaluated corn’s performance in a four-year furrow irrigated rotation with cotton at Stoneville, MS. The experimental design was a randomized complete block with a split-plot arrangement of treatments replicated eight times. Whole plots were cropping sequences assigned at random. Cropping sequences were continuous cotton, continuous corn, corn-cotton-corn-cotton, or cotton-corncorn-cotton. Four adapted corn hybrids and cotton cultivars were grown as subplots beginning in 2000 to 2003. Corn grain yields were greater following cotton than continous corn in 2001 (169 bu/acre vs. 160 bu/acre) and 2002 (126 bu/acre vs. 117 bu/acre). Grain yields from continuous corn differed among years but with no consistency. Hybrids differed in yield among all years but no consistency in these data was noted. Test weights for continuous corn differed among years but were not below the requirement for US No. 2 yellow corn. Weights of 100 kernels did not differ among years or treatments. Economics dictate cropping sequences but corn can benefit from following cotton in rotation.

Ear Leaf Photosynthesis and Related Parameters of Transgenic and Non-GMO Maize Hybrids

Hybrid maize (Zea mays L.) through transgenics now includes δ-endotoxins for insect control and tolerance to the herbicides glyphosate and glufosinate. Some hybrids have multiple transgenic traits as part of their genotype (stacked gene). Limited information is available on how these traits alone affect (net assimilation rate; µmol CO2 m−2 s−1) and related physiological parameters. A two-year, two-location, irrigated experiment comparing four stacked gene, four glyphosate tolerant, and two non-GMO hybrids for ear leaf , (stomatal conductance; mol H2O m−2 s−1), Em (transpiration; mol H2O m−2 s−1), IWUE (intrinsic water use efficiency; ), and Ci (intercellular [CO2] µmol CO2 mol air−1) was completed at Stoneville, MS, in 2012. Data were collected at growth stages R1 (anthesis) and R2 (early kernel filling) using a Li-Cor LI-6400XT set at 355 μmol mol−1 CO2 with a flow rate of 500 μmol s−1 and a 6400-02 light source set at 87.5% full sunlight. Measurements were made between 08:30 h and 11:30 h CST, within 48 h of 25 ha mm irrigation and ≥33.0% cloud cover. Transgenic traits did not influence the physiological parameters of , , Em, IWUE, or Ci during the critical growth stages of R1 or R2.

Comparisons of Herbicide Treated and Cultivated Herbicide-Resistant Corn

Four glyphosate resistant corn (Zea mays L.) hybrids, a glufosinate-ammonium resistant hybrid, and a conventional atrazine resistant hybrid gown at Stoneville, MS in 2005, 2006, and 2007 with furrow irrigation were treated with their respective herbicides and their growth, yield, and mycotoxin incidence were compared with untreated cultivated plots. Leaf area index (LAI) and dry matter accumulation (DMA) were collected on a weekly basis beginning at growth stage V3 and terminating at anthesis. Crop growth rates (CRGs) and relative growth rates (RGRs) were calculated. Plots were later harvested, yield and yield component data collected, and kernel samples analyzed for aflatoxin and fumonisin. Leaf area index, DMA, CRG, and RGR were not different among the herbicide treated plots and from those that were cultivated. Curves for LAI and DMA were similar to those previously reported. Aflatoxin and fumonisin were relatively low in all plots. Herbicide application or the lack thereof had no negative impact on the incidence of kernel contamination by these two mycotoxins. Herbicides, especially glyphosate on resistant hybrids, have no negative effects on corn yields or kernel quality in corn produced in a humid subtropical environment.