Assessing corn recovery from early season nutrient stress under different soil moisture regimes.

Corn (Zea mays) biomass accumulation and nutrient uptake by the six-leaf collar (V6) growth stage are low, and therefore, synchronizing nutrient supply with crop demand could potentially minimize nutrient loss and improve nutrient use efficiency. Knowledge of corn's response to nutrient stress in the early growth stages could inform such nutrient management. Field studies were conducted to assess corn recovery from when no fertilizer application is made until the V6 growth stage, and thereafter, applying fertilizer rates as those in non-stressed conditions. The early season nutrient stress and non-stress conditions received the same amount of nutrients. As the availability of nutrients for plant uptake is largely dependent on soil moisture, corn recovery from the early season nutrient stress was assessed under different soil moisture regimes induced via irrigation scheduling at 50% and 80% field capacity under overhead and subsurface drip irrigation (SSDI) systems. Peanut (Arachis hypogaea) was the previous crop under all conditions, and the fields were under cereal rye (Secale cereale) cover crop prior to planting corn. At the V6 growth stage, the nutrient concentrations of the early season-stressed crops, except for copper, were above the minimum threshold of sufficiency ranges reported for corn. However, the crops showed poor growth, with biomass accumulation being reduced by over 50% compared to non-stressed crops. Also, the uptake of all nutrients was significantly lower under the early season nutrient stress conditions. The recovery of corn from the early season nutrient stress was low. Compared to non-stress conditions, the early season nutrient stress caused 1.58 Mg ha-1 to 3.4 Mg ha-1 yield reduction. The percent yield reduction under the SSDI system was 37.6-38.2% and that under the overhead irrigation system was 11.7-13%. The high yield reduction from the early season nutrient stress under the SSDI system was because of water stress conditions in the topsoil soil layer. The findings of the study suggest ample nutrient supply in the early season growth stage is critical for corn production, and thus, further studies are recommended to determine the optimum nutrient supply for corn at the initial growth stages.

Nutrient Sufficiency Ranges for Corn at the Early Growth Stage: Implications for Nutrient Management.

Growers rely on nutrient sufficiency ranges (NSRs) after plant tissue analysis to inform timely nutrient management decisions. The NSRs are typically established from survey studies across multiple locations, which could be confounded by several abiotic and biotic factors. We conducted field studies in 2020, 2021, and 2022 to validate the lower thresholds of the NSRs for corn (Zea mays) at the early growth stage as reported in the Southern Cooperative Series Bulletin #394. We induced various corn nutritional levels by making different nutrient application rates. If the NSRs are valid, samples within the same replication that satisfy the NSRs of all nutrients should have similar biomass accumulation. The results showed that the NSRs were not valid under the conditions tested. In total, 47.6% of the samples satisfied all the lower thresholds of the NSRs, and 25.4% of those samples had relative biomass <50%, with relative biomass even as low as 24.2% observed. Moreover, 9.6% of the total samples had P and Cu levels that failed to meet the lower threshold but still had relative biomass ≥75%. The findings highlight the sensitivity of corn to nutrient imbalance and the need to optimize nutrient diagnostic methods at the early growth stage.

Deciphering drought-induced metabolic responses and regulation in developing maize kernels.

Drought stress conditions decrease maize growth and yield, and aggravate preharvest aflatoxin contamination. While several studies have been performed on mature kernels responding to drought stress, the metabolic profiles of developing kernels are not as well characterized, particularly in germplasm with contrasting resistance to both drought and mycotoxin contamination. Here, following screening for drought tolerance, a drought-sensitive line, B73, and a drought-tolerant line, Lo964, were selected and stressed beginning at 14 days after pollination. Developing kernels were sampled 7 and 14 days after drought induction (DAI) from both stressed and irrigated plants. Comparative biochemical and metabolomic analyses profiled 409 differentially accumulated metabolites. Multivariate statistics and pathway analyses showed that drought stress induced an accumulation of simple sugars and polyunsaturated fatty acids and a decrease in amines, polyamines and dipeptides in B73. Conversely, sphingolipid, sterol, phenylpropanoid and dipeptide metabolites accumulated in Lo964 under drought stress. Drought stress also resulted in the greater accumulation of reactive oxygen species (ROS) and aflatoxin in kernels of B73 in comparison with Lo964 implying a correlation in their production. Overall, field drought treatments disordered a cascade of normal metabolic programming during development of maize kernels and subsequently caused oxidative stress. The glutathione and urea cycles along with the metabolism of carbohydrates and lipids for osmoprotection, membrane maintenance and antioxidant protection were central among the drought stress responses observed in developing kernels. These results also provide novel targets to enhance host drought tolerance and disease resistance through the use of biotechnologies such as transgenics and genome editing.

Responses of Aspergillus flavus to Oxidative Stress Are Related to Fungal Development Regulator, Antioxidant Enzyme, and Secondary Metabolite Biosynthetic Gene Expression.

The infection of maize and peanut with Aspergillus flavus and subsequent contamination with aflatoxin pose a threat to global food safety and human health, and is exacerbated by drought stress. Drought stress-responding compounds such as reactive oxygen species (ROS) are associated with fungal stress responsive signaling and secondary metabolite production, and can stimulate the production of aflatoxin by A. flavus in vitro. These secondary metabolites have been shown to possess diverse functions in soil-borne fungi including antibiosis, competitive inhibition of other microbes, and abiotic stress alleviation. Previously, we observed that isolates of A. flavus showed differences in oxidative stress tolerance which correlated with their aflatoxin production capabilities. In order to better understand these isolate-specific oxidative stress responses, we examined the transcriptional responses of field isolates of A. flavus with varying levels of aflatoxin production (NRRL3357, AF13, and Tox4) to H2O2-induced oxidative stress using an RNA sequencing approach. These isolates were cultured in an aflatoxin-production conducive medium amended with various levels of H2O2. Whole transcriptomes were sequenced using an Illumina HiSeq platform with an average of 40.43 million filtered paired-end reads generated for each sample. The obtained transcriptomes were then used for differential expression, gene ontology, pathway, and co-expression analyses. Isolates which produced higher levels of aflatoxin tended to exhibit fewer differentially expressed genes than isolates with lower levels of production. Genes found to be differentially expressed in response to increasing oxidative stress included antioxidant enzymes, primary metabolism components, antibiosis-related genes, and secondary metabolite biosynthetic components specifically for aflatoxin, aflatrem, and kojic acid. The expression of fungal development-related genes including aminobenzoate degradation genes and conidiation regulators were found to be regulated in response to increasing stress. Aflatoxin biosynthetic genes and antioxidant enzyme genes were also found to be co-expressed and highly correlated with fungal biomass under stress. This suggests that these secondary metabolites may be produced as part of coordinated oxidative stress responses in A. flavus along with antioxidant enzyme gene expression and developmental regulation.

Stress Sensitivity Is Associated with Differential Accumulation of Reactive Oxygen and Nitrogen Species in Maize Genotypes with Contrasting Levels of Drought Tolerance.

Drought stress decreases crop growth, yield, and can further exacerbate pre-harvest aflatoxin contamination. Tolerance and adaptation to drought stress is an important trait of agricultural crops like maize. However, maize genotypes with contrasting drought tolerances have been shown to possess both common and genotype-specific adaptations to cope with drought stress. In this research, the physiological and metabolic response patterns in the leaves of maize seedlings subjected to drought stress were investigated using six maize genotypes including: A638, B73, Grace-E5, Lo964, Lo1016, and Va35. During drought treatments, drought-sensitive maize seedlings displayed more severe symptoms such as chlorosis and wilting, exhibited significant decreases in photosynthetic parameters, and accumulated significantly more reactive oxygen species (ROS) and reactive nitrogen species (RNS) than tolerant genotypes. Sensitive genotypes also showed rapid increases in enzyme activities involved in ROS and RNS metabolism. However, the measured antioxidant enzyme activities were higher in the tolerant genotypes than in the sensitive genotypes in which increased rapidly following drought stress. The results suggest that drought stress causes differential responses to oxidative and nitrosative stress in maize genotypes with tolerant genotypes with slower reaction and less ROS and RNS production than sensitive ones. These differential patterns may be utilized as potential biological markers for use in marker assisted breeding.

Protein profiles reveal diverse responsive signaling pathways in kernels of two maize inbred lines with contrasting drought sensitivity.

Drought stress is a major factor that contributes to disease susceptibility and yield loss in agricultural crops. To identify drought responsive proteins and explore metabolic pathways involved in maize tolerance to drought stress, two maize lines (B73 and Lo964) with contrasting drought sensitivity were examined. The treatments of drought and well water were applied at 14 days after pollination (DAP), and protein profiles were investigated in developing kernels (35 DAP) using iTRAQ (isobaric tags for relative and absolute quantitation). Proteomic analysis showed that 70 and 36 proteins were significantly altered in their expression under drought treatments in B73 and Lo964, respectively. The numbers and levels of differentially expressed proteins were generally higher in the sensitive genotype, B73, implying an increased sensitivity to drought given the function of the observed differentially expressed proteins, such as redox homeostasis, cell rescue/defense, hormone regulation and protein biosynthesis and degradation. Lo964 possessed a more stable status with fewer differentially expressed proteins. However, B73 seems to rapidly initiate signaling pathways in response to drought through adjusting diverse defense pathways. These changes in protein expression allow for the production of a drought stress-responsive network in maize kernels.

Environmental influences on maize-Aspergillus flavus interactions and aflatoxin production.

Since the early 1960s, the fungal pathogen Aspergillus flavus (Link ex Fr.) has been the focus of intensive research due to the production of carcinogenic and highly toxic secondary metabolites collectively known as aflatoxins following pre-harvest colonization of crops. Given this recurrent problem and the occurrence of a severe aflatoxin outbreak in maize (Zea mays L.), particularly in the Southeast U.S. in the 1977 growing season, a significant research effort has been put forth to determine the nature of the interaction occurring between aflatoxin production, A. flavus, environment and its various hosts before harvest. Many studies have investigated this interaction at the genetic, transcript, and protein levels, and in terms of fungal biology at either pre- or post-harvest time points. Later experiments have indicated that the interaction and overall resistance phenotype of the host is a quantitative trait with a relatively low heritability. In addition, a high degree of environmental interaction has been noted, particularly with sources of abiotic stress for either the host or the fungus such as drought or heat stresses. Here, we review the history of research into this complex interaction and propose future directions for elucidating the relationship between resistance and susceptibility to A. flavus colonization, abiotic stress, and its relationship to oxidative stress in which aflatoxin production may function as a form of antioxidant protection to the producing fungus.