Role of the molecular structure of humified organic matter in rice plant response to environmental lead pollution.

Humified organic matter has been shown to decrease Pb toxicity in plants. However, there are still gaps in our understanding of the mechanism by which this phenomenon occurs. In this study, we aimed to assess the ability of humic substances (HSs), humic acids (HAs), and fulvic acids (FAs) to enhance defense mechanisms in rice plants under lead (Pb)-stressed conditions. HS fractions were isolated from vermicompost using the chemical fractionation methodology established by the International Humic Substances Society. These fractions were characterized by solid-state NMR and FTIR. Chemometric analysis was used to compare humic structures and correlate them with bioactivity. Three treatments were tested to evaluate the protective effect of humic fractions on rice plants. The first experiment involved the application of humic fractions along with Pb. The second comprised pretreatment with humic fractions followed by subsequent exposure to Pb stress. The third experiment involved Pb stress and subsequent treatment with humic fractions. The root morphology and components of the antioxidative defense system were evaluated and quantified. The results showed that HS + Pb, HA + Pb, and FA + Pb treatment preserved root growth and reduced the levels of O2- and malondialdehyde (MDA) in the roots by up to 5% and 2%, respectively. Pretreatment of the plants with humic fractions promoted the maintenance of root growth and reduced the contents of O2-, H2O2, and MDA by up to 48%, 22%, and 20%, respectively. Combined application of humic fractions and Pb reduced the Pb content in plant tissues by up to 60%, while pretreatment reduced it by up to 80%. The protective capacity of humic fractions is related to the presence of peptides, lignin, and carbohydrate fragments in their molecular structures. These results suggest that products could be developed that can mitigate the adverse effects of heavy metals on agricultural crops.

Spectroscopic-chemometric modeling of 80 humic acids confirms the structural pattern identity of humified organic matter despite different formation environments.

The structure of humic substances (HSs) and the humification process are critical topics for understanding the dynamics of carbon on the planet. This study aimed to assess the structural patterns of 80 humic acid (HA) samples isolated from different soils, namely, Histosols, Ferralsols, Cambisols, Mollisols, Planosols and vermicompost, by spectroscopic characterization using solid-state 13C nuclear magnetic resonance cross-polarization/magic angle spinning combined with chemometric techniques. All 80 HAs had a similar structural pattern, regardless of their source of origin, but they had different relative quantities of organic C species. The different structural amounts of the various organic C fractions generated different properties in each of the HAs. This explains why there were similarities in the HS functions but why the intensities of these functions varied among the samples from the different soil types and environments, confirming that HSs are a group of compounds with a structural identity distinct from the molecules that give rise to them. There appears to be no single definition for the humification process; therefore, for the soils from each source of origin, a specific humification process occurs that depends on the characteristics of the local environment. Humification can be understood as a process that is similar to a chemical reaction, where the key factor that determines the formation of the products is the structural characteristics of the reactants (organic substrates deposited in the soil). The degree to which the reaction progresses is governed by the reaction conditions (chemical, physical, and biological properties of the soil). The structural patterns for HSs obtained in this study justify the existence of HSs structured as self-assembled, hydrophilic and hydrophobic domains that, under certain conditions, can undergo transformations, altering the balance of organic carbon in the environment.

Environmental implications of the organic matter structure for white-rot fungus Pleurotus eryngii growth in a tropical climate.

White-rot fungi (Pleurotus eryngii) are decomposers of lignocellulosic substrates. The relationship between the structure of humified organic matter and P. eryngii growth, is poorly understood. This study aimed to evaluate the relationship between the growth and development of white-rot fungi (P. eryngii) in two structurally different sources of humified organic matter. Fungus growth and development (mycelium diameter, fresh and dry mycelium mass, mycelium density, and biological yield) were evaluated in experiments with the application of humic substances (HS) extracted from vermicompost (VC) and peat. Both HS were characterized by CP/MAS 13C NMR spectroscopy associated with chemometrics analysis. The HS present different structural characteristics, with those extracted from VC having a predominance of functionalized C-aliphatics (carbohydrates), low hydrophobicity, and a 90% proportion of cellulose/hemicellulose carbon in the composition. HS extracted from peat have a predominance of C-aromatics (lignin fragments), higher hydrophobicity, and a proportion of lignin carbon of up to 80%. The results showed that P. eryngii growth is dependent on the C-cellulosic and C-lignin balance. HS extracted from lignin-rich peat regulates the fungus growth at initial times and sometimes inhibits the biological performance. The highly cellulosic HS from VC regulate the fungus growth at later times and its biological performance.

Influence of humic acid structure on the accumulation of oxyfluorfen in tropical soils of mountain agroecosystems.

Herbicide application is a practice commonly used in agricultural systems because it is an efficient method of weed control. An inherent characteristic of some herbicides used in mountain agriculture, such as oxyfluorfen, is high adsorption to soil organic matter (SOM). Thus, intensive management that changes the quantity and quality of SOM, such as soil tillage and the massive application of organic fertilizers such as poultry litter, may favor soil contamination by this herbicide and alter its dynamics in the environment. Therefore, this study aimed to characterize the structures of humic substances (HSs) in the soil of forest areas and areas with intensive production of vegetables, relating them to the accumulation of the herbicide oxyfluorfen in tropical mountain agroecosystems. Organic carbon content was quantified in HSs, humic acid (HAs) were structurally characterized by CP/MAS 13C-NMR spectroscopy, and the oxyfluorfen molecule was detected and quantified using the QuEChERS residue detection method with subsequent analysis by LC-MS/MS. Oxyfluorfen was not detected in the forest areas, but it was detected in the vegetable growing areas at points with the lowest slope and high contents of organic matter and clay, with values of up to 0.13 mg kg-1. The intensification in the SOM mineralization process, promoted by the intensive management adopted in the vegetable growing areas, resulted in a 16.46% reduction in COT, a 58.84% reduction in the carbon content in the form of SH and a reduction in the structures that give recalcitrance to the HA molecule (CAlkyl-H,R, CCOO-H,R, CAromatic-H,R, and CAromatic-O) when compared to those values in the forest area, presenting HAs with more aliphatic and labile properties. Thus, due to the structural characteristics of the HAs in the vegetable production areas, the herbicide oxyfluorfen showed a close relationship with the more aliphatic oxygenated structures, namely, CAlkyl-O,N, CAlkyl-O and CAlkyl-di-O.

Humic acids induce a eustress state via photosynthesis and nitrogen metabolism leading to a root growth improvement in rice plants.

Chemical eustressors induce a eustress state "positive stress" increasing the resistance and improve the plant growth. The potentiality of humic acids (HA) to act as a eustressor has been scarcely explored. The present study aims to evaluate how HA with different structural characteristics induce differently, a eustress state in rice plants through the regulation of photosynthesis. The photosynthetic performance index showed an initial eustress state in plant by HA application characterized by reduction in photosynthesis followed by an increase in photosynthetic efficiency. The HA as a chemical eustressor triggering changes in plant metabolism indicate that the interaction of HA with root system induces a roots growth stimulus preceded by an initial positive stress. The eustress caused by HA is differentiated and is related to its chemical-physics characteristics. The HAVC, with a predominance of CAlkyl-(O,N), CAlkyl-di-O, CAromatic-O structures and greater polarity, stimulated the accumulation of N-NO3- and of soluble sugars in the sheath, increase carbohydrates content in the root and the root emission, resulting in higher total biomass production. The HASOIL, with a predominance of CCOOH-(H,R), CAlkyl-O, CAromatic-H,R structures and greater hydrophobicity caused a decrease in N-NH4+ and N-amine. The HARN, with a predominance of CAlkyl-O, CAlkyl-H,R, and CO, characterized by average polarity, caused an increase in photosynthetic pigment and N-NH4+ content. These results are keys to understand that quality of soil organic matter is related to plant development and that HA are efficient proxies for elucidate its function in natural environments.

Fire lead to disturbance on organic carbon under sugarcane cultivation but is recovered by amendment with vinasse.

Sugarcane burning has been widely practiced in Brazil and worldwide. In the long term, this farming practice can cause soil erosion, reduction in organic carbon (OC) and consequently, changes in the structure of soil organic matter (SOM). Such changes may be difficult to reverse. This study aimed to assess the medium- and long-term effects of sugarcane burning on SOM characteristics, both in terms of quantity and structural quality and evaluate the application of vinasse as a strategy to attenuate fire-induced changes in burned soil. The experiment was conducted in a 50-year-old sugarcane field on soils classed as Cambissolo Háplico (Inceptisol). Four plots were sampled: a) burning of sugarcane for harvest for 37 years (SCB37); b) renewal of the sugarcane field and burning for harvest for 3 years (SCB3); c) renewal of the sugarcane field without burning for harvest for 3 years (SCWB), and d) renewal of the sugarcane field and burning for harvest with the application of vinasse for 3 years (SCV). Chemical and physical characterization of SOM was performed by solid-state spectroscopy (UV-vis, ATR-FTIR e 13C NMR CP/MAS) and chemometric techniques. The results showed that sugarcane burning drastically impacts SOM content and its chemical structure, however, the application of vinasse preserves and restores the soil from the fire effects. Content of soil OC, particulate OC, mineral-associated OC, humic acid, humin and light fraction OM that were affected by fire, had an increase and recovery of contents by the vinasse application. Solid state spectroscopy showed that labile structures were lost in humic acids (HA) by fire and recalcitrant structures were preserved. The application of vinasse incorporated fragments of lipids and carbohydrates in HA structure. Burning sugar cane straw affects the integrity of soil organic matter but can be restored by applying vinasse.

Relation between changes in organic matter structure of poultry litter and heavy metals solubility during composting.

Poultry litter (PL) is widely used as fertilizer because of its rich N, P and Ca content. When PL is applied to previously untreated soil, it is a potential contaminant. Composting is an alternative for stabilizing organic and mineral components. This study aimed to elucidate the structural changes and its influence on the solubility of heavy metals in poultry litter during the first 30 d of composting, which is the period when the most intense transformations occur. For this analysis, the transformation dynamics of the organic structures and the availability of mineral elements were studied via spectroscopic characterization, total heavy metal content determination and chemical fractionation at three composting times (0, 15, and 30 d). During composting, the material's aromaticity increased, while its aliphaticity decreased, and the hydrophobicity index increased as the polarity decreased. These results indicate that during the first 30 d, PL composting occurs via transformation of the most labile structures (carbohydrate, peptide and fatty acid fragments), thereby preserving the most stable and least functionalized structures. Composting increased the concentrations of Cu, Cr, Pb and Zn and the transformation of CAlk-O and CAlk-di-O associated with peptides and carbohydrates and favored solubilizing and leaching a water-soluble fraction rich in these compounds. The labile fractions of Fe and the humified organic matter fractions of Cu, Fe, Mn, Zn, and Al increased. The structural changes reduced heavy metal solubility, thus indicating that after 30 d of composting, the heavy metal contamination risk is low.

Carbon input and the structural quality of soil organic matter as a function of agricultural management in a tropical climate region of Brazil.

Carbon (C) stabilization and the quality of soil organic matter (SOM) in a tropical climate are key aspects regulating carbon dioxide emissions and maintaining the C cycle. Soil management influences the accumulation of C, regulating the balance between mineralization and/or the humification of SOM. This study aimed to quantify inputs of different chemical forms of C into soil and to evaluate the structural chemical characteristics of humified SOM. Four management systems were established: Forest (F), Pasture (P), Conventional tillage (T), and No-tillage (NT). Total organic carbon (TOC) and nitrogen (TN) by depth, chemical forms of organic matter input, and spectroscopic characterization of SOM in the form of humic acids (HA) were analyzed. The results obtaining by PCA-13C NMR show that the forest accumulated a high amount of C on the surface (surpassing 20 Mg ha-1), favoring the formation of aliphatic HA (CAlkyl-H,R; CAlkyl-O,N; CAlkyl-O). In the NT management that increases biomass in the soil (14 Mg ha-1), the mineralization process occurred to a greater extent, allowing HA to form with a predominance of aromatic structures (CArm-H,R and CArm-O,N). The PCA-FTIR analysis showed that the P system contributed to the formation of similar HA to those under F management. The T management system incorporated the least TOC and TN, with different HA types being formed in these soils than what was found in other managements. Thus, minimally managed and more stabilized systems in tropical climates form HA of structural and compositional similarity, regardless of the nature of C (C3 or C4). In contrast, soils subjected to agricultural uses that promote higher or lower C inputs, form HA that are structurally different from P and F. This study demonstrates the need for developing experiments for model building to elucidate the relationships among C input, management type, and the formation of humic substances.

Dark septate endophyte decreases stress on rice plants

Abstract Abiotic stress is one of the major limiting factors for plant development and productivity, which makes it important to identify microorganisms capable of increasing plant tolerance to stress. Dark septate endophytes can be symbionts of plants. In the present study, we evaluated the ability of dark septate endophytes isolates to reduce the effects of water stress in the rice varieties Nipponbare and Piauí. The experiments were performed under gnotobiotic conditions, and the water stress was induced with PEG. Four dark septate endophytes were isolated from the roots of wild rice (Oryza glumaepatula) collected from the Brazilian Amazon. Plant height as well as shoot and root fresh and dry matter were measured. Leaf protein concentrations and antioxidant enzyme activity were also estimated. The dark septate endophytes were grown in vitro in Petri dishes containing culture medium; they exhibited different levels of tolerance to salinity and water stress. The two rice varieties tested responded differently to inoculation with dark septate endophytes. Endophytes promoted rice plant growth both in the presence and in the absence of a water deficit. Decreased oxidative stress in plants in response to inoculation was observed in nearly all inoculated treatments, as indicated by the decrease in antioxidant enzyme activity. Dark septate endophytes fungi were shown to increase the tolerance of rice plants to stress caused by water deficiency.

Dark septate endophyte decreases stress on rice plants.

Abiotic stress is one of the major limiting factors for plant development and productivity, which makes it important to identify microorganisms capable of increasing plant tolerance to stress. Dark septate endophytes can be symbionts of plants. In the present study, we evaluated the ability of dark septate endophytes isolates to reduce the effects of water stress in the rice varieties Nipponbare and Piauí. The experiments were performed under gnotobiotic conditions, and the water stress was induced with PEG. Four dark septate endophytes were isolated from the roots of wild rice (Oryza glumaepatula) collected from the Brazilian Amazon. Plant height as well as shoot and root fresh and dry matter were measured. Leaf protein concentrations and antioxidant enzyme activity were also estimated. The dark septate endophytes were grown in vitro in Petri dishes containing culture medium; they exhibited different levels of tolerance to salinity and water stress. The two rice varieties tested responded differently to inoculation with dark septate endophytes. Endophytes promoted rice plant growth both in the presence and in the absence of a water deficit. Decreased oxidative stress in plants in response to inoculation was observed in nearly all inoculated treatments, as indicated by the decrease in antioxidant enzyme activity. Dark septate endophytes fungi were shown to increase the tolerance of rice plants to stress caused by water deficiency.

Involvement of Hormone- and ROS-Signaling Pathways in the Beneficial Action of Humic Substances on Plants Growing under Normal and Stressing Conditions.

The importance of soil humus in soil fertility has been well established many years ago. However, the knowledge about the whole mechanisms by which humic molecules in the rhizosphere improve plant growth remains partial and rather fragmentary. In this review we discuss the relationships between two main signaling pathway families that are affected by humic substances within the plant: one directly related to hormonal action and the other related to reactive oxygen species (ROS). In this sense, our aims are to try the integration of all these events in a more comprehensive model and underline some points in the model that remain unclear and deserve further research.

Root-Shoot Signaling crosstalk involved in the shoot growth promoting action of rhizospheric humic acids.

Numerous studies have shown the ability of humic substances to improve plant development. This action is normally reflected in an enhancement of crop yields and quality. However, the mechanisms responsible for this action of humic substances remain rather unknown. Our studies have shown that the shoot promoting action of sedimentary humic acids is dependent of its ability to increase root hydraulic conductivity through signaling pathways related to ABA, which in turn is affected in roots by humic acids in an IAA-NO dependent way. Furthermore, these studies also indicate that the primary action of humic acids in roots might also be physical, resulting from a transient mild stress caused by humic acids associated with a fouling-cleaning cycle of wall cell pores. Finally the role of alternative signal molecules, such as ROS, and corresponding signaling pathways are also discussed and modeled in the context of the above-mentioned framework.

Vermicompost humic acids modulate the accumulation and metabolism of ROS in rice plants.

This work aims to determine the reactive oxygen species (ROS) accumulation, gene expression, anti-oxidant enzyme activity, and derived effects on membrane lipid peroxidation and certain stress markers (proline and malondialdehyde-MDA) in the roots of unstressed and PEG-stressed rice plants associated with vermicompost humic acid (VCHA) application. The results show that the application of VCHA to the roots of unstressed rice plants caused a slight but significant increase in root ROS accumulation and the gene expression and activity of the major anti-oxidant enzymes (superoxide dismutase and peroxidase). This action did not have negative effects on root development, and an increase in both root growth and root proliferation occurred. However, the root proline and MDA concentrations and the root permeability results indicate the development of a type of mild stress associated with VCHA application. When VCHA was applied to PEG-stressed plants, a clear alleviation of the inhibition in root development linked to PEG-mediated osmotic stress was observed. This was associated with a reduction in root ROS production and anti-oxidant enzymatic activity caused by osmotic stress. This alleviation of stress caused by VCHA was also reflected as a reduction in the PEG-mediated concentration of MDA in the root as well as root permeability. In summary, the beneficial action of VCHA on the root development of unstressed or PEG-stressed rice plants clearly involves the modulation of ROS accumulation in roots.

Structure-Property-Function Relationship in Humic Substances to Explain the Biological Activity in Plants.

Knowledge of the structure-property-function relationship of humic substances (HSs) is key for understanding their role in soil. Despite progress, studies on this topic are still under discussion. We analyzed 37 humic fractions with respect to their isotopic composition, structural characteristics, and properties responsible for stimulating plant root parameters. We showed that regardless of the source of origin of the carbon (C3 or C4), soil-extracted HSs and humic acids (HAs) are structurally similar to each other. The more labile and functionalized HS fraction is responsible for root emission, whereas the more recalcitrant and less functionalized HA fraction is related to root growth. Labile structures promote root stimulation at lower concentrations, while recalcitrant structures require higher concentrations to promote a similar stimulus. These findings show that lability and recalcitrance, which are derived properties of humic fractions, are related to the type and intensity of their bioactivity. In summary, the comparison of humic fractions allowed a better understanding of the relationship between the source of origin of plant carbon and the structure, properties, and type and intensity of the bioactivity of HSs in plants. In this study, scientific concepts are unified and the basis for the agronomic use of HSs is established.