Daniel Basílio Zandonadi

Chemical composition and bioactivity properties of size-fractions separated from a vermicompost humic acid.

Preparative high performance size-exclusion chromatography (HPSEC) was applied to humic acids (HA) extracted from vermicompost in order to separate humic matter of different molecular dimension and evaluate the relationship between chemical properties of size-fractions (SF) and their effects on plant root growth. Molecular dimensions of components in humic SF was further achieved by diffusion-ordered nuclear magnetic resonance spectroscopy (DOSY-NMR) based on diffusion coefficients (D), while carbon distribution was evaluated by solid state (CP/MAS) (13)C NMR. Seedlings of maize and Arabidopsis were treated with different concentrations of SF to evaluate root growth. Six different SF were obtained and their carbohydrate-like content and alkyl chain length decreased with decreasing molecular size. Progressive reduction of aromatic carbon was also observed with decreasing molecular size of separated fractions. Diffusion-ordered spectroscopy (DOSY) spectra showed that SF were composed of complex mixtures of aliphatic, aromatic and carbohydrates constituents that could be separated on the basis of their diffusion. All SF promoted root growth in Arabidopsis and maize seedlings but the effects differed according to molecular size and plant species. In Arabidopsis seedlings, the bulk HA and its SF revealed a classical large auxin-like exogenous response, i.e.: shortened the principal root axis and induced lateral roots, while the effects in maize corresponded to low auxin-like levels, as suggested by enhanced principal axis length and induction of lateral roots. The reduction of humic heterogeneity obtained in HPSEC separated size-fractions suggested that their physiological influence on root growth and architecture was less an effect of their size than their content of specific bioactive molecules. However, these molecules may be dynamically released from humic superstructures and exert their bioactivity when weaker is the humic conformational stability as that obtained in the separated size-fractions.

Organic matter quality in a soil cultivated with perennial herbaceous legumes

Using herbaceous legumes in agricultural systems yields great quantities of plant residues, allowing changes in soil organic matter quality and content over the years. This study was conducted on an Ultisol, at Seropedica, RJ, Brazil, to evaluate the effects of different perennial herbaceous legumes on soil organic matter quality. A factorial scheme with three replications was used to evaluate the species: forage groundnut cv. BR-14951 (Arachis pintoi), tropical kudzu (Pueraria phaseoloides), and siratro (Macroptilium atropurpureum). After the first cut, each plot was divided into two subplots; plants were cut and left on the soil surface or cut and removed. Soil samples of a closed area covered by spontaneous vegetation (mainly C3 plants) or by Panicum maximum were also analysed. Samples were collected from two layers (0-5 and 5-10 cm), processed for the fractionation of organic matter and the evaluation of structural characteristics of humic acids (HA). Evaluated legumes did not change total organic carbon contents, but promoted HA accumulation in the superficial soil layer. Humic acids may be used as indicators of the management effects on soil organic fractions, because there was significant incorporation of carbon and nitrogen derived from the legume residues, even for the short experimentation time (28 months). Residue management did not modify quantitative aspects of the distribution of the humified organic matter, but promoted, however, a higher condensation degree of humic acids evaluated by the elementary composition, IR and fluorescence spectroscopy.

Estímulo no crescimento e na hidrólise de ATP em raízes de alface tratadas com humatos de vermicomposto: II - Efeito da fonte de vermicomposto

Um dos fatores mais limitantes para a producao de vermicomposto e a disponibilidade de esterco. Neste trabalho, foi avaliado o efeito da substituicao parcial do esterco por bagaco de cana e por residuos de leguminosa (Gliricidia sepium) na vermicompostagem sobre a qualidade do vermicomposto e sobre a bioatividade dos humatos, avaliadas por meio da analise do crescimento radicular e da atividade das bombas de H+ isoladas de raizes de alface. A substituicao do esterco por bagaco de cana e por residuos de leguminosas nao acarretou prejuizo as caracteristicas quimicas dos vermicompostos. No entanto, os humatos isolados dos diferentes vermicompostos apresentaram caracteristicas quimicas distintas, tais como: acidez e propriedades oticas distintas. Os humatos produzidos a partir de esterco de bovino e da mistura esterco bovino + bagaco proporcionaram maiores estimulos no crescimento radicular das plantas de alface, sendo os mais indicados para uso na forma soluvel. A inclusao de residuos de leguminosas no processo de vermicompostagem produziu humatos sem efeito sobre o desenvolvimento das raizes de alface.

Distribuição de massa molecular de ácidos húmicos e promoção do crescimento radicular

Humic acids (HA) directly stimulate several physiological processes that promote plant growth, particularly of the root system. Knowledge about the chemical nature and the role of HA in the effects observed under biofertilization and biostimulation is essential to develop HA-based biological resources. The aim of this study was to evaluate a possible relationship between the distribution of the apparent molecular weight of HA isolated from vermicompost and the response in root growth promotion. HA sub-fractions were obtained through preparative chromatography by size exclusion using Sephadex gel G-50 gel (CGE). The preparative process was validated by high-performance size-exclusion chromatography (HPSEC). The five sub-fractions were tested at different concentrations (0; 0.0001; 0.001; 0.003; 0.005; and 0.001 mol L-1 C) for their capacity to stimulate root growth of Arabidopisis thaliana (ecotype col 4) seedlings. For maize seedlings (Zea mays hybrid UENF 506-6) a rate of 0.002 mol L-1 C was used. The quadratic model described the relationship between root growth induction and HA dose in Arabidopsis; 0.00511 mol L-1 C was the average inflection point. At the optimum concentration, a significant negative correlation between molecular weight distribution and the number of lateral roots induced in A. thaliana was observed. However, other root traits, e.g., area and length, were not influenced by the apparent molecular weight of fractions. An increase in the number of mitotic and lateral root emission was observed for maize seedlings, both for the treatment with HA as well as with the sub-fractions. The H+-ATPase activity of plasma membrane was significantly affected by HA, although differently in the sub-fractions. The stimulating activity of root growth seems to be more related to the chemical structure of humic substances than to the distribution of molecular weight of the humic aggregates.

Humic substances isolated from residues of sugar cane industry as root growth promoter

Substâncias promotoras do crescimento vegetal sao amplamente utilizadas na agricultura moderna. Existem varios produtos no mercado, muitos dos quais sao substâncias humicas isoladas de diferentes fontes. A torta de filtro, um residuo da producao do acucar, e uma fonte rica e renovavel de materia orgânica e essas caracteristicas a tornam uma possivel fonte de substâncias promotoras do crescimento vegetal. Acidos humicos (AH) da torta de filtro foram caracterizados, e foi avaliado seu efeito como promotor de crescimento radicular. As caracteristicas quimicas dos AH foram avaliadas por meio da composicao elementar, grupos funcionais acidos, relacao E4/E6 e espectroscopia de infravermelho. A atividade biologica dos AH foi acessada avaliando-se a arquitetura radicular e a atividade da H+-ATPase de membrana plasmatica. O desenvolvimento de raizes laterais foi diretamente relacionado ao estimulo da atividade da H+-ATPase. A habilidade dos AH em promover o desenvolvimento radicular indica que AH extraidos da torta de filtro podem ser utilizados como estimuladores do crescimento de plantas.

Plant proton pumps as markers of biostimulant action

A standard protocol to evaluate the effects of biostimulants on plant physiology is still lacking. The proton pumps present in the vacuolar and plasma membranes are the primary agents responsible for the regulation of the electrochemical gradient that energizes the nutrient uptake system and acid growth mechanism of plant cells. In this study, two of these enzymes were characterized as biochemical markers of biostimulant activity. A simple and fast protocol based on the degree of root acidification using a pH sensitive dye and the Micro-Tom tomato as a plant model is proposed as an efficient methodology to prove the efficacy of biostimulants that are claimed to improve nutrient acquisition and root growth. The results agree with the data from more conventional, expensive and time-consuming proton pump assays. A direct correlation was found between plasmalemma proton-adenosine triphosphatase (H+-ATPase) activation and the amount of rhizosphere acidification observed in the bromocresol gel. Moreover, roots of the diageotropica (dgt) Micro-Tom plants, defective in auxin responses, barely acidify bromocresol purple gel even in the presence of indole-3-acetic acid (IAA, 1 μM). The biostimulant TEA (vermicompost water extract, 25 %) enhances proton extrusion by 40 % in wild type (WT) plants, but no effect was induced in dgt plants. These results reinforce the notion that the class of biostimulant known as humic substances stimulates plant proton pumps and promotes root growth by exerting an auxin-like bioactivity and establish the usefulness of an economically and technically feasible assay to certify this kind of biostimulant.

Short-Term Changes in Fertility Attributes and Soil Organic Matter Caused by the Addition of EM Bokashis in Two Tropical Soils

The present work aimed to evaluate the behavior of ten fertility attributes of soil organic matter physical fractions and total organic carbon upon addition of three EM Bokashis to a Rhodic Ferralsol (FRr) and a Dystric Cambisol (CMd). An experiment was carried out in greenhouse in which the soils were placed into plastic trays and cultivated with tomato. A completely randomized design was used with four repetitions and factorial scheme of 2 × 3

Compost biofortification with diazotrophic and P‐solubilizing bacteria improves maturation process and P‐availability

BACKGROUND Phosphorus-containing fertilizers play an important role in tropical agriculture owing to the well documented shortage of plant-available P in soils. Traditional P fertilizer production is based on chemical processing of insoluble rock phosphate (RP), which includes an acid treatment at high temperature. Processing the RP increases fertilizer costs, making it unavailable for undercapitalized and typically family-based farmers. Biotechnological methods have been proposed as an alternative to increase phosphate availability in RP. In this study, Burkholderia silvatlantica and Herbaspirillum seropedicae were co-inoculated into an RP-enriched compost with the aim of determining the effects of this technology on the levels of phosphatase activities and release of plant-available P. RESULTS Inoculation of both microorganisms resulted in higher organic matter decomposition and higher humic acid formation in composting. Herbaspirillum seropedicae was the most promising microorganism for the production of acid and alkaline phosphatase enzymes. Both microorganisms presented potential to increase the supply of P from poorly soluble sources owing to increased levels of water-soluble P and citric acid P. CONCLUSION Burkholderia silvatlantica and H. seropedicae in RP-enriched compost may represent an important biotechnological tool to reduce the overall time required for composting and increase the supply of P from poorly soluble sources.

Organic matter pools and nutrient cycling in different coffee production systems in the Brazilian Cerrado

Agroforestry and organic systems have been used to reduce the negative effects that conventional coffee cultivation has on soils. In this work, 13C-CPMAS-NMR, Fourier transform infrared spectroscopy, elemental composition, classical humus fractionation and the soil fertility status were used to evaluate the impact of these three systems on a Latosol from the Brazilian Cerrado. Continuous input of tree residues promoted changes to the soil organic matter with increase in total organic carbon, humic acids (HA) and light organic matter, mainly in the topsoil. Available P and cation exchange capacity were also increased and the acidity status decreased in the agroforestry system. Moreover, HA from the agroforestry were enriched in O-alkyl C, O-di-alkyl C and alkyl C groups and the organic system resulted in HA richer in carboxyl groups. The conventional system resulted in greater aromatic and methoxyl participation, and lower phenol groups. HA from the agroforestry system were richer in easily degradable structures and the chemical fractionation demonstrated a decrease in both recalcitrant fractions, allowing for a more conservative and sustainable management of soil fertility. The modifications were not as evident in the organic system, probably due to the low organic fertilizer input.