Abstract The use of native species’ stem cuttings in riparian forests disturbed by silting could be a promising low-tech alternative for restoration practitioners in riverscape rehabilitation. In this study, we evaluated the vegetative propagation of Amazonian native plants (Buchenavia parviflora, Euterpe oleracea, Ficus insipida and Socratea exorrhiza) with the addition of a bio-fertilizer, and humic and fulvic acids in soil disturbed by human-induced silting. We found that F. insipida and B. parviflora were able to grow and showed high survival percentage with the development of leaves, buds, and roots; even in nutrient deficient and clayey soils. We also found that the frequency of application and the concentration of the organic additives did not show significant influence on plants’ development. Thus, advance in situ tests with both species could be an interesting step to contribute to riverine ecosystems restoration practices.

ABSTRACT 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.

Since the beginning of Human civilization, the soil organic matter has been used as plant growth promoter and/or regulator. Indeed, early in plant science history, even before the auxin concept has been established, the term "auximones" was coined to describe plant growth promoting humic acids derived from peat. Despite of this, until the end of the 20th century, humic substances remained as some of the most neglected environment signals in plant physiology research. However, this scenario has changed in last decade with the discovery that the major systems of energy transduction of the plant cell membranes, the proton pumps, can be tightly orchestrated by humic substances just as elicited by a hormonal signaling. Differential activations of both plasma membrane (PM H+-ATPase) and vacuolar pumps (V-ATPase and H+-PPase) are modulated by humic substances triggering ion signatures related to specific patterns of plant growth and development. Phytohormones have been found to be associated with this humus bioactivity, and nitric oxide acting as a second messenger in a signaling pathway in which plants can sense the soil environment to cope with specific conditions. In this review, we discuss some of the most influential data available in literature, which have shaped this underexplored interface between the chemistry of the organic matter and the plant physiology. The key role of organic matter in the sustainable agriculture will also be highlighted from a biochemical perspective of the plant cell responses to biofertilization, specially in tropical environments.

Substâncias promotoras do crescimento vegetal são amplamente utilizadas na agricultura moderna. Existem vários produtos no mercado, muitos dos quais são substâncias húmicas isoladas de diferentes fontes. A torta de filtro, um resíduo da produção do açúcar, é uma fonte rica e renovável de matéria orgânica e essas características a tornam uma possível fonte de substâncias promotoras do crescimento vegetal. Ácidos húmicos (AH) da torta de filtro foram caracterizados, e foi avaliado seu efeito como promotor de crescimento radicular. As características químicas dos AH foram avaliadas por meio da composição elementar, grupos funcionais ácidos, relação E4/E6 e espectroscopia de infravermelho. A atividade biológica dos AH foi acessada avaliando-se a arquitetura radicular e a atividade da H+-ATPase de membrana plasmática. O desenvolvimento de raízes laterais foi diretamente relacionado ao estímulo da atividade da H+-ATPase. A habilidade dos AH em promover o desenvolvimento radicular indica que AH extraídos da torta de filtro podem ser utilizados como estimuladores do crescimento de plantas.

Plant growth promoting substances are widely used in modern agriculture. Several products in the market are humic substances isolated from different sources. The filter cake, a residue of sugar production, is a rich and renewable source of organic matter and these characteristics place the filter cake as a possible source of plant growth promoting substances. Humic acids (HA) from filter cake were characterized, and their effects as root growth promoters were evaluated. Chemical features of the HA were evaluated through elemental composition, acidic functional groups, E4/E6 ratio and infrared spectroscopy analyzes. The biological activity of the HA was assessed using root architecture parameters and the P-type H+-ATPase activity. The lateral root development was directly related to the stimulation of plasma membrane ATPase activity. The ability of HA to promote root development indicate that HA from filter cake can be used as environmental plant growth stimulators.

Os ácidos húmicos (AH) estimulam diretamente vários processos fisiológicos que promovem o crescimento vegetal, especialmente do sistema radicular. O conhecimento da natureza química e do papel de AH na expressão de efeitos biofertilizantes e bioestimulantes é fundamental para o desenvolvimento de insumos biológicos à base de ácidos húmicos. O objetivo deste trabalho foi avaliar uma possível relação entre a distribuição de massa molecular aparente de AH isolados de vermicomposto e a magnitude de resposta na promoção do crescimento radicular. Para isso, foram obtidas subfrações (SF) dos AH por meio de cromatografia preparativa por exclusão de tamanho em gel de Sephadex G-50 (CGE). O processo preparativo foi validado pelo uso da cromatografia líquida de alta performance por exclusão de tamanho (HPSEC). As cinco frações obtidas foram testadas em diferentes concentrações (0; 0,0001; 0,001; 0,003; 0,005; e 0,01 mol L-1 de C) quanto à sua capacidade de estimular o crescimento radicular de plantas de Arabidopsis thaliana (ecótipo col. 4). Para plantas de milho (Zea mays hyb. UENF 506-6) foi utilizada a dose de 0,002 mol L-1 de C. O modelo quadrático descreveu a indução do crescimento radicular de Arabidopsis e a dose de AH, sendo 0,00511 mol L-1 de C o ponto de inflexão médio. Na concentração ótima, foi observada correlação inversa e significativa entre distribuição de massa molecular e indução do número de raízes laterais em Arabidopsis. No entanto, outros atributos, como área e comprimento radicular, não sofreram influência da massa molecular aparente. Em plântulas de milho, foi observado aumento no número de sítios de mitose e raízes laterais tanto no tratamento com AH como com suas subfrações. A atividade da H+-ATPase de membrana plasmática foi significativamente alterada pelo AH, porém não ocorreu o mesmo com todas as subfrações. A atividade de estimulação do crescimento radicular parece estar mais relacionada com a estrutura química das substâncias húmicas do que com a distribuição de massa molecular dos agregados húmicos.

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.

Um dos fatores mais limitantes para a produção de vermicomposto é a disponibilidade de esterco. Neste trabalho, foi avaliado o efeito da substituição parcial do esterco por bagaço de cana e por resíduos de leguminosa (Gliricidia sepium) na vermicompostagem sobre a qualidade do vermicomposto e sobre a bioatividade dos humatos, avaliadas por meio da análise do crescimento radicular e da atividade das bombas de H+ isoladas de raízes de alface. A substituição do esterco por bagaço de cana e por resíduos de leguminosas não acarretou prejuízo às características químicas dos vermicompostos. No entanto, os humatos isolados dos diferentes vermicompostos apresentaram características químicas distintas, tais como: acidez e propriedades óticas distintas. Os humatos produzidos a partir de esterco de bovino e da mistura esterco bovino + bagaço proporcionaram maiores estímulos no crescimento radicular das plantas de alface, sendo os mais indicados para uso na forma solúvel. A inclusão de resíduos de leguminosas no processo de vermicompostagem produziu humatos sem efeito sobre o desenvolvimento das raízes de alface.

Cattle manure availability is one of the most limiting factors for vermicompost production. The effects of the partial substitution of manure with sugarcane bagasse or residues of Gliricidia sepium on the quality of vermicomposts and the bioactivity of their humates were evaluated by analyzing the root growth and H+-ATPase activity of lettuce seedling roots. The substitutions of manure by sugar cane bagasse and legume cover crops residues did not affect the chemical properties of humates. Humates isolated from the different vermicomposts, however, presented different chemical characteristics, such as acidity and distinct optical properties. Humates isolated from manure and manure + bagasse enhanced growth; these are more adequate for use in soluble form. Humates isolated from vermicompost with legume substitution were ineffective in promoting lettuce root growth.

O vermicomposto contém uma concentração elevada de substâncias húmicas e já é bem conhecido o efeito do seu uso sobre as propriedades do solo. No entanto, a ação direta das substâncias húmicas sobre o metabolismo das plantas é menos conhecida. O objetivo deste trabalho foi avaliar o uso de humatos extraídos de vermicomposto de esterco de curral com KOH 0,1 mol L-1 sobre o desenvolvimento e metabolismo de ATP em plântulas de alface. Após a germinação, plântulas de alface foram tratadas com os humatos em concentrações que variaram de 0 a 100 mg L-1 de C, durante quinze dias. Foram avaliados o crescimento da raiz e a atividade das bombas de H+ isoladas da fração microssomal do sistema radicular. Foi observado aumento na matéria fresca e seca do sistema radicular, bem como no número de sítios de mitose, raízes emergidas do eixo principal, na área e no comprimento radiculares, com o uso do humato na concentração de 25 mg L-1 de C. Também foi observado, nessa concentração, aumento significativo na hidrólise de ATP pelas bombas de H+, responsáveis pela geração de energia necessária à absorção de íons e pelo crescimento celular.

Vermicomposts present a high content of humic substances and their effects on soil properties are well-documented. However, direct effects of humic substances on plant metabolism are less known. The aim of this work was to evaluate the use of humates isolated from cattle manure vermicompost with 0.1 mol L-1 KOH on root growth and ATP metabolism of lettuce seedlings. After germination, lettuce seedlings were treated with different humate concentrations (0 to 100 mg L-1 of C) for 15 days. The root growth and the H+-ATPase activity of root cell microsomes isolated by differential centrifugation were evaluated. The humate treatment, at the concentration of 25 mg L-1 of C, increased fresh and dry root mass as well as root area and total length. Also there was a large increase in the number of root mitotic sites and emerged lateral roots in addition to a significant increase of microsomal ATP hydrolysis. The data suggest that the vermicompost bioactivity could be related, at least in part, to the humate-induced activation of the plant cell H+ pumps, which are responsible for generating the energy for nutrient absorption and cell growth.

O uso de leguminosas herbáceas em sistemas agrícolas permite o aporte de quantidades expressivas de fitomassa, possibilitando alterações no teor e na qualidade da matéria orgânica do solo ao longo dos anos. Este trabalho avalia a qualidade da matéria orgânica de um Argissolo Vermelho Amarelo localizado em Seropédica, RJ, e cultivado com diferentes espécies de leguminosas herbáceas perenes: amendoim forrageiro cv. BR-14951 (Arachis pintoi), cudzu tropical (Pueraria phaseoloides) e siratro (Macroptilium atropurpureum),num delineamento fatorial com três repetições, considerando a manutenção da parte aérea depois do corte na parcela ou a retirada da parte aérea da área experimental. Foram também analisadas amostras de uma área adjacente com vegetação espontânea (mata de capoeira) e capim colonião (Panicum maximum). As amostras de solo foram coletadas em duas profundidades (0-5 e 5-10 cm), realizando-se o fracionamento da matéria orgânica e a avaliação das características estruturais dos ácidos húmicos. As leguminosas avaliadas não alteraram o conteúdo de carbono orgânico total, mas promoveram acúmulo de ácidos húmicos (AH) na camada superficial. Os AH podem ser utilizados como compostos indicadores dos efeitos do manejo sobre a fração orgânica do solo, pois apesar do pouco tempo de implementação do experimento (28 meses) foi verificada incorporação significativa de carbono e nitrogênio provenientes dos resíduos orgânicos das leguminosas. O manejo dos resíduos vegetais não alterou aspectos quantitativos da distribuição de matéria orgânica humificada, mas conferiu maior grau de condensação aos ácidos húmicos avaliados pela análise da composição elementar, espectroscopia de IV e de fluorescência.

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 Seropédica, 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.