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.

O objetivo do presente trabalho foi correlacionar a atividade da redutase do nitrato e a eficiência fotoquímica máxima do fotossistema II (FSII), expressa pela razão F V/F M (F V = fluorescência variável e F M = fluorescência máxima), em mamoeiro (Carica papaya L.) cv. Tainung 01 e Sunrise Solo 72/12 em condições de campo. O potencial fotoquímico do FSII foi medido in situ em folhas adaptadas ao escuro. Depois, nas mesmas folhas, foi medida a atividade da enzima. Não houve diferença significativa entre a eficiência fotoquímica máxima do FSII entre as cultivares Tainung 01 e Sunrise Solo 72/12, porém a atividade da redutase do nitrato foi notoriamente maior na primeira. A atividade da redutase do nitrato foi altamente correlacionada à eficiência fotoquímica máxima do FS II tanto na cv. Tainung 01 (coeficiente de correlação r= 0,740 e coeficiente de determinação r²= 0,706) quanto na Sunrise Solo 72/12 (coeficiente de correlação r= 0,960 e coeficiente de determinação r²= 0,945). Esses resultados sugerem que há uma correlação entre a fluorescência da clorofila a e a atividade da redutase do nitrato nessas plantas.

The objective of the present work was to connect the nitrate reductase activity and the photochemical efficiency of the photosystem II (PSII), measured as F V/F M, in papaya (Carica papaya L.) cv. Tainung 01 and Sunrise Solo 72/12 in field conditions. The photochemical potential of PSII was measured in situ on attached leaves that had been dark adapted, prior to the fluorescence measurement. After, in same leaves, the enzyme activity was measured. There was not significant difference among the photochemical efficiency of PSII between cv. Tainung 01 and Sunrise Solo 72/12, even so, the activity of the enzyme was markedly higher in cv. Tainung 01 than in cv. Sunrise Solo 72/12. The nitrate redutase activity was highly correlated with the photochemical efficiency of the PSII in both cv. Tainung 01 (coefficient of correlation r= 0,740 and coefficient of determination r²= 0,706) and Sunrise Solo 72/12 (coefficient of correlation r= 0,960 and coefficient of determination r²= 0,945). These results suggest that there is a correlation between chlorophyll a fluorescence and nitrate reductase activity in these plants.

O objetivo deste trabalho foi avaliar a atividade da pectinametilesterase (PME) em mamão (Carica papaya L.), cv. Sunrise Solo e Tainung, e relacionar suas alterações com a perda de firmeza da polpa durante o amadurecimento. Os frutos foram coletados diariamente em uma lavoura comercial em Aracruz - ES, e armazenados sob temperatura ambiente (aproximadamente, 25ºC) até o período das análises. A perda da firmeza da polpa dos frutos das cultivares iniciou-se no 4º dia após a colheita (DAC); entretanto, a cv. Sunrise Solo apresentou uma redução mais acentuada em relação à cv. Tainung. A atividade da PME na cv. Sunrise Solo foi alta no 1º DAC e reduziu gradativamente até o 4º DAC, quando atingiu seu menor valor. No 5º DAC, a atividade aumentou, permanecendo praticamente inalterada até o 8º DAC. Na cv. Tainung, a atividade aumentou até o 3º DAC e decresceu no 4º DAC. No 5º DAC, sua atividade aumentou novamente, atingindo seu maior valor no 6º DAC. A partir daí, a atividade da enzima reduziu-se significativamente, mantendo o mesmo valor até o último dia de análise (8º DAC). O coeficiente de correlação obtido entre a atividade da PME e a firmeza da polpa dos frutos da cv. Sunrise Solo, após o 4º DAC, foi de R = - 0,84, indicando que, durante esse período, a firmeza da polpa dos frutos diminuiu à medida que a atividade da PME aumentou. Por outro lado, não houve correlação significativa entre essas duas características na cv. Tainung.

The objective of this work was to evaluate the pectinmethylesterase (PME) activity in papaya (Carica papaya L.) fruits cv. Sunrise Solo and Tainung, and to correlate its changes with the pulp firmness loss during fruit ripening. Fruits were collected daily from a commercial farm localized in Aracruz - ES, and stored under room temperature (approximately 25 °C) until the period of analysis. The pulp firmness loss of the fruits in both cultivars began at the 4th day after harvest (DAH); however, the cv. Sunrise Solo presented highest reduction than the cv. Tainung. The PME activity of cv. Sunrise Solo was higher at the 1st DAH, and decreased slowly at the 4th DAH, reaching its smallest value. At the 5th DAH the activity increased, remaining practically the same at the 8th DAH. In the cv. Tainung, the PME activity increased until the 3rd DAH and decreased at the 4th DAH. In the 5th DAH, its activity increased again reaching its higher value at the 6th DAH. From this on, the enzyme activity decreased significantly, maintaining the same value until the last day of analysis (8th DAH). The correlation coefficient obtained between the PME activity and the pulp firmness in the cv. Sunrise Solo after the 4th DAH was R = - 0,84, indicating that during this period, the pulp firmness of the fruits decreased when the PME activity increased. On the other hand, there was no significant correlation between these two characteristics in the cv. Tainung.