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.

Auxin governs dynamic cellular processes involved at several stages of plant growth and development. In this review, we discuss the mechanisms employed by auxin in light of recent scientific advances, with a focus on synthetic auxins as herbicides and synthetic auxin resistance mechanisms. Two auxin receptors were reported. The plasma membrane receptor ABP1 (Auxin Binding Protein 1) alters the structure and arrangement of actin filaments and microtubules, leading to plant epinasty and reducing peroxisomes and mitochondria mobility in the cell environment. The second auxin receptor is the gene transcription pathway regulated by the SCFTir/AFB ubiquitination complex, which destroys transcription repressor proteins that interrupt Auxin Response Factor (ARF) activation. As a result mRNA related with Abscisic Acid (ABA) and ethylene are transcribed, producing high quantities of theses hormones. Their associated action leads to high production of Reactive Oxygen Species (ROS), leading to tissue and plant death. Recently, another ubiquitination pathway which is described as a new auxin signaling route is the F-box protein S-Phase Kinase-Associated Protein 2A (SKP2A). It is active in cell division regulation and there is evidence that auxin herbicides can deregulate the SKP2A pathway, which leads to severe defects in plant development. In this discussion, we propose that SFCSKP2A auxin binding site alteration could be a new auxinic herbicide resistance mechanism, a concept which may contribute to the current progress in plant biology in its quest to clarify the many questions that still surround auxin herbicide mechanisms of action and the mechanisms of weed resistance.

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.

Variedades ou híbridos de tomateiro utilizados para produção de molhos e ketchups costumam ter hábito determinado, enquanto a maioria dos genótipos utilizados na produção para mesa (consumo in natura) possuem hábito indeterminado. Além de influenciar no manejo da cultura, o hábito de crescimento pode alterar parâmetros de produtividade, como o teor de sólidos solúveis totais (SST) nos frutos. O hábito de crescimento é controlado principalmente pelo gene SELF-PRUNING (SP), que é um dos componentes de uma pequena família gênica da qual faz parte também o gene SINGLE FLOWER TRUSS (SFT), atualmente considerado um dos componentes do tão buscado "florígeno". O entendimento da função bioquímica e o efeito fisiológico de tais genes em interação com o ambiente e outros genes (epistasia) possibilita a manipulação de parâmetros como precocidade e SST. Além disso, fornece subsídios para compreender a base genética do crescimento semideterminado, que combina vantagens do hábito determinado e indeterminado, podendo ser usado pelos melhoristas de plantas para o desenvolvimento de novas cultivares.

Hybrids or open pollinated tomato cultivars used for sauces and ketchups production usually has determinate growth habit, while most of the genotypes used in the production to salads (in natura consumption) has indeterminate growth habit. Additionally, growth habit can have influence on culture management, productivity and total soluble solids (TSS) in fruits. The growth habit is mainly controlled by the gene SELF-PRUNING (SP), which is a component of a small gene family which is also part of the gene SINGLE FLOWER TRUSS (SFT), currently considered one of the components so sought after 'florigin'. Understanding the biochemical function and physiological effect of such genes in interaction with the environment and other genes (epistasis), allows the manipulation of parameters such as precocity and TSS. It also provides subsidies to understand the genetic basis of semideterminate growth, which combines the advantages of determinate and indeterminate habit and can be used for plant breeders to development of new cultivars.

As turfeiras constituem uma reserva importante de C humificado nos ecossistemas terrestres. O interesse pelo uso de substâncias húmicas como agentes promotores do crescimento vegetal vem crescendo continuamente. O objetivo deste trabalho foi avaliar a bioatividade das substâncias húmicas alcalino solúveis (SH), ácidos húmicos (AH) e fúlvicos (AF) isolados de turfeiras com matéria orgânica em diferentes estádios de decomposição da matéria orgânica (sáprica, hêmica ou fíbrica) localizadas na Serra do Espinhaço Meridional, MG. Foram obtidas curvas de dose-resposta para o número de raízes emergidas do eixo principal em plântulas de tomateiro. Os resultados mostraram que os AH foram mais bioativos (maior indução de raízes laterais com menor dose) enquanto a maioria dos AF não promoveu o crescimento radicular. Tanto SH como AH estimularam o aparecimento de pelos radiculares. Em baixas concentrações, SH e AH induziram o aparecimento de pelos próximos à coifa, típico de plantas com desbalanço hormonal. O uso do tomateiro transgênico com gene repórter DR5::GUS permitiu a observação da indução de rotas de sinalização ligadas à ação das auxinas na promoção do crescimento radicular pelos AH.

Peats are an important reserve of humified carbon in terrestrial ecosystems. The interest in the use of humic substances as plant growth promoters is continuously increasing. The objective of this study was to evaluate the bioactivity of alkaline soluble humic substances (HS), humic (HA) and fulvic acids (FA) isolated from peats with different decomposition stages of organic matter (sapric, fibric and hemic) in the Serra do Espinhaço Meridional, state of Minas Gerais. Dose-response curves were established for the number of lateral roots growing from the main plant axis of tomato seedlings. The bioactivity of HA was greatest (highest response in lateral roots at lowest concentration) while FA did not intensify root growth. Both HS and HA stimulated root hair formation. At low concentrations, HS and HA induced root hair formation near the root cap, a typical hormonal imbalance effect in plants. Transgenic tomato with reporter gene DR5::GUS allowed the observation that the auxin-related signalling pathway was involved in root growth promotion by HA.

As micorrizas arbusculares (MAs) são associações simbióticas mutualistas entre fungos do filo Glomeromycota e a maioria das plantas terrestres. A formação e o funcionamento das MAs depende de um complexo processo de troca de sinais, que resulta em mudanças no metabolismo dos simbiontes e na diferenciação de uma interface simbiótica no interior das células das raízes. Os mecanismos que regulam a formação das MAs são pouco conhecidos, mas sabe-se que a concentração de fosfato (P) na planta é um fator determinante para o desenvolvimento da simbiose. A disponibilidade de P na planta pode afetar o balanço de açúcares e de fitormônios (FHs), além da expressão de genes de defesa vegetal. Com o advento da genômica e proteômica, vários genes essenciais para o desenvolvimento das MAs já foram identificados e seus mecanismos de regulação estão sendo estudados. Até o presente, sabe-se que as plantas secretam substâncias que estimulam a germinação de esporos e o crescimento de fungos micorrízicos arbusculares (FMAs). Há evidências também de que os FMAs sintetizam moléculas sinalizadoras, que são reconhecidas pelas plantas hospedeiras. Pelo menos três genes são essenciais para o reconhecimento dessa molécula e a transdução do sinal molecular. Discutem-se os papéis desses genes e os possíveis mecanismos que regulam sua expressão, bem como os papéis dos FHs na regulação de MAs são discutidos.

Arbuscular mycorrhizae (AM) are mutualistic symbiotic associations between fungi of the phylum Glomeromycota and most terrestrial plants. The formation and functioning of AM depend on a complex signal exchange process, which ultimately results in shifts in the metabolism of the symbionts and differentiation of a symbiotic interface in cortical root cells. The mechanisms regulating AM development are not well understood, but it is known that phosphate (P) concentration in plants plays a key role in this process. Plant P concentration may affect the balance of sugars and phytohormones (PH), as well as the expression of plant defense-related genes. With the advent of genomics and proteomics, several genes essential for the development of AM have been identified and their regulation mechanisms are being elucidated. Hitherto, it is known that plants secrete several compounds that stimulate spore germination and the growth of arbuscular mycorrhizal fungi (AMF). There is also evidence that AMF synthesize molecules that are recognized by the host plants. At least three genes are essential for the recognition of this molecule and the transduction of the molecular signal. The roles of these genes and the possible mechanisms regulating their expression, as well as the role of PH in controlling AM development are discussed in this review.

Os hormônios vegetais são possíveis reguladores chave do desenvolvimento de micorrizas arbusculares (MAS). Contudo, seus papéis em MA são pouco conhecidos. No presente estudo, foram utilizados mutantes em cinco classes hormonais introgredidos em uma única cultivar (cv. Micro-Tom) de tomateiro (Lycopersicon esculentum Mill. Syn Solanum lycopersicum L.) para determinar seus efeitos no desenvolvimento de MA e expressão de genes relacionados à defesa (quitinases e b-1,3-glucanases) em raízes. Sob condição de baixo P, os mutantes epinastic (epi) e Never ripe (Nr), os quais são super produtores e pouco sensíveis a etileno, respectivamente, tiveram a colonização intra-radicular por Glomus clarum inibida quando comparada com o controle Micro-Tom (MT). Não se observou alterações significativas na colonização fúngica nos mutantes afetando outras classes hormonais. Sob condição de baixo P, o nível de transcritos codificando uma quitinase básica de classe I (chi9) foi mais elevado em raízes micorrizadas dos mutantes epi e Nr, quando comparado com o controle MT. Em contraste, o nível de transcritos de uma b-1,3-glucanase ácida da classe III (TomPR-Q'a) em raízes micorrizadas do mutante epi foi significativamente menor que em raízes micorrizadas de MT. A colonização de raízes no mutante epi foi acompanhada por várias alterações na morfologia fúngica, quando comparada com o controle MT. Os resultados sugerem que o etileno pode desempenhar um importante papel controlando o crescimento fúngico intra-radicular nas MAS.

Plant hormones are likely key regulators of arbuscular mycorrhizae (AM) development. However, their roles in AM are not well known. Here mutants in five hormone classes introgressed in a single tomato (Lycopersicon esculentum Mill. Syn Solanum lycopersicum L.) background (cv. Micro-Tom) were used to determine their effects on AM development and the expression of defense-related genes (chitinases and b-1,3-glucanases) in roots. Under low P conditions, mutant epinastic (epi) and Never ripe (Nr), ethylene overproducer and low sensitivity, respectively, had the intraradical colonization by Glomus clarum highly inhibited, as compared to the control Micro-Tom (MT). No significant alterations in fungal colonization were observed in mutants affecting other hormone classes. Under low P conditions, the steady state levels of transcripts encoding a class I basic chitinase (chi9) were higher in mycorrhizal epi and Nr mutant roots as compared to MT controls. In contrast the steady state levels of a class III acidic b-1,3-glucanase (TomPR-Q'a) transcripts in mycorrhizal epi mutant roots were significantly lower than in mycorrhizal MT roots. Root colonization in epi mutants was accompanied by several alterations in fungal morphology, as compared to root colonization in MT controls. The data suggest that ethylene may play an important role in controlling intraradical arbuscular mycorrhizal fungal growth.

O nanismo em plantas de bananeira micropropagada, representa uma das formas mais comum de alteração genética no subgrupo Cavendish. Rizomas de plantas normais e anãs de bananeira cv. Grande Naine, incubados durante 5 dias na presença de [³H]-L-triptofano (Trp), [³H]-ácido 3-indolil acético e giberelina (GA3) foram utilizados para o estudo da biossíntese de ácido 3-indolil acético e ácido 3-indolil acético amida e dos níveis endógenos de citocininas totais (Cks) e ácido 3-indolil acético livre. Os teores endógenos de ácido 3-indolil acético e de seus conjugados éster e amida foram obtidos de plantas anãs e normais incubadas durante 30 dias na presença de GA3, ácido 3-indolil acético e L- e D-triptofano. A aplicação de [³H]-L- triptofano resultou na obtenção de quantidades mais elevadas de radioatividade nas plantas normais, no tempo de retenção coincidente com o de ácido 3-indolil acético, ácido 3-indolil acético-aspartato, ácido 3-indolil acético-glicina e ácido 3-indolil acético-Alanina, valores estes superiores aos observados nas plantas anãs. A ocorrência de maior quantidade de ácido 3-indolil acético radioativo e de formas amida nas plantas anãs, foi verificada nos rizomas tratados com [³H]-L-triptofano e GA3 simultaneamente. Os níveis endógenos de Cks totais foram praticamente iguais em ambos materiais, sendo os teores de ácido 3-indolil acético nas plantas normais cerca de 1,5 vezes superiores aos das plantas anãs. Os tratamentos com triptofano e GA3 favoreceram incremento nos níveis de ácido 3-indolil acético livre nas plantas anãs, paralelamente a uma diminuição nos teores de ácido 3-indolil acético éster e amida.

Nanism is one of the most frequent type of mutant in micropropagated banana plants from the Cavendish subgroup. The present study aimed at studying some of the hormone factors involved in this type of mutation. Rhizomes from normal and dwarf plants from the cultivar Grand Naine were incubated for 5 d in the presence of [³H]-L-tryptophan, [³H]-indole-3-acetic acid and gibberellin, to quantify the endogenous levels of indole-3-acetic acid-ester, indole-3-acetic acid-amide, free indole-3-acetic acid, and cytokinins. The endogenous levels of indole-3-acetic acid and its ester- and amide-conjugated forms were measured in normal and dwarf plants incubated for 30 d with gibberellin, indole-3-acetic acid, and L- and D-tryptophan. In normal plants, the use of [³H]-L-tryptophan resulted in higher levels of radioactivity in the retention times corresponding to indole-3-acetic acid, indole-3-acetic acid-aspartate, indole-3-acetic acid-glycine and indole-3-acetic acid-alanine. These values were higher than those observed in dwarf plants. Higher quantities of radioactive indole-3-acetic acid and of amide-forms in dwarf plants occurred in rhizomes treated with [³H]-L-tryptophan and gibberellin simultaneously. The endogenous levels of total cytokinins were the same in both materials, while the levels of indole-3-acetic acid in normal plants were 1.5 times higher than in dwarf plants. Moreover, in these ones application of tryptophan and gibberellin coincided in an increase in the levels of free indole-3-acetic acid in dwarf plants and to a decrease in the levels of indole-3-acetic acid-ester and indole-3-acetic acid-amide.