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Mucor circinelloides is an opportunistic dimorphic pathogen, with the dimorphic process controlled in parts by fermentative and oxidative metabolisms, which lead to yeast or mycelial growth, respectively. Dimorphic transition is important for pathogenesis since the mycelium represents the virulent morphology. We previously reported that the deletion of arl1 or arl2 stimulate anaerobic germination in M. circinelloides, suggesting an augmented fermentative metabolism. In the present study, we demonstrate that the heterokaryon Δarl1(+)(−) and homokaryon Δarl2 strains contain low number of mitochondria, which possibly results in a dysfunctional oxidative metabolism, marked by a low oxygen consumption in glucose and poor growth in glycerol as the unique carbon source. This dysfunction is compensated for by an increase in the glycolysis and fermentation in aerobic conditions, demonstrating growth kinetics similar to that in the wild-type strain. Moreover, as a consequence a high fermentative activity, the Δarl1(+)(−) and Δarl2 strains possibly increased the yeast cell growth during low oxygen concentrations in presence of glucose. To the best of our knowledge, this is the first study to demonstrate the control of members of Arf family on the mitochondrial population in a Mucor species.

The plant hormone ethylene induces auxin biosynthesis and transport and modulates root growth and branching. However, its function on root stem cells and the identity of interacting factors for the control of meristem activity remains unclear. Genetic analysis for primary root growth in wild-type (WT) Arabidopsis thaliana seedlings and ethylene-related mutants showed that the loss-of-function of CONSTITUTIVE TRIPLE RESPONSE1 (CTR1) inhibits cell division and elongation. This phenotype is associated with an increase in the expression of the auxin transporter PIN2 and a drastic decrease in the expression of key factors for stem cell niche maintenance such as PLETHORA1, SHORT ROOT and SCARECROW. While the root stem cell niche is affected in ctr1 mutants, its maintenance is severely compromised in the ctr1-1eir1-1(pin2) double mutant, in which an evident loss of proliferative capacity of the meristematic cells leads to a fully differentiated root meristem shortly after germination. Root traits affected in ctr1-1 mutants could be restored in ctr1-1ein2-1 double mutants. These results reveal that ethylene perception via CTR1 and EIN2 in the root modulates the proliferative capacity of root stem cells via affecting the expression of genes involved in the two major pathways, AUX-PIN-PLT and SCR-SHR, which are key factors for proper root stem cell niche maintenance.

The MEDIATOR complex influences the transcription of genes acting as a RNA pol II co-activator. The MED16 subunit has been related to low phosphate sensing in roots, but how it influences the overall plant growth and root development remains unknown. In this study, we compared the root growth of Arabidopsis wild-type (WT), and two alleles of MED16 (med16-2 and med16-3) mutants in vitro. The MED16 loss-of-function seedlings showed longer primary roots with higher cell division capacity of meristematic cells, and an increased number of lateral roots than WT plants, which correlated with improved biomass accumulation. The auxin response reported by DR5:GFP fluorescence was comparable in WT and med16-2 root tips, but strongly decreased in pericycle cells and lateral root primordia in the mutants. Dose-response analysis supplementing indole-3-acetic acid (IAA), or the auxin transport inhibitor N-1-naphthylphthalamic acid (NPA), indicated normal responses to auxin in the med16-2 and med16-3 mutants regarding primary root growth and lateral root formation, but strong resistance to NPA in primary roots, which could be correlated with cell division and elongation. Expression analysis of pPIN1::PIN1::GFP, pPIN3::PIN3::GFP, pIAA14:GUS, pIAA28:GUS and 35S:MED16-GFP suggests that MED16 could mediate auxin signaling. Our data imply that an altered auxin response in the med16 mutants is not necessarily deleterious for overall growth and developmental patterning and may instead directly regulate basic cellular programmes.

N,N-dimethyl-hexadecylamine (DMHDA) is released as part of volatile blends emitted by plant probiotic bacteria and affects root architecture, defense and nutrition of plants. Here, we investigated the changes in gene expression of transcription factors responsible of maintenance of the root stem cell niche and jasmonic acid signaling in Arabidopsis seedlings in response to this volatile. Concentrations of DMHDA that repress primary root growth were found to alter cell size and division augmenting cell tissue layers in the meristem and causing root widening. DMHDA triggered the division of quiescent center cells, which correlated with repression of SHORT ROOT (SHR), SCARECROW (SCR), and PLETHORA 1 (PLT1) proteins and induction of WUSCHEL-RELATED HOMEOBOX 5 (WOX5) transcription factor. Interestingly, an activation of the expression of the jasmonic acid-related reporter genes JAZ1/TIFY10A-GFP and JAZ10pro::JAZ10-GFP suggests that the halted growth of the primary root inversely correlated with expression patterns underlying the defense reaction, which may be of adaptive importance to protect roots against biotic stress. Our data help to unravel the gene expression signatures upon sensing of a highly active bacterial volatile in Arabidopsis seedlings.

Sucrose is synthesized from UDP-Glc and Fru-6-phosphate via the activity of sucrose-phosphate synthase (SPS) enzymes, which produce Suc-6-phosphate. Suc-6-phosphate is rapidly dephosphorylated by phosphatases to produce Suc and inorganic phosphate. Arabidopsis has four sps genes encoding SPS enzymes. Of these enzymes, AtSPS1F and AtSPS2F have been grouped with other dicotyledonous SPS enzymes, while AtSPS3F and AtSPS4F are included in groups with both dicotyledonous and monocotyledonous SPS enzymes. In this work, we generated Arabidopsis thaliana transformants containing the promoter region of each sps gene fused to gfp::uidA reporter genes. A detailed characterization of expression conferred by the sps promoters in organs and tissues was performed. We observed expression of AtSPS1F, AtSPS2F and AtSPS3F in the columella roots of the plants that support sucrose synthesis. Hence, these findings support the idea that sucrose synthesis occurs in the columella cells, and suggests that sucrose has a role in this tissue. In addition, the expression of AtSPS4F was identified in embryos and suggests its participation in this developmental stage. Quantitative transcriptional analysis of A. thaliana plants grown in media with different osmotic potential showed that AtSPS2F and AtSPS4F respond to osmotic stress.

Folic acid is a precursor of tetrahydrofolate (vitamin B9), which is an essential cofactor in most organisms, acting as a carrier for one-carbon units in enzymatic reactions. In this work, we employed pharmacological, genetic and confocal imaging strategies to unravel the signaling mechanism by which folic acid modulates root growth and development. Folic acid supplementation inhibits primary root elongation and induces lateral root formation in a concentration-dependent manner. An analysis of the expression of cell cycle genes pCycD6;1:GFP and CycB1:uidA, and cell expansion Exp7:uidA showed that folic acid promotes cell division but prevented cell elongation, and this correlated with altered expression of auxin-responsive DR5:GFP gene, and PIN1:PIN1:GFP, PIN3:PIN3:GFP, and PIN7:PIN7:GFP auxin transporters at the columella and vasculature of primary roots, whereas mutants defective in auxin signaling (tir1/afb1/afb2 [receptors], slr1 [repressor] and arf7/arf19 [transcription factors]) were less sensitive to folic acid induced primary root shortening and lateral root proliferation. Comparison of growth of WT and TARGET OF RAPAMYCIN (TOR) antisense lines indicates that folic acid acts by an alternative mechanism to this central regulator. Thus, folic acid modulation of root architecture involves auxin and acts independently of the TOR kinase to influence basic cellular programs.

Amino acids serve as structural monomers for protein synthesis and are considered important biostimulants for plants. In this report, the effects of all 20-L amino acids in Arabidopsis primary root growth were evaluated. 15 amino acids inhibited growth, being l-leucine (l-Leu), l-lysine (l-Lys), l-tryptophan (l-Trp), and l-glutamate (l-Glu) the most active, which repressed both cell division and elongation in primary roots. Comparisons of DR5:GFP expression and growth of WT Arabidopsis seedlings and several auxin response mutants including slr, axr1 and axr2 single mutants, arf7/arf19 double mutant and tir1/afb2/afb3 triple mutant, treated with inhibitory concentrations of l-Glu, l-Leu, l-Lys and l-Trp revealed gene-dependent, specific changes in auxin response. In addition, l- isomers of Glu, Leu and Lys, but not l-Trp diminished the GFP fluorescence of pPIN1::PIN1:GFP, pPIN2::PIN2:GFP, pPIN3::PIN3:GFP and pPIN7::PIN7:GFP constructs in root tips. MPK6 activity in roots was enhanced by amino acid treatment, being greater in response to l-Trp while mpk6 mutants supported cell division and elongation at high doses of l-Glu, l-Leu, l-Lys and l-Trp. We conclude that independently of their auxin modulating properties, amino acids signals converge in MPK6 to alter the Arabidopsis primary root growth.

Phosphate (Pi) is a critical macronutrient for the biochemical and molecular functions of cells. Under Pi limitation, plants manifest adaptive strategies to increase Pi scavenging. However, the molecular mechanisms underlying these responses remain unclear and only a few components have been identified to date. Here, we show that under Pi-scarcity, the MEDIATOR (MED) transcriptional co-activator, through its MED16 subunit, plays a critical role in Arabidopsis root system architecture remodeling, including primary root growth, lateral root formation, and root hair development. Transcriptomic and genetic analyses further revealed that MED16, via a direct interaction with SENSITIVE TO PROTON RHIZOTOXICITY 1 (STOP1), is required for the transcriptional activation of ALUMINUM ACTIVATED MALATE TRANSPORT 1 (ALMT1), a membrane permease, to increase malate exudation in response to low Pi. Our results unravel a critical transcriptional component for understanding how plant cells orchestrate gene expression with the STOP1-ALMT1 module for survival under deficiency of a critical macronutrient.

Strawberries are one of the most popular fruits in the world. The strawberry production process includes generation of plants through in vitro germination of achenes, micropropagation, and production under greenhouse conditions. In this study, we tested the effect of the plant biostimulant bacteria, Arthrobacter agilis UMCV2 and Bacillus methylotrophicus M4-96, on those processes. Volatile compounds produced by A. agilis UMCV2 stimulated achene germination while B. methylotrophicus M4-96 induced germination through production of diffusible compounds. Growth of in vitro strawberry plants inoculated with A. agilis UMCV2 or exposed to their volatile compounds was also stimulated. Plants inoculated in vitro with A. agilis UMCV2 and B. methylotrophicus M4-96, and then cultured in a greenhouse for 10 months had a 42 % higher fruit yield than un-inoculated controls. Confocal microscopy, used to assess fluorescent in situ hybridization, showed that both bacteria were present in the leaf mesophyll of inoculated plants, but not in control plants. Our results showed that endophytic bacteria, such as A. agilis UMCV2 and B. methylotrophicus M4-96, can be inoculated into micropropagated strawberry plants and colonize the plant tissues, thereby increasing the yield and fruit quality under greenhouse conditions through different mechanisms of plant biostimulation.

The aim of this ex vivo study was to evaluate the adherence capacity of Streptococcus mutans after being exposed to three different sizes of silver nanoparticles on healthy human dental enamel. Three different sizes of silver nanoparticles (9.3, 21.3 and 98 nm) were prepared, characterized and an adherence testing was performed to evaluate their anti-adherence activity on a reference strain of S. mutans on healthy dental enamel surfaces. Colony-Forming Unit count was made for adherence test and light microscopy, atomic force microscopy and scanning electron microscopy were used to compare qualitative characteristics of S. mutans. 9.3 nm and 21.3 nm groups did not show differences between them but statistical differences were found when 9.3 nm and 21.3 nm groups were compared with 98 nm and negative control groups (p

Dental caries is a common infectious disease in humans all over the world and is considered a public health problem with an incidence and prevalence still high. The principal pathogen associated with the development of dental caries is Streptococcus mutans (S. mutans), a microorganism that has been widely studied. A new approach to treat dental caries is the use of silver nanoparticles (SNP) due to their antibacterial effect. It has been reported the effectiveness of SNP against S. mutans, but sensibility of S. mutans serotypes to SNP is still unknown. The aim of this study is to analyze the inhibitory effect of three different sizes of silver nanoparticles on S. mutans obtained from clinical isolates and one reference bacteria strain, determining sensibility differences between serotypes, which were determined by using PCR analysis. Silver nanoparticles with sizes around 9.3, 21.3 and 98 nm were prepared, characterized and used to find, with a microdilution method, the minimum inhibitory concentration (MIC) on S. mutans. SNP showed bactericidal effect on all strains tested with statistical differences between smaller nanoparticles and larger nanoparticles, 9.3 nm SNP showed a higher antibacterial activity than 21.3 and 98 nm SNP; however, sensibility was different among all clinical strains (p

Dental caries is a very infectious disease; in humans, 95% of the worldwide population is affected. The microorganism associated with dental caries is Streptococcus mutans (S. mutans). Although several mechanisms for its control have been used, its prevalence and incidence are still high. New alternatives are silver nanoparticles due to their antibacterial effect. In this work, we determined the antibacterial effect of silver nanoparticles on S. mutans. Three sizes of silver nanoparticles were used to find minimum inhibitory concentrations (MIC) in S. mutans using reference and clinical stocks. Kruskal–Wallis and U of Mann–Whitney statistical tests were applied. We found bactericidal effect for the three groups, with significant statistical differences between them. Our results agree with those already reported by several authors. This study concludes that silver nanoparticles present antibacterial activity on S. mutans and this property is better when the particle size is diminished.