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Biomedical subjects

Jorge M Vivanco

Publications and source records attributed to Jorge M Vivanco.

At least 19 recordsLinked to original sources

The floral volatile, methyl benzoate, from snapdragon (Antirrhinum majus) triggers phytotoxic effects in Arabidopsis thaliana.

Previously it has been shown that the floral scent of snapdragon flowers consists of a relatively simple mixture of volatile organic compounds (VOCs). These compounds are thought to be involved in the attraction of pollinators; however, little is known about their effect on other organisms, such as neighboring plants. Here, we report that VOCs from snapdragon flowers inhibit Arabidopsis root growth. Out of the three major snapdragon floral volatiles, myrcene, (E)-beta-ocimene, and methyl benzoate (MB), MB was found to be primarily responsible for the inhibition of root growth. Ten micromoles MB reduced root length by 72.6%. We employed a microarray approach to identify the MB target genes in Arabidopsis that were responsible for the root growth inhibition phenotype in response to MB. These analyses showed that MB treatment affected 1.33% of global gene expression, including cytokinin, auxin and other plant-hormone-related genes, and genes related to seed germination processes in Arabidopsis. Accordingly, the root growth of cytokinin (cre1) and auxin (axr1) response mutants was less affected than that of the wild type by the volatile compound: roots of the treated mutants were reduced by 45.1 and 56.2%, respectively, relative to untreated control mutants.

Antirrhinum↗

Can plant biochemistry contribute to understanding of invasion ecology?

Ecologists have long searched for an explanation as to why some plant invaders become much more dominant in their naturalized range than in their native range, and, accordingly, several non-exclusive ecological hypotheses have been proposed. Recently, a biochemical explanation was proposed--the "novel weapons hypothesis"--based on findings that Centaurea diffusa and Centaurea maculosa produce bioactive compounds (weapons) that are more active against naïve plant species in the introduced range than against co-evolved species in the native range. In this Opinion article, we revise and expand this biochemical hypothesis and discuss experimental and conceptual advances and limitations.

Adaptation, Physiological↗

Effect of transporters on the secretion of phytochemicals by the roots of Arabidopsis thaliana.

Root exudation, the process by which plants secrete compounds into the soil, is becoming accepted as a communicative process that determines organismal interactions in the rhizosphere. However, the mechanistic processes involved in the root exudation of phytochemicals have not been elucidated; traditionally, exudation has been regarded as a passive process. There is evidence that transporters in plants (and other organisms) have been involved in the movement of chemicals across different membranes. Here, we describe the involvement of different transporters in root exudation of phytochemicals by employing a pharmacological approach. We used a range of concentrations of several compounds known to inhibit different transporters, including potassium cyanide, orthovanadate, quinidine, glibenclamide, nifedipine and verapamil, to examine the effects of transporter inhibition on root exudation profiles in Arabidopsis. Generally, the exudation profile of phenolic compounds in 18-day-old plants shows more than 15 major phytochemicals. In contrast, the inhibitors listed above caused differences in the secretion of specific compounds. For instance, nifedipine and verapamil completely inhibited the exudation of the phytochemicals with molecular masses of 142 and 294, respectively. These results highlight that root exudation of phytochemicals is an active process controlled at the biochemical level and that different transporters may be involved in this root-specific mechanism.

ATP-Binding Cassette Transporters↗

Ribosome-inactivating proteins in edible plants and purification and characterization of a new ribosome-inactivating protein from Cucurbita moschata.

The basic protein fraction of tissue extracts from 40 edible plants inhibited cell-free protein synthesis and released adenine from herring sperm DNA, thus having adenine glycosylase activity. This suggested the presence of ribosome-inactivating proteins (RIPs) in the plant extracts. This indication was further strengthened by the presence of the two activities after a partial chromatographic purification of three extracts, including that from Lycopersicon esculentum (tomato), which had very low activity. From the extract of Cucurbita moschata (pumpkin), the most active one, a glycoprotein of 30,665 Da was purified which had the properties of a RIP, in that (i) it inhibited protein synthesis by a rabbit reticulocyte lysate with IC50 (concentration giving 50% inhibition) 0.035 nM (1.08 ng ml(-1)) and by HeLa, HT29 and JM cells with IC50 in the 100 nM range, (ii) deadenylated hsDNA and other polynucleotidic substrates, and (iii) depurinated yeast rRNA at a concentration of 0.1 ng ml(-1), all values being comparable to those of other RIPs. The C. moschata RIP gave a weak cross-reaction only with an antiserum against dianthin 32, but not with antisera against other RIPs, and had superoxide dismutase, antifungal and antibacterial activities.

Amino Acid Sequence↗

Oxalate contributes to the resistance of Gaillardia grandiflora and Lupinus sericeus to a phytotoxin produced by Centaurea maculosa.

Centaurea maculosa Lam. is a noxious weed in western North America that produces a phytotoxin, (+/-)-catechin, which is thought to contribute to its invasiveness. Areas invaded by C. maculosa often result in monocultures of the weed, however; in some areas, North American natives stand their ground against C. maculosa and show varying degrees of resistance to its phytotoxin. Two of these resistant native species, Lupinus sericeus Pursh and Gaillardia grandiflora Van Houtte, were found to secrete increased amounts of oxalate in response to catechin exposure. Mechanistically, we found that oxalate works exogenously by blocking generation of reactive oxygen species in susceptible plants and reducing oxidative damage generated in response to catechin. Furthermore, field experiments show that L. sericeus indirectly facilitates native grasses in grasslands invaded by C. maculosa, and this facilitation can be correlated with the presence of oxalate in soil. Addition of exogenous oxalate to native grasses and Arabidopsis thaliana (L.) Heynh grown in vitro alleviated the phytotoxic effects of catechin, supporting the field experiments and suggesting that root-secreted oxalate may also act as a chemical facilitator for plant species that do not secrete the compound.

Antioxidants↗

The role of root exudates in rhizosphere interactions with plants and other organisms.

The rhizosphere encompasses the millimeters of soil surrounding a plant root where complex biological and ecological processes occur. This review describes recent advances in elucidating the role of root exudates in interactions between plant roots and other plants, microbes, and nematodes present in the rhizosphere. Evidence indicating that root exudates may take part in the signaling events that initiate the execution of these interactions is also presented. Various positive and negative plant-plant and plant-microbe interactions are highlighted and described from the molecular to the ecosystem scale. Furthermore, methodologies to address these interactions under laboratory conditions are presented.

Host-Parasite Interactions↗

Isolation and purification of ribosome-inactivating proteins.

Ribosome-inactivating proteins (RIPs) are cytotoxic N-glycosidases identified in plants, fungi, and bacteria. RIPs inhibit protein synthesis by virtue of their enzymatic activity, selectively cleaving a specific adenine residue from a highly conserved, surface-exposed, stem-loop (S/R loop) structure in the 28S rRNA of ribosomes. Some RIPs also exhibit a number of other enzymatic activities such as RNase, DNase, phospholipase, and superoxide dismutase (SOD). RIPs are considered to be plant defense-related proteins as they are able to inhibit the multiplication and growth of several pathogenic virus, fungi, and bacteria either alone or in conjugation with other defense-related proteins. The mechanism of inhibitory activity of RIPs against fungal pathogens seems to be by directly inhibiting fungal growth rather than depurinating host plant ribosomes and causing cell death as previously envisaged. This chapter describes the protocol used to isolate and purify RIPs from plant tissues.

Glycoside Hydrolases↗

Soil nematodes mediate positive interactions between legume plants and rhizobium bacteria.

Symbiosis between legume species and rhizobia results in the sequestration of atmospheric nitrogen into ammonium, and the early mechanisms involved in this symbiosis have become a model for plant-microbe interactions and thus highly amenable for agricultural applications. The working model for this interaction states that the symbiosis is the outcome of a chemical/molecular dialogue initiated by flavonoids produced by the roots of legumes and released into the soil as exudates, which specifically induce the synthesis of nodulation factors in rhizobia that initiate the nodulation process. Here, we argue that other organisms, such as the soil nematode Caenorhabditis elegans, also mediate the interaction between roots and rhizobia in a positive way, leading to nodulation. We report that C. elegans transfers the rhizobium species Sinorhizobium meliloti to the roots of the legume Medicago truncatula in response to plant-released volatiles that attract the nematode. These findings reveal a biologically-relevant and largely unknown interaction in the rhizosphere that is multitrophic and may control the initiation of the symbiosis.

Animals↗

Analysis of natural flavonoids by microchip-micellar electrokinetic chromatography with pulsed amperometric detection.

Catechins (catechin and other derivatives) are naturally occurring flavonoids present in a number of plants and foods. They are also part of numerous nutraceutical formulations because they are believed to have antioxidant, cancer chemo-preventative, anti-inflammatory and antimicrobial properties. The determination of catechins has traditionally been performed by HPLC. However, this methodology is both time and sample intensive and generates large amounts of organic solvent waste. In the current report, an application of MEKC using a PDMS microchip is presented for the analysis of catechins. The system uses pulsed amperometric detection for direct analysis of important naturally occurring catechins. The effect of pH, surfactant concentration, detection potential and signal stability were analyzed. Linear relationships were found between the concentration and peak current, with good stability and limits of detection of 8 [micro sign]M for catechin, epigallocatechin gallate and epicatechin, and 14 [micro sign]M for epicatechin gallate. Optimum conditions were applied to the detection of selected catechins in a commercially available green tea extract nutraceutical and the results were compared to HPLC analysis. The analysis using microchip micellar electrokinetic chromatography and pulsed amperometric detection was completed in 4.5 min, 10 times faster than the HPLC analysis.

Catechin↗

Mediation of pathogen resistance by exudation of antimicrobials from roots.

Most plant species are resistant to most potential pathogens. It is not known why most plant-microbe interactions do not lead to disease, although recent work indicates that this basic disease resistance is multi-factorial. Here we show that the exudation of root-derived antimicrobial metabolites by Arabidopsis thaliana confers tissue-specific resistance to a wide range of bacterial pathogens. However, a Pseudomonas syringae strain that is both at least partly resistant to these compounds and capable of blocking their synthesis/exudation is able to infect the roots and cause disease. We also show that the ability of this P. syringae strain to block antimicrobial exudation is dependent on the type III secretory system.

Anti-Infective Agents↗

Bacterial expression and enzymatic activity analysis of ME1, a ribosome-inactivating protein from Mirabilis expansa.

Ribosome-inactivating proteins (RIPs) are toxic proteins synthesized by many plants and some bacteria, that specifically depurinate the 28S RNA and thus interrupt protein translation. RIPs hold broad interest because of their potential use as plant defense factors against pathogens. However, study of the activity of type I RIPs has been hampered since their expression in Escherichia coli has typically been toxic to the model system. Mirabilis expansa, an Andean root crop, produces a type I RIP called ME1 in large quantities in its storage roots. In this study, the cDNA sequence of ME1 was used to successfully express the recombinant ME1 protein in E. coli. The production of recombinant ME1 in E. coli was confirmed by Western blot analysis using anti-ME1 antibodies. The studies with fluorescence-labeled ME1 showed that ME1 can enter bacteria and be distributed in the cytoplasm uniformly, indicating its ability to access the protein synthesis machinery of the bacteria. The recombinant enzyme was active and depurinated yeast ribosomes. However, both native and recombinant ME1 proteins failed to depurinate the E. coli ribosomes, explaining the non-toxicity of recombinant ME1 to E. coli. Structural modeling of ME1 showed that it has folding patterns similar to other RIPs, indicating that ME1 and PAP, which share a similar folding pattern, can show different substrate specificity towards E. coli ribosomes. The results presented here are very significant, as few reports are available in the area of bacterial interaction with type I RIPs.

Base Sequence↗

Staphylococcus aureus pathogenicity on Arabidopsis thaliana is mediated either by a direct effect of salicylic acid on the pathogen or by SA-dependent, NPR1-independent host responses.

Staphylococcus aureus is a ubiquitous gram-positive bacterium that can cause superficial to serious systemic infections in animals and humans. Here we report the development of a plant infection model to study the pathogenesis of this bacterium. Three global regulatory mutants, RN6911 (agr-), ALC 488 (sarA-) ALC 842 (sarA-/agr-) and an alpha-toxin mutant defective in biofilm formation (DU1090) which are attenuated in animal pathogenesis, were also attenuated in their ability to infect plants, suggesting that these regulators that mediate synthesis of virulence factors essential for animal pathogenesis are also required for plant pathogenesis. Further, using Arabidopsis plants altered in defense responses such as the transgenic lines NahG [defective in salicylic acid (SA) accumulation], and 35S-LOX2- (defective in jasmonic acid production and hyper-accumulator of SA), and mutants ics1 (depleted in SA accumulation), and npr1-1 (non-expressor of pathogenesis-related protein) we show that resistance of Arabidopsis to typical plant pathogens and the animal pathogen S. aureus is conserved and is mediated by SA. The data presented here suggest that Arabidopsis thaliana resistance to S. aureus is mediated either by a direct effect of SA on the pathogen, specifically one that affects the attachment/aggregate formation on the root surface and reduces the pathogen's virulence, or by SA-dependent, NPR1-independent host responses.

Arabidopsis↗

Enterococcus faecalis mammalian virulence-related factors exhibit potent pathogenicity in the Arabidopsis thaliana plant model.

Some pathogenic bacteria belong to a large, diverse group of species capable of infecting plants, animals, and humans. Enterococcus faecalis is an opportunistic human pathogen capable of infecting patients with a deficient immune system. Here we report that three E. faecalis strains (FA-2-2, V583, and OG1RF) are capable of infecting the leaves and roots of the model plant species Arabidopsis thaliana, causing plant mortality 7 days postinoculation. We found that E. faecalis pathogenesis in A. thaliana leaves is determined by the following series of events: attachment to leaf surface, entry through stomata or wounds, and colonization in intercellular spaces, leading to rotting and to the disruption of plant cell wall and membrane structures. The three E. faecalis strains colonize the roots of A. thaliana by forming a mosaic of large clusters of live bacteria on the root surface, as observed by scanning electron microscopy, phase-contrast microscopy, and fluorescence microscopy. To dissect the involvement of mammalian virulence-related factors in plant pathogenicity, we tested E. faecalis mutant strains DeltafsrA (TX5240), DeltafsrB (TX5266), DeltafsrC (TX5242), DeltagelE (TX5264), and DeltasprE (TX5243), which correspond to virulence factors involved in pathogenesis in different animal models. Two E. faecalis virulence-related factors that play an important role in mammalian and nematode models of infection, a putative quorum-sensing system (DeltafsrB) and serine protease (DeltasprE), were also found to be important for plant pathogenesis. The development of an E. faecalis-A. thaliana model system could potentially be used to circumvent certain inherent limitations that an animal model imposes on the identification and study of virulence factors. Furthermore, our study suggests an evolutionary crossover of virulence factors in plant, animal, and nematode pathogenesis.

Arabidopsis↗

The N-glycosidase activity of the ribosome-inactivating protein ME1 targets single-stranded regions of nucleic acids independent of sequence or structural motifs.

ME(1), a type I ribosome-inactivating protein (RIP), belongs to a family of enzymes long believed to possess rRNA N-glycosidase activity directed solely at the universally conserved residue A4324 in the sarcin/ricin loop of large eukaryotic and prokaryotic rRNAs. We have investigated the effect of modifying the structure of nonribosomal RNA substrates on their interaction with ME(1) and other RIPs. ME(1) was shown to depurinate a variety of partially denatured nucleic acids, randomly removing adenine residues from single-stranded regions and, to a lesser extent, guanine residues from wobble base-pairs in hairpin stems. A defined sequence motif was not required for recognition of non-paired adenosines and cleavage of the N-glycosidic bond. Substrate recognition and ME(1) activity appeared to depend on the physical availability of nucleotides, and denaturation of nucleic acid substrates increased their interaction with ME(1). Pretreatment of mRNA at 75 degrees C rather than 60 degrees C, for example, lowered the apparent K(D) from 87.1 to 73.9 nm, making it more vulnerable to depurination by RIPs. Exposure to ME(1) in vitro completely abolished the infectivity of partially denatured RNA transcripts of the potato spindle tuber viroid, suggesting that RIPs may target invading nucleic acids before they reach host ribosomes in vivo. Our data suggest that the extensive folding of many potential substrates interferes with their ability to interact with RIPs, thereby blocking their inactivation by ME(1) (or other RIPs).

Adenine↗

Phytotoxic and antimicrobial activities of catechin derivatives.

(+/-)-Catechin is a potent phytotoxin, with the phytotoxicity due entirely to the (-)-catechin enantiomer. (+)-Catechin, but not the (-)-enantiomer, has antibacterial and antifungal activities. Tetramethoxy, pentaacetoxy, and cyclic derivatives of (+/-)-catechin retained phytotoxicity. The results indicate that antioxidant properties of catechins are not a determining factor for phytotoxicity. A similar conclusion was reached for the antimicrobial properties. Centaurea maculosa (spotted knapweed) exudes (+/-)-catechin from its roots, but the flavanol is not re-absorbed and hence the weed is not affected. The much less polar tetramethoxy derivative may, however, be absorbed and hence be able to cause toxicity. Because of the combination of phytotoxicity and antimicrobial activity, (+/-)-catechin could be a useful natural herbicide and antimicrobial.

Anti-Bacterial Agents↗

Biochemical and physiological mechanisms mediated by allelochemicals.

Allelochemistry, the production and release of toxic chemicals produced by one species that affect a receiving susceptible species, has been the subject of diverse degrees of scientific enquiry. Recent advances in plant biology have permitted the revamp of allelochemistry as a biologically and ecologically sound explanation for plant invasion and plant-plant communication in the rhizosphere. Recent progress has been made in understanding the biochemical and molecular changes that are induced by allelochemicals in susceptible plant species, and the complex mechanisms that are used by allelochemical-resistant plants to defend against this toxic insult.

Models, Chemical↗

How plants communicate using the underground information superhighway.

The rhizosphere is a densely populated area in which plant roots must compete with invading root systems of neighboring plants for space, water, and mineral nutrients, and with other soil-borne organisms, including bacteria and fungi. Root-root and root-microbe communications are continuous occurrences in this biologically active soil zone. How do roots manage to simultaneously communicate with neighboring plants, and with symbiotic and pathogenic organisms within this crowded rhizosphere? Increasing evidence suggests that root exudates might initiate and manipulate biological and physical interactions between roots and soil organisms, and thus play an active role in root-root and root-microbe communication.

Communication↗