Leslie A. Weston

The role of root exudates and allelochemicals in the rhizosphere

Plant roots serve a multitude of functions in the plant including anchorage, provision of nutrients and water, and production of exudates with growth regulatory properties. The root–soil interface, or rhizosphere, is the site of greatest activity within the soil matrix. Within this matrix, roots affect soil structure, aeration and biological activity as they are the major source of organic inputs into the rhizosphere, and are also responsible for depletion of large supplies of inorganic compounds. Roots are very complicated morphologically and physiologically, and their metabolites are often released in large quantities into the soil rhizosphere from living root hairs or fibrous root systems. Root exudates containing root-specific metabolites have critical ecological impacts on soil macro and microbiota as well as on the whole plant itself. Through the exudation of a wide variety of compounds, roots impact the soil microbial community in their immediate vicinity, influence resistance to pests, support beneficial symbioses, alter the chemical and physical properties of the soil, and inhibit the growth of competing plant species. In this review, we outline recent research on root exudation and the role of allelochemicals in the rhizosphere by studying the case of three plants that have been shown to produce allelopathic root exudates: black walnut, wheat and sorghum

Weed and Crop Allelopathy

Allelopathy can be defined as an important mechanism of plant interference mediated by the addition of plant-produced secondary products to the soil rhizosphere. Allelochemicals are present in all types of plants and tissues and are released into the soil rhizosphere by a variety of mechanisms, including decomposition of residues, volatilization, and root exudation. Allelochemical structures and modes of action are diverse and may offer potential for the development of future herbicides. We have focused our review on a variety of weed and crop species that establish some form of potent allelopathic interference, either with other crops or weeds, in agricultural settings, in the managed landscape, or in naturalized settings. Recent research suggests that allelopathic properties can render one species more invasive to native species and thus potentially detrimental to both agricultural and naturalized settings. In contrast, allelopathic crops offer strong potential for the development of cultivars that are more highly weed suppressive in managed settings. Both environmental and genotypic effects impact allelochemical production and release over time. A new challenge that exists for future plant scientists is to generate additional information on allelochemical mechanisms of release, selectivity and persistence, mode of action, and genetic regulation. In this manner, we can further protect plant biodiversity and enhance weed management strategies in a variety of ecosystems. Referee: Dr. Steve Weller, Purdue University, Dept. of Horticulture, West Lafayette, IN 47907

Biological Activity of Resveratrol, a Stilbenic Compound from Grapevines, Against Botrytis cinerea, the Causal Agent for Gray Mold

The biological activity of resveratrol, a stilbenic compound synthesized by grapevines in response to various stresses, was reevaluated against Botrytis cinerea using a novel in vitro system that enabled direct observation of the fungus with an inverted microscope. We determined that 90 μg resveratrol/ml reduced germination of B. cinerea conidia by ca. 50%. Moreover, resveratrol was shown to significantly reduce mycelial growth of B. cinerea at concentrations ranging from 60 to 140 μg/ml. Exposure to resveratrol at concentrations ranging from 60 to 140 μg/ml resulted in cytological changes in B. cinerea, such as production of secondary or tertiary germ tubes by conidia, cytoplasmic granulations, protoplasmic retractation in the hyphal tip cells, and formation of curved germ tubes. These data reinforce the role played by this compound in the B. cinerea–grapevine interaction.

Mode of Action, Localization of Production, Chemical Nature, and Activity of Sorgoleone: A Potent PSII Inhibitor in Sorghum spp. Root Exudates1

The root exudates produced by sorghums contain a biologically active constituent known as sorgoleone. Seven sorghum accessions were evaluated for their exudate components. Except for johnsongrass, which yielded 14.8 mg root exudate/g fresh root wt, sorghum accessions consistently yielded approximately 2 mg root exudate/g fresh root wt. Exudates contained four to six major components, with sorgoleone being the major component (> 85%). Three-dimensional structure analysis was performed to further characterize sorgoleones mode of action. These studies indicated that sorgoleone required about half the amount of free energy (493.8 kcal/mol) compared to plastoquinone (895.3 kcal/mol) to dock into the QB-binding site of the photosystem II complex of higher plants. Light, cryo-scanning, and transmission electron microscopy were utilized in an attempt to identify the cellular location of root exudate production. From the ultrastructure analysis, it is clear that exudate is being produced in the root hairs and being deposited between the plasmalemma and cell wall. The exact manufacturing and transport mechanism of the root exudate is still unclear. Studies were also conducted on sorgoleones soil persistence and soil activity. Soil impregnated with sorgoleone had activity against a number of plant species. Recovery rates of sorgoleone-impregnated soil ranged from 85% after 1 h to 45% after 24 h. Growth reduction of 9 14-d-old weed species was observed with foliar applications of sorgoleone. Nomenclature: Sorgoleone (2-hydroxy-5-methoxy-3-[(8′Z,11′Z)-8′,11′,14′-pentadecatriene]-p-hydroquinone); common purslane, Portulaca oleracea L. POROL; common ragweed, Ambrosia artemisiifolia L. AMBEL; cress, Lepidium sativum L. ;ns3 LEPSA; giant foxtail, Setaria faberi Herrm. SETFA; johnsongrass, Sorghum halepense (L.) Pers. SORHA; lambsquarters, Chenopodium album L. CHEAL; large crabgrass, Digitaria sanguinalis (L.) Scop. DIGSA; lettuce, Lactuca sativa L.; nightshade, Solanum spp.; purple photosynthetic bacterium, Rhodopseudomonas viridis; redroot pigweed, Amaranthus retroflexus L. AMARE; sicklepod, Cassia obtusifolia L. CASOB; spinach, Spinacea oleracea; shattercane, Sorghum bicolor (L.) Moensch SORVU; sorghum, S. bicolor (L.) Moensch SORVU; sudex, S. bicolor × Sorghum sudanense; sweet sorghum, S. bicolor ‘Della’; SX-15 and SX-17, S. bicolor × S. sudanense; 8446 and 855-F, S. bicolor (L.) Moensch SORVU; tomato, Lycopersicon esculentum L.; velvetleaf, Abutilon theophrasti Medicus ABUTH. Additional index words: Sorgoleone, root hairs, SORVU, SORHA. Abbreviations: ER, endoplasmic reticulum; HPLC, high-pressure liquid chromatography; PSII, photosystem II; QB, quinone binding; SEM, scanning electron microscopy; TEM, transmission electron microscopy; TLC, thin-layer chromatography; 3D, three dimensional; UV, ultraviolet.

Flavonoids: Their Structure, Biosynthesis and Role in the Rhizosphere, Including Allelopathy

Flavonoids are biologically active low molecular weight secondary metabolites that are produced by plants, with over 10,000 structural variants now reported. Due to their physical and biochemical properties, they interact with many diverse targets in subcellular locations to elicit various activities in microbes, plants, and animals. In plants, flavonoids play important roles in transport of auxin, root and shoot development, pollination, modulation of reactive oxygen species, and signalling of symbiotic bacteria in the legume Rhizobium symbiosis. In addition, they possess antibacterial, antifungal, antiviral, and anticancer activities. In the plant, flavonoids are transported within and between plant tissues and cells, and are specifically released into the rhizosphere by roots where they are involved in plant/plant interactions or allelopathy. Released by root exudation or tissue degradation over time, both aglycones and glycosides of flavonoids are found in soil solutions and root exudates. Although the relative role of flavonoids in allelopathic interference has been less well-characterized than that of some secondary metabolites, we present classic examples of their involvement in autotoxicity and allelopathy. We also describe their activity and fate in the soil rhizosphere in selected examples involving pasture legumes, cereal crops, and ferns. Potential research directions for further elucidation of the specific role of flavonoids in soil rhizosphere interactions are considered.

Grass roots chemistry: meta-Tyrosine, an herbicidal nonprotein amino acid

Fine fescue grasses displace neighboring plants by depositing large quantities of an aqueous phytotoxic root exudate in the soil rhizosphere. Via activity-guided fractionation, we have isolated and identified the nonprotein amino acid m-tyrosine as the major active component. m-Tyrosine is significantly more phytotoxic than its structural isomers o- and p-tyrosine. We show that m-tyrosine exposure results in growth inhibition for a wide range of plant species and propose that the release of this nonprotein amino acid interferes with root development of competing plants. Acid hydrolysis of total root protein from Arabidopsis thaliana showed incorporation of m-tyrosine, suggesting this as a possible mechanism of phytotoxicity. m-Tyrosine inhibition of A. thaliana root growth is counteracted by exogenous addition of protein amino acids, with phenylalanine having the most significant effect. The discovery of m-tyrosine, as well as a further understanding of its mode(s) of action, could lead to the development of biorational approaches to weed control.

Evaluation of root exudates of seven sorghum accessions

Seven sorghum accessions were evaluated quantitatively and qualitatively for the composition of their root exudates. Utilizing a unique capillary mat growing system, root exudates were collected from all sorghum accessions. Exudates were subjected to TLC and HPLC analysis to evaluate their chemical composition. Within each sorghum accession, variation existed in the amount of exudate produced and the chemical constituents of each exudate. Sorgoleone was the predominant constituent identified in each accessions exudate. Other closely related compounds, including 5-ethoxysorgoleone, 2,5-dimethoxysorgoleone, three other minor components (MW = 364, 388, and 402), and one unidentified component comprised the minor constituents of the root exudate. Our past work has shown that several of these compounds have potent phytotoxic activity as photosystem II inhibitors, thereby lending further support to the concept that Sorghum spp. are allelopathic and weed suppressive.

Are Laboratory Bioassays for Allelopathy Suitable for Prediction of Field Responses

One concern often voiced by researchers of allelopathic interactions is that many laboratory bioassays do not adequately predict the responses observed in field situations. The questions that arise are: (1) What criteria should be implemented to design ecologically relevant bioassays? (2) What species (crops or weeds) are involved in the interaction? (3) Are we investigating allelopathy of debris/residues or interactions involving living plants? (4) Which plant indicator species are actually cohabiting with the species under investigation? and (5) What are appropriate experimental controls? It is difficult to design a bioassay that can be used to examine responses in all species. In fact, each bioassay must be designed specifically to assess species interactions after careful consideration of growth habit, biotic characteristics, and ecophysiological factors. The objective of this paper is to discuss the significance of bioassays designed to study a particular aspect of allelopathy. We conclude that through a laboratory bioassay we can not demonstrate that allelopathy is operational in natural settings. An investigator should consider allelopathy as one component in a multifaceted approach to ecology and address key questions to determine the relevance of a particular assay. Due to the complexity of field interactions and responses, one can only hope to predict and describe some of the cause-and-effect relationships observed in a field setting. An accurate assessment of these main effects will prove invaluable in directing the focus of future research emphasis.

ISOLATION AND CHARACTERIZATION OF ALLELOPATHIC VOLATILES FROM MUGWORT (Artemisia vulgaris)

Several volatile allelochemicals were identified and characterized from fresh leaf tissue of three distinct populations of the invasive perennial weed, mugwort (Artemisia vulgaris). A unique bioassay was used to demonstrate the release of volatile allelochemicals from leaf tissues. Leaf volatiles were trapped and analyzed via gas chromatography coupled with mass spectrometry. Some of the components identified were terpenes, including camphor, eucalyptol, α-pinene, and β-pinene. Those commercially available were tested individually to determine their phytotoxicity. Concentrations of detectable volatiles differed in both absolute and relative proportions among the mugwort populations. The three mugwort populations consisted of a taller, highly branched population (ITH-1); a shorter, lesser-branched population (ITH-2) (both grown from rhizome fragments from managed landscapes); and a population grown from seed with lobed leaves (VT). Considerable interspecific variation existed in leaf morphology and leaf surface chemistry. Bioassays revealed that none of the individual monoterpenes could account for the observed phytotoxicity imparted by total leaf volatiles, suggesting a synergistic effect or activity of a component not tested. Despite inability to detect a single dominant phytotoxic compound, decreases in total terpene concentration with increase in leaf age correlated with decreases in phytotoxicity. The presence of bioactive terpenoids in leaf surface chemistry of younger mugwort tissue suggests a potential role for terpenoids in mugwort establishment and proliferation in introduced habitats.

Allelopathic potential of sorghum-sudangrass hybrid (sudex)

Experiments were conducted under controlled conditions to investigate the apparent allelopathic effects of sudex [Sorghum bicolor (L.) Moench ×Sorghum Sudanese (P.) Stapf, cv. FFR 201] on weed and vegetable species. Allelopathic potential, as measured by radicle elongation of herbaceous indicator species, decreased with increasing sudex age. Greatest potential allelopathic activity of sudex shoot tissue was observed when sudex was collected at 7 days of age. Small-seeded broadleaf species were more inhibited in the presence of sudex shoot tissue than were grass species. Two major phytoinhibitors were isolated from aqueous extracts of sudex shoot material by partitioning with diethyl ether, followed by thin-layer and liquid column chromatography. Phytoinhibitors were identified asp-hydroxybenzoic acid andp-hydroxybenzaldehyde, potentially the enzymatic breakdown products of the cyanogenic glycoside dhurrin. The I50 values of these compounds using a cress (Lepidium sativum L.) seed bioassay were 140 and 113 μg/ml for the acid and aldehyde, respectively. Sudex tissue collected at 7 days of age possessed a greater percentage of these phytoinhibitors on a per gram basis than did older sudex tissue. As sudex tissue age increased, the percentage ofp-hydroxybenzaldehyde in ether extracts of tissue also increased, while the percentage ofp-hydroxybenzoic acid decreased.